A Lanata

Official Journal of International Quarterly Publication
ISSN 1608-2281
Original Journal Name: Asian Journal of Drug Metabolism and Pharmacokinetics (2001-2005)
Asian Journal of
Pharmacodynamics and
Pharmacokinetics
Academic Editor-in-Chief
Chang-Xiao Liu and Yuichi Sugiyama
Volume 9 Number 1
March 2009
Hong Kong Medical Publisher
Asian Journal of Pharmacodynamics and Pharmacokinetics

An International Quarterly Publication
ISSN 1608-2281
NR 3880/169/00
http://www.hktmc.com/ChineseMedia/Magazine/Medicine/ajdmpk
Publisher President
Kwong Cheung Ku Hong Kong Medical Publisher 24/F Yuen Long Trading Center, 99-109 Castle Peak Road, Yuen
Long, Hong Kong, China
Academic Editor-in-chief
Chang-Xiao Liu Tianjin Institute of Pharmaceutical Research, 308 An-Shan West Road, Tianjin 300193, China
Yuichi Sugiyama Department of Molecular Pharmacokinetics, The University of Tokyo, Tokyo, 113-0033, Japan
Assistant Academic Editor-in-Chief

Da-Fang Zhong Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 646 Songtao Road Shanghai,
201203, China
Guang-Hua Du Institute of Materia Medica, Chinese Academy of Medical Sciences, Beijing, 100050, China
Qiang Zhang School of Pharmaceutical Sciences, Peking University, Beijing, 100083, China
Publishing Editor-in-chief
Annie Ning Editor-in-chief of Hong Kong Medical Publisher
Hui Qing Shi Vice-editor-in-chief of Hong Kong Medical Publisher
Members of Editorial Committee

Henning
H.
Blume
(Oberursel,
Terry D. Lindstrom (Indiana, USA)
Chang-Koo Shim (Seoul, KOREA)
Chang-Xiao Liu (Tianjin, CHINA)
Guang-Ji Wang (Nanjing, CHINA)
Zheng-Min Chen (Tianjin, CHINA)
Xiao-Dong Liu (Nanjing, CHINA)
Hui Wang (Wuhan, CHINA)
Moses SS Chow (Hong Kong, CHINA)
Ke-Xin Liu (Dalian, CHINA)
Ming-Wei Wang (Shanghai, CHINA)
Guang-Hua Du (Beijing, CHINA)
Jian-Shi Lou (Tianjin, CHINA)
Zhi-Min Wang (Beijing, CHINA)
Paul Fawcett (Otago, NEW ZEALAND)
Takeo Murakawa (Osaka, JAPAN)
Yi-Tao Wang (Macau, CHINA)
Jack Gao (Carborough, CANADA)
Terumichi Nakagawa (Kyoto, JAPAN)
Guang-Li Wei (Tianjin, CHINA)
GERMANY)
Mei-Yu Geng (QingdaoCHINA
Charles H. Nightingale (Hartford, USA)
Chun-Fu Wu (ShenyangCHINA)
Patrick M. Gennissel (Paris, FRANCE)
Inotsume Nobuo (Hokkaido, JAPAN)
Jinn Wu
Kun Han (Chungbuk, KOREA)
Richard ZM Qian (Hong Kong, CHINA)
Fu-Ming Xie ( Indiana, USA)
Guo-Zhu Han (Dalian, CHINA)
Madhubala Rentala (New Delhi,
Guo-Wang Xu (Dalian, CHINA)
Pei Hu (Beijing, CHINA)
INDIA)
(New Jersey, USA)
Ming Xue (Beijing, CHINA)
Zhuo-Han Hu (Shanghai, CHINA)
Jin-Xiu Ruan (Beijing, CHINA)
Bao-Feng Yang (HarbinCHINA)
Xi Huang (Chengdu, CHINA)
Guo-Wei Sang (Beijing, CHINA)
Zong-Hui Yuan (Wuhan, CHINA)
Jun-Yan Hong (New Jersey, USA)
Xiang-Guo Shi (Boston, USA)
Fan-Dian Zeng (Wuhan, CHINA)
Li-Ya Ju (Charenton le Pont, FRANCE)
Duan-Yun Si (Tianjin, CHINA)
Su Zeng (Hangzhou, CHINA)
Devarakonda R. Krishna (Kakatiya,
Chang Koo Shim (Seoul, KOREA)
Qiang Zhang (Beijing, CHINA)
Yuichi Sugiyama (Tokyo, JAPAN)
Jiang Zheng (SeattleUSA)
June Lee (NIH, USA)
Ding-Feng Su (Shanghai, CHINA)
Da-Fang Zhong (Shanghai, CHINA)
Jian-Guo Li
INDIA)
Nikolaus Sucher (Hong Kong, CHINA)
Hong-Hao Zhou (Changsha, CHINA)
Lin-Lin Li (Xinjiang, CHINA)
Yong-Da Sun (Bradford, UK)
Yao-Wei Zhu (PA, USA)
Zhi-Bin Lin (Beijing, CHINA)
Hui-Qing Shi (Hong Kong, CHINA)
Zhu Zhu (Beijing, CHINA)
(Wilmington, USA)

(An International Quarterly Publication)
Volume 9
Number 1
March
2009
Content
3
Ethical Guideline to Authors, Editors and Reviewers
Special Repart
5
Introduction to Professor Leslie Z. Benet and his Lecture in China: Predicting Drug Absorption and
Disposition Using a Biopharmaceutics Drug Disposition Classification System
Review Papers
11
Hai-Yu Xu , Tie-Jun Zhang , Xue-Yu Zhu,Yu-Bo Li. Recent advance on chemical compositions and
pharmacodynamic and pharmacokinetic studies of Rhizoma Coptidis
27
Tie-Feng Cheng, Yong-Da Sun, Duan-Yun SiChang-Xiao Liu. Attention on research of

pharmacology and toxicology of nanomedicines
Research Papers
51
Rui Zhang, Benjie Wang, Hengli Zhao, Chunmin Wei, Guiyan Yuan, Ruichen Guo. Tissue
58
63
distribution of Curcumol in rats after intravenous injection of zedoary turmeric oil fat emulsion
Sama Venkatesh, Yanadaiah JP, Zareen N, Madhava Reddy B, Ramesh M. Antinociceptive effect
of Aerva lanata ethanolic extract in mice: A possible mechanism
Run Li, Zong-Peng Zhang, Yi-Hong Tian. Therapeutic effect and mechanism for anti-fibrosis of
polyhydroxysilbene of Rhizoma Scirpi in hepatic fibrosis rats
71
Xiao-Pu Nie, Wen-Yuan Gao, Pei-Gen Xiao. Changes of the adenosine content in single and mixed
decoctions of Gualou-xiebai-baijiu Decoction
Information and News

10
26
50
50
77
78
Publication News Chinese Herbal Medicines

Tianjin Centre for Drug Safety Evaluation and Research
Publication News Drug Evaluation Research
The 3rd Asian Pacific ISSX Meeting
Successful the 2nd AsianPacific Regional ISSX Meeting
Information for authors
Ethical Guideline. Asian Journal of Pharacodynamics and Pharmacokinetics 2009;9(1):3-4
Asian Journal of
Pharmacokinetics
ISSN
1608-2281
Copyright by Hong Kong Medical Publisher

Publisher Homepage: www.hktmc.com
Ethical Guideline to Authors, Editors and Reviewers

originated.
6. The authors should reveal to the editor any
potential conflict of interest. The authors should
ensure that no contractual relations or proprietary
considerations exist that would affect the
publication of information in a submitted
manuscript.
7. The co-authors of a paper should be all those
persons who have made significant scientific
contributions to the investigation reported and share
duty and accountability for the results. Other
contributions should be indicated in a footnote or in
an acknowledgments section.
8. The author who submits a manuscript for
publication takes the responsibility of having
included as co-authors all persons appropriate and
none inappropriate.
9. The submitting author should have sent each
living co-author a draft copy of the manuscript and
have obtained the co-authors assent to the
co-authorship of it.
10. It is improper for an author to submit
manuscripts describe essentially the same research
in more than one journal for primary publication,
unless it is a resubmission of a manuscript rejected
for or withdrawn from publication.
Drug products are specific goods with safety

and effectiveness in medical health case. All of
researchers (authors), reviewers and editors must
abide by medical ethical obligation, and also must
deter to ethical obligation for publication. These
guidelines are offered as ethical behavior standards.
We now present a set of ethical guidelines for
persons engaged in the publication of drug research,
specifically, for editors, authors, and reviewers. We
believe that the guidelines offered are understood
and subscribed to by the pharmaceutical research
scientists. They may be helpful to those who are
related to editors, authors, and reviewers of journal
publication.
Ethical Obligation of Authors

1. An authors key obligation is to present an
accurate account of the research paper.
2. An author should recognize that journal
space is precious resource created at considerable
cost. Therefore, an author has an obligation to use it
widely and economically.
3. An author should cite those publications that
are influential in determining the nature of the
reported study work and that will guide the reader
to the earlier work quickly and that are essential for
understanding the present investigation.
4. An author is obligated to perform a literature
search to find, and then cite, the original
publications that describe the current research,
closely related citation to sources should also be
made when a non-author supplied these.
5. An author should identify the source of all
information quoted or offered. Information obtained
privately, ad in conversation, should not be used in
the authors work without explicit permission from
the investigators with whom the information is
Ethical Obligation of Editors

1. An editor should respect the intellectual
independence of authors.
2. An editor should give unbiased
consideration to all manuscripts submitted, and
judge each on its merits without regard to race,
religion, nationality, sex, seniority, or institutional
affiliation of authors.
3. An editor should consider manuscripts
submitted for publication with reasonable speed.
3
Ethical Guideline. Asian Journal of Pharacodynamics and Pharmacokinetics 2009;9(1):3-4
4. An editor should not disclose any

information about a manuscript under consideration
to anyone other than those from whom professional
advice is sought.
5. Editorial responsibility and authority for any
manuscript authored by an editor and submitted to
the journal should be distributed to some other
qualified editors of the journal. The editorial
consideration of the manuscript in any way or form
by the author-editor would constitute a conflict of
interest, and is therefore improper.
6. The sole responsibility for acceptance or
rejection of a manuscript rests with the editor.
Responsible and prudent exercise of the duty
normally requires that editor seek advice from
reviewers, chosen for their expertise and good
judgment, as to the quality and reliability of
manuscript submitted for publication.
7. The editor and members of the editors staff
should not disclose any information about a
manuscript under consideration to anyone other
than those from whom professional advice is sought.
The editor and members of the editors staff may
disclose or publish manuscript titles and author
names of papers that have been accepted for
publication.
8. Unpublished information, or interpretations
disclosed in a submitted manuscript should not be
used in an editors own research without the consent
of author.
9. An author may request that the editor not
choose certain reviewers in consideration of a
manuscript.
10. When a manuscript is closely related to the
current or past research of an editor as to create a
conflict of interest, the editor should arrange some
other qualified person to take editorial responsibility
for that manuscript.
with high scientific and literary standards. A

reviewer
should
respect
the
intellectual
independence of the authors.
3. A chosen reviewer who feels inadequately
qualified to judge the research reported in a
manuscript should return it to editor.
4. A reviewer should explain and support their
judgment adequately so that editors and authors
may understand the basis of their comments.
5. A reviewer should treat a manuscript sent for
review as a confidential document. It should neither
be shown to nor be discussed with other expect, in
special cases, to persons from whom specific advice
may be sought. In that event, the identifications of
those consulted should be disclosed to the editor.
6. A reviewer should act promptly, submitting
a report in a timely manner. If a reviewer receives a
manuscript at a time when circumstances preclude
prompt attention to it, the unreviewed manuscript
should be returned immediately to the editor.
7. A reviewer should be alert to failure of
authors to cite relevant work by other persons,
bearing in mind that compliantly that the reviewer
own research was insufficiently cited may seem
self-serving. A reviewer should call the editors
attention any substantial similarity between the
manuscript under consideration and any published
paper and any manuscript submitted concurrently to
other journals.
8. A reviewer should be sensitive to the
appearance of a conflict of interest when the
manuscript under review is closely related to the
reviewers work in progress or published. If in
doubt, the reviewer should return the manuscript
promptly without review, advising the editor of the
conflict of interest or bias.
9. A reviewer should not evaluate a manuscript
authored or co-authored by a person with whom the
reviewer has a personal or professional connection
if the relationship would bias judgment of the
manuscript.
10. A reviewer should not use or disclose
unpublished
information,
arguments,
or
interpretations contained in a manuscript under
consideration.
11. The review of submitted manuscript may
sometimes justify criticism, even severe criticism,
from a reviewer.
Ethical Obligation of Reviewers

1 As the reviewing of manuscripts is an
essential step in journal publication, therefore in the
operation of the reviewers, every reviewer has an
obligation to do a fair share of reviewing.
2. A reviewer of a manuscript should judge
objectively the quality of the manuscript, of its
experimental and theoretical work, of its
interpretations and its exposition, in accordance
Special Report. Asian Journal of Pharmacodynamics and Pharmacokinetics 2009; 9(1):5-9
Asian Journal of
Pharmacokinetics
ISSN
1608-2281

Special report
Introduction to Professor Leslie Z. Benet and his Lecture in China:
Predicting Drug Absorption and Disposition Using a Biopharmaceutics
Drug Disposition Classification System
Pharmaceutical Scientists, the Rawls-Palmer
Progress in Medicine award of the American
Society for Pharmacology and Therapeutics, the
Higuchi Research Prize of the American
Pharmacists Association and the Host-Madsen
Medal of the International Pharmaceutical
Federation (FIP). Dr. Benet formerly served as
Chair of the Pharmacology Study Section and the
Pharmacological Sciences Review Committee for
the NIH, the FDA Center for Biologics (CBER)
Peer Review Committee, the FDA Expert Panel on
Individual Bioequivalence, the Board of
Pharmaceutical Sciences of the FIP, the Organizing
Committee for the Millenial World Congress of
Pharmaceutical Sciences, the Congressionally
mandated IOM/NRC Committee on Accelerating
the Research, Development and Acquisition of
Medical Countermeasures Against Biological
Warfare Agents and as a member of the FDA
Generic Drugs Advisory Committee and the FDA
Science Board. He was the Founder and first
President of the American Association of
Pharmaceutical Scientists that now numbers more
than 13,000 members. Dr. Benets research interests,
more than 490 scientific publications, 7 books and
11 patents are in the areas of pharmacokinetics,
biopharmaceutics,
drug
delivery
and
pharmacodynamics.
Professor Leslie Z. Benet visited Tianjin

Institute of Pharmaceutical Research, China on
Wednesday January 7, 2009. During the visitation,
Professor
Benet
made
an
academic
lecture/presentation titled Predicting Drug
Absorption
and
Disposition
Using
a
Biopharmaceutics Drug Disposition Classification
System to scientists from Tianjin State Laboratory
of Pharmacokinetics and Pharmacodynamics.
Introduction to Professor Leslie Z. Benet
Professor Leslie Z. Benet, Ph.D., Distinguished

Clinical Research Lecturer and chairman of the
department
of
biopharmaceutical
sciences
(1978-1998), University of California, San
Francisco (UCSF), has received international
recognition for his work related to pharmacokinetics
and pharmacodynamics. Dr. Benet with six
honorary doctorates is recognized with the highest
scientific awards of nine professional societies,
including the Distinguished Pharmaceutical
Scientist award of the American Association of
Lecture: Predicting Drug Absorption and

Disposition Using a Biopharmaceutics Drug
Disposition Classification System
The Biopharmaceutics classification system
5
drug substance from its degradation products.

The permeability class of a drug substance can
be determined in human subjects using mass
balance, absolute BA, or intestinal perfusion
approaches: 1. Pharmacokinetic Studies in Humans:
a. Mass Balance Studies b. Absolute Bioavailability
Studies; 2. Intestinal Permeability Methods: The
following methods can be used to determine the
permeability of a drug substance from the
gastrointestinal tract: (1) in vivo intestinal perfusion
studies in humans; (2) in vivo or in situ intestinal
perfusion studies using suitable animal models; (3)
in vitro permeation studies using excised human or
animal intestinal tissues; or (4) in vitro permeation
studies across a monolayer of cultured epithelial
cells; 3. Instability in the Gastrointestinal Tract:
determining the extent of absorption in humans
based on mass balance studies using total
radioactivity in urine does not take into
consideration the extent of degradation of a drug in
the gastrointestinal fluid prior to intestinal
membrane permeation.
Dissolution testing should be carried out in
USP Apparatus I at 100 rpm or Apparatus II at 50
rpm using 900 mL of the following dissolution
media: (1) 0.1 N HCl or Simulated Gastric Fluid
USP without enzymes; (2) a pH 4.5 buffer; and (3) a
pH 6.8 buffer or Simulated Intestinal Fluid USP
without enzymes. For capsules and tablets with
gelatin coating, Simulated Gastric and Intestinal
Fluids USP (with enzymes) can be used. When
comparing the test and reference products,
dissolution profiles should be compared using a
similarity factor (f2). The similarity factor is a
logarithmic reciprocal square root transformation of
the sum of squared error and is a measurement of
the similarity in the percent (%) of dissolution
between the two curves. Two dissolution profiles
are considered similar when the f2 value is 50.
By understanding the relationship between a
drugs absorption, solubility, and dissolution
characteristics, it is possible to define situations
when in vitro dissolution data can provide a
surrogate
for
in
vivo
bioequivalence
assessments,that is to find under which
circumstances dissolution testing can be prognostic
for in vivo performance.
The Biopharmaceutics classification system

(BCS) has been one of the most significant
prognostic tools created to promote product
development in recent years. It is a scientific
framework for classifying drug substances based on
their aqueous solubility and intestinal permeability
characteristics, which will substantially facilitate
drug product selection and approval process for a
large group of drug candidates. The goal of the BCS
is to function as a tool for developing in vitro
dissolution specifications for drug products that are
predictive of their in vivo performance.
According to the BCS, drug substances are
classified as follows:
Class 1: High Solubility-High Permeability:
generally very well-absorbed compounds
Class 2: Low Solubility-High Permeability:
exhibit dissolution rate-limited absorption
Class 3: High Solubility-Low Permeability:
exhibit permeability rate-limited absorption
Class 4: Low Solubility-Low Permeability:
very poor oral bioavailability.
The Class Boundaries:
z A drug substance is considered HIGHLY
SOLUBLE when the highest dose strength is
soluble in250 ml water over a pH range of 1 to
7.5.
z A drug substance is considered HIGHLY
PERMEABLE when the extent of absorption in
humans is determined to be 90% of an
administered dose, based on mass-balance or in
comparison to an intravenous reference dose.
z A drug product is considered to be
RAPIDLY DISSOLVING when 85% of the
labeled amount of drug substance dissolves within
30 minutes using USP apparatus I or II in a volume
of 900 ml buffer solutions.
The pH-solubility profile of the test drug
substance should be determined in aqueous media
with a pH in the range of 1-7.5 using traditional
shake-flask method as well as acid or base titration
methods. A sufficient number of pH conditions
should be evaluated to accurately define the
pH-solubility profile. Concentration of the drug
substance in selected buffers (or pH conditions)
should be determined using a validated
stability-indicating assay that can distinguish the
6
definition of extensive metabolism be pushed

to 70% metabolism of an oral dose in vivo in
humans while the poor metabolism be defined
as 50% of the dose be excreted unchanged. Use
of the more stringent BDDCS metabolism criteria
versus the BCS permeability characterization
resulted in only 10 compounds requiring
reclassification and allowed for the inclusion of
an additional 38 drugs/compounds.
The
Biopharmaceutics
Drug
Disposition
Classification System (BDDCS)
The development of the BCS was a major step
in bringing rational science to regulation, allowing
waivers of in vivo bioavailability and
bioequivalence testing (biowaiver) of immediate
release dosage forms for high-solubility,
highpermeability drugs when such drug products
also exhibited rapid dissolution. This application of
science yielded a decrease in the regulatory burden.
When the BCS was first developed there was only a
nascent understanding of the importance of drug
transporters to bioavailability. However, as pointed
out here, for Class 1 compounds neither efflux nor
absorptive transporters should influence oral
bioavailability, and meal effects on Fextent should
be negligible.
The Biopharmaceutics Drug Disposition
Classification System (BDDCS) replaces the
permeability criteria with the major route of
elimination because of the belief that it is easier and
less ambiguous to determine the assignment of
BDDCS for marketed drugs based on the extent of
metabolism than using permeability (i.e. extent of
absorption) in BCS assignments. Designation of the
major route of drug elimination as part or instead of
the permeability criteria would reduce the
regulatory burden for many more Class 1
compounds, would eliminate the ambiguity and
difficulty in determining 90% (or 85%) absorption
for Classes 1 and 2 compounds, and would allow
predictability of absorption and disposition
characteristics of drugs in all four BDDCS.
BDDCS Class compounds would then be
designated as:
Class 1: high solubility, extensive metabolism.
Waiver of in vivo bioequivalence studies for
BDDCS Class 1 drugs would still require rapid
dissolution.
Class 2: poor solubility, extensive metabolism
Class 3: high solubility, poor metabolism
Class 4: low solubility, poor metabolism.
When initially proposed, extensive
metabolism was defined as 50% metabolism of
an oral dose in vivo in humans. Further
consideration of this parameter designation led to
the realization that there are very few
drugs/compounds that are intermediately
metabolized. It is now proposed that the
Predicting Drug Absorption and Disposition

The BDDCS was developed to allow prediction
of in vivo pharmacokinetic performance of drug
products. It may be useful in predicting overall drug
disposition, including routes of drug elimination
and the effects of efflux and absorptive transporters
on oral drug absorption; when transporter-enzyme
interplay will yield clinically significant effects (e.g.,
low bioavailability and drug-drug interactions); the
direction, mechanism, and importance of food
effects; and transporter effects on postabsorption
systemic drug concentrations following oral and
intravenous dosing. These predictions are supported
by a series of studies investigating the effect of
transporter inhibition and induction on drug
metabolism.
Major Routes of Drug Elimination:
Class 1 and Class 2: compounds are eliminated
primarily via metabolism;
Class 3 and Class 4: compounds are primarily
eliminated unchanged into the urine and bile.
For the 130 drugs/compounds gathered from
the literature, only 13 of the substances do not have
readily
accessible,
critically
evaluated
pharmacokinetic parameters. If you know the
intestinal absorption (or more likely a surrogate as
Caco-2 permeability) of an NME, you can predict
whether the major route of elimination of the NME
will be metabolism.
Note that the permeability parameter does not
predict the ability for the NME to enter the liver/
hepatocytes (since a number of non-metabolized
Classes 3 & 4 compounds will be excreted in the
bile), but rather the access to the metabolic enzymes
within the hepatocytes.
Oral
7
Dosing
and
the
Predictability
of
Class 3: highly soluble, low permeability, poorly

metabolized drugs.
Uptake transporters will be important for

intestinal absorption and liver entry for these poor
permeability drugs.
However, once these poorly permeable

drugs get into the enterocyte or the hepatocyte
efflux transporter effects can occur.
Class 4: poorly soluble, low permeability,
poorly metabolized drugs
Oral bioavailability is minimal and
transporter effects could be relevant.
Transporter Effects:
Since extent of metabolism correctly predicts
high vs low intestinal permeability for at least 33 of
35 drugs , where human permeability
measurements exist. Benet and co-workers propose
the following: We recommend that regulatory
agencies add the extent of drug metabolism (i.e.,
90% metabolized) as an alternate method for the
extent of drug absorption (i.e., 90% absorbed) in
defining Class 1 drugs suitable for a waiver of in
vivo studies of bioequivalence.
Class 1: highly soluble, high permeability,
extensively metabolized drugs
Transporter effects will be minimal in the

intestine and the liver.
Even compounds like verapamil that can

be
shown
in
certain
cellular
systems
(MDR1-MDCK) to be a substrate of P-gp will
exhibit no clinically significant P-gp substrate
effects in the gut and liver.
A major proposition (and probably the

primary advance in knowledge) of BDDCS is that
Class 1 drugs are not substrates for transporters in
the intestine and liver. (but not at the BBB and the
kidney)
Class 2: poorly soluble, highly permeable,
extensively metabolized drugs
Efflux transporter effects will be

important in the intestine and the liver.
In the intestine efflux transporter

enzyme (CYP 3A4 and UGTs) interplay can
markedly affect oral bioavailability.
In the liver the efflux transporter-enzyme

interplay will yield counteractive effects to that seen
in the intestine.
Uptake transporters can be important for

the liver but not the intestine.
Following oral dosing, major significant

interactions will occur for Class 2 drugs that are
substrates for intestinal enzymes (e.g. CYP3A,
UGTs) and efflux transporters (e.g. P-gp, MRP2,
BCRP). Since concomitant inhibition of the
intestinal enzyme and the efflux transporter both
lead to less gut metabolism that synergistically
increase systemic AUC. It is not surprising that
drugs removed from the market due to drug-drug
interactions predominate for orally dosed drugs that
are substrates for CYP3A and P-gp.
Food Effects (High-Fat Meals):

It is well-known that food can influence drug
bioavailability, both increasing and decreasing the
extent of availability (Fextent) and the rate of
availability.
Class 1: High-fat meals will have no significant
effect on Fextent for Class 1 compounds.
Because complete absorption may be

expected for high solubility/high permeability
compounds, and as noted previously, no transporter
drug interactions would be expected for Class 1
compounds.
However, high-fat meals may delay

stomach emptying and therefore cause an increase
in peak time.
Class 2: High-fat meals will increase Fextent for
Class 2 compounds
Due to inhibition of efflux transporters in

the intestine and additional solubilization of drug in
the intestinal lumen (e.g., micelle formation).
Peak time could decrease due to

inhibition of efflux cycling or increase due to
slowing of stomach emptying;
Formulation changes that markedly

increase the solubility of Class 2 compounds will
decrease or eliminate the high-fat meal effects for
these drugs.
Class 3: High-fat meals will decrease Fextent for
Class 3 compounds
Due to inhibition of uptake transporters

in the intestine.
Class 4: For Class 4 compounds, it is difficult to
predict what will occur.
Postabsorption
8
Effects
and
Intravenous
P-glycoprotein, MRP2, BCRP).
The enzyme-efflux transporter interplay

that is so important in the intestine will not be as
significant in the liver (and the kidney) due to the
reverse order in which drug molecules encounter
the two proteins.
Inhibition of hepatic uptake transporters

can lead to significantly increased systemic drug
concentrations for Class 2 compounds
Class 3 and 4: Uptake transporter, efflux
transporter and uptake-efflux transporter interplay.
Inhibition of hepatic and renal uptake
transporters can lead to significant increases in the
systemic concentration of Classes 3 and 4
compounds.
Dosing:
For intravenous dosing, drug concentrations at
the eliminating organ will always be relatively low
due to the diluting effects of volume of distribution,
as compared to concentrations of drug in the
intestine. Therefore, saturation of transporters (and
enzymes) will be minimal, if at all, and solubility
considerations will be unimportant when
measurable systemic concentrations of the drug are
achieved.
High extraction ratio drugs, where

clearance approaches blood flow, are mainly limited
to Class 1 compounds.
Post intestinal absorption and following

intravenous dosing, both uptake and efflux
transporters can be important determinants of the
disposition for Classes 2, 3, and 4 compounds.
Biliary secretion of parent drug can be an

important component of disposition for Classes 3
and 4 compounds.
Renal elimination of Classes 3 and 4

compounds can be affected by both uptake and
efflux transporters.
Conclusions
The interplay between transporters, both influx
and efflux, and metabolic enzymes in the intestine
and the liver could differ depending on the drugs
solubility and permeability characteristics as
reflected in the Biopharmaceutical Classification
System (BCS). Although BCS has had a marked
effect in decreasing the regulatory burden by
allowing a waiver of in vivo bioequivalence studies
for a limited number of Class 1 drugs, little
predictive use has been made of Classes 2, 3, and 4
in the BCS categorization. In general, BCS Classes
1 and 2 are highly metabolized, whereas BCS
Classes 3 and 4 drugs are primarily excreted
unchanged via the biliary or renal routes. Therefore,
changing the permeability component to a route of
elimination component in a Biopharmaceutics Drug
Disposition Classification System (BDDCS) will
facilitate predictions, markedly expand the number
of Class 1 drugs eligible for waiver of in vivo
bioequivalence studies, and provide new insight. It
may be easier to determine classification based on
major routes of elimination than upon permeability;
an extent of metabolism criterion for waiver of in
vivo bioequivalence studies; and how predictive
algorithms may be developed using only in vitro or
in silico methods to facilitate class assignment in
BDDCS. A further advantage of BDDCS is that a
preliminary class assignment for NMEs may be
obtained from a metabolism measure in human
hepatocytes, prior to in vivo studies in humans.
Understanding transporter-enzyme interactions in
terms of the permeability and solubility of drug
Potential Drug-Drug Interactions Predicted

by BDDCS:
Class 1: Only metabolic in the intestine and
liver
Drug-drug interactions for Class 1

compounds will be primarily metabolic, with
transporter-enzyme interplay only becoming
important for those drugs where high permeability
is a result of rapid transporter uptake rather than
high Log P.
Class 2: Metabolic, efflux transporter and efflux
transporter-enzyme interplay in the
intestine.
Metabolic,
uptake
transporter, efflux transporter and
transporter-enzyme interplay in liver.
Drug-drug interactions are not limited to

enzymatic processes but can frequently be mediated
by transporter interactions and often involve
transporter-enzyme interplay for Class 2
compounds.
Following oral dosing, major significant

interactions will occur for Class 2 drugs that are
substrates for both intestinal enzymes (e.g., CYP3A,
UGTs) and intestinal apical efflux transporters (e.g.,
9
expected differences in vivo. Phenotype-genotype

discordance (as well as changes in the relationship
as a function of disease states) may be explained by
the effects of transporters on metabolic clearance;
Obtaining a realistic perspective of new
techniques (e.g., predictive ADME, QSAR,
microdosing, systems biology). If validation of the
technique primarily uses Class 1 drugs, there is no
assurance that the technique will work for NMEs.
compounds offers the potential for predicting:

1.
Major routes of elimination;
2.
Transporter effects of drug absorption;
3.
Food (High Fat Meal) effects;
4.
Transporter effects on post absorption
systemic levels and after i.v. dosing;
5.
Enzyme transporter interplay;
6.
Drug-drug interaction potential and its
relationship to enzyme-transporter interplay;
7.
Previously unexplained effects of renal
disease on hepatic metabolism that can result from
accumulation of substances (toxins) in renal failure
that modify hepatic uptake and efflux transporters;
8.
The translation of pharmacogenetic
differences in metabolic enzymes (genetic
polymorphisms) that do not always result in the
It may be easier to determine classification based on

major routes of elimination than upon permeability; an
extent of metabolism criterion for waiver of in vivo
bioequivalence studies; and how predictive algorithms
may be developed using only in vitro or in silico methods
to facilitate class assignment in BDDCS.
(Gu Y, Si DY and Liu CX)
Publication News
Chinese Herbal Medicines

Chinese Herbal Medicines, an academic journal in English edition, has been approved by the State Press
and Publication Administration in December 2008.
Approval Journal number is CN 12-1410/R. Chinese Herbal Medicines will officially be published in 2009,
in China.
Sponsored by Tianjin Institute of Pharmaceutical Research and Institute of Medicinal Plants, Chinese
Academy of Medical Sciences. Published by Chinese herbal medicines magazine. Edited by Editorial
Committee of Chinese herbal medicines
Chinese Herbal Medicines, an international journal, is the official publication Sponsored by Tianjin
Institute of Pharmaceutical Research and Institute of Medicinal Plants, Chinese Academy of Medical
Sciences. The Journals purposes are to provide a forum for the studies on Chinese herbal medicines,
traditional medicines and natural products. The Journal will accept the following contributions:
original research articles; review papers, short communications; letters to the editor; book reviews;
conference announcements and news. The journal covers the wide range from the research to
development and application of traditional medicines, herbal medicines and natural products, which
including medicinal resource, phytochemical, pharmacological, toxicological, pharmacokinetic and
therapeutic studies of active ingredients and complex formulations in experimental and clinical trials.
Original articles in English are published.
10
Xu HY et al. Asian Journal of Pharmacodynamics and Pharmacokinetics 2009; 9(1):11-25
Asian Journal of
Pharmacokinetics
ISSN
1608-2281

Recent advance on chemical compositions and pharmacodynamic and

pharmacokinetic studies of Rhizoma Coptidis
Hai-Yu Xu 1,3, Tie-Jun Zhang 2 , Xue-Yu Zhu 2,Yu-Bo Li1,3
1
Tianjin State Key Laboratory of Pharmacidymanamics and Pharmcokinetics, Tianjin Institute of
Pharmaceutical Research, Tianjin 300193, China
2
Tianjin Research Center of Modern Chinese Medicines, Tianjin Institute of Pharmaceutical Research,
Tianjin 300193, China
3
Tianjin University of Traditional Chinese Medicine, Tianjing, 300193, China.
Abstract
Rhizoma Coptidis has long been used for the treatment of gastrointestinal disorders, pyretic
toxicity, abdominal typhus, bacillary dysentery, Swelling of throat in traditional oriental
medicine in china. Recent studies found that its active constituents are alkaloids comprising
berberine, coptisine, palmaine, jatrorrhizine etc. And also Rhizoma Coptidis has been proved to
have extensive pharmacological activities including anti-cancer, anti-microbial activity,
hypoglycemic activity, LDL-lowering activity, anti-inflammatory potential and antioxygen and
free radicals scavenging etc.. In order to improve its therapeutic value and reduce its side effects,
it is necessary to study the relationship between its activity and pharmacokinetics in vitro and in
vivo. Some studies have been showed that berberine, an important ingredient in Coptis chinensis
Franch displays a linear pharmacokinetic phenomenon and has poor bioavailability. This article
summarizes chemical compositions, pharmacological actions and pharmacokinetics of Rhizoma
Coptidis.
Key words
Rhizoma Coptidis; chemical constituents; antimicrobial activity; hypoglycemic Activity;

anticancer action; anti-inflammatory; antioxygen; cardiovascular action; free radicals
scavenging; pharmacokinetics
Article history
Received 22 June 2008;
Accepted 30 December 2008
Publication data
Pages: 15;
Figures: 4;
Corresponding author
Professor Zhang Tie-Jun, Tianjin Institute of Pharmaceutical Research,308 An-Shan West Road, Tianjin,
Tables: 0;
300193, China.
References: 58;
Paper ID: 1608-2281-2009-0901011-15
E-mail: tiezheng4@sina.com.
biological activities. Rhizoma Coptidis, the dried root

and rhizome of Huang-lian named in Chinese, has
long been used for the treatment of gastrointestinal
disorders, pyretic toxicity, abdominal typhus,
bacillary dysentery, Swelling of throat in traditional
oriental medicine. Ethnopharmacologically, the
property and taste of Rhizoma Coptidis are bitter and
Introduction
Herbs have been widely employed as important
remedies all over the world[1]. Progress in science and
technology in recent decades has been made possibly
not only to isolate and to characterize the biologically
active constituents of herbs, but also to evaluate their
11
both protoberberine and quaternary-amine alkaloid.

the content of berberine is approximately 5 percent
to 8 percent ,which is the most constituent of the
total alkaloids. In addition, Rhizoma Coptidis may
contain magnoflorine, ferulaic acid (Fig 4) and so
on[3].
cold; the meridians are into heart, liver, stomach and

large intestine. The functions are to clear heat and dry
dampness, and to reduce fire and dispel toxins. It is
used to eliminate heat in the heart for insomnia,
fidget, delirium due to high fever, inflammation of
the month and tongue, and to stop diarrhea for acute
enteritis and dysentery.[2] It can be devided into 3
spicies which are Coptis chinensis Franch, (Fig 1,
www.newdruginfo.com) Coptis deltoidea C.Y.
Cheng et Hsiao and C. teeta Wall. Rhizoma Coptidis
(Fig 2, www.zgycsc.com) is thought as high grade
which has good effect and little toxin, recorded in
TCM Classic Shennong Bencaojing.
Recent in the chemical Constituents and
pharmacological studies,it indicate that the total
alkaloids is thought as main active ingredients and
berberine is the highest in the total alkaloids of
Rhizoma Coptidis. Rhizoma Coptidis and berberine
have been proved to have extensive pharmacological
activities, such as antibacterial effect, antiinflammatory effects, antioxygen and free radicals
scavenging ect. and new pharmacological activities,
such as hypoglycemic and hypolipidemic effects,
anticancer and effects on cardiovascular system etc..
In order to improve its therapeutic value and reduce
its side effects, it is necessary to study the
relationship
between
its
activity
and
pharmacokinetics in vitro and in vivo. Some studies
have been showed that berberine ,an important
ingredient in Coptis chinensis Franch displays a
linear pharmacokinetic phenomenon and have poor
bioavailability.
In this review paper, we collected some
information
about
chemical
Fig 1. Coptis chinensis Franch
Fig 2. Rhizoma Coptidis
instistuents,
pharmacological actions and pharmacokinetics of
Kobayashi[4] et al had reported that inhibitors of
Rhizoma Coptidis. The introduced information on

Rhizoma
Coptidis
will
help
us
to
topoismerase I and topoismerase II were separated
know
from Rhizoma Coptidis which could duplicate,
pharmacological actions, the mechanism of TCM,
transcribe and recombine DNA in the cell. Aqueous
secondary exploitation for Rhizoma Coptidis and its
extract
be value for studies on TCM
of
Rhizoma
Coptidis
may
make
cleavable-complex of DNA Topoismerase steadily in

mammal. Both epiberberine and groenlandicine are
Chemical Constituents
active constitutents of topoismerase I mediating DNA

cleavage and berberrubine is a specific inductor of
In Rhizoma Coptidis, major chemical

constituents are several alkaloids such as
berberine,coptisine,
palmaine,
jatrorrhizine,
worenine, epiberberine (Fig 3) which of all are
topoismeras I mediating DNA cleavage. Hirano[4] et

al had separated lariciresinol and anti-ferulaic
acid-duplet hydroxyl-benzene-ethyl ester.
12
In mixed decoction of Rhizoma Coptidis and
combination of Rhizoma Coptidis with Fructus
Radix et Rhizoma Glycyrrhizae, the formers main
Evodiae,the dissolution rate of jateorhizine, coptisine,
constituent
chemical
palmatine and berberine decreased obviously. At the
constituents of Radix et Rhizoma Glycyrrhizae
same time, the dissolution rate lineared correlation
disappear
chemical
with the proportion of Rhizoma Coptidis. It could be
compound could be found in the process . Studing
concluded that Rhizoma Coptidis Alkaloids reacted
on content variation of chemical constituents during
with Fructus Evodiaes Flavonoids[7].
may
be
completely.
remained,
But
no
but
new
[6]
Berberine
Coptisine
Palmatine
Jatrorrhizine
Worenine
Epiberberine
Fig 3. Chemical structures of berberine,coptisine, palmatine, jatrorrhizine, worenine and epiberberine
Magnoflorine
Ferulaic acid
Fig 4. Chemical structures of magnoflorine and ferulaic acid
pharmacological effects including inhibition of

adipocyte differentiation, anti-cancer effects,
anti-microbial effects, LDL-lowering effects and
anti-inflammatory potential[8].
Pharmacological Actions
In Traditional Chinese Medicine, Coptidis
Rhizoma has the effects of suppressing fever,
dispelling dampness and deintoxication. Through
modern pharmacological studies, it has been proved
to have many pharmacological effects including
antimicrobial activity, hypoglycemic activity,
anti-inflammatory, antioxygen and free radicals
scavenging and anticancer effects etc..Berberine, the
major component of this herb, has also many
Antimicrobial Activity
Coptis and berberine showed wide inhibitory
actionon bafungi prorozoon and virus in vitro and in
vivo.[2] Berberine was found to be active against a
number of Gram-positive and Gram-negative bacteria,
such as Stephylococcus aureus, S. hemolylicus,
13
purging fire for removing toxin. These pharmacologic

actions attribute to antibacterial action which can
cure infectious diseases. For example, Zuo-Jin-Wan,
a Chinese medical formula including Rhizoma
Coptidis and Fructus Evodiae, is used to cure
gastrointestinal disorders, such as peptic ulcer,
chronic gastritis, functional dyspepsia etc., which are
correlated to Zuo-Jin-Wan on anti-Hp action[13].
Salmonella typhsa, Shigella dysenterial, S.

paradysenterial, and so on. Berberine sulfate showed
to be bacterial to vibrio chloral and and bacteriostatic
to Stephylococcus aureus at concentrations of 35 and
50 gmL-1, respectively. It showed bacteriadal
activity against Stephylococcus aureus at 50-500
gml-1 in culture medium. Berberine sulfate in
concentrations 10-25gml-1 inhibited the growth of
11 fugni. Oral administration of berberine sulfate at
dose of 35-700 mgkg-1 was effective in treating
candicans infections of the intestine in mice.[9]
Rhizoma Coptidis has strong antibacterial action
and broad spectrum antibacterial activities, including
Gram-positive and Gram-negative bacteria, aerobe
and anaerobe. The biologically active constituent of
C. chinensis extract is characterized as the
isoquinoline
alkaloids,such
as
berberine,
palmatine,jateorhizine,
coptisine
etc..
Those
constituents are potent inhibitors of sortase (a
bacterial surface protein anchoring transpeptidase
from Staphylococcus aureus ATCC 6538p), with an
IC50 value of 8.7 gmL-1 and have antibacterial
activity against Gram-positive bacteria with a
minimum inhibitory concentration (MIC) in the range
of 50400 gmL-1. Among the four isoquinoline
alkaloids tested, berberine chloride had the strongest
inhibitory activity[10]. George Tegos[11] et al have
reported that the activities of berberine were
considerably against the gram-positive bacterium
Staphylococcus aureus and Bacillus megaterium and
that disabling of the MDRs (multidrug resistance
pumps ) in gram-negative species leaded to a striking
increase in antimicrobial activity. Gram-negative
bacteria had an effective permeability barrier,
comprised of the outer membrane, which restricts the
penetration of amphipathic compounds, and
multidrug resistance pumps (MDRs), which extrude
toxins across this barrier. Direct measurement of the
uptake of berberine, a model plant antimicrobial,
confirmed that disabling of the MDRs strongly
increases the level of penetration of berberine into the
cells of gram-negative bacteria.
Rhizoma Coptidis have long been used for the
treatment of gastrointestinal disorders, insomnia,
refractoriness dental ulcer, tinnitus in traditional
oriental medicine[12]. According to the Basic Theory
of Chinese Traditional Medicine, it has the efficacy
of Clearing away heat, depriving evil wetness and
Hypoglycemic Activity
Rhizome Coptidis and its major constituent
berberine
have
good
hypoglycemic
and
hypolipidemic effects. The potential glucoselowering effect of berberine was noted when it was
used for diarrhea in diabetic patients. In vitro and in
vivo studies have showed its effects on
hyperglycemia and dyslipidemia. Zhang Y [14] et al
have reported that in the berberine group, fasting and
postload plasma glucose decreased from 7.0 0.8 to
5.6 0.9 and from 12.0 2.7 to 8.9 2.8 mML-1,
HbA1c from 7.5 1.0% to 6.6 0.7%, triglyceride
from 2.51 2.04 to 1.611.10 mML-1, total
cholesterol from 5.310.98 to 4.350.96 mML-1, and
low-density lipoprotein-cholesterol from 3.23 0.81
to 2.55 0.77 mML-1, with all parameters differing
from placebo significantly. The glucose disposal rate
was increased after berberine treatment. Mild to
moderate constipation was observed in five
participants in the berberine group. So berberine is
effective and safe in the treatment of type 2 diabetes
and dyslipidemia. In one clinical study, 60 patients
with type 2 diabetes were treated with berberine for
13 months, and 90% of patients showed
improvement in their clinical symptoms[15].
Recently, it has been reported that activating the
AMP-activated protein kinase (AMPK) pathway is
the underlying mechanism for berberine improving
insulin resistance, lowering blood sugar, and
correcting lipid metabolism disorders[16-19].
Berberine can reduce body weight and lipid
levels and improve insulin action .Berberine acutely
activates AMPK activity, and induces a variety of
metabolic effects consistent with
AMPK
activation. These include activation of GLUT4
translocation; increasing phosphorylation of AMPK,
ACC, and p38 MAPK; reducing lipid content in
adipocytes; increasing expression of genes involved
14
may result in reduced retention of chemotherapeutic

agents[24].
Of course, berberine also can inhibit colon
carcinoma cell growth. Li etc. thinked that the
inhibition of cell growth by huanglian was associated
with up to 8-fold suppression of cyclin B1 protein.
This resulted in complete inhibition of cdc2 kinase
activity and accumulation of cells in G2. Therefore,
the effect of huanglian on inhibiting tumor growth
seems to be mediated by the selective suppression of
cyclin B1, which results in the inhibition of cdc2
kinase activity. Inhibition of cyclin dependent kinase
(cdk) activity is emerging as an attractive target for
cancer chemotherapy[25]. Fukuda etc. used a
beta-galactosidase reporter gene system and found
berberine, an isoquinoline alkaloid effectively
inhibiting COX-2 transcriptional activity in colon
cancer cells in a dose- and time-dependent manner at
concentrations higher than 0.3 M[26]; Berberine also
inhibited arylamine Nacetyltransferase (NAT)
activity in a human colon tumor (adenocarcinoma)
cell line in a dose-dependent manner[27].
In conclusion, Rhizome Coptidis can inhibit
tumour cells growth through different mechanisms of
action. These results indicate that traditional Chinese
herbs may represent a new source of agents in cancer
therapy. Furthermore,it have many advantages such
as high safetyaffluent resourceslow prices,little
side effects ETC. So anti-tumor action of Rhizome
Coptidis in clinic will have broad and bright prospect.
in lipid oxidation; and decreasing expression of genes

involved in lipid synthesis [20].
Berberine and other AMPK agonists, such as
metformin,have no effect on blood glucose. The
chronic effects involving changes in gene expression,
which likely contribute to reducing fat cell
differentiation
and
increasing
mitochondrial
biogenesis, again contribute to lipid lowering, fat
mass reducing, and insulin sensitivity improving. A
significant reduction in the expression of lipogenic
genes in adipose tissue following treatment with
berberine either in vitro or in vivo. Moreover,
berberine inhibit adipocyte differentiation probably
by inhibiting PPAR activity which is an important
transcriptional regulator of adipogenesis in 3T3-L1
cells[21, 22].
Anticancer action
Berberine is the anti-tumor constituent of
Rhizome Coptidis and can inhibit different
tumorscell growth including colon carcinoma, liver
tumor, carcinoma of bladder and leukemia etc.. The
anti-tumor mechanism of berberine is different
according to different tumors.
Both hepatovirus and environmental pollution
are two main causative agents of liver tumor. And
nitrosamine is main material of environmental
pollution, produced by product, N-methylN-nitro-N-nitroguanidine which is larvaceous
carcinogenic factor in stomach acid environment.
Berberine
could
significantly
inhibit
the
carcinogenesis induced by 20-methylcholanthrene
(200 g0.1ml-1 per a mouse) or N-nitrosodiethylamine (NDEA; 0.02% NDEA in distilled
water, 2.5 ml to each an animal by gavage, five days
a week for 20 weeks) in a dose-dependent manner.
Administration of berberine (0.5, 2.5 or 5.0 mgkg-1)
could significantly reduce the incidence of tumour in
animals after an injection of 20-methylcholanthrene
and increase their life span compared with the
control[23]. At the same time, berberine also reduced
cell proliferation and alpha-fetoprotein expression in
human hepatoma HepG2 cells. Multidrug resistance
transporter (pgp-170) is known to be overexpressed
in Hep G2 cells. Lin etc. found that berberine could
modulate the expression and function of pgp-170 in
hepatoma cells and other hepatoma cell lines, which
suggest that treatment of tumor cells with berberine
Anti-Inflammatory action
Rhizome
Coptidis
demonstrate
anti-inflammatory effects in various experimental models.
Berberine, which has strong anti-inflammatory
effects [28-30], is a major active constituent in Rhizoma
Coptidis. Recently studies show that berberine could
inhibit active chronic inflammation, delayed type
hypersensitivity and experimental ulcerative colitis
etc..
The mechanism of anti-inflammatory activity
have not yet been clarified completely. But a lot of
studies focused on the following aspects. Firstly,
berberine could exhibite the inhibition for the
adhesion of leukocyte and endothelial cell. Leukocyte
trafficking plays an important role in inflammatory
reaction.He Yu[31] et al studied the simulative
inflammatory reaction state in vivo through inducing
15
strong superoxide- and hydroxyl radical-scavenging

activity, but exerted only a slight pro-oxidant effect[36].
In Coptis chinensis, berberine, jatrorrhizine, and
magnoflorine behaved antioxidant activities and
antiradical activities. Jatrorrhizine and magnoflorine
showed better activities than berberine not bearing
any readily abstractable hydrogen on its skeleton. The
former two showed antiperoxidative efficiency in
DOPC liposomal membrane comparable to that of an
effective scavenger of peroxyl radicalsstobadine. It
could be concluded that the favorable antioxidant
features of the hydroxylated alkaloids were most
probably ensured by the combination of reasonably
high antiradical reactivity with high lipophilicity. [37]
human poly morphonulear leucocytes(PMN) and

endothelial cells adhesion increasing by IL-1TNF
and the results showed that berberine inhibited not
only the adhesion of endothelial cell induced by IL-1
and TNF together with PMN but also the adhesion of
PMN induced by TNF together with endothelial cell.
Secondly, berberine could regulate transcription
factors related to inflammatory diseases. Bong-Hyuk
Choi et al reported that BBR reduced the mRNA
expression level of inflammation factors such as TNF,
IL-6, CRP and HP. The mRNA expression level of
HP is most significantly reduced by BBR treatments
among these. The mRNA expression levels of other
inflammatory factor are decreased by 39%, 17% and
19%, respectively[32]. Thirdly,berberine could also
inhibite inflammatory cytokine. Kuo CL et al studied
that the anti-inflammatory mechanism of berberine is
mediated through COX-2 regulation and found that
berberine induced effect occurred rapidly (3 h) as a
result of reduced COX-2 protein, but not enzyme
activity. These anti-inflammatory effects paralleled to
the in vivo results where berberine pretreatment of
Wistar rat inhibited the production of exudates and
PGE2 in carrageenan induced air pouch[33].
Effects on cardiovascular system

The antiarrythmic action of Coptis and the role
of berberine was assessed in various animal modes.
Berberine was shown to be able to raise the VFT of
the normal and ischemic myocardium in aneshetized
rats. Experiments on isolated papillary muscle and
right atrium preparations from guinea pig leart
indicated that berberine has positive inotropic and
negative effects.[2]
To observe the effects of active ingredients
from Chinese drugs for activating blood circulation
and detoxicating, including notoginseng saponins,
Coptis chinensis, giant knotweed rhizome and
rhubarb, on blood lipids and inflammatory reaction of
aortic atherosclerotic plaques in ApoE knockout mice.
ApoE knockout mice were fed with high-fat diet for
26 weeks, then they were randomized into 6 groups,
the untreated model group and the test groups treated
with various test drugs respectively. After ending the
13 weeks of treatment, all the mice were sacrificed
with their blood lipids detected, and their heart and
aorta were taken out to make slices with paraffin
embedding. Four sections from aortic root of each
mouse were chosen to measure and calculate the
percentage of lipid core (LC) in the total area of
plaque (TP) and the lipid/collagen ratio (L/C) in the
plaque by HE and Movat staining respectively, and
the mean value of the four sections was taken for
analysis. The expressions of granulocyte-macrophage
colony-stimulating factor (GM-CSF) and tumor
necrosis factor- (TNF-) in mice's aorta root were
determined by immunohistochemical staining as well.
After being treated for 13 weeks, either the
Antioxygen and Free Radicals Scavenging

Activating oxides which is produced by
sunshine, ultraviolet radiation, ionizing radiation,
chemical reaction and metabolism is very important
reason of pathologic demages,such as DNA demage,
cancer and cell ageing. So antioxygen agents play
very important roles in treatment with these diseases,
especially crude drugs are in well with people
because of strong antioxygen and low toxicity.
Schinella G R studied the antioxidant properties of
twenty medical herbs used in the traditional
Mediterranean and Chinese medicine and founded
that Coptis chinensis at a concentration of 100
microg/ml exhibited the highest scavenging activity
on the superoxide radical[34]. Moreover, many
efficacies of Coptis chinensis such as checking
diarrhea, deintoxication, anti-inflammatory etc. were
possiblly concerned with antixoygen and free radicals
scavening[35].
The aqueous extracts of Coptis chinensis
exhibited the highest potency in inhibiting rat
erythrocyte hemolysis and lipid peroxidation in rat
kidney and brain homogenates and also demonstrated
16
platelet-derived growth factor (PDGF) and Cyclin D1

expression in a transcriptional level. However,
berberine significantly attenuated MEK/ERK
activation and downstream target (Egr-1, c-Fos,
Cyclin D1 and PDGF-A) expression after mechanic
injury in vitro. Their study showed that berberine
blocked injury-induced SMC regrowth by
inactivation of ERK/Egr-1 signaling pathway thereby
preventing early signaling induced by injury in vitro.
The anti-proliferative properties of berberine may be
useful in treating disorders due to inappropriate SMC
growth. [39]
Berberine was considered to be useful in
treating some diseases of the cardiovascular system,
such as hypertension and chronic heart failure (CHF).
Hong Y et al investigated the inhibitory effect of
berberine on experimental cardiac hypertrophy,
which is regarded as a risk factor of CHF and other
heart diseases. Forty-two male SD rats were divided
into four groups: age-matched control, aortic banding
model, berberine-treated group and captopril-treated
group. Cardiac hypertrophy was induced by
suprarenal abdominal aorta constriction. These drugs
were orally administered for 8 weeks starting from 4
weeks after surgery at dosage of berberine 10 mgkg-1
and captopril 50 mgkg-1. Blood pressure (BP) was
measured four times during the period of the
experiment, and hemodynamic parameters, cardiac
index, cell size of left ventricular myocardium and
total protein of left ventricular tissue were detected 8
weeks after treatment with these drugs. The data from
the present study showed that: The BP of the aorta
banded rats was increased compared with those of the
normal (P < 0.001) and the age-matched control rats
(P< 0.001), and berberine showed no significant
effect on it; after 8 weeks of treatment with berberine,
the elevated left ventricular end diastolic pressure
(LVEDP) was slightly decreased compared with the
aortic banded rats. Meanwhile, the maximum rates of
contraction and relaxation ( dp/dtmax) was
increased (P < 0.05) and the time to reach the point
of maximum rate from beginning of contraction
(t-dp/dt) was shortened (P < 0.01), indicating that the
functions of heart, both contraction and relaxation,
were improved; cardiac growth was inhibited by
treatment with berberine. Both whole heart and left
ventricular weight were notably decreased compared
with the banded rats (P < 0.05 and P < 0.01); and the
percentage of LC in TP and the L/C ratio was

significantly lower in all the test drug treated groups
than those in the model group, respectively (P < 0.01),
especially prominent in the group treated with giant
knotweed rhizome. Although lowering of the two
indexes presented in all of the groups treated by
notoginseng saponins, Coptis chinensis and giant
knotweed rhizome, significant difference still
presented between giant knotweed rhizome treated
group vs notoginseng saponins and Coptis chinensis
treated group (P < 0.05). As for the expressions of
GM-CSF and TNF-, in comparing with the
untreated model group, significant decreasing of the
TNF- showed only in the rhubarb treated group,
while that of GM-CSF could be found in all the test
drug treated groups (P < 0.05). All the four drugs
tested in the recommended dosage can stabilize the
vulnerable plaques in ApoE knockout mice by
improving the constitution of plaque, among them,
giant knotweed rhizome and rhubarb, the drugs
possess both the actions of activating blood
circulation and detoxicating, show more significant
effect, and their mechanisms may be related to their
actions in regulating lipid metabolism and inhibiting
inflammatory reaction.[38]
We known that vascular smooth muscle cell
(SMC) proliferation plays an important role in the
pathogenesis of atherosclerosis and post-angioplasty
restenosis. Berberine is a well-known component of
Coptis chinensis, and is capable of inhibiting SMC
contraction and proliferation, yet the exact
mechanism is unknown. Therefore, Liang KW et al
investigated the effect of berberine on SMC growth
after mechanic injury in vitro. DNA synthesis and
cell proliferation assay were performed to show that
berberine inhibited serum-stimulated rat aortic SMC
growth in a concentration-dependent manner.
Mechanical injury with sterile pipette tip stimulated
the regrowth of SMCs. Treatment with berberine
prevented the regrowth and migration of SMCs into
the denuded trauma zone. Western blot analysis
showed
that
activation
of
the
MEK1/2
(mitogen-activated protein kinase1/2), extracellular
signal-regulated kinase (ERK), and up-regulation of
early growth response gene (Egr-1), c-Fos and Cyclin
D1 were observed sequentially after mechanic injury
in vitro. Semi-quantitative reverse-transcription PCR
assay further confirmed the increase of Egr-1, c-Fos,
17
berberine. However, no significant concentration

dependency for the permeability (Papp) of berberine
was observed for B-A transport over a concentration
range of 5-300 M, indicating that the Km value of
berberine for the carrier system is greater than 300
M. Well-documented P-glycoprotein (P-gp)
substrates such as verapamil, daunomycin, and
rhodamine123 inhibited the B-A flux of berberine,
whereas tetraethylammonium and taurocholate did
not, suggesting that P-gp is involved in the transport.
For the case of daunomycin, the B-A flux, but not the
apical-to-basolateral flux, was significantly increased
after pretreatment of the cell monolayers with
berberine. In addition, the uptake of 1 microM
daunomycin into Caco-2 cells was decreased as a
result of this pretreatment. These results suggest that
the repeated administration of berberine may
up-regulate P-gp functions in Caco-2 cells. If this
occurs in the gastrointestinal epithelial cells, the
repeated administration of berberine may reduce the
gastrointestinal absorption of P-gp substrates
including chemotherapeutic agents such as
daunomycin.[42]
Berberine is an important ingredient in Coptis
chinensis Franch but has been shown to have poor
bioavailability in the dog. The aim of this study was
to use the P-glycoprotein (P-glycoprotein) inhibitors
cyclosporin A, verapamil and the monoclonal
antibody C219 in in vivo and in vitro models of
intestinal absorption to determine the role of
P-glycoprotein in berberine absorption. In the rat
recirculating perfusion model, berberine absorption
was improved 6-times by P-glycoprotein inhibitors.
In the rat everted intestinal sac model, berberine
serosal-to-mucosal transport was significantly
decreased by cyclosporin A. In Ussing-type chambers,
the rate of serosal-to-mucosal transport across rat
ileum was 3 times greater than in the reverse
direction and was significantly decreased by
cyclosporin A. In Caco-2 cells, berberine uptake was
significantly increased by P-glycoprotein inhibitors
and by monoclonal antibody C219. P-glycoprotein
appears to contribute to the poor intestinal absorption
of berberine which suggests P-glycoprotein inhibitors
could be of therapeutic value by improving its
bioavailability.[43]
In order to investigate the detailed
pharmacokinetics of berberine and its mechanisms of
cell size of left ventricular myocardium was

significantly reduced (P < 0.001) and the total protein
of left ventricular tissue was slightly down-regulated
by treatment with berberine. These results suggest
that berberine can improve abnormal cardiac function
and can prevent the development of left ventricular
hypertrophy induced by pressure-overload. This
indicates that it may have therapeutic potential in the
treatment of CHF.[40]
Pharmacokinetics studies
It has been reported that berberine is valuable
for long-term treatment of ventricular premature
beats (VPBs) and leads to a decrease in mortality for
patients with congestive heart failure (CHF). In order
to improve its therapeutic value and reduce its side
effects, it is necessary to study the relationship
between its activity and plasma concentration in
patients with CHF. Patients with CHF were treated
with conventional therapy for 2 weeks. Immediately
after the data from a dynamic electrocardiogram
(DCG) and left ventricular ejection fraction (LVEF)
were obtained, 1.2 gd-1 of oral berberine was given.
After 2 weeks of berberine therapy, the DCG data
and LVEF were reassessed and the plasma berberine
concentration was measured by HPLC. Plasma
samples were pretreated by extraction with
chloroform. Berberine in all samples was determined
using a mu Bondapak C18 column, a mobile phase of
acetonitrile: 0.02 molL-1 phosphoric acid (45:55, v/v),
and a UV detector at 346 nm. The mean recovery was
96.5%. The linear range was 40-1600 ngmL-1. The
detection limit for berberine in plasma was 0. 4 ng.
The decrease in frequency and complexity of VPBs
and the increase in LVEF in patients with plasma
berberine concentrations higher than 0.11 mgL-1
were more significant than at concentrations lower
than 0.11 mgL-1 (P< 0.01 vs P < 0.05).[41]
In the study to investigate the mechanisms by
which berberine is transported in the secretory and
absorptive directions across Caco-2 cell monolayers,
the basolateral-to-apical (B-A) flux was 30-fold
greater than the apical-to-basolateral flux and
temperature dependent (i.e., drastic decrease at 4
degrees C compared with 37 degrees C). The above
results suggest the involvement of a carrier-mediated
active transport mechanism for the B-A transport of
18
determined and the accuracy of intra- and inter-day

assays from rat samples were less than 7% RSD.
Following intravenous administration of C. rhizoma
extract at a dose of 10.2 mg/kg containing 3 mg/kg
berberine, berberine in the plasma eliminated rapidly
t1/2 =1.13 h). However, berberine in the hippocampus
increased rapidly t1/2=0.215 h), peaked at 3.67 h with
a concentration of 272 ngg-1, and had a slow
elimination rate t1/2 =12.0 h), which suggests that
berberine could have a direct action on neuron and
accumulate in the hippocampus. This study first
showed the pharmacokinetic characteristics of
berberine in rat hippocampus and the kinetic
characteristics of berberine are dissimilar in the
hippocampus and plasma.[45]
To study the effects of berberine (BBR) on the
blood concentration and pharmacokinetics of
cyclosporin A (CsA) in renal-transplant recipients., a
randomized and controlled clinical trial was carried
out in 52 renal-transplant recipients were treated with
CsA and 0.2 g BBR three times daily for 3 months,
while another 52 subjects received CsA without BBR
co-administration. Blood trough concentration of
CsA and biochemistry indexes for hepatic and renal
functions were determined. For the pharmacokinetic
study, six renal-transplant recipients were included
with a 3-mg/kg dosage of CsA twice daily before and
after oral co-administration of 0.2 g BBR three times
daily for 12 days. The trough blood concentrations
and the ratios of concentration/dose of CsA in the
BBR-treated group increased by 88.9% and 98.4%,
respectively, compared with those at baseline (P <
0.05). As for the BBR-free group, they rose by 64.5%
and 69.4%, respectively, relative to those at baseline
(P < 0.01). Nevertheless, the final blood concentrations and the ratios of concentration/dose of CsA
in BBR-treated patients were still 29.3% and 27.8%,
respectively, higher than those in BBR-free patients
(P < 0.05). No significant effects on liver or renal
functions were observed under co-administration of
BBR. After co-administration of BBR in 6 patients
for 12 days, the mean AUC of CsA was increased by
34.5% (P < 0.05). The mean time taken to reach the
peak blood concentration (Tmax) and the mean
half-life (t1/2) of CsA were increased by 1.7 h and 2.7
h, respectively (P < 0.05). The average percentage
increases in the steady-state drug concentration (Css)
and minimum blood concentration (Cmin) were 34.5%
hepatobiliary excretion, an in vivo microdialysis

coupled with HPLC was performed. In the control
group, rats received berberine alone; in the
drug-treated group, 10 min before berberine
administration, the rats were injected with
cyclosporin A (CsA), a P-gp inhibitor; quinidine,
both organic cation transport (OCT) and P-gp
inhibitors; SKF-525A (proadifen), a cytochrome
P450 inhibitor; and probenecid to inhibit the
glucuronidation. The results indicate that berberine
displays a linear pharmacokinetic phenomenon in the
dosage range from 10 to 20 mgkg-1, since a
proportional increase in the area under the
concentration-time curve (AUC) of berberine was
observed in this dosage range. Moreover, berberine
was processed through hepatobiliary excretion
against a concentration gradient based on the
bile-to-blood distribution ratio (AUCbile/AUCblood);
the active berberine efflux might be affected by P-gp
and OCT since coadministration of berberine and
CsA or quinidine at the same dosage of 10 mgkg-1
significantly decreased the berberine amount in bile.
In addition, berberine was metabolized in the liver
with phase I demethylation and phase II
glucuronidation,
as
identified
by
liquid
chromatography/tandem mass spectrometry. Also, the
phase I metabolism of berberine was partially
reduced by SKF-525A treatment, but the phase II
glucuronidation of berberine was not obviously
affected by probenecid under the present study
design.[44]
In order to investigate the pharmacokinetics of
berberine in Coptidis rhizoma extract in rat
hippocampus and plasma, a simple and accurate
high-performance liquid chromatography method was
employed in this study. Berberine was determined
using a Hypersil C18 column with an isocratic mobile
phase of acetonitrile-0.05 M potassium dihydrogen
phosphate (containing 0.5% triethylamine, pH 3.0)
and with UV detection at 236 nm. The lower limit of
quantification for berberine in both hippocampus and
plasma was 24 ngml-1, and the lowest concentrations
of berberine determined in rat hippocampus and
plasma samples were 30.7 ngmL-1 at 48 h and 38.5
ngmL-1 at 4 h, respectively. The calibration curve for
berberine was linear over the concentration range
24--6000 ngmL-1. At this concentration range, the
overall recoveries (90.6--94.2%) for berberine were
19
metabolites and conjugates were evidently excreted

into the duodenum as bile. The pharmacokinetics of
Ber and the four metabolites were determined in
conventional and pseudo germ-free rats (treated with
antibiotics) after p.o. administration with 40 mgkg -1
Ber. The AUC0-limt and mean transit time values of
the metabolites significantly differed between
conventional and pseudo germ-free rats. The amounts
of metabolites were remarkably reduced in the
pseudo germ-free rats, whereas levels of Ber did not
obviously differ between the two groups. The
intestinal flora did not exert significant metabolic
activity against Ber and its metabolites, but it played
a significant role in the enterohepatic circulation of
metabolites. In this sense, the liver and intestinal
bacteria participate in the metabolism and disposition
of Ber in vivo.[48]
To investigate whether baicalin and berberine
affects the transport of nimodipine (NMD) across the
blood-brain barrier (BBB). Primary-cultured, rat
brain microvascular endothelial cells (rBMEC) were
used as an in vitro model of the BBB. When cells
became confluent, the steady-state uptake of NMD by
rBMEC with or without baicalin and berberine was
measured. The effects of baicalin and berberine on
the efflux of NMD from rBMEC were also studied.
Baicalin (2-5 gmL-1) increased the uptake of NMD,
and baicalin (10-20 gmL-1) decreased the uptake.
The steady-state uptake of NMD was higher than that
of control group in the presence of 0.01-1 gmL-1
berberine, but was lower in the presence of 2-10
gmL-1 berberine. Their results indicated that the
bidirectional effect of baicalin and berberine on the
uptake of NMD by rBMEC was found. Higher
concentration showed an inhibitory effect, and lower
concentration demonstrated an increasing effect.[49]
Hong ZY et al[50-57], School of Pharmacy,
Second Military Medical University , have studied
the pharmacokinetics of tetrahydropalmatine (THP).
They investigated the stereoselective pharmacokinetic process of THP in rats. The concentrations of
tetrahydropalmatine enantiomers in rat plasma were
determined by coupled achiral and chiral HPLC
method. The differences in plasma concentrations
and pharmacokinetic parameters between the two
enantiomers were compared by paired t-test. The
plasma levels of l-THP were always higher than those
of d-THP in eight rats. There was significant
and 88.3%, respectively (P < 0.05). In addition, the

average percentage decrease in CL/F was 40.4% (P <
0.05) and the peak-to-through fluctuation index was
significantly reduced (P < 0.01). The study showed
that BBR can markedly elevate the blood
concentration of CsA in renal-transplant recipients in
both clinical and pharmacokinetic studies. This
combination may allow a reduction of the CsA
dosage. The mechanism for this interaction is most
likely explained by inhibition of CYP3A4 by BBR in
the liver and/or small intestine.[46]
A rapid and sensitive LC-MS/MS method was
developed to simultaneously determine berberine,
palmatine and jatrorrhizine in rat plasma. After
mixing
with
the
internal
standard
(IS)
tetrahydropalmatine, plasma samples were pretreated
by protein precipitation with acetonitrile-methanol
(1:2, v/v). Chromatographic separation was carried
out on a C18 column using a mixture of water
(containing 0.1% formic acid) and acetonitrile (30:70,
v/v) as mobile phase. The detection was performed
by selected reaction monitoring (SRM) mode via
electrospray ionization (ESI) source operating in the
positive ionization mode. The method was linear over
the concentration range of 1.0-250.0 ngmL-1 for all
components. The intra- and inter-day precision values
were less than 14.6% and the deviations were within
4.0%. The fully validated LC-MS/MS method has
been successfully applied to the pharmacokinetic
study of berberine, palmatine and jatrorrhizine in rat
plasma after oral administration of coptis-evodia herb
couple. Three peaks were observed in both individual
and mean plasma-concentration curves of berberine,
palmatine and jatrorrhizine, which may be attributed
to distribution re-absorption and enterohepatic
circulation.[47]
Berberine (Ber) and its main metabolites were
identified and quantified using liquid chromatography/electrospray
ionization/ion
trap
mass
spectrometry. Rat plasma contained the main
metabolites, berberrubine, thalifendine, demethyleneberberine, and jatrorrhizine, as free and
glucuronide conjugates after p.o. Ber administration.
Moreover, the original drug, the four main
metabolites, and their glucuronide conjugates were
all detected in liver tissues after 0.5 h and in bile
samples 1 h after p.o. Ber administration. Therefore,
the metabolic site seemed to be the liver, and the
20
plasma and other brain regions. The tissue

distribution studies also revealed significant
differences between the two enantiomers in all tissues
except the lung. The highest concentrations of both
enantiomers were found in the liver. The (-)/(+)-THP
ratios in six brain regions and other tissues were
consistent with that observed in plasma indicating
that the stereoselective disposition of THP in rat brain
and other tissues reflects the situation in plasma.[52]
THP is the active component in Rhizoma
corydalis and the medicine Yuanhu-Baizhi (YB),
which consists of Rhizoma corydalis and Radix
angelicae dahuricae. The aim of this work was to
compare pharmacokinetic features of THP
enantiomers in rats dosed with racemic THP
(rac-THP), Rhizoma corydalis, or YB extracts. A
single dose of rac-THP (5 mgkg-1) or extracts of
Rhizoma corydalis and YB (both equivalent to 5
mgkg-1 of rac-THP) was given orally to three groups
of Sprague-Dawley rats, respectively. Blood samples
were
collected
periodically
and
plasma
concentrations of THP enantiomers were determined
using an achiral-chiral HPLC method previously
reported, with some modifications. The Cmax ratio
(-/+) of THP was 2.91, 1.38, and 1.19, and the AUC0
approximately infinity) ratio (-/+) of THP was 2.84, 1.50, and
1.35 in rats after dosed with rac-THP, extracts of
Rhizoma corydalis and YB, respectively. The mean
AUC0 approximately infinity and Cmax of (+)-THP dosed with
YB extracts were 0.652 0.30 ghml-1 and 0.148
0.09 gml-1, significantly higher (P < 0.05) than
those dosed with rac-THP and Rhizoma corydalis
extracts. The mean AUC0 approximately infinity and Tmax of
rac-THP dosed with YB extracts were 1.500 0.56
ghml-1 and 2.12 1.1 h, significantly higher (P <
0.05) than those dosed with rac-THP or Rhizoma
corydalis extracts. These findings suggested the
stereoselectivity in pharmacokinetics of THP
enantiomers in rats was decreased when dosed in
plant form, while the AUC0-approximately infinity of
rac-THP increased when YB extracts were dosed,
confirming the compatibility in drug combination of
Rhizoma corydalis and Radix angelicae dahuricae.[53]
The main objective of this study was to
determine the brain pharmacokinetics and tissue
distribution of THP enantiomers in rats after oral
administration of racemic THP (rac-THP). Rats (5
animals/group/per time) were given a single oral dose
difference between the main pharmacokinetic

parameters of the two enantiomers. THP showed
significant stereoselective pharmacokinetics in rats
after an ig dose of the racemate.[50]
A selective chiral HPLC method coupled with
achiral column was developed and validated to
separate and quantify THP enantiomers in dog
plasma. Chromatography was accomplished by two
steps: (1) racemic THP was separated from biological
matrix and collected on a Kromasil C18 column (150
mmx4.6 mm, 5m) with the mobile phase
acetonitrile-0.1% phosphoric acid solution, adjusted
with triethylamine to pH 6.15 (47:53); (2)
enantiomeric separation was performed on a
Chiralcel OJ-H column (250 mmx4.6 mm, 5m) with
the mobile phase anhydrous ethanol. The detection
wavelength was set at 230 nm. (+)-THP and (-)-THP
were separated with a resolution factor (Rs) of at
least 1.6 and a separation factor (alpha) greater than
1.29. Linear calibration curves were obtained over
the range of 0.025-4 gmL-1 in plasma for each of
(+)-THP and (-)-THP (R2>0.999) with a limit of
detection (LOD) of 0.005 gmL-1 and the recovery
was greater than 88% for each enantiomer. The
method was used to determine the pharmacokinetics
of THP enantiomers after oral administration of
racemic THP. The results presented herein showed
the stereoselective disposition kinetics of THP in
dogs and were a further contribution to the
understanding of the kinetic behavior of THP
analogues.[51]
The main objective of this study was to
determine the brain pharmacokinetics and tissue
distribution of THP enantiomers in rats after oral
administration of racemic THP (rac-THP). Rats (5
animals/group/per time) were given a single oral dose
of rac-THP and killed after different post-treatment
times. The concentrations of THP enantiomers in
plasma, cortex, cerebellum, diencephalon, brain stem,
striatum and hippocampus were measured using a
validated chiral HPLC method coupled with an
achiral column. The pharmacokinetic profiles of the
two enantiomers in six brain regions were
significantly different. The peak concentrations (Cmax)
and AUC0-infinity values of the (-)-enantiomer were
significantly greater than the corresponding values
for the (+)-enantiomer while the striatum contained
the highest peak concentrations compared with the
21
(+)-THP and (-)-THP (R2>0.999) with a limit of

detection (LOD) of 0.005 gmL-1 and the recovery
was greater than 88% for each enantiomer. The
relative standard deviation (RSD) and relative error
values were less than 10% at upper and lower
concentrations. The method was used to determine
the pharmacokinetics of THP enantiomers after oral
administration of racemic THP. The results presented
herein showed the stereoselective disposition kinetics
of THP in dogs and were a further contribution to the
understanding of the kinetic behavior of THP
analogues.[56]
THP is a racemic mixture which contains 50%
of the (+) and 50% of (-) enantiomer. The (-)
enantiomer accounts for most of the analgesic effects.
Plasma concentrations of THP enantiomers were
analyzed by chiral HPLCon a Chiralcel OJ column
with quantification by UV at 230 nm. The method
was used to determine the pharmacokinetics of THP
enantiomers in rats and dogs after oral administration
of rac-THP or (-)-THP. The pharmacokinetic profiles
of the two enantiomers after dosing with rac-THP
were significantly different both in rats and dogs. The
mean Cmax and AUC0-infinity values in rats were 1.93
0.36 gml-1 and 6.65 2.34 ghml-1 for the (-)
enantiomer, and 1.11 0.25 gmL-1 and 2.03 0.45
ghml-1 for the (+) enantiomer. The mean Cmax and
AUC0-infinity in dogs were 1.60 0.81gmL-1 and
9.88 2.58 ghmL-1 for the (-) enantiomer, while
0.36 0.21 gmL-1 and 1.22 0.40 ghml-1 for the
(+) enantiomer. The rac-THP at 40 mgkg-1 and
(-)-THP at 20 mg/kg had very similar plasma
concentration-time profiles, and Cmax, AUC0-infinity,
and t1/2 of the (-) enantiomer in both rats and dogs,
indicating that the two treatments were equivalent
with respect to the pharmacokinetic properties of the
(-) enantiomer.[57]
of rac-THP and killed after different post-treatment

times. The concentrations of THP enantiomers in
plasma, cortex, cerebellum, diencephalon, brain stem,
striatum and hippocampus were measured using a
validated chiral HPLC method coupled with an
achiral column. The pharmacokinetic profiles of the
two enantiomers in six brain regions were
significantly different. The peak concentrations (Cmax)
and AUC0-infinity values of the (-)-enantiomer were
significantly greater than the corresponding values
for the (+)-enantiomer while the striatum contained
the highest peak concentrations compared with the
plasma and other brain regions. The tissue
distribution studies also revealed significant
differences between the two enantiomers in all tissues
except the lung. The highest concentrations of both
enantiomers were found in the liver. The (-)/(+)-THP
ratios in six brain regions and other tissues were
consistent with that observed in plasma indicating
that the stereoselective disposition of THP in rat brain
and other tissues reflects the situation in plasma.[54]
To investigate the stereoselective pharmacokinetic
process of THP in rats, the concentrations of THP
enantiomers in rat plasma were determined by
coupled achiral and chiral HPLC method. The plasma
levels of l-THP were always higher than those of
d-THP in eight rats. There was significant difference
between the main pharmacokinetic parameters of the
two enantiomers. THP showed significant
stereoselective pharmacokinetics in rats after an ig
dose of the racemate.[55]
A selective chiral HPLC method coupled with
achiral column was developed and validated to
separate and quantify THP-enantiomers in dog
plasma. Chromatography was accomplished by two
steps: (1) racemic THP was separated from biological
matrix and collected on a Kromasil C18 column (150
mmx4.6 mm, 5 m) with the mobile phase
acetonitrile-0.1% phosphoric acid solution, adjusted
with triethylamine to pH 6.15 (47:53); (2)
enantiomeric separation was performed on a
Chiralcel OJ-H column (250 mmx4.6 mm, 5 m)
with the mobile phase anhydrous ethanol. The
detection wavelength was set at 230 nm. (+)-THP and
(-)-THP were separated with a resolution factor (Rs)
of at least 1.6 and a separation factor greater than
1.29. Linear calibration curves were obtained over
the range of 0.025-4 gmL-1 in plasma for each of
Application of Rhizoma Coptidis

Rhizoma Coptidis is used in traditional Chinese
medicine for long history in China. The earliest
record was listed in Shengnong Bencaojing. Five to
seven year old roots or rhizomes of Coptis chinensis
Franch, Coptis deltoidea C.Y. Cheng et Hsiao or C.
teeta Wall are the medicinal use in in traditional
Chinese medicine.
22
Rhizoma Coptidis is a very useful herb in

traditional oriental medicine. Cheng[58] did statistics
of 13 reciping books before the Song Dynasty in
which Chinese medicinal formulaes including
Rhizoma Coptidis were approximate 5 percentage of
all. There are many known formulaes having
continued to be used until now such as Zuojinwan
(including Rhizoma Coptidis and Fructus Evodiae, 6
1)Fanzuojinwan (including Rhizoma Coptidis and
Fructus Evodiae 1 6), Coptidis Decoction for
Detoxification(including Rhizoma Coptidis, Radix
Scutellariae, Cortex Phellodendri and Gardenia
jasminoides Ellis).
Most chemical constituents of Rhizoma Coptidis
had been found and total alkoids had been thinked as
main active components espcially berberine.
Pharmacological action of Coptis chinensis and
berberine had been studied systematically.Both in
Traditional Chinese and Western Medicine,berberine
was widly applied to treat with infectious diseases
including not only mainly gastrointestinal bacterial
infection but also respiratory infection and
dermatologic infection. Recently, new pharmacologic
action such as anticancer, hypoglycemic activity
improving function of the circulatory system are
more and more paid attention to by people. With the
development of the living standard of the people and
aging of societal population, the incidence of
diabetes cardiovascular disease and hypertension
were increasing and both Coptis chinensis and
berberine have good efficacy to treat with these
diseases. Bescause of extensive pharmacologic
effects and affluent resource of Coptis chinensis in
China , it is worth doing more in-depth research on
its chemical composition and pharmacologic action
and making full use of it.
2.
Liu CX, Xiao PG and Li DP. Modern Research and

Application of Chinese Medicinal Plants. Hong Kong, Hong
Kong Medical Publisher. 2000; 296-300
3.
Kuang HX. Chinese Medicine Chemistry. Beijing, China

Press of Traditional Chinese Medicine,2003.
4.
Kobayashi Y,Yamashita Y,Fujii N,Takaboshi K,Kawakami

T,Kawamura M,Mizukami T,Nakano H.In-hibitors of DNA
toposiomeraes I and II isolated from the Coptis Rhizomes.
Planta Med. 1995; 61(5):414-417.
5.
Xu N. The Scavenger of Free Radical is seprated from

Rhizoma Coptidis.Foreign Medical Science Traditional
Chinese Medicine. 1998; 20(6):30-30.
6.
Li SJ, Wang F, Zhao JJ ,Chen H, Wu JG.. Study on the

Interactions
between
Glycyrrhiza.Guang
Rhizoma
Pu
Xue
Coptidis
Yu
and
Guang
Radix
Pu
Fen
Xi.2007;27(4):730-734.
7.
Peng MX, Wu YJ, Cheng YY. Study on the Solubility

Changes of Rhizoma CoptidisMain Chemical Constistuents
Influenced by the Proportion of Rhizoma Coptidis and
Fructus Evodiae. China Journal of Chinese Materia Medica.
2003; 28(7):629-631.
8.
Bong-Hyuk Choi, In-Sook Ahn, Yu-Hee Kim, Ji-Won Park,

So-Young
Lee,
Chang-Kee
Hyun,Myoung-Sool
Do.
Berberine reduces the expression of adipogenic enzymes

and
inflammatory
molecules
of
3T3-L1
adipocyte.
Experimental and Molecular Medicine. 2006;38(6):599-605.

9.
Tang WC, and Erirenbrand G. Chinese Drugs of Plant
10.
Kim SH, Shin DS, Oh MN, Chung SC, Lee JS, Oh KB.
Origin. Berlin, Springer-Verlag, 1992; 361-372.

Inhibition of the Bacterial Surface Protein Anchoring
Transpeptidase
Sortase
by
Isoquinoline
Alkaloids.
Bioscience Biotechnology and Biochemistry. 2004; 68

(2) :421-424.
11.
George Tegos, Frank R. Stermitz, Lomovskaya O, Lewis K.

Multidrug Pump Inhibitors Uncover Remarkable Activity of
Plant Antimicrobials. Antimicrob Agents Chemother. 2002;
46(10): 3133-3141.
12.
Chen J. The clinical application of Pinelliae Decoction for

Purging
Stomach-Fire.Tianjin
Traditional
Chinese
Medicine.1985;(5):37.
13.
Zhou H, Li P. Summerise on Anti-HP Infection with

Propolis Zuojinwei Trilink. Journal of Yunnan College of
Traditional Chinese Medicine. 2006; 29(6):53-55.
14.
Acknowledgement
This project was
supported by National Key Technologies R&D
Program of China in the Eleventhh Five-year Plan
under
Grant
No.
2006BAI06A01
and
No.2007BAI41B06.
Zhang Y, Li X, Zou D, Liu W, Yang J, Zhu N, Huo L, Wang

M,Hong J, Wu P, Ren G, Ning G . Treatment of type 2
diabetes and dyslipidemia with the natural plant alkaloid
berberine. J Clin Endocrinol Metab. 2008; 93(7):2559-65.
15.
Ni YX. Therapeutic effect of berberine on 60 patients with

type II diabetes mellitus and experimental research. Zhong
Xi Yi Jie He Za Zhi.1988; 8:711-713.
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Deng Y, Liao Q, Li S, Bi K, Pan B, Xie Z. Simultaneous

determination of berberine, palmatine and jatrorrhizine by
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fates
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pharmacokinetics
and
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germ-free
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2006; 27(3):111-7.
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in dogs. J Chromatogr B Analyt Technol Biomed Life Sci.
Hong ZY, Fan GR, Chai YF, Yin XP, Wen J, Wu YT.
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pharmacokinetic study of tetrahydropalmatine enantiomers
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pharmacokinetics
and
tetrahydropalmatine
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tissue
distribution
in
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after
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including Rhizoma Coptidis. Chinese Traditional Patent
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oral
administration of the racemate. Biopharm Drug Dispos.
25
Tianjin Centre for Drug Safety Evaluation and

Research
Good Laboratory Practice certification
Tianjin Centre for Drug Safety Evaluation and
built up a perfect quality-assured system. They also agreed
Research affiliated in Tianjin Institute of Pharmaceutical
that the studies on the safety evaluation, the arrangement,
Research has already passed the Good Laboratory Practice
and the operation in the Centre could be carried on more
(GLP) certification by State Food and Drug Administration
smoothly and better under the desirable GLP standard.
(SFDA), which has filled the gap in Tianjin municipality
In the light of the comments of Mr.Xianglong DAI
since there was no institution for GLP safety evaluation
and Mr.Dongliang YANG, etc., the mayors and leaders of
before.
Tianjin municipality, Tianjin Centre for Drug Safety
SFDA designated the experts who are engaged in the
Evaluation and Research has been built and listed in the
on-the-spot inspection for the Centre according to the latest
first-approval item in the support of Tianjin Special Fund
revision of GLP certification method (Tentative Version).
for Technical Innovation. In Tianjin, the Centre has set up
The inspection team members learnt the reports on the GLP
the
construction and checked the complete soft and hard
(GAP-GLP-GCP-GMP), become an essential technical
conditions as well as the reference materials of non-clinical
platform in the innovative system of novel drugs, and will
safety evaluation for drug experiments. Based on the GLP
make the innovatory potential of novel drugs promote
standard, all the experts confirmed that the Centre has
greatly. Also it will provide a technical prop for the
modern
integrated
pharmaceutical
Industry
already performed 493 items of various experiments for
sustainable developing in modern pharmaceutical Industry
safety evaluation among 228 novel drugs, drafted and
and the establishment of China International Innovation
carried out 744 standard operating procedures (SOP), and
Zone for Biomedicine.
26
Cheng TF et al. Asian Journal of Pharmacodynamics and Pharmacokinetics 2009; 9(1):27-49
Asian Journal of
Pharmacokinetics
ISSN
1608-2281

Attention on research of pharmacology and toxicology of

nanomedicines
Tie-Feng Cheng1, 2, Yong-Da Sun3,4,, Duan-Yun Si2,3Chang-Xiao Liu2,3
1
Key Laboratory for Special Functional Materials, Henan University, Kaifeng, 475001, China
2
Research Center of New Drug Evaluation, The State Key Laboratories of Pharmacodynamics and
Pharmacokinetics, Tianjin Institute of Pharmaceutical Research, Tianjin 300193, China
3
Research Center of Biological Evaluation of Nanopharmaceuticals, China National Academy of
Nanotechnology and Engineering, Tianjin, 300457, China
4.
Tianjin Crystec Pharmaceutical technology Ltd, Tianjin, 300457, China
Abstract
In the 21st century, nanoscience and nanotechnology obtains the world attention due to this
revolutionary theory and technical features. Nanoscience and nanotechnology cover the theory
and technology of physics, chemistry, medicine, material science, biomedical engineering and
biology, therefore, they have no less contribution to science and technology as biotechnology
and information technology. Recent years have witnessed the rapid development of Chinas
nanoscience and nanotechnology with widespread influence. It was attended by scientists of the
world. Research, development and application of nanotechnology research in China can be
summed up in three characteristics: the first, China government in support of sustainable
development; the second, significant academic achievements, and the third, a clear consensus on
sustainable development for nanoscience and nanotechnology research and development. In this
review paper, we discussed the pharmacology and toxicology of nanomedicines, and presented
some issues on research and development and application of nanomedicines in the future.
Key words
Nanoscience; nanotechnology; nanomedicines; pharmacology; toxicology; China; sustainable

development; academic achievements
Article history
Received 26 December 2008;
Publication data
Pages: 23;
Tables: 2;
Accepted 27 February 2009
Figures: 1;
References: 47;
Paper ID: 1608-2281-2009-0901027-23
Professor Chang-Xiao Liu, Tianjin Institute of Pharmaceutical Research, 308 An-Shan West Road, Tianjin,
300193, China.
E-mail: liuchangxiao@vip.163.com.
and nanotechnology, as a newly emerging leading-edge
Introduction
discipline, cover many fields such as physics, chemistry,
Nanoscience and nanotechnology has attracted
medicine, material science, biomedical engineering and
full attention of scientists around the world due to the
biology, therefore, the rapid development of nanoscience
breakthrough theory and technical feature. Nanoscience
and nanotechnology has contributed more and more deep

27
Frederick, National Institutes of Health, USA, and

Professor Michael M. Gottesman (National Cancer
Institute, National Institutes of Health, USA), as
co-chairmen of this meeting, lead scientists from
China and USA to discussed nanoscience,
nanotechnology and nanomedicines for cancer
treatment. An emerging field that takes full
advantage of expertise and research approaches
from such academic disciplines as nanotechnology,
biology,
chemistry,
physics,
medicine,
pharmaceutics and public health. No single
discipline can deal with the new field characterized
with a lot of interdisciplinary and comprehensive
studies. It is both a topic at the cutting-edge of
science development and an important social issue
closely related to peoples health and environment,
offering unlimited opportunities for innovation. The
meeting was focused on nanomedicines and
nanotechnology for cancer treatment, environmental
health of nanotechnology and its safety, and the
strategy
and
policy
for
nanotechnology
development. Central topics: (1) Molecular Basis of
Nanomedicine, (2) Development of Smart
Nanoparticles, biomarker and targeted delivery for
cancer therapy and imaging, (3) Nanotechnology :
Path to the clinic promises and hurdles and (4) The
molecular basis for engineered nanomaterial
interactions with human health and the environment.
The 4 topics provided exciting information for
nanomedicine researches in basis and clinical
research strategy for further development of
nanomedicines.[3]
knowledge to other discipline as biotechnology and

information technology information technology.[1]
Recently, nanotechnology proves its diverse

applications to take up the international market in
areas of biomedicine, informatics, energy resource,
astronautics, oceanography and national defense,
and so on. These applications offer huge economic
and technological potentialities. Current advances in
nanoscience and nanotechnology have led to the
development of the new field of nanomedicine,
which includes many applications of nanomaterials
and nanodevices for diagnostic and therapeutic
purposes. At the Third Annual Meeting of the
American Academy of Nanomedicine held at the
University of California San Diego, in San Diego,
USA, during September 7-8, 2007. The discussion
was
focused
on
successful
translational
nanomedicine: from bench to bedside. There were
four keynote lectures and eight scientific
symposiums in this meeting. The researchers and
investigators reported the results and process of
current nanomedicine research and approaches to
clinical applications. The meeting provided exciting
information for nanomedicine clinical-related
researches and strategy for further development of
nanomedicine research which will be benefits to
clinical practice.[2]
Recent years have witnessed the rapid
development of Chinas nano-science and
technology with widespread influence. It was
attended by scientists of the world. On January
2008, a proposal to convene a Sino-US symposium
on nanomedicine and nanobiology was jointly made
by Dr. Elias Zerhouni (Director of the National
Institutes of Health, USA), Dr. John E. Niederhuber
(Director of the National Cancer Institute, USA and
Dr. Samuel Wilson (Director of the National
Institute of Environmental Health Sciences, USA).
To integrate outstanding research forces in China
and carry out exchanges with scholars of the world,
in particular US, the Xiangshan Science Conference
(the 331st Xiangshan Science Conference) on
nanotechnology and nanomedicine for cancer
treatment held from 21 to 24 October in Beijing,
China. Professor Zhao YL (Chinese National
Research Center of Nanosceince), Professor Robert
P. Blumenthal (Center for Cancer Research
Nanobiology ProgramNational Cancer Institute
Research and development of nanoscience

and nanomedicines in China
Now
in
China,
nanoscience
and
nanotechnology become ever more consequential in
our lives, we in the scientific community need to
better inform and educate the public about the
transformations this new nano era is likely to bring.
Among the fields that have enjoyed particularly
rapid development in China in the past decade are
nanoscience and nanotechnology. These terms refer
to the growing knowledge base and technical
framework for understanding and manipulating
matter on nanometer scale ranging from the atomic
to the cellular. Like many other countries, we in
28
From 1990 to 2005 alone, over 1200 projects were

implemented. In addition, during this period, NSFC
approved nearly 1000 grants for small-scale projects
in related areas. With so much going on in
nano-related R&D in so many different places in
China, we created in 2000 the National Steering
Committee for Nanoscience and Nanotechnology to
oversee national policy and planning in these
arenas.[4]
Moving forward in nanoscience and
nanotechnology requires a particularly wide
spectrum of skills and knowledge. The demand for
multidisciplinary
research
platforms
with
components assembled from academia and industry
and that also have educational functions has become
especially strong in recent years. According to
incomplete statistics, more than 50 universities, 50
institutes and over 300 industry enterprises in China
have engaged in nanoscience and nanotechnology
research and development, with the involvement of
more than 3000 researchers across China. To move
forward and become more competitive in
nanoscience and nanotechnology, China needs to
continue to expand its now-limited research
infrastructure. In some areas, such as nanoscale
devices with novel electronic and optoelectronic
features, efforts to consolidate resources to tackle
key technological issues are under way. Efforts
have also been made to pursue industrial-scale
production of nanomaterials, such as CNTs,
polymeric nanocomposites, and nanoparticle
materials, with the intention of opening up
opportunities for new businesses to sprout and grow.
The nanoscience and nanotechnology community in
China has made remarkable advances across the
research and development spectrum, from
fundamental scientific research to studies into the
potential
societal
implications
of
new
nanotechnologies. China still has a long way to go
to improve the overall competitiveness of its
nanoscience and nanotechnology enterprise.[4]
During the Ninth Five-year Plan period
China expect that the development of nanoscience

and nanotechnology will greatly affect many areas
of scientific research and industrial development,
and many aspects of everyday life.[4]
Research, development and application of
nanotechnology research in China can be summed
up in three characteristics: the first, the government
in support of sustainable development; the second,
significant academic achievements, and the third, a
clear consensus on nano-innovation.
The government in support of sustainable
development
When the concept of nanoscience and
nanotechnology was first introduced in the 1980s, it
was received favorably in China. The initial interest
was in part stimulated by the development of new
tools and techniques for observing materials on the
nanoscale, especially scanning probe microscopes
(SPMs). Soon after the concept began trickling
through the scientific ranks, the Chinese Academy
of Sciences (CAS), the National Natural Science
Foundation of China (NSFC), and the State Science
and Technology Commission (SSTC)/ the Ministry
of Science and Technology (MOST) began funding
nanoscience-related work and activities. China also
has helped those who work in nanoscience and
nanotechnology to develop their sense of being part
of a new research and development community.
Since 1990, for example, dozens of international
and domestic conferences in the field have been
held in China. These conferences addressed a wide
range of topics in nanoscience and nanotechnology
and attracted wide attention and public interest. In
the 1990s, support for the development of
nanoscience and nanotechnology increased
substantially, largely through several major
initiatives. In 1990, for example, SSTC launched
the nearly decade-long "Climbing Up" project on
nanomaterial science. In 1999, MOST started a
national basic research project (973 Plan) entitled
"Nanomaterial and Nanostructure" and has been
funding basic research on nanomaterials, such as
nanotubes, ever since. China National High
Technology Plan(863 Plan), which encompasses
many categories of technology, has included a
series of projects for nanomaterial applications.
(1996-2000), the national 863 Plan supported by

China government starts the projects of improving
nanobiotechnology; during the Tenth Five-year
period (2001-2005) national 863 and 973
Plans
29
and
National
Natural
science
made
nanoscience, nanotechnology, and nanomedicine

studies
as
priority
subjects
to
support
A clear consensus on nano-innovation

Facing on the arduous in nanoscience and
nanotechnology research, and the risks of
nanopharmaceutical industry, we think in this area
should pay attention on four-oriented development,
according to China's national conditions. The first is
the practice research-oriented, combining basic and
application. The second is to set up different
professional disciplines for the research bases and
to strengthen the efficacy, safety, and the
industrialization, and feasibility study of
nanomedicines in order to ensure sustainable
development. The third, focus on solving the
challenging problem of the difficult implementation,
and
breakthroughs
in
nanoscience
and
nanotechnology. The fourth, the complexity in
research and development of the new technologies
requires to support with long-term development,
and to know the risks for technological
transformation to industrialization.
by
government. During the Eleventh Five-year Plan

period (2006-2010), the state is increased support
for nanoscience and nanotechnology research, the
annual input on
research,
one
billions of funds to carry out

of
three
subjects
of
nanopharmaceuticals are listed of the research plan.

Significant academic achievements
The scientific output of Chinese nanoscientists
is becoming ever more significant. According to the
Scientific Citation Index, CAS ranked fourth in the
world in total number of citations among those
institutions and universities that published more
than 100 nanotechnology papers from 1992 to 2002.
Another recent analysis of nanoscience productivity
around the world ranked China at the top for the
first 8 months of 2004. This should not give the
Chinese research community reason to be overly
optimistic, however. The volume of published
papers and total number of citations is only one
indicator of the value of research. Another is the
impact, or the number of citations per paper. From
2001 to 2003, the number of citations per
nanotechnology paper published by scientists in the
United States, Germany, Japan, and China was
about 6.56, 4.54, 3.7, and 2.28, respectively.
Since 2006, Chinese basic research papers on
nanoscience and nanotechnology and total number
of citations have become the world's second largest,
behind only the United States. According to
statistical data from www.cnki.net (2004-2008),
Chinese scholars published a large number of
nano-page research thesis in Chinese academic
journals (as shown in Table 1).
Nanotoxilogy and Nanopharmacology

Nanotechnology is a newly fashionable field
but in the world of drug development it is certainly
not new. Nanotechnology has a vital role to play in
realizing cost-effective diagnostic, therapeutic and
prevent tools. The applications of nanotechnology
for treatment, diagnosis, monitoring and control of
biological systems have recently been referred to as
nanomedicine. The nanocarriers have been made of
safe materials, including synthetic biodegradable
polymers, lipids and polysaccharides. Nanomedicines (nanopharmaceuticals) are the convergence of nanotechnology and biotechnology and an
important
component
of
nanotechnology.
Application of nanotechnology is just started in
traditional Chinese medicines.
Nanopharmaceuticals or "Nanomedicines" can
be developed either as drug delivery systems or
biologically active drug products. They comprise
nanometre size scale complex systems, consisting
of at least two components, one of which being the
active ingredient. Drug delivery is an
interdisciplinary area of research that aims at
making the administration of complex new drugs
feasible, as well as adding critical value to the drugs
that are currently in the market. At present, one of
Table 1. Papers published in Chinese journals from

2004 to 2008
Year
Nanoscience and
nanotechnology
Nanomedicine
2004
2005
2006
2007
2008
179
226
280
169
235
11
22
41
18
7
30
staff safety and waste management in environment.

The development of in vitro models of testicular
toxicity may provide important tools for
investigating specific mechanisms of toxicity in the
testis. Although various systems have been reported,
their application in toxicological studies has been
limited by the poor ability to replicate the complex
biochemical, molecular, and functional interactions
observed in the testis. In vitro models have been
established, and some of them have tried to
reproduce the complex interactions that take place
between the different germ cells. These models are
limited by the poor viability of freshly isolated germ
cells. So the development of a germ-line stem cell is
of great interest. After previous studies to develop
an immortalized cell line[6-8] with promising
application in the study of testis toxicity. In vivo
system for evaluation on safety and toxicology is
very importance. This evidence of physiologically
significant histopathological changes clearly
indicates the potential of these nanomaterials for
human toxicity at realistic doses.
Nanoscale materials are seeming application in
direct interventions to improve public health both
through therapeutic strategies and environmental
remediation. Recent years have seen the emergence
of nano-engineered drug delivery strategies.
Approval of abraxane, a nano-formulation of taxol
for the treatment of breast cancer, was received by
Food and Drug Administration (FDA), USA. This
protein nano-bead conjugated pharmaceutical has
increased water solubility allowing for elimination
of the toxicity associated with the solvent vehicle
and improved therapeutic index. The benefit of
abraxane relies on the nanoscale formulation rather
than on the emergent properties of the
nanomaterials as a therapeutic modality.[9] Powers et
al pointed
out
that basis nanoparticle
characterization techniques are discussed, along
with some of the issues and implications associated
with measuring nanoparticle properties and their
interactions with biological systems. Recommendations
regarding
how
to
approach
nanomaterial characterization include using proper
sampling and measurement techniques, forming
multidisciplinary teams, and making measurements
as close to the biological action point as possible.[10]
The science of toxicology has provided the
the most attractive areas of research in drug delivery

is the design of nanomedicines consisting of
nanosystems that are able to deliver drugs to the
right place, at appropriate times. The goal of the
present article is to review the advances we have
made in the development and characterization of
nanosystems intended to be used as drug carriers for
mucosal administration. These nanocarriers are able
to protect the associated drug against degradation
and facilitate its transport across critical and
specific barriers. Some are further able to release
the associated drug to the target tissue in a
controlled manner. These nanocarriers have been
made of safe materials, including synthetic
biodegradable polymers, lipids and polysaccharides.
The change in the physicochemical and structural
properties of engineered nanosized materials with a
decrease in size could be responsible for a number
of material interactions that could lead to
toxicological effects. At present, scientists must
accept that it is still very early in the toxicological
evaluation for nanomaterials and nanomedicines,
and few data on the safety and toxicity. The safety
evaluation of nanomedicines includes workforce
exposure limits in manufacturing process,
environment impact with general impact and to
patients after administration and safety for human
use, such as depends on route of administration,
dose and dosing frequency, as well as safety in drug
delivery relates to toxicity of drug payload. The
biomedical evaluation of nanomedicines includes
biodistribution, metabolic fate, Persistance of
non-degradable systems, Specific therapeutic issues
and immunogenicity. We must pay an attention on
the relative issues of nanomedicines with human
health and safety and toxicity to develop the
evaluation methods of nanoproducts and make
nanotechnology play a great role in the progress in
nanotechnology and medicines and medicine
engineering.[5]
Evaluation on safety and toxicology of

nanomedicines
The toxicology of nanomedicines used in
device manufacture should be considered during
their entire life cycle at stages of manufacture and
preclinical and clinical development, consumer and
31
directed toward understanding and creating

improved materials, devices, and systems that
exploit these properties. In a review, Thomas et al
reviewed that a limited subset of products that
contain nanoscale materials, assess the available
data for evaluating the consume exposures and
potential hazard associated with these products, and
discuss the capacity of US regulatory agencies to
address the potential risks associated with these
products.[11] Some of the potential impacts of
dermal exposure to nanoscale materials include the
following: (1) enhanced amount and depth of
penetration of active ingredients in cosmetic into
the skin resulting in increased activity, (2)
ingredients that are chemically unstable in air and
light (as retinal and vitamin E) may be more readily
used in topical products following encapsulation in
nanoparticles, and (3) and timed release of
ingredients may become more feasible in topical
products and could allow for improved
effectiveness equivalent to current controlled
release orally administered drugs.
foundation for understanding and studying the

interactions between chemical drugs and biology.
While the use of nanomaterials, nanomedicines/
nanopharmaceuticals is rather new in the
commercial products, the philosophical basis for
performing the toxicological evaluation of these
products is not expected to be different form other
chemical drugs.
At present, scientists must accept that it is still
very early in the toxicological evaluation for
nanomaterials, nanomedicines/nanopharmaceuticals,
and few data on the safety and toxicity. The basic
tenet of study designed to develop a study system of
toxic effects of nanomaterials, nanomedicines/
nanopharmaceuticals on biological systems is to
understand the physico-chemical characteristics of
nanomaterials, nanomedicines. Therefore, the
approach to addressing the safety and toxicity of
these products will best be conducted via
multidisciplinary terms. Many traditional methods
and approaches will likely be applicable to study of
nanomaterials,nanomedicines/nanopharmaceuticals.
Nanotechnology research and development is
Table 2. The biomedical evaluation of nanomedicines

Evaluation terms
Evaluation contents
Biodistribution
Metabolic fate
Immunogenicity
Persistance of non-degradable systems
Biocompatibility
Whole organism, cellular level

Absorption, distribution, metabolism and excretion
IgG/IgM production, cytokine induction
Possibility of lysosomal storage disease
Biological environment and toxicology and adverse
effect to patients
Therapeutic index of nanomedicines and its delivery
systems in drug delivery relates to toxicity pf drug
payload
Specific therapeutic issues
play a significant role. It should be realized that the

animal bioassays presently used in toxicity testing
are model systems for the prediction of toxicity in
humans or the environment. In the last few decades
new technologies and new knowledge have become
available. This development is the result of
intensive fundamental toxicological research and the
implementation of new methods and technologies.[12]
The biomedical evaluation of nanomedicines
includes biodistribution, metabolic fate, persistance
Due to the nanotechnology combines with

biotechnology, a newly emerging cross-disciplinary
field nanobiotechnology, this becomes the new
developing area. As the research and application of
nanotechnology, studying and understanding the
complex relationship between nanomaterials/
nanomedicines and biological system will show
special important to environmental, human health
and safety. Criticism of the use of laboratory
animals for the safety testing of chemicals is
increasing, in society as a whole and also in the
scientific world. This criticism is not only limited to
ethical concerns, but scientific considerations also
of non-degradable systems, Specific therapeutic issues
and immunogenicity (Table 2).[1]
32
delivery vehicles, and other useful biological tools.

The unprecedented freedom to design and modify
nanomaterials to target cells, chaperone drugs,
image biomolecular processes, sense and signal
molecular responses to therapeutic agents, and guide
surgical procedures is the fundamental capability
offered by nanotechnology, which promises to
impact drug development, medical diagnostics, and
clinical applications profoundly (Fig 1).[14]
Intraperitoneal injection of [Gd@C82(OH)22]n

nanoparticles decreased activities of enzymes
associated with the metabolism of reactive oxygen
species (ROS) in the tumor-bearing mice. Several
physiologically relevant ROS were directly
scavenged by nanoparticles, and lipid peroxidation
was inhibited in this study. [Gd@C82(OH)22]n
nanoparticles significantly reduced the electron spin
resonance
(ESR)
signal
of
the
stable
2,2-diphenyl-1-picryhydrazyl radical measured by
ESR spectroscopy. Like-wise, studies using ESR
with
spin-trapping
demonstrated
efficient
scavenging of superoxide radical anion, hydroxyl
radical,
and
singlet
oxygen
(1O2)
by
[Gd@C82(OH)22]n nanoparticles. In vitro studies
using liposomes prepared from bovine liver
phosphatidylcholine revealed that nanoparticles also
had a strong inhibitory effect on lipid peroxidation.
Consistent with their ability to scavenge ROS and
inhibit lipid peroxidation, we determined that
[Gd@C82(OH)22]n nanoparticles also protected cells
subjected in vitro to oxidative stress. Studies using
human lung adenocarcinoma cells or rat brain
capillary endothelial cells demonstrated that
[Gd@C82(OH)22]n nanoparticles reduced H2O2induced ROS formation and mitochondrial damage.
[Gd@C82(OH)22]n nanoparticles efficiently inhibited the growth of malignant tumors in vivo. In
summary, the results obtained in this study reveal
antitumor
activities
of
[Gd@C82(OH)22]n
nanoparticles in vitro and in vivo. Because ROS are
known to be implicated in the etiology of a wide
range of human diseases, including cancer, the
present findings demonstrate that the potent
inhibition of [Gd@C82(OH)22]n nanoparticles on
tumor growth likely relates with typical capacity of
scavenging reactive oxygen species.[13]
Fig 1. Medical applications of nanotechnology. The

size and tailorability of nanoparticles may lea
to their widespread use in a variety of medical
applications.[14]
Engineered nanomaterials are at the leading

edge of the rapidly developing nanosciences and are
founding an important class of new materials with
specific physicochemical properties different from
bulk materials with the same compositions. The
potential for nanomaterials is rapidly expanding
with novel applications constantly being explored in
different areas. The unique size-dependent
properties of nanomaterials make them very
attractive
for
pharmaceutical
applications.
Investigations of physical, chemical and biological
properties of engineered nanomaterials have yielded
valuable information. Cytotoxic effects of certain
engineered nanomaterials towards malignant cells
form the basis for one aspect of nanomedicine. It is
inferred that size, three dimensional shape,
hydrophobicity and electronic configurations make
them an appealing subject in medicinal chemistry.
Their unique structure coupled with immense scope
for derivatization forms a base for exciting
developments in therapeutics. This review article
addresses the fate of absorption, distribution,
metabolism and excretion (ADME) of engineered
nanoparticles in vitro and in vivo. It updates the
distinctive methodology used for studying the
biopharmaceutics of nanoparticles. This review
addresses the future potential and safety concerns
and genotoxicity of nanoparticle formulations in
Evaluation on pharmacology of
nanomedicines
Nanotechnology manifests itself in a wide
range of materials that can be useful to medical
application. Virtually all of these materials have
been designed with chemically modifiable surfaces
to attach a variety of legends that can turn these
nanomaterials into biosensors, molecular-scale
fluorescent tags, imaging agents, targeted molecular
33
dendrimers (in various sizes, surface substituents,

and net charges) and inorganic nanoparticles,
properties of both of which can be individually
modified and optimized. In this study we examine
effects of size and surface charge on the behavior of
Au-dendrimer CNDs in mouse tumor models.
Quantitative biodistribution and excretion analyses
including 5-nm and 22-nm positive surface, 5-nm
and 11-nm negative surface, and a 5-nm neutral
surface CNDs were carried out in the B16 mouse
melanoma tumor model system. Results seen with
the 22-nm CND in the B16 melanoma model were
corroborated in a prostate cancer mouse tumor
model system. Quantitative in vivo studies confirm
the importance of charge and show for the first time
the importance of size in affecting CND
biodistribution and excretion. Interestingly, CNDs
of different size and/or surface charge had high
levels of uptake (selective targeting) to certain
organs without specific targeting moieties placed on
their surfaces. Researchers conclude that size and
charge greatly affect biodistribution of CNDs.
These findings have significance for the design of
all particle-based nanodevices for medical uses. The
observed organ selectivity may make these
nanodevices exciting for several targeted medical
applications.[17]
general. It particularly emphasizes the effects of

nanoparticles on metabolic enzymes as well as the
parenteral or inhalation administration routes of
nanoparticle formulations. This paper illustrates the
potential of nanomedicine by discussing
biopharmaceutics of fullerene derivatives and their
suitability for diagnostic and therapeutic purposes.
Future direction is discussed as well.[15]
With the rapid development of quantum dot
(QD) technology, water-soluble QDs have the
prospect of being used as a biological probe for
specific diagnoses, but their biological behaviors in
vivo are little known. Our recent in vivo studies
concentrated on the bio-kinetics of QDs coated by
hydroxyl group modified silica networks (the QDs
are 21.32.0 nm in diameter and have maximal
emission at 570 nm). Male ICR mice were
intravenously given the water-soluble QDs with a
single dose of 5 nmol/mouse. Inductively coupled
plasma-mass spectrometry was used to measure the
(111)Cd content to indicate the concentration of
QDs in plasma, organs, and excretion samples
collected at predetermined time intervals.
Meanwhile, the distribution and aggregation state of
QDs in tissues were also investigated by
pathological
examination
and
differential
centrifugation. The plasma half-life and clearance
of QDs were 19.83.2 h and 57.39.2 mlh-1kg-1,
respectively. The liver and kidney were the main
target organs for QDs. The QDs metabolized in
three paths depending on their distinct aggregated
states in vivo. A fraction of free QDs, maintaining
their original form, could be filtered by glomerular
capillaries and excreted via urine as small
molecules within five days. Most QDs bound to
protein and aggregated into larger particles that
were metabolized in the liver and excreted via feces
in vivo. After five days, 8.6% of the injected dose of
aggregated QDs still remained in hepatic tissue and
it was difficult for this fraction to clear.[16]
There is growing interest in developing
tissue-specific multifunctional drug delivery
systems with the ability to diagnose or treat several
diseases. One class of such agents, composite
nanodevices
(CNDs),
is
multifunctional
nanomaterials with several potential medical uses,
including cancer imaging and therapy. Nanosized
metal-dendrimer CNDs consist of poly(amidoamine)
Study on responses of Ferric oxide

nanoparticles:
Ferric oxide (Fe2O3) nanoparticles are of
considerable
interest
for
application
in
nanotechnology related fields. However, as iron
being a highly redox-active transition metal, the
safety of iron nanomaterials need to be further
studied. In this study, the size, dose and time
dependent of Fe2O3 nanoparticle on pulmonary and
coagulation system have been studied after
intratracheal instillation. The Fe2O3 nanoparticles
with mean diameters of 22 and 280 nm, respectively,
were intratracheally instilled to male Sprague
Dawley rats at low (0.8 mgkg-1) and high (20
mg/kg) doses. The toxic effects were monitored in
the post-instilled 1, 7 and 30 days. Our results
showed that the Fe2O3 nanoparticle exposure could
induce oxidative stress in lung. Alveolar
macrophage (AM) over-loading of phagocytosed
nanoparticle by high dose treatment had occurred,
34
Misfolding and self-assembly of proteins in

nanoaggregates of different sizes and morphologies
(nanoensembles, primarily nanofilaments and
nanorings) is a complex phenomenon that can be
facilitated, impeded, or prevented by interactions
with various intracellular metabolites, intracellular
nanomachines controlling protein folding, and
interactions with other proteins. A fundamental
understanding of molecular processes leading to
misfolding and self-aggregation of proteins
involved in various neurodegenerative diseases will
provide important information to help identify
appropriate therapeutic routes to control these
processes. An elevated propensity of misfolded
protein conformation in solution to aggregate with
the formation of various morphologies impedes the
use of traditional physiochemical approaches for
studies of misfolded conformations of proteins.
Kransnoslobodtsev et al tethered the protein
molecules to surfaces to prevent aggregation and,
with force spectroscopy using an atomic force
microscopy, probed the interaction between protein
molecules depending on their conformations.
Research results show that formation of filamentous
aggregates is facilitated at pH values corresponding
to the maximum of rupture forces. They report here
on development of a novel surface chemistry for
anchoring of amyloid (A) peptides at their
N-terminal moieties. The use of the site-specific
immobilization procedure allowed us to measure the
rupture of A-A contacts at the single-molecule
level. The rupture of these contacts is accompanied
by the extension of the peptide chain detected by a
characteristic elastomechanical component of the
force-distance curves. Potential applications of
nanomechanical studies for understanding the
mechanisms of development of protein misfolding
diseases are discussed.[20]
while the non-phagocytosed particles were found

entering into alveolar epithelial in day 1 after
exposure. Several inflammatory reactions including
inflammatory and immune cells increase, clinical
pathological changes: follicular hyperplasia, protein
effusion, pulmonary capillary vessel hyperaemia
and alveolar lipoproteinosis in lung were observed.
The sustain burden of particles in AM and
epithelium cells has caused lung emphysema and
pro-sign of lung fibrosis. At the post-instilled day
30, the typical coagulation parameters, prothrombin
time (PT) and activated partial thromboplastin time
(APTT) in blood of low dose 22 nm-Fe2O3 treated
rats were significantly longer than the controls. We
concluded that both of the two-sized Fe2O3 particle
intratracheal exposure could induce lung injury.
Comparing with the submicron-sized Fe2O3 particle,
the nano-sized Fe2O3 particle may increase
microvascular permeability and cell lysis in lung
epitheliums and disturb blood coagulation
parameters significantly.[18]
Superparamagnetic iron oxide nanoparticles
(SPIONs) are applied in stem cell labeling because
of their high magnetic susceptibility as compared
with ordinary paramagnetic species, their low
toxicity, and their ease of magnetic manipulation.
The present work is the study of CD133+ stem cell
labeling by SPIONs coupled to a specific antibody
(AC133), resulting in the antigenic labeling of the
CD133+ stem cell, and a method was developed for
the quantification of the SPION content per cell,
necessary for molecular imaging optimization. Flow
cytometry analysis established the efficiency of the
selection process and helped determine that the
CD133 cells selected by chromatographic affinity
express the transmembrane glycoprotein CD133.
The presence of antibodies coupled to the SPION,
expressed in the cell membrane, was observed by
transmission electron microscopy. Quantification of
the SPION concentration in the marked cells using
the ferromagnetic resonance technique resulted in a
value of 1.70 1013 mol iron (9.5 pg) or 7.0 106
nanoparticles per cell (the measurement was carried
out in a volume of 2 L containing about 6.16 105
pg iron, equivalent to 4.5 1011 SPIONs).[19]
Research and Application of Carbon

Nanotubes
A good representative of this fast-moving field
is the family of nanomaterials known as carbon
nanotubes (CNTs). These all-carbon tubes are just a
few nanometers in diameter, which makes them
comparable in girth to DNA molecules, and come in
either singlewalled varieties or multiwalled varieties
with a nesting of carbon shells resembling the
Mechanisms of development of protein

misfolding diseases
35
angiotensin II type 1 receptor by AFM with

functionalized tip is introduced in this article. Some
prospective methods to improve the imaging
resolution are also discussed.[22]
CNTs are nanodevices with important potential
applications in biomedicine such as drug and gene
delivery. Brain diseases with no current therapy
could be candidates for CNT-based therapies. Little
is known about toxicity of CNTs and of their
dispersion factors in the brain. Reaearchers show
that multiwall CNTs (MWCNTs) coated with
Pluronic F127 (PF127) surfactant can be injected in
the mouse cerebral cortex without causing
degeneration of the neurons surrounding the site of
injection. They also show that, contrary to previous
reports on lack of PF127 toxicity on cultured cell
lines, concentrations of PF127 as low as 0.01% can
induce apoptosis of mouse primary cortical neurons
in vitro within 24 hours. However, the presence of
MWCNTs can avoid PF127-induced apoptosis.
These results suggest that PF127-coated MWCNTs
do not induce apoptosis of cortical neurons.
Moreover, the presence of MWCNTs can reduce
PF127 toxicity.[23]
Interactions of multiwalled carbon nanotubes
(MWCNTs) with human epidermal keratinocytes
(HEKs) were studied with respect to the effect of
surfactant on dispersion of MWCNT aggregates and
cytotoxicity. Our earlier studies had shown that the
unmodified MWCNTs were localized within the
cytoplasmic vacuoles of HEKs and elicited an
inflammatory response. However, MWCNTs in
solution tend to aggregate and, therefore, cells are
exposed to large MWCNT aggregates. The purpose
of this study was to find a surfactant that prevents
the formation of large aggregates of MWCNTs
without being toxic to the HEKs. HEKs were
exposed to serial dilutions (10% to 0.1%) of L61,
L92, and F127 Pluronic and 20 or 60 Tween for 24
hours. HEK viability, proportional to surfactant
concentration, ranged from 27.1% to 98.5% with
Pluronic F127; viability with the other surfactants
was less than 10%. Surfactants dispersed and
reduced MWCNT aggregation in medium.
MWCNTs at 0.4 mgml-1 in 5% or 1% Pluronic
F127 were incubated with HEKs and assayed for
interleukin 8 (IL-8). MWCNTs were cytotoxic to
HEKs independent of surfactant exposure. In
structure of a retractable antenna. CNTs are

nanodevices with important potential applications in
biomedicine such as drug and gene delivery.
Recognition
of
functionalization
of
nanotubes
Current advances in nanotechnology have led
to the development of the new field of
nanomedicine, which includes many applications of
nanomaterials and nanodevices for diagnostic and
therapeutic purposes. The same unique physical and
chemical properties that make nanomaterials so
attractive may be associated with their potentially
calamitous effects on cells and tissues. The recent
study on nanomedicine and nanotoxicology
published by Kagan et al demonstrated that
aspiration of single-walled CNTs elicited an
unusual inflammatory response in the lungs of
exposed mice with a very early switch from the
acute inflammatory phase to fibrogenic events
resulting in pulmonary deposition of collagen and
elastin. This was accompanied by a characteristic
change in the production and release of
proinflammatory to anti-inflammatory profibrogenic cytokines, decline in pulmonary function, and
enhanced susceptibility to infection. Chemically
unmodified (nonfunctionalized) CNTs are not
effectively
recognized
by
macrophages.
Functionalization of nanotubes results in their
increased recognition by macrophages and is thus
used for the delivery of nanoparticles to
macrophages and other immune cells to improve the
quality of diagnostic and imaging techniques as
well as for enhancement of the therapeutic
effectiveness of drugs. These observations on
differences in recognition of nanoparticles by
macrophages have important implications in the
relationship between the potentially toxic health
effects of nanomaterials and their applications in the
field of nanomedicine.[21] Although membrane
proteins consist of a substantial amount of the
human genome and are the main drug targets, the
study of cell membrane proteins in situ is
complicated by the technical limitations. The recent
development of atomic force microscopy (AFM)
opens a new way to study the functions of cell
membrane proteins in situ at the single-molecule
level. A detailed procedure for investigation of
36
opportunities for treating both skin and systemic

infections.[26]
contrast, MWCNT-induced IL-8 release was

reduced when exposed to 1% or 5% Pluronic F127
(P <0 .05). However, both MWCNTs and surfactant,
alone or in combination, increased IL-8 release
compared with control exposures at 12 and 24 hours.
These results suggest that the surfactant- MWCNT
interaction is more complex than simple dispersion
alone and should be investigated to determine the
mode of interaction.[24]
Tsuneo Urisu et al have developed two basic
technologies for fabrication of supported planar
lipid bilayer membrane ion channel biosensors: a
defect-free lipid bilayer formation on the substrate
surface with electrode pores and a patterning
technique for the hydrophobic self-assembledmonolayer to form the guard ring that reduces the
lipid bilayer edge-leak current. The importance of
the supported-membrane structure to achieve low
noise and high-speed performance is suggested on
the basis of the observed relation between the
single-ion-channel current noise and the pore
size.[25]
Pegylated liposomal doxorubicin

According to the American Cancer Society in
2006, an estimated 20180 new cases of ovarian
cancer will be diagnosed in the US in 2006.
Approximately 15310 of these women will die of
this disease. The vast majority will present with
advanced disease and will require chemotherapy,
and the majority of these will relapse. Safe,
effective cancer treatments are needed for relapsed
ovarian cancer. Goals include improving symptoms,
enhancing quality of life, and prolonging survival.
Currently, in the USA, the initial treatment consists
of maximal surgical debulking followed by
carboplatin and taxane chemotherapy. When the
disease recurs, the patient and physician are
presented with a host of chemotherapy options. One
drug that is increasingly being used is pegylated
liposomal doxorubicin.[27].
Encapsulation in polyethylene glycol-coated
(pegylated; Stealth) liposomes alters the
pharmacokinetic characteristics, and hence the
safety and tolerability profile, of doxorubicin.
Pegylated liposomal doxorubicin administered as a
single agent is generally well tolerated. Grade III/IV
leucopenia,
stomatitis
and
palmar-plantar
erythrodysaesthesia affected 16, 6 and 18% of solid
tumour patients, respectively. Other adverse effects
included nausea and vomiting and alopecia. Acute
hypersensitivity infusion reactions have been
reported in up to 9% of patients in some studies.
Recently published data from a phase II trial in
patients with refractory ovarian cancer showed no
alopecia or cardiotoxicity and little nausea and
vomiting after pegylated liposomal doxorubicin.
Unlike free doxorubicin, pegylated liposomal
doxorubicin is not a vesicant. Preliminary data, not
yet confirmed in comparative studies, suggest that
the pegylated liposomal formulation may be less
cardiotoxic than free doxorubicin. Mucositis,
however, appears to be increased. Studies to
determine optimal dosing schedules and safety of
pegylated liposomal doxorubicin in combination
with other agents are ongoing.[28]
Polyacrylate nanoparticle emulsions

Greenhalgh et al have recently reported on a
new nanomedicine containing antibiotic-conjugated
polyacrylate nanoparticles, which has shown
activity against methicillin-resistant Staphylococcus
aureus (MRSA) in vitro and no cytotoxicity toward
human dermal cells. The water-based nanoparticle
emulsion is capable of solubilizing lipophilic
antibiotics for systemic administration, and the
nanoparticle drug delivery vehicle has shown
protective properties for antibiotics from hydrolytic
cleavage by bacterial penicillinases, thus
rejuvenating the drug's activity against resistant
microbes such as MRSA. In Greenhalgh et al report,
the first in vivo study of this penicillin-conjugated
nanoparticle emulsion in determining toxicological
responses initiated upon systemic and topical
application in a murine model. Favorable results
were observed in vivo upon both routes of
administration and, when topically applied to a
dermal abrasion model, the emulsion enhanced
wound healing by an average of 3 to 5 days. This
study suggests that polyacrylate nanoparticlecontaining emulsions may afford promising
37
preferential uptake and decreased clearance of the

drug delivery system, increasing the exposure of the
tumor to the drug. When the liposome does leave
the intravascular compartment, in normal tissues it
is Phase II single-agent studies In a subsequent
Phase II study, evaluated 79 better-defined patients
all of whom were platinum and taxane refractory.
Eighty-five percent of the patients had received
more than 2 prior chemotherapy regimens. These
doubly refractory patients were treated with 50
mg/m2 of pegylated liposomal doxorubicin every 4
weeks. Fourteen partial responses and 1 complete
response were reported for an overall response rate
of 16.9%. The median time to response was 15
weeks. The median progression-free survival for all
patients treated in this study was 19.3 weeks (range
0.786 weeks). In addition, 36 patients (57%) were
classified as having stable disease, and achieved a
median progression-free survival of 21.9 weeks.
This was one of the first studies to show that disease
stabilization in recurrent ovarian cancer is of
clinical benefit. All patients reported at least 1
adverse event, but the majority were grade 1 or 2.
Asthenia and palmar-plantar erythrodysesthesia
(PPE) were seen in 41.6%. Only 1 patient
experienced any cardiac complications, and there
were no treatment-related deaths. This study
demonstrated that pegylated liposomal doxorubicin
was useful in this drug-resistant setting, and
associated with no life-threatening toxicities.
In China, Liang W et al research results on
doxorubicin- containing PEG-PE micelles are an
important
contribution
to
nanomedicine
development (which is called nanoparticles carry
chemotherapy drug deeper into solid tumors).
Editorial members, Dreher MR and Chilkoti A in J
Natl Cancer Inst get a high evaluation for their
research. Solid tumors account for more than 85%
of cancer mortality. To obtain nutrients for growth
and to metastasize, cancer cells in solid tumors must
grow around existing vessels or stimulate formation
of new blood vessels. These new vessels are
abnormal in structure and characterized by leakage,
tortuousness, dilation, and a haphazard pattern of
interconnection. Tumor structure and blood flow
hinder the treatment of solid tumors. To reach
cancer cells in optimal quantity, a therapeutic agent
must pass through an imperfect blood vasculature to
Pegylated liposomal doxorubicin is a

formulation of doxorubicin in which the molecule
itself is packaged in a liposome made of various
lipids with an outer coating of polyethylene glycol.
Liposomal technology is being used in increasing
amounts in the therapy of a variety of cancers,
including ovarian cancers. A reviews written by
Green et al on the mechanistic actions of this
formulation, the Phase II and Phase III data that
helped define the role of pegylated liposomal
doxorubicin in recurrent ovarian cancer, as well as a
discussion of some of the side-effects and their
management. [27] Pegylated liposomal doxorubicin
is one of a new class of drug formulations. The
doxorubicin molecules in pegylated liposomal
doxorubicin are encapsulated in a bilayer sphere of
lipids. This vesicle is then surrounded by a dense
layer of polyethylene glycol (PEG), hence the name
pegylated liposomal doxorubicin. The size of the
liposomes, approximately 100 nm, prevents them
from entering tissues with tight capillary junctions,
such as the heart and gastrointestinal tract, as well
as selectively depositing the liposome into the
tumor. In contrast to normal vessels, the vessels of
the
tumor
are
tortuous,
dilated,
have
morphologically abnormal endothelial cells, and are
leaky due to large spaces between pericytes. The
study on mechanism of action exhibited that these
physical characteristics allow more extravasation of
the vesicles into the tumor, thus encouraging more
deposition of the chemotherapy agent into the tumor.
The PEG coating on the liposome creates a
hydrophilic layer around the liposome that buffers
the liposome wall from the surrounding milieu. This
decreases proteins from binding to the lipid bilayer.
These proteins act as opsonins, attracting
foreign particles that in turn activate the
mononuclear phagocytic cells. This leads to break
down of the liposome and release of the drug.
Therefore, the PEG coating on the liposome
increases the longevity of the liposome. Pegylated
liposomal doxorubicin was cleared via the
lymphatic system and returned to the circulation. In
tumor tissue, however, there are no lymphatics.
Therefore, when the liposome is deposited it
remains for a longer time. This allows a higher dose
of doxorubicin to be released in the tumor, and a
lower dose in normal tissue. Collectively, there is
38
patient. In platinum-sensitive patients, pegylated

liposomal doxorubicin also produced a survival
advantage. Results from prospective and
retrospective studies further demonstrate the
improved cardiac safety of pegylated liposomal
doxorubicin
compared
to
conventional
anthracyclines. Based on survival and toxicity
advantages and a once-monthly administration
schedule, pegylated liposomal doxorubicin is the
first-choice nonplatinum agent for relapsed ovarian
cancer. Pegylated liposomal doxorubicin may also
have clinical application in combination regimens
for platinum-sensitive ovarian cancer, as
consolidation/maintenance therapy for ovarian
cancer, as a component of first-line therapy for
ovarian cancer, and in the treatment of other
gynecologic malignancies. Future clinical trials will
further define and maximize the role of pegylated
liposomal doxorubicin in the treatment of ovarian
cancer and other gynecologic malignancies.[32]
the tumor, cross vessel walls into the interstitium

and penetrate multiple layers of solid tumor cells.
Recent studies have demonstrated that poor
penetration and limited distribution of doxorubicin
in solid tumors are the main causes of its
inadequacy as a chemotherapeutic agent.
Encapsulation of doxorubicin into PEG-PE micelles
increased its accumulation and penetration in
tumors in terms of both the percentage of cells that
were reached by the drug and the intracellular levels
that were attained. This increased accumulation and
penetration can be attributed to the efficient
internalization of the drug-containing micelles by
the endocytotic cell uptake mechanism and
enhanced permeability and retention of tumors with
leaky vasculature. High intracellular retention is
especially important because doxorubicin must be
internalized to be effective in tumor therapy. The
doxorubicin- containing PEG-PE micelles had
greatly increased antitumor activity in both
subcutaneous and lung metastatic LLC tumor
models compared with free doxorubicin. However,
mice treated micelle- encapsulated doxorubicin
showed fewer signs of toxicity than those treated
with free doxorubicin. This drug packaging
technology may provide a new strategy for design
of cancer therapies.[29,30] At our laboratory, studied
nanoparticle of doxorubicin eliminate the
accumulation in tissues of tumor-bearing mice.
Compared with general doxorubicin preparation,
which is a marketed product, nanoparticle micelle
of doxorubicin has the similar pharmacokinetics in
the tissue, and the similar concentrations in the
tumor tissue. Howerever, the accumulation of
doxorubicin in the heart, spleen, kidney, lung,
tumor, muscle and skin decreased significantly after
three intravenous injections, showing that the
nano-micelle can accumulatew the elimilation of
doxorubicin in most tissues. It is deduced that the
study was effects of doxorubicin after clinical use
may be reduced significantly. [31]
Pegylated liposomal doxorubicin is effective
and well tolerated in relapsed ovarian cancer. When
compared with topotecan in a phase III randomized
trial, pegylated liposomal doxorubicin showed
several advantages: improved quality of life, fewer
severe adverse events, fewer dose modifications,
less hematologic support, and lower total cost per
Doxorubicin nanoparticles
A novel hyaluronic acid-poly(ethylene
glycol)-poly(lactide-co-glycolide) (HA-PEG-PLGA)
copolymer was synthesized and characterized by
infrared and nuclear magnetic resonance
spectroscopy. The nanoparticles of doxorubicin
(DOX)-loaded HA-PEG-PLGA were prepared and
compared with monomethoxy (polyethylene glycol)
(MPEG)-PLGA nanoparticles. Nanoparticles were
prepared using drug-to-polymer ratios of 1:1 to 1:3.
Drug-to-polymer ratio of 1:1 is considered the
optimum formulation on the basis of low particle
size and high entrapment efficiency. The optimized
nanoparticles were characterized for morphology,
particle size measurements, differential scanning
calorimetry, x-ray diffractometer measurement,
drug content, hemolytic toxicity, subacute toxicity,
and in vitro DOX release. The in vitro DOX release
study was performed at pH 7.4 using a dialysis
membrane. HA-PEG-PLGA nanoparticles were
able to sustain the release for up to 15 days. The
tissue distribution studies were performed with
DOX-loaded HA-PEG-PLGA and MPEG-PLGA
nanoparticles after intravenous (IV) injection in
Ehrlich ascites tumorbearing mice. The tissue
distribution studies showed a higher concentration
of DOX in the tumor as compared with
39
protein as a typical membrane protein. Landscape

phage peptides specific for specific tumors can be
obtained by affinity selection, and purified fusion
coat proteins can be assimilated into liposomes to
obtain specific drug-loaded nanocarriers. As a
paradigm
for
inceptive
experiments,
a
streptavidin-specific phage peptide selected from a
landscape phage library was incorporated into
100-nm liposomes. Targeting of liposomes was
proved by their specific binding to streptavidincoated beads.[35]
MPEG-PLGA nanoparticles. The in vivo tumor

inhibition study was also performed after IV
injection
of
DOX-loaded
HA-PEG-PLGA
nanoparticles up to 15 days. DOX-loaded
HA-PEG-PLGA nanoparticles were able to deliver
a higher amount of DOX as compared with
MPEG-PLGA nanoparticles. The DOX-loaded
HA-PEG-PLGA nanoparticles reduced tumor
volume
significantly
as
compared
with
MPEG-PLGA nanoparticles.[33] Chitosan, PCEP
(poly{[(cholesteryl oxocarbonylamido ethyl) methyl
bis(ethylene) ammonium iodide] ethyl phosphate}),
and magnetic nanoparticles (MNPs) were evaluated
for the safe delivery of genes in the eye. Prow et al
studied ocular nanoparticle toxicity and transfection
of the retina and retinal pigment epithelium. Rabbits
were injected with nanoparticles either intravitreally
(IV) or subretinally (SR) and sacrificed 7 days later.
Eyes were grossly evaluated for retinal pigment
epithelium abnormalities, retinal degeneration, and
inflammation. All eyes were cryopreserved and
sectioned for analysis of toxicity and expression of
either enhanced green or red fluorescent proteins.
All of the nanoparticles were able to transfect cells
in vitro and in vivo. IV chitosan showed
inflammation in 12/13 eyes, whereas IV PCEP and
IV MNPs were not inflammatory and did not induce
retinal pathology. SR PCEP was nontoxic in the
majority of cases but yielded poor transfection,
whereas SR MNPs were nontoxic and yielded good
transfection. Therefore, researchers concluded that
the best nanoparticle evaluated in vivo was the least
toxic nanoparticle tested, the MNP.[34]
Drug
Loading
and
Release
From
Biodegradable Microcapsules
Microcapsules made of biopolymers are of
both scientific and technological interest and have
many potential applications in medicine, including
their use as controlled drug delivery devices. The
present study makes use of the electrostatic
interaction between polycations and polyanions to
form a multilayered microcapsule shell and also to
control the loading and release of charged drug
molecules inside the microcapsule. Micron-sized
calcium carbonate (CaCO3) particles were
synthesized and integrated with chondroitin sulfate
(CS) through a reaction between sodium carbonate
and calcium nitrate tetrahydrate solutions suspended
with CS macromolecules. Oppositely charged
biopolymers were alternately deposited onto the
synthesized
particles
using
electrostatic
layer-by-layer self-assembly, and glutaraldehyde
was introduced to cross-link the multilayered shell
structure. Microcapsules integrated with CS inside
the multilayered shells were obtained after
decomposition of the CaCO3 templates. The
integration of a matrix (i.e., CS) permitted the
subsequent selective control of drug loading and
release. The CS-integrated microcapsules were
loaded with a model drug, bovine serum albumin
labeled with fluorescein isothiocyanate (FITC-BSA),
and it was shown that pH was an effective means of
controlling the loading and release of FITC-BSA.
Such CS-integrated microcapsules may be used for
controlled localized drug delivery as biodegradable
devices, which have advantages in reducing
systemic side effects and increasing drug efficacy.
Liposomes targeted by fusion phage

proteins
Targeting of nanocarriers has long been sought
after to improve the therapeutic indices of
anticancer drugs. Jayanna et al provide the proof
of principle for a novel approach of nanocarrier
targeting through their fusion with target-specific
phage coat proteins. The source of the targeted
phage coat proteins are landscape phage
librariescollections of recombinant filamentous
phages with foreign random peptides fused to all
4000 copies of the major coat protein. Prashanth et
al exploit in our approach the intrinsic
physicochemical properties of the phage major coat
[36]
40
long as 24 hours after application, thus apparently a

suitable inert carrier for ophthalmic drug delivery.
Amphotericin Bintercalated liposomes

Nanotechnology in drug delivery is a rapidly
expanding field. Nanosized liposomal preparations
are already in use for efficient drug delivery with
better therapeutic indices. Existing methods of
liposome preparation are limited by problems of
scale-up, difficulty in controlling size, and
intercalation efficiency. Here researchers prepare
amphotericin Bintercalated liposomes by a novel
process where amphotericin B and purified
phosphatidyl choline are solubilized in suitable
solvent and precipitated in supercritical fluid carbon
dioxide (known as a gas antisolvent technique), to
obtain microsized particles that are subsequently
introduced into a buffer solution. The morphology
of liposomes was characterized through a
phase-contrast microscope, and the particle size
distribution studied by laser technique showed
nanosize with a narrow range of size distribution
(between 0.5 and 15 m) and a higher intercalation
efficiency. In vitro studies conducted using
Aspergillus fumigatus (MTCC 870) strain proved to
be efficient in the retardation of the growth of the
organism.[37]
[38]
Paclitaxel nanoparticles
Karmali et al have used tumor-homing
peptides to target abraxane, a clinically approved
paclitaxel-albumin nanoparticle, to tumors in mice.
The targeting was accomplished with two peptides,
CREKA and LyP-1 (CGNKRTRGC). Fluorescein
(FAM)-labeled CREKA-abraxane, when injected
intravenously into mice bearing MDA-MB-435
human cancer xenografts, accumulated in tumor
blood vessels, forming aggregates that contained
red blood cells and fibrin. FAM-LyP-1-abraxane
co-localized with extravascular islands expressing
its receptor, p32. Self-assembled mixed micelles
carrying the homing peptide and the label on
different subunits accumulated in the same areas of
tumors as LyP-1-abraxane, showing that Lyp-1 can
deliver intact nanoparticles into extravascular sites.
Untargeted, FAM-abraxane was detected in the
form of a faint meshwork in tumor interstitium.
LyP-1-abraxane produced a statistically highly
significant inhibition of tumor growth compared
with untargeted abraxane. These results show that
nanoparticles can be effectively targeted into
extravascular tumor tissue and that targeting can
enhance the activity of a therapeutic nanoparticle.
Diclofenac-loaded biopolymeric nanosuspensions

Polymeric nanoparticle suspensions (NS) were
prepared from poly(lactide-co- glycolide) and
poly(lactide-co-glycolide-leucine) {poly[Lac (GlcLeu)]} biodegradable polymers and loaded with
diclofenac sodium (DS), with the aim of improving
the ocular availability of the drug. NS were
prepared by emulsion and solvent evaporation
technique and characterized on the basis of
physicochemical properties, stability, and drug
release features. The nanoparticle system showed an
interesting size distribution suitable for ophthalmic
application. Stability tests (as long as 6 months'
storage at 5C or at 25C/60% relative humidity) or
freeze-drying were carried out to optimize a suitable
pharmaceutical preparation. In vitro release tests
showed a extended-release profile of DS from the
nanoparticles. To verify the absence of irritation
toward the ocular structures, blank NS were applied
to rabbit eye and a modified Draize test performed.
Polymer nanoparticles seemed to be devoid of any
irritant effect on cornea, iris, and conjunctiva for as
[39]
Nanoatropine sulfate dry powder inhaler

The work of Raisuddin Ali et al was to develop,
characterize, and carry out a clinical trial with
nanoatropine sulfate (nano-AS) dry powder inhaler
(DPI), because this route may offer several
advantages over the conventional intramuscular
route as an emergency treatment, including ease of
administration and more rapid bioavailability.
Different batches of nanoparticles of AS were
produced using variants of nanoprecipitation
method. The influence of the process parameters,
such as the types and quantity of solvent and
nonsolvent, the stirring speed, the solventto-nonsolvent volume ratio, and the drug
concentration, was investigated. The methodology
resulted in optimally sized particles. Bulk properties
of the particles made by the chosen methodology
41
adenocarcinomas and in 5/34 (15%) lymph node

metastases. In contrast, normal squamous and
cardiac mucosa, as well as noninvasive Barrett
lesions, failed to label with mesothelin. Mesothelin
was expressed in the esophageal adenocarcinoma
cell line JH-EsoAd1 but not in primary human
esophageal epithelial cells. Anti-mesothelin
antibodyconjugated CdSe/CDS/ ZnS quantum rods
were synthesized, and confocal bioimaging
confirmed robust binding to JH-EsoAd1 cells.
Anti-mesothelin antibody conjugated nanoparticles can be useful for the diagnosis and therapy
of mesothelin-overexpressing esophageal adenocarcinomas. [42]
were evaluated. A clinical trial was conducted in six

healthy individuals using a single DPI capsule
containing 6 mg nano-AS DPI in lactose. Early
blood bioavailability and atropinization pattern
confirmed its value as a potential replacement to
parenteral atropine in field conditions. The
formulation seems to have the advantage of early
therapeutic drug concentration in blood due to
absorption through the lungs as well as sustained
action due to absorption from the gut of the
remaining portion of the drug. [40]
Drug delivery of siRNA therapeutics
A
review
by
Daniela Reischl
and
Andreas Zimmer focuses on different pathways for
siRNA delivery and summarizes recent progress
made in the use of vector-based siRNA technology.
Gene therapy is a promising tool for the treatment
of human diseases that cannot be cured by rational
therapies. The major limitation for the use of small
interfering RNA (siRNA), both in vitro and in vivo,
is the inability of naked siRNA to passively diffuse
through cellular membranes due to the strong
anionic charge of the phosphate backbone and
consequent electrostatic repulsion from the anionic
cell membrane surface. Therefore, the primary
success of siRNA applications depends on suitable
vectors to deliver therapeutic genes. Cellular
entrance is further limited by the size of the applied
siRNA molecule. Multiple delivery pathways, both
viral and nonviral, have been developed to bypass
these problems and have been successfully used to
gain access to the intracellular environment in vitro
and in vivo, and to induce RNA interference
(RNAi). [41]
Research and development of nanomedicines

in the future
Nanotechnology will alter our relationship with
molecules and matter profoundly. Research on
productive nanosystems will eventually develop
programmable, molecular-scale systems that make
other useful nano-structured materials and devices.
These systems will enable a new manufacturing base
that can produce both small and large objects
precisely and inexpensively. Nano-risk research is
conducted by agencies that oversee health and
environmental regulations. Nanotechnology, dealing
with functional structures and materials smaller than
100nm, is emerging as a truly interdisciplinary research
area spanning several traditional scientific disciplines. In
keeping with the growing trend, there is a strong need for a
platform to share original research related to applications of
nanotechnology in biomedical fields. At the hearing,
leaders of the Nanotechnology Environmental and

Health Implications working group, an interagency
panel that coordinates federal funding on health and
environmental risks of nanotechnology, released a
long-overdue report outlining research needed to
buttress regulation of products in the field. There is
far less agreement on how that money should be
spent and coordinated. Research on Nanotech
environmental health and safety in government
agencies, academic institutions, and industry is being
performed in an ad hoc fashion according to
individual priorities. Yet the vast majority of
nanotoxicology studies focus on those materials,
while ignoring broad classes of other materials
Translational implications for diagnosis and

therapy: Esophageal adenocarcinoma arises in the
backdrop of Barrett metaplasia-dysplasia sequence,
with the vast majority of patients presenting with
late-stage malignancy. Mesothelin, a glycophosphatidylinositol-anchored protein, is aberrantly
overexpressed on the surface of many solid cancers.
Mesothelin expression was assessed in esophageal
tissue microarrays encompassing the entire
histological
spectrum
of
Barrett-associated
dysplasia
and
adenocarcinoma.
Mesothelin
expression was observed in 24/84 (29%) of invasive
42
already on the market.[43]

The nanomedicine research is a goal and needs a
long-term plan, which is to quantitatively
characterize the molecular-scale components, or
nanomachinery, of living cells and to precisely
control and manipulate these molecular and
supramolecular assemblies in living cells to improve
human health. Nanomedicine will exploit and build
upon other research findings in nanotechnology and
apply it to the study of molecular systems in living
cells that contain a multitude of nanoscale structures,
such as membrane transporters, processes such as
self-assembly of proteinnucleic acid complexes, and
nanomachines such as molecular motors. The
benefits of nanomedicine include dramatically
expanded knowledge of the human genome, a greater
understanding of the pathophysiology of specific
diseases at the molecular scale, more specific
treatment of diseases, and the ability to understand
the dynamic behavior of dysfunctional cellular
machinery in
the
context of the total cell
machinery.[44] Robert A and Freitas Jr given an
overview of this rapidly expanding and exciting
nanomedicine field. Over the next 5 to 10 years,
nanomedicine will address many important medical
problems by using nanoscale-structured materials and
simple nanodevices that can be manufactured today.
Many approaches to nanomedicine being pursued
today are already close enough to fruition that it is
fair to say that their successful development is almost
inevitable, and their subsequent incorporation into
valuable medical diagnostics or clinical therapeutics
is highly likely and may occur very soon.[45]
The science of nanomedicine exploits and builds
upon novel research findings in nanotechnology,
biology, and medicine; it unifies the efforts of
scientists, engineers, and physicians determined to
apply their latest research results to translational and
clinical medicine by developing novel approaches
and a better understanding of solutions to
health-relatedissues, ultimately improving the quality
of life. The last few years have seen unprecedented
advances in the field of biology. The decoding of the
human genome coupled with improving gene
transfection technologies offer great opportunities for
treating illnesses. In analysis and diagnosis,
lab-on-a-chip methods have surpassed earlier ex-vivo
and in-vivo detection methods while also aiding
toxicology efforts. In medicine, improvements in

targeted drug delivery, imaging, and therapy have led
to such successful interventions in cancer therapies.
[46]
Although there are only a few FDA-approved

nanopharmaceuticals on the market today, these
formulations are already impacting medicine and
promise to alter healthcare. Based on their ability to
reduce time-to-market, extend the economic life of
proprietary drugs and create additional revenue
streams, nanopharmaceuticals should greatly impact
medical practice and healthcare. However, if this is to
happen effectively, there are a few key biological
requirements for nanopharmaceuticals to fulfill: (1)
they must exhibit stealth qualities to evade
macrophage attack and the immune response; (2) be
nontoxic and traceable; (3) display effective
pharmacokinetic properties; (4) be biodegradable
following systemic administration through any route
(but the polymer must protect the embedded active);
and (5) they must be selective to be effective in
targeting specific tissue sites. Srikumaran Melethil
(Chair and Professor of Pharmaceutical Sciences at
the University of Findlay, Findlay, OH) discussed the
metabolic fate of nanopharmaceuticals upon delivery
to the human body, and presented pharmacokinetic
data relating to numerous nanoparticulate drugs and
highlighted the critical role of the FDA in
nanomedicine. According to him, further knowledge
of how the human body transports, distributes and
clears nanoparticles via the vascular and lymphatic
systems (i.e.,biodistribution of nanoparticles) is also
needed to get a handle on metabolic and toxicity
issues. Nanomedicine will eventually become an
integral part of mainstream medicine and a standard
in the drug industry. For example, the market impact
of nanopharmaceuticals on the pharmaceutical and
biotech industries is already being felt. However, for
nanomedicine to be a viable commercial entity,
desperately needed reforms to overhaul the PTO
along with clearer regulatory guidelines and safety
standards from federal agencies such as the FDA will
be needed.
Ethical question of nanomedicine is an important
issue. Ginger Gruters (The President's Council on
Bioethics, Washington, DC) presented on ethical
considerations that are likely to play a significant role
in nanomedicine, and stated that, as with other
43
biomedical advances coming before it, nanomedicine

will face significant ethical challenges as it moves
from proof-of-concept to the clinic. Along the
way,ethical questions regarding social justice,
privacy and confidentiality, long-term risks and
benefits, and human enhancement are certain to
arise.[47]
important
papers
published
in
other
journals,
commentaries, book reviews, correspondence, and articles

about the broader nanotechnology picture funding,
commercialization, ethical and social issues, and so on. In
this way, the journal aims to be the voice of the
worldwide nanoscience and nanotechnology community.
Nature Nanotechnology offers readers and authors high
visibility, access to a broad readership, high standards of
copy editing and production, rigorous peer review, rapid
Appurtenances:
Introduction to Journals on Nano
publication, and independence from academic societies

and other vested interests.
Nature Nanotechnology
Nature
is
Nanotechnology
multidisciplinary
Journal of Nanoscience and Nanotechnology
journal that publishes papers of the highest quality and
Journal of Nanoscience and Nanotechnology (JNN)
and
is an international and multidisciplinary peer-reviewed
nanotechnology. The journal covers research into the
journal with a wide-ranging coverage, consolidating
design, characterization and production of structures,
research activities in all areas of nanoscience and
devices and systems that involve the manipulation and
nanotechnology into a single and unique reference source.
control of materials and phenomena at atomic, molecular
JNN is the first cross-disciplinary journal to publish
and
and
original full research articles, rapid communications of
top-down approaches - and combinations of the two - are
important new scientific and technological findings,
covered. Nature Nanotechnology also encourages the
timely state-of-the-art reviews with author's photo and
exchange of ideas between chemists, physicists, material
short biography, and current research news encompassing
scientists, biomedical researchers, engineers and other
the fundamental and applied research in all disciplines of
researchers who are active at the frontiers of this diverse
science, engineering and medicine. Topics covered in the
and multidisciplinary field. Coverage extends from basic
journal
research in physics, chemistry and biology, including
Nanoscale Materials, Nanofabrication and Processing of
computational work and simulations, through to the
Nanoscale Materials and Device; Atomic and Nanoscale
significance
in
all
macromolecular
areas
of
scales.
nanoscience
Both
bottom-up
include:
Synthesis
Nanostructured
Characterization
applications in a wide range of industrial sectors
Bio-assemblies; Nanoprobes, Properties of Nanoscale
(including
Materials, Nanocatalysis; Nanocomposites, Nanoparticles,
technology,
medicine,
Functional
Nanocrystalline
and environmental technologies). Organic, inorganic and
Superlattices, Quantum Dots, Quantum Wires, Quantum
hybrid materials are all covered. Research areas covered
Wells, Nanoscale Thin Films ; Fullerenes, Nanotubes,
in the journal include: Carbon nanotubes and fullerenes,
Nanorods, Molecular Wires, Molecular Nanotechnology;
Computational nanotechnology, Electronic properties and
Supramolecules,
devices,
Environmental,
health
machines
self-assembly,
Nanobiotechnology,
Nanomagnetism
Nanomedicine,
and
and
Nanometrology
and
Dendrimers,
Nanoclusters;
Self-Assemblies,
safety
Low-dimension Structures; Nanophysics, Nanoelectronics,
Molecular
Nano-Optics, Nanomagnetism and Nanodevices; Atomic
Nanofluidics,
Manipulation, Computational Nanotechnology, Molecular
and
motors,
spintronics,
and
and
manufacturing, high-performance materials, and energy
issues,Molecular
Materials,
Materials
and
development of new devices and technologies for

information
of
of
Nanoscience;
Nanomaterials,
Nanochips,
Nano-integration,
instrumentation,
Nanosensors
Nanofluidics,
and
Nanomachining;
Nanoparticles, Nanosensors and other devices, NEMS,
Structure Analysis at Atomic, Molecular, and Nanometer
Organicinorganic nanostructures, Photonic structures
range;
and devices, Quantum information, Structural properties,
Applications
Surface patterning and imaging and Synthesis and
Nanobiotechnology, Biochemical Assemblies, BioMEMS,
processing.
Biomimetic
Nanorobotics,
of
Nanotribology,
Nanostructured
Materials
Nanoscale
and
Materials
Genomics,
Novel
and
DNA
Sequencing, Nanomedicines, Drug Delivery, Biomedical
Nature Nanotechnology also publishes review
Nanotechnolog.
articles, news and views, research highlights about

44

made in nanoscience and nanotechnology and the future
predictions for this extraordinary technology.
Nano
NANO is an international peer-reviewed journal for
nanoscience and nanotechnology that presents forefront
Journal of Nanotechnology Online
fundamental research and new emerging topics. It
The Online Journal of Nanotechnology is based on a
features timely scientific reports of new results and
free access publishing model, coupled with what is
technical breakthroughs and also contains interesting
believed to be a unique development in the field of
review articles about recent hot issues. NANO provides an
scientific publishing the distribution of journal revenue
ideal forum for presenting original reports of theoretical
between the authors, peer reviewers and site operators
and experimental nanoscience and nanotechnology
(OARS). The revenue received from the journal related
research.
include:
advertising and sponsorship will be distributed according
nanomaterials including nano-related biomaterials, new
Research
areas
of
interest
to the following general criteria: Authors receive a
phenomena and newly developed characterization tools,
revenue share of 50% of the related revenue their
fabrication methods including by self-assembly, device
contributions attract. Peer reviewers receive a revenue
applications, and numerical simulation, modeling, and
share of 20%. The site administrators receive a revenue
theory.
share of 30%. This revenue share will apply throughout

the on-line published life of the individual article or paper.
The Online Journal of Nanotechnology papers will
Nano Letters
Nano Letters reports on fundamental research in all
benefit from being hosted on the AZoNano.com website
branches of the theory and practice of nanoscience and
and database platform as they will take advantage of the
nanotechnology, providing rapid disclosure of the key
existing AZoNano.com search tools. These search tools
elements of a study, publishing preliminary, experimental,
make it very easy for site visitors to locate nanotech
and theoretical results on the physical, chemical, and
information which directly relates to their research areas,
biological
applications and industrial sectors.
phenomena,
along
with
processes
and
applications of structures within the nanoscale range.

Among the areas of interest the journal covers are:
Journal of Nano Education
Synthesis and processing of organic, inorganic, and
The Journal of Nano Education (JNE) is a
hybrid nanosized materials by physical, chemical, and
peer-reviewed international journal that aims to provide
biological methods; Modeling and simulation of synthetic,
the most complete and reliable source of information on
assembly, and interaction processes; Characterization of
current developments in nanoscale science, technology,
size-dependant
and
engineering, and medical education. JNE publishes a
application of novel nanostructures and nanodevices The
comprehensive range of articles including topics in the
Nano Letters manuscript submission process is fully
following
electronic, to ensure the rapid publication of results.
engineering, and medical education at the K-12,
Manuscripts should be submitted via our secure Web site.
undergraduate and graduate levels (formal and informal,
Manuscripts submitted by hardcopy mail or by e-mail will
including public outreach and dissemination activities);
not be processed. Introduction Nano Letters invites
K-12
original reports of fundamental research in all branches of
development; Scientific and technological literacy/public
the
understanding
theory
properties;
and
practice
and
of
Realization
nanoscience
and
areas:
science
Nanoscale
teacher
of
science,
education
nanoscale
and
science,
technology,
professional
technology,
engineering, and medicine; Curriculum development and
nanotechnology.
assessment; Social and ethical issues associated with

nanoscale science, technology, engineering, and medical
Journal of Nano Research
research;
Journal of Nano Research (J Nano R) is a
Workforce
preparation
(professional
and
multidisciplinary peer-reviewed journal, which publishes
vocational); National and state science standards and their
high quality work on ALL aspects of nanoscience and
relationships to the goals of nanoeducation initiatives
nanotechnology. Currently, it stands alone in serving the
worldwide; Current nanoscale science, technology,
global nano community in providing up-to-date
engineering, and medical education research; Other
information on all developments and progresses being
pertinent
45
areas
of
interest
to
nanoscale
science,

technology, engineering, and medical researchers &
thermodynamics thought experiments, wear, and much
educators. JNE also will serve as a forum for commentary
more.
and debate on related issues.
Journal of Biomedical Nanotechnology

Journal of
Nanoscience
Computational
and
Journal of Biomedical Nanotechnology (JBN) is
Theoretical
being created as an international peer-reviewed periodical

Theoretical
that covers applications of nanotechnology in all fields of
Nanoscience is an international peer-reviewed journal
life sciences. JBN publishes original full papers and
with a wide-ranging coverage, consolidates research
timely state-of-the-art reviews with author's photo and
activities in all aspects of computational and theoretical
biography, and short communications encompassing the
nanoscience into a single reference source. This journal
fundamental and applied research aspects. To speed up
offers scientists and engineers peer-reviewed research
the reviewing process, we will provide on-line refereeing
papers in all aspects of computational and theoretical
of all articles submitted in electronic form. Authors
nanoscience and nanotechnology in chemistry, physics,
receive these benefits: Electronic submission of articles,
materials science, engineering and biology to publish
Fast reviewing, Rapid times to publication, No page
original full papers and timely state-of-the-art reviews
charges, Free color where justified, Distinguished
and short communications encompassing the fundamental
editorial board
and applied research. Topics include: Assemblers, basic
editions. Topics include: Broadly speaking, Journal of
physics, biological systems, biochemical systems, bionics,
Biomedical Nanotechnology covers applications of
biophysics, CAD, carbon systems, cellular mechanisms,
nanotechnology in biotechnology, medicine, biosciences,
chaotic systems, circuits, clusters, cluster systems,
and all other related fields of life sciences. The coverage
complex aggregates, computer codes, crystal growth, data
includes applications of nanotechnology in all fields of
analysis, defined chain length molecules, devices,
life sciences, all kinds of nanoscale biomaterials,
diffusion processes, DNA, drug design, dynamics,
biomimetics of biological materials and machines,
electronics, electronic properties, enzyme reactivity and
nanoprobes,
biocompatible
reactions, equation of state, friction, computational
bioengineered
materials,
genomics, gene technology, genetics, holistic views,
biopolymers,
information theory, interactions, ion channelling, kinetics,
nanocomposites, biological macromolecules, proteins,
macromolecules, molecular interactions, large scale
enzymes,
simulations,
Journal
of
Computational
liquids,
and
and Available in print and online
surfaces,
polypeptides,
organic-inorganic
kinases,
functional
bioceramics,
hybrid
biomaterials,
phosphatases,
DNA-based
crystals,
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nanostructures, molecular assemblies, biomolecules, cells,
manufacturing,
liquid
many-particle
and glycans, biochips, microarrays, biocompatibility
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References
digesting the foreign consciousness, so that publishers can

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construct this journal with a high impact factor in the

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Publication News
Drug Evaluation Research
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Evaluation Research will officially be published in 2009, in China. Sponsored by China
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Drug Evaluation Research, a national journal, is an official publication of Chinese
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studies on the academic and technological evaluation and research of drugs including chemical
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The 3rd Asian Pacific ISSX Meeting

May 10 - 12, 2009
The Imperial Queen's Park Hotel| Bangkok, Thailand
Understanding Xenobiotics for Better Drug Development and Therapy
The 3rd Asian Pacific Regional Meeting of the International Society for the Study of
Xenobiotics, it is our pleasure to invite you to the meeting which will be held May 10 12, 2009
in Bangkok, Thailand. he theme of this meeting is Understanding Xenobiotics for Better Drug
Development and Therapy. Symposia will address topics including Drug Safety and
Development, the Exposure, Disposition and Health Effects of Arsenic, Genome-Based
Technologies for Drug Toxicity Assessment, Drug Metabolizing Enzymes involved in Activation
and Detoxification, and much more including a symposium on the Development of Herbal
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by the faculty of Pharmaceutical Sciences at Chulalongkorn University in Bangkok, Thailand.
50
Zhang R et al. Asian Journal of Pharmacodynamics and Pharmacokinetics 2009;9(1):51-57
Asian Journal of
Pharmacokinetics
ISSN
1608-2281

Tissue distribution of Curcumol in rats after intravenous injection

of zedoary turmeric oil fat emulsion
Rui Zhang1, Benjie Wang1, Hengli Zhao2, Chunmin Wei1, Guiyan Yuan1, Ruichen Guo1*
1
Institute of Clinical Pharmacology, Qilu Hospital of Shandong University, 107 Wenhua West Road, Jinan,
250012, China
2
Department of Pharmacy, The Second Hospital of Shandong University,247 Beiyuan Road, Jinan, 250033,
China
Abstract
Aim To determine the concentration of curcumol in different tissues and investigate the tissue
distribution of curcumol in rats after intravenous injection of zedoary turmeric oil fat emulsion.
Methods Wister rats were intravenously injected with a dose of 10.0 mgkg-1 zedoary turmeric
oil fat emulsions. The tissue samples (heart, liver, lung, kidney and brain) were collected at
scheduled times. Curcumol was extracted with chloroform/isopropyl alcohol (95:5, v/v) from
tissue homogenates and separated on a C18 column with a mobile phase of acetonitrile (0.3M
ammonium acetate containing 0.1% formic acid)/water (0.1% formic acid) (95:5, v/v). Detection
was carried out by positive elevtrospray ionization (ESI) in multiple reactions monitoring
(MRM) mode of 254.3219.4 (m/z) for curcumol and 220.3128.1 (m/z) for ornidazole (I.S.),
respectively. Results Curcumol was distributed to liver, kidney, brain, heart, lung 10 min after
intravenous injection and obtained the maximum concentration of 108.8565.91, 105.1942.92,
92.3817.63, 82.9638.06, 10.012.97 gg-1, respectively. Conclusions The method was
successfully applied in the tissue distribution study. The results showed that the curcumol was
markedly decreased after 0.5 h and almost eliminated at 5 h after administration.
Key words
Curcumol; tissue distribution; LC-MS/MS
Article history
Received 6 August 2008;
Publication data
Pages: 7 ;
Tables: 4;
Accepted 30 December 2008
Figures: 4 ;
References: 10 ;
Paper ID 1608-2281-2009-0901051-07
Prof. Ruichen GuoInstitute of Clinical Pharmacology, Qilu Hospital of Shandong University, 107 Wenhua
West Road, Jinan, 250012, China
Tel: +86 531 82169636; Fax: +86 531 86109975 E-mail address:
grc7636@126.com
antivirus, antiinflammation, antibacterial, and so on.

Curcumol (content was about 4.12%) is an important
ingredient with antibiosis and antiviral effects in
zedoary turmeric oil[2-4], which can inhibited the
growth of Bacterium typhi, Bacterium coli,
respiratory syncytial virus (RSV), bastard measles,
chickenpox virus, and so on [5].
Introduction
Zedoary turmeric oil is the essential oil derived
from the rhizome of Chinese medicinal herb
Curcuma phaeocaulis Valeton, C. kwangsinensis S.G.
Lee et C. F. Liang and C. wenyujin Y. H. Chen et C.
Ling[1]. It possesses the activities of antitumor,
51
China). Ornidazole (internal standard, >99.6%) was

donated by the Institute of Wuhan Jingxi Chemical
Production (Hubei, China). Acetonitrile (Lot No:
044263), formic acid (Lot No: 406014), chloroform
(Lot No: 506023) and isopropyl alcohol (Lot No:
503027) were chromatographic grade from TEDIA
Company (USA); ammonium acetate (Lot No:
20050516) was analytical grade from Tianjin Tianda
Chemistry Industry Factory (Tianjin, China).
The pharmacokinetic and tissue distribution

studies were important for clinical application. These
studies of curcumol by isotope tracer method was
reported[6], it was sensitive enough to determine the
low concentration of curcumol in biotic sample, but
the radioactivity were the sum of 3H labeled original
and its metabolites, which can not exactly illustrate
the disposition process in vivo. In order to illustrate
the disposal process of curcumol in vivo more exactly,
other analysis method should be developed. Several
quantitative method of curcumol in medical material
and preparations such as RP-HPLC[1], infrared
spectrophotometry[7], thin-layer scanning method[8]
and vapor phase chromatography[9] were reported.
These methods were not sensitive enough to
quantitate the curcumol in biological specimen. We
had
developed
a
high-performance
liquid
chromatography coupled with tandem mass
spectrometry
quantitative
detection
method
(HPLC-MS/MS) to determine the curcumol in plasma
and the method had been applied in pharmacokinetics
study in Beagle dogs successfully[10]. In this study,
the established method was validated in tissue
samples and used to investigate tissue distribution of
curcumol after intravenous injection zedoary turmeric
oil fat emulsion in rats. The chemical structure of
curcumol is shown in Fig 1.
Animal
Male and female Wister rats (20020 g) were
purchased from the Laboratory Animal Center of
Shandong University(Jinan, China) .
Apparatus
The LC-MS/MS system include a 1100 Series
HPLC (Agilent company, USA) and API 4000
MS/MS (Applied Biosystem, USA); AX-205
Electronic Balance (METTLER TOLEDO Instrument
Shanghai Company); XW-80A Vortex (Shanghai
Jingke Industry Company); SORVALL Biofuge
PRIMO centrifuge (Kendro Company,USA).
Chromatographic Condition
Separation was carried out on a DiamonsilTM C18
column (150mm4.6mm, 5m) (Dikma Technologies,
China) and eluted with a mobile phase of acetonitrile
(0.3M ammonium acetate containing 0.1% formic
acid-water (0.1% formic acid) (95:5, v/v). The flow
rate was 1.0 mLmin-1. 40L of extracted samples
was injected for analysis.
CH2
O
CH3
OH
CH(CH3) 2
Mass spectrometric condition
The ESI source was set to positive ion mode

with the following parameters: ion source
temperature 550 C, spray voltage 5500 V, sheath
gas 70 psi and auxiliary gas 40 psi, the collision
gas was argon at a pressure of 12.5 V (curcumol)
and 20 V (ornidazole, I.S.) for collision-induced
dissociation (CID). Quantification was performed
using multiple reaction monitoring (MRM) of
254.3219.4
(m/z)
for
curcumol
and
220.3128.1 (m/z) for ornidazole (I.S.),
respectively.
Fig 1. Chemical structure of curcumol
Materials and methods

Chemicals and materials
Zedoary turmeric oil fat emulsion (Lot No:
20051213, 200mg/100ml) was supplied by the
Research Institute of Shandong Juren Biology
Medicinal Technology (Shandong, China). Curcumol
standard (Lot No: 100185-200405, 99.1%) was
obtained from National Institute for the Control of
Pharmaceutical and Biological Products (Beijing,
Preparation of stock and working standard

solutions
52
A stock solutions of curcumol at 100 gmL-1

and ornidazol at 50 gmL-1 were prepared by
dissolving appropriate amounts standards in mobile
phase, respectively. The stock solution of curcumol
was diluted subsequently to obtain a series working
standard solutions. The stock solution of I.S. was
further diluted to a working concentration of
0.05gmL-1. All solutions were stored at 4 C.
phase was moved and evaporated to dryness under

gentle nitrogen stream in water-bath at 37 C. The
residues were reconstituted with 100 L mobile phase
and 40 L was injected into the HPLC/MS/ MS
system.
Tissue distribution study
Rats were acclimated for at least 1 week before
experiments. Thirty Wister rats were randomized into
six groups and intravenously injected with a single
dose of 10.0 mgkg-1 zedoary turmeric oil fat
emulsions. Then five rats per group were sacrificed at
0.17, 0.5, 1.0, 2.0, 3.0 and 5.0 h after administration
and tissue samples (heart, liver, lung, kidney and
brain) were collected, respectively. Tissue samples
were rinsed individually with 0.9% sodium chloride
solution, blotted with filter paper, and stored
immediately at -20 C until analysis. The
experimental procedures were carried out in
accordance with the Standard Operating Procedure
for Animal Experimentation of Shandong University
(Shandong, China).
Preparation of calibration and quality control

Samples (QC)
Wister rats without administration were
sacrificed and tissues were collected. Blank tissue
homogenate was spiked by appropriate concentration
of working standard solution to yield final
concentrations between 0.25-50 ngmL-1 of curcumol
in tissue homogenate. Quality control samples were
prepared in the same way at three concentration of
1.0, 10, 40ngmL-1.
Sample preparation
The tissue samples were accurately weighted
and cut into pieces and homogenated after adding the
appropriate amount of 0.9% sodium chloride solution
(3 mLg-1 tissue). 1.0 mL tissue homogenate were
transferred to a centrifuge tube, ornidazole (15 L,
0.05 gmL-1) and sodium hydroxide solutions (100
L, 0.1M) were added and well-mixed. Then 5ml of
chloroform / isopropyl alcohol (95:5, v/v) was added.
The mixture was vortex-mixed for 2min and
centrifuged at 4000 rmin-1 for 5min. The organic
Results
Method validation
Specificity Five batches of different blank tissue
homogenate were analyzed on HPLC-MS/MS to
assess specificity. Typical chromatograms in kidney
and heart were shown in Fig 2 and 3.
Fig 2. Chromatograms of blank kidney (A), blank kidney spiked with curcumol and I.S.
(B), kidney sample spiked with I.S. (C)
1: ornidazole; 2: Curcumol
53
Fig 3. Chromatograms of blank heart (A), blank heart spiked with curcumol(B), kidney samples (C) 1: Curcumol
Table 1. The standard curve equation of curcumol in five tissues (n=5)

tissue
regression equation
linear range (ngmL-1)
R2
heart
Y=4890C+1740
0.5-50.0
0.9968
liver
Y=4230C+3530
0.25-50.0
0.9990
lung
Y=8510C-1240
0.25-25.0
0.9967
kidney
Y=0.075C-0.00532
0.5-25.0
0.9988
brain
Y=3970C-729
0.5-50.0
0.9970
Table 2. Precision and accuracy data for curcumol in tissues (n=5, meanSD)
Tissue
Intra-day
QC
-1
Heart
Liver
Lung
Kidney
Brain
-1
Inter-day
-1
( ngmL )
C( ngmL )
RSD (%)
RE (%)
C( ngmL )
RSD (%)
RE (%)
1.0
0.960.05
5.3
-4.4
0.970.05
5.3
-3.4
10
9.750.17
1.7
-2.5
9.700.26
2.7
-3.0
40
39.790.61
1.5
-0.5
39.560.71
1.8
-1.1
0.5
0.470.03
6.5
-6.4
0.480.03
5.5
-4.3
5.0
4.920.11
2.2
-1.5
4.920.11
2.3
-1.7
40
39.870.62
1.6
-0.3
39.610.71
1.8
-1.0
0.5
0.480.02
3.3
-4.0
0.480.02
4.8
-3.9
4.0
3.910.10
2.7
-2.4
3.940.09
2.2
-1.4
20
19.630.41
2.1
-1.8
19.660.36
1.8
-1.7
1.0
0.970.03
3.1
-2.8
0.970.03
3.5
-2.9
5.0
4.950.06
1.3
-1.0
4.950.08
1.7
-1.0
20
19.860.42
2.1
-0.7
19.740.41
2.1
-1.3
1.0
0.970.04
4.3
-3.2
0.97.04
4.6
-2.8
5.0
4.930.07
1.3
-1.4
4.950.08
1.6
-1.0
40
39.640.44
1.1
-0.9
39.720.87
2.2
-0.7
54

Table 3.
25C,4h
freeze/thaw*
-20C, 1day
-20C, 7days
Stability of curcumol in liver (n=5, meanSD)
QC( ngmL-1)
C( ngmL-1)
RSD (%)
RE (%)
0.5
0.490.01
2.0
-2.1
5.0
4.930.06
1.2
-1.3
40
38.770.69
1.8
-3.1
0.5
0.490.01
2.2
-2.6
5.0
4.920.07
1.4
-1.6
40
39.410.62
1.6
-1.5
0.5
0.480.01
1.9
-4.1
5.0
4.900.07
1.5
-1.9
40
38.560.77
2.0
-3.6
0.5
0.480.01
2.1
-3.8
5.0
4.800.05
1.0
-4.1
40
37.680.39
1.0
-5.8
* After three freeze/thaw cycles at -20C
Calibration and LOQ Five sets of calibration

curves were prepared over the concentration range as
shown in Table 1. The linear regression of the curve
for the peak area ratio (Y) versus concentration(C)
was plotted. The R2 values for the standard curves are
also listed in Table 1. The calibration curve of
curcumol in five tissues showed good linearity. All
coefficients of correlation were between 0.9967 and
0.9990. The limit of quantitation (LOQ) in liver and
lung was 0.25 ngmL-1. For other tissues, LOQ was
0.5 ngmL-1. The limit of quantitation for all samples
was sufficient for tissue distribution study.
Stability The stability of curcumol in tissue

homogenate was studied by analyzing of QC samples.
The samples, which were placed at room temperature
(25 C) for 4h, freezing/thawing for three cycles,
stored at -20 C for 1day and 7days, were
investigated. Results of stability in liver were shown
in Table 3. The stabilities of curcumol in other tissues
were investigated in the same way, the data were not
shown. Curcumol was found to be stable (RE
within8.2%) at different conditions. No significant
degradation occurred during extraction and storage
processes.
Extraction recovery The extraction recovery

was calculated at three QC levels. QC samples were
treated as Sample Preparation. The extraction
recovery was obtained by comparing the curcumol
peak area in tissue samples with those found by direct
injection of corresponding standard solutions. The
mean extraction recovery for all tissues was higher
than 42.3%.
Tissue distribution
Curcumol was distributed to liver, kidneys, brain,
heart, lungs 10 min after intravenous injection and
obtained
the
maximum
concentration
of
108.8565.91, 105.1942.92, 92.3817.63, 82.96
38.06, 10.012.97 gg-1, respectively. The concentrations of curcumol in various tissues of rats at
scheduled time after intravenous injection of 10
mgkg-1 zedoary turmeric oil fat emulsion were
presented in Table 4 and tissues concentration-time
column profiles of curcumol were shown in Fig 4.
Precision and Accuracy The precision and

accuracy of method were assessed by intra-day and
inter-day RSD (relative standard deviation) and RE
(relative error). The precision was less than 6.5% and
the accuracy was less than 6.4%. The results were
shown in Table 2.
Discussion
55
Zedoary turmeric oil fat emulsion was a new

preparation of zedoary turmeric oil. The
pharmacokinetic and tissue distribution results of
animal were necessary to its clinical application.
Zedoary turmeric oil was a multicomponent drug. We
always took the active component as the target

analyte to study its disposition process in vivo.
Curcumol was selected in this study for its high
content and effects.
Table 4 . Mean tissue concentrations of curcumol in rats at scheduled time after intravenous injection of
zedoary turmeric oil fat emulsion(gg-1n=5, meanSD
Time
Heart
Liver
Lung
Kidney
Brain
0.17h
82.9638.06
108.8565.91
10.012.97
105.1942.92
92.3817.63
0.5h
21.0212.36
32.6640.21
3.991.25
40.6618.32
35.326.7
1h
16.2019.99
23.0119.54
1.660.3
26.3516.11
18.8510.42
2h
2.651.76
10.814.16
1.560.39
12.769.71
4.762.34
3h
2.250.68
9.417.88
0.730.49
7.265.39
3.312.06
5h
5.13.02
0.820.2
4.744.05
2.611.32
ND: not detected
120
concentration(g/g)
100
0.17h
80
0.5h
1h
60
2h
3h
40
5h
20
0
Heart
Liver
Lung
Kidney
Brain
Fig 4. Tissue distribution profiles of curcumol at scheduled time after intravenous injection
of zedoary turmeric oil fat emulsion
The quantitation methods of five tissues were

different in this study. The results of specificity in
method validation showed that no distinguish
interference from endogenous compounds was
observed to curcumol in all tissues, but ornidazole
I.S. was interfered in most tissues except kidney.
So internal standard method was selected to
determine the curcumol in kidney, and external
standard method was used to other tissues.

As listed in Table 4, curcumol was detected at
10 min after intravenous injection and undetected in
heart or lower concentrations detected in other tissues
at 5h after administration, which indicated that
curcumol was distributed rapidly and no long-term
accumulation in tissues. It can be seen from Fig 3,
that curcumol in all tissues showed similar dynamic
56
change, the maximum concentration was at 10 min

and markedly decreased at 30min, then declined with
time. The highest level of curcumol appeared in the
liver, followed by kidney, brain, heart, and the lowest
level of curcumol appeared in the lung. These
implied that curcumol were mainly metabolized and
eliminated in the liver and kidney. The higher
concentration in the brain and heart indicated that
curcumol can pass through the blood-brain barrier,
which attributed to its application in cardiovascular
and cerebrovascular disease. The lowest concentration in lung perhaps called in a question in the use
of disease of respiratory system, especially
pneumonia.
The satisfactory outcome was obtained in
present study, but only five main tissues of rats were
investigated, it could not supply the distribution
information of curcumol in vivo completely, so
experiments including more tissues or experiments in
other more superior animals should be studied.
1.
Wu YC, Liu TY, Jiang LG, Bai ZZ. Determination of

curcumol and germacrone in zedoary turmeric oil by
RP-HPLC. Infomation of traditional Chinese medicine 2004;
21(4):64-65.
2.
Xia Q, Zhao KJ, Huang ZG, Zhang P, Dong TT, Li SP, Tsim
KW. Molecular genetic and chemical assessment of
Rhizoma Curcumae in China. J Agric Food Chem 2005;
53(15): 6019-6026.
3.
Yang FQ, Li SP, Chen Y, Lao SC, Wang YT, Dong TT,
Tsim KW. Identification and quantitation of eleven
sesquiterpenes in three species of Curcuma rhizomes by
pressurized liquid extraction and gas chromatography -mass
spectrometry. J Pharm Biomed Anal 2005; 39(3-4):552-558.
4.
Zhou X, Li ZW, Wang DP, Liang GY, Peng BX. Study on

fingerprint of volatile oil of Curcumol wenyujin by GC-MS.
China Journal of Chinese Materia 2004; 29(12):1138-1141.
5.
Zhao Y, Yang R G, Luo M. Progress in pharmacological

action and clinical application of zedoary turmeric oil. J
Practical Tradit Chin internal med 2006; 20: 125-126.
6.
Su CY, Liu JY, Xu HX, et al. The metabolism of

3
H-curcumol in normal rats and tumor-bearing mice. Acta
Pharm Sin 1980; 15:257-262.

7.
Yang SD, Chen JM, Chen YH. Determination of curcumol

in zedoary turmeric oil of Curcumol wenyujin. Acta Pharm
Sin 1979; 1: 356-360.
Conclusion
8.
Tian
SJ,
Liang
WF.
Study
of
zedoary
turmeric
oilDetermination of curcumol and curdione in Curcumol
The tissue distribution of curcumol after

intravenous injection of zedoary turmeric oil fat
emulsion was investigated in the present study. The
established LC-MS/MS method was validated and
applied in determination of curcumol in tissues. The
results showed that curcumol was distributed rapidly
and no long-term accumulation in tissues.
wenyujin with thin-layer scanning method. Chin J Pharm

Anal 1985; 5: 136.
9.
Gu XM, Yang Y. Determination of curcumol in zedoary

turmeric
oil
and
peppermint
oil
by
vapor
phase
chromatography. Chin J Pharm Anal 1982; 2: 75.

10.
Zhang R, Wang BJ, Zhao HL, et al. Determination of

curcumol in plasma by HPLC-MS/MS method and its
pharmacokinetics in Beagle Dogs. Acta Pharmaceutica
Sinica 2007; 42(8):973-977.
References
57
Venkatesh S et al. Asian Journal of Pharmacodynamics and Pharmacokinetics 2009;9(1):58-62
Asian Journal of
Pharmacokinetics
ISSN
1608-2281
Antinociceptive effect of Aerva lanata ethanolic extract in mice:

A possible mechanism
Sama Venkatesh1*, Yanadaiah JP1, Zareen N1, Madhava Reddy B1, Ramesh M2
1
G. Pulla Reddy College of Pharmacy, Mehdipatnam, Hyderabad-500 028, India
Jubilant Innovation, Yeshwantapur, Bangalore-560 022, India

Abstract
Aim Aerva lanata Juss is traditionally claimed to be useful in the treatment of urolithiasis,
strangury, diabetes, headache and pains. In the present study, 80% aqueous ethanolic extract of
(the dried aerial parts of) A. lanata Juss is investigated for antinociceptive activity. Methods
Male Swiss Albino mice were used to test the antinociceptive activity on acetic acid-induced
writhing and hot plate test, at an oral dose of 50 and 100 mgkg-1. The aspirin and morphine
served as standards. Results indicate that the ethanolic extract has produced a significant
(P<0.001) dose dependent analgesic activity in both the test models. The activity is comparable
with aspirin and morphine. The nonselective opioid receptor antagonist naloxone has no effect
on the activity of A. lanata. Conclusion The observed antinociceptive activity may be through
peripheral pain receptors and not by central opioid receptors.
Key words
Aerva lanata Juss; ethanolic extract; antinociceptive activity; analgesic activity; naloxone; mice
Article history
Received
Publication data
Pages:5 Tables: 2 Figures: 0 References: 21

Paper ID 1608-2281-2009-0901058-05
Dr. Sama Venkatesh, Professor, G. Pulla Reddy College of Pharmacy, Mehdipatnam, Hyderabad,
India-500 028. Phone: 91-40-23517222; Fax: 91-40-23515513.
E. mail:
venkateshsama@hotmail.com
7 January 2009;
Accepted 20 February2009
weed in fields and waste places [1]. In traditional

medicine the plant is claimed to be useful in cough,
urolithiasis, diabetes and demulcent. In Ayurveda, A.
lanata is reported to be useful in strangury (slow and
painful discharge of urine) and headache [2, 3].
The presence of alkaloids - methyl aervine,
aervoside, aervoline[4], flavonoid glycosides[5],
-sitosterol, campesterol, chrysin[6] and hentriacontane, -amyrin and buteolin[7] were reported.
Previously it was reported that the alcoholic extract
of A. lanata aerial parts possesses significant diuretic,
anti-inflammatory[8], anti-diabetic[9], antimicrobial [10],
cytotoxic properties[11] and hepatotoxic activity[12].
Introduction
A large proportion of the Indian population for
their physical and psychological health needs depend
on traditional systems of medicine. The Indian
traditional system of medicine, especially Ayurveda
has put forward a number of therapeutic claims on
plant drugs. However it is important to conduct
thorough investigation of as many traditionally used
medicinal plants with reference to modern system of
medicine. Aerva lanata Juss. (Amaranthaceae)
locally known as Pindikunda is an erect; prostrate
under shrub occurs throughout India as a common
58
Committee approved all the procedures for investigating experimental pain in conscious animals [15].
The ethanolic extract of whole plant of A. lanata was

reported to possess nephroprotective activity and in
the treatment of acute renal injury caused by
nephrotoxins like cisplatin and gentamicin [13]. To the
best of our knowledge no report is available on the
analgesic activity of A. lanata. In view of the fact that
the plant is used by the traditional healers to alleviate
headache among the natives, the present study was
undertaken to verify the claim and evaluate the
antinociceptive property of the aerial parts of A.
lanata with the aim of developing a natural analgesic
agent.
Acetic acid-induced abdominal writhing test

The Siegmund et al [16] technique modified by
Koster et al[17] was adopted to assess the
antinociceptive activity in pre-screened mice. Over
night (16 h) fasted mice were divided into six groups
of six animals each. Group-1 served as a control
group, which received 0.5% CMC (carboxy methyl
cellulose) in water.
Group-2 and 3 received
ethanolic extract of A. lanata at a dose of 50 and 100
mgkg-1, respectively as a fine suspension in 0.5%
CMC, orally. Group-4 animals served as positive
control animals received acetyl salicylic acid (100
mgkg-1, po). In an attempt to investigate the
participation of opioid system in the antinociceptive
effect of this plant, separated groups of mice were
pretreated with non selective opioid receptor
antagonist, naloxone (5 mgkg-1, ip), which was
injected 15 min before the administration of extract
(100 mgkg-1, p.o) and acetyl salicylic acid (100
mgkg-1, p.o) for Group-5 and 6, respectively. After
30 min administration, all the animals were given an
intraperitoneal (i.p) injection of 0.6% acetic acid
(volume of injection 0.1 ml/10 g) and number of
writhes produced in these animals was recorded for
30 min.
Material and Methods

Plant Material
Aerva lanata Juss. aerial parts were collected
from Surampet, Karimnagar district, Andhra Pradesh,
India and authenticated by Dr. Prabhakar Reddy,
Taxonomist, Department of Botany, Osmania
University, Hyderabad. A voucher specimen
(AVL-12-2006) is being maintained in the
Department of Pharmacognosy and Phytochemistry,
G. Pulla Reddy College of Pharmacy, Hyderabad,
Andhra Pradesh, India. The aerial parts were cut, air
dried and grounded into powder.
Preparation of Extract
Dried aerial parts powder (650 g) was extracted
with 80% aqueous ethanol by maceration for seven
days. The concentrated aqueous ethanolic extract (23
g; % yield 3.53) was tested for qualitative
phytoconstitiuents and indicated the presence of
alkaloids, steroids and/or triterpenoids, flavonoids
and their glycosides and tannins [14], which
corroborated with the earlier findings.
Hot-plate test
The method of Eddy and Leimback[18] and
Hosseinzabeh et al[19] was employed. The
temperature of hot plate (Eddys hot plate, Dolphin,
Mumbai, India) was maintained at 55 0.2C.
Animals were placed into the Perspex square on the
heated surface, and the time between placement and
licking of paws and jumping was recorded as
response latency. Over night (16 h) fasted mice were
divided into six groups of ten animals each. Control
animals were treated with 0.5% CMC (Group-1),
while morphine (5 mgkg-1, p.o) was used as positive
control (Group-4). Aqueous ethanolic extract of A.
lanata was administered orally, at dose of 50 and 100
mgkg-1 as a fine 0.5% CMC suspension (Group-2
and 3, respectively). The opioid receptor antagonist
naloxone (5 mgkg-1, i.p) was also tested along with
oral administration of ethanolic extract (100 mgkg-1,
Group-5) and morphine (5 mgkg-1, Group-6). All
Animals
Male Swiss Albino mice (25-30 g) were used
throughout the experiment. They were maintained
under standard environmental conditions. Animals
had free access to feed and tap water ad libitum
during the quarantine period. The animal
experimentation was carried according to the
Committee for the Purpose of Control and
Supervision of Experimentation on Animals
(CPCSEA) guidelines and Institutional Animal Ethics
59
substances were administered 30 min before the

beginning of experiment. The reaction time was
measured before and at 30, 60, 90, 120 and 180 min
after substance administration. The latency period of
40 sec was defined as complete analysis and

measurement was terminated if the latency exceeded
latency period to avoid injury.
Table 1. Effect of A. lanata ethanolic extract on the acetic acid-induced abdominal writhing in mice (Mean S.E.M, n = 6)
Dose
(mgkg-1)
Number of writhes
% Inhibition
Control
---
75.35 1.73
---
Aq ethanolic ext.
50
38.5 2.67*
48.90
Aq. ethanolic ext.
100
21.74 0.89*
71.14
Acetyl salicylic acid (positive control)
100
22.72 1.59*
69.84
Aq. ethanolic ext. + Naloxone (ip)
100 + 5
25.8 1.11*
65.75
Acetyl salicylic acid + Naloxone (ip)
100 + 5
28.16 1.07*
62.62
Group
Treatment
*p<0.001 vs control
Table 2.
Group
Effect of A. lanata ethanolic extract on the hot plate test in mice (Mean S.E.M, n=10)
Treatment
Dose
Latency time (min)

-1
( mgkg )
1
2
Control
Aq.
ethanolic
ext.
Aq.
ethanolic
ext.
Morphine
(positive
control)
Aq.
ethanolic
ext.
+
Naloxone
(ip)
Morphine +
Naloxone
(ip)
a
30
60
90
120
180
8.0
0.51
8.5
0.61
9.1
0.47
8.4
0.52
8.5
0.49
8.8
0.44
50
10.7
0.97
13.7
0.76a
14.7
0.74a
13.1 0.64a
17.8
0.89b
18.9
1.08b
100
9.6
0.45
12.9
0.52a
19.1
0.49b
22.1
0.72c
26.3
0.32c
29.7
1.11c
9.3
0.47
21.6
1.89c
40.0
1.37c
39.6
1.49c
36.4
1.51c
31.6
2.10c
8.5
0.56
18.9
0.68b
29.4
0.55c
22.9
0.58c
28
0.61c
30.8
0.77c
10.9
0.52
14.6
0.47b
13.4
0.95a
13
0.73a
9.0
0.33
9.6
0.63
---
100 + 5
5+5
p<0.05, b p< 0.01,c p< 0.001 vs control
60
the scientific basis for the utilization of A. lanata in

the treatment of pain in traditional medicine. The test
extracts and the standard drugs were administered
orally as it is simple and physiological. The
antinociceptive effect of A. lanata ethanolic extract
has shown a significant dose dependent reduction in
the number of abdominal writhes compared to the
control values. Strong inhibition was observed at a
dose of 100 mgkg-1 and the activity is comparable
with that of acetyl salicylic acid. In addition,
pretreatment with non selective opioid receptor
antagonist naloxone has not antagonized the A. lanata
(100 mgkg-1) and acetyl salicylic acid induced
analgesic activity. The % inhibition is 65 and 62 of
extract and acetyl salicylic acid, respectively in
naloxone treated animals.
Despite the significant results obtained, the
abdominal writhing test method is reported to be a
less selective model. It has been reported that acetic
acid acts indirectly by releasing endogenous
mediators which stimulate neurons that are sensitive
to centrally acting agents such as narcotics[20]. The
hot plate test was employed to verify if the extract
could show central antinociceptive effect as the test is
a specific central antinociceptive test [21]. In the
present study it was demonstrated that the animals
treated with ethanolic extract of A. lanata showed
increase in their response latency period in
comparison with the control group. The
antinociceptive activity of A. lanata was not
antagonized by pretreatment with naloxone, whereas
it significantly decreased the latency period in the
morphine induced antinociception.
Statistical Analysis
The results were expressed as mean S.E.M.
The differences between experimental groups were
compared by one-way ANOVA (control versus
treatment by Bonferronis method; using Jandal
Scientific, Sigmastat statistical software, version 1.0)
and were considered statistically significant when P <
0.05.
Results
Effect of acetic acid- induced writhing test
Oral administration of A. lanata ethanolic
extract (50 and 100 mgkg-1) has produced dose
dependent significant (P<0.001) antinociceptive
activity in acetic acid induced writhing test with 48
and 71% inhibition, respectively (Table 1). The
activity produced by acetyl salicylic acid is
comparable with the activity produced by A. lanata at
a dose of 100 mgkg-1. The administration of
naloxone along with A. lanata (100 mg/kg) and
aspirin demonstrated significant antinociceptive
effects. The results indicate naloxone has no effect on
the activity of A. lanata and acetyl salicylic acid
(Table 1).
Effect on hot plate test
The results presented in Table-2 show the time
course of antinociception produced by
A. lanata
-1
(50 and 100 mgkg ).
Oral administration of
ethanolic extract of A. lanata resulted in a significant
and dose dependent prolongation of the response
latency in the hot plate test. The prolongation of
latency period was observed with the time course of
test. Morphine sulphate (5 mgkg-1) significantly
(P<0.001) increased the response latency period of
mice with maximum effect obtained 1 hr after
treatment. However the activity of A. lanata is
comparable with morphine at 180 min. In addition,
the administration of naloxone demonstrated
significant reversal of the antinociceptive effect of
morphine and had no effect on the activity of
ethanolic extract of A. lanata (100 mgkg-1).
Conclusion
The results of the present investigation indicate
that pretreatment of animals with naloxone has not
antagonized the analgesic activity of A. lanata extract
in both acetic acid-induced writhing and hot plate test.
Since naloxone, is a classical non selective opioid
receptor antagonist, it was not able to alter the A.
lanata induced analgesic activity in both models.
Thus the observed significant antinociceptive activity
may have resulted from antinociception of peripheral
receptors and not by central opioid receptors. Though
the exact mechanism of action is not known at this
stage, the present experimental results conclude that
Discussion
The results presented here may help to establish
61
aqueous ethanolic extract of A. lanata is a potential

analgesic agent.
11.
solid tumor induced by DLA cells in mice.

12.
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Kirtikar KR, Basu BD. Indian Medicinal Plants. 2nd edn, Vol.
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Kokate CK. Practical Pharmacognosy. 4th ed, India:
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Biol Med 1957; 95: 729-31.
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Chopra RN, Nayar SL, Chopra IC, Glossary of Indian
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Zapesochnaya GG, Pervykh LN, Kurkin VA. A study of
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Pervykh LN, Karasartov BS, Zapesochnaya GG. A study
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Flavonoid
20.
Chandra S, Shastry MS. Chemical constituents of Aerva
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Salmami
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Collier HO, Dinen LC, Johnson CA, Schneider C. The

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effect of fentanyl and other morphine-like analgesics on
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the warm water induced tail withdrawal reflex in rats.
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alcoholic extracts of Aerva lanata Juss. in rats.
H,
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Acknowledgements
The authors wish to
thank All India Council of Technical Education, New
Delhi, India financial support. The authors also wish
to thank management of the college for providing
research facilities.
3.
Nevin KG, Vijayammal PL. Effect of Aerva lanata on
Chowdhury D, Sayeed A, Islam A, Shah Alam Bhuiyan M,

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Li R et al. Asian Journal of Pharmacodynamics and Pharmacokinetics 2009; 9(1):63-70
Asian Journal of
Pharmacokinetics
ISSN
1608-2281

Therapeutic effect and mechanism for anti-fibrosis of

polyhydroxysilbene of Rhizoma Scirpi in hepatic fibrosis rats
Run Li, Zong-Peng Zhang, Yi-Hong Tian
Research Center for New Drug Evaluation, Tianjin State Key Laboratory of Pharmacokinetics and
Pharmacodynamics, Tianjin Institute of Pharmaceutical Research, Tianjin 300193, China
Abstract
Aim To study therapeutic effect and mechanism for anti-fibrosis of polyhydroxysilbene of

Rhizoma Scirpi in hepatic fibrosis rats. Methods Fluidextract contained polyhydroxystilbene
(PHS), The radix of Spcirpus yagara Ohwi. Was extracted with ethyl acetatethe therapeutic
effect and mechanism on hepatic fibrosis is studied in this article. The test assay included serum
levels of ALT, AST, TP, ALB and the levels of HA, LN, MDA, SOD and hapetic tissue
pathomorphological examination. Results In animal experiment, study showed that the
levels of ALT, AST in serum and the levels of HA, LN, MDA in rats liver in treatment groups is
significant lower than that in pathologic control group(model group, group B).Compared with
rats in pathologic control groupthe levels of SOD significant increased in the groups rats
treated with Fluidextract or malotilate.In vitro, hepatic microsomes of rats were prepared by
centrifugation. NADPH/Vit-C or cysteine/FeSO4 reaction system is used to induce the
production of MDA in rats liver microsomes. Conclusion The fluidextract suggested a better
antioxidant effect.A dose-response relationship was found between the antioxidant effect and
concentration of the fluidextract in the two reaction systems.The antioxidant effect may be one
of the cardinal mechanisms in the therapeutic effect of extract of Spcirpus yagara Ohwi to
hepatic fibrosis.
Key words
Spcirpus yagara Ohwi, polyhydroxystilbene, Hepatic fibrosis, lipid peroxidation, Hepatic

microsomes
Article history
Publication data
Received 9 December 2006; Accepted 21 July 2008

Pages: 8; Tables: 5 ; Figures: 7; References:18; Paper ID 1608-2281-2009-09010063-08
Professor Zong-Peng Zhang, Research Center for New Drug Evaluation, Tianjin State Key
Laboratory of Pharmacokinetics and Pharmacodynamics, Tianjin Institute of Pharmaceutical
Research, Tianjin 300193, China. E-mail: zhang88985@163.com
development of irreversible cirrhosis in patents[1-3].

There are various kinds of chronic liver injuries all
over the world, causing great affection to patents.
Therefore, searches for effective ways to inhibit
Introduction
Hepatic fibrosis is a common pathological
process of chronic hepatic disease, leading to the
63
fibrosis and prevent the development of cirrhosis

are great significance[4]. Chinese traditional and
herbal medicines, which are well known for the
long history of prevent therapy of various diseases
with low cost and few side effects, have particular
potentials in the treatment of hepatic fibrosis[5-11].
Rhizoma Sparganii, which is a dried radix of
Sparganium stoloniferum Buch.-Ham. (Family:
Sparganiaceae) possesses wide pharmacological
activities and is a commonly used traditional Chinese
medicine for the treatment of chronic hepatitis in
clinics of traditional medicine. Xiao and his
co-workers reported that the Rhizoma Sparganii can
protent hepetic cells, alleviate degeneration and
necrosis, recover hepatic cell structure and liver
function, and reduce the proliferation of fibrous tissue
in the rat immunohepatic fibrosis model. Rhizoma
Spcirpi, which is a dried radix of Spcirpus yagara
Ohwi (family: Scirpusaceae) (Fig 1, www.
zwy.csuft.edu.cn is also sued as a substitute for
Rhizoma Sparganii in traditional Chinese medicine[15].
In chemical ingredients, the two herbs are different.
Polyhydroxystilbene (3, 3, 4, 5-tetrahydroxystilbene),
scirpusia A and B, veroeratrol, betulin and betulinic
acid were found from Spciepus yagara and were not
found
from
Sparganium
stoloniferum.
Polyhydroxystilbene is a main chemical ingredient of
Spciepus yagara and its pharmacological activity on
inhibition of lever fibrosis was not investigated.
Fig 1.

The extract and drug
Polyhydroxystilbene
fluidextract
(PHS)
extracted from plant materimals of Spcirpus yagara
Ohwi. Rhizome with ethyl acetate is
Yellowish-brown.
Eevery
gram
of
polyhydroxystibene fluidextract is equivalent to 300
g crude materials.As a positive control drug
malotilate tablets are purchased from Shanxi Yabao
Pharmaceutical Co.Ltd., China.
Animals
Wister rats, male and female, with body weight
from 250g to 300gsupplied by laboratory animal
unit of Tianjin Institute of Pharmaceutical Research.
Instruments and Reagents
The centrifugeVITALAB Selectra 2 automatic
biochemistry analyzer and ELISA were used in
experiment.The biochemistry assay kits and the
quality control serums are provided by Zhongsheng
Beikong Bio-technology and Science Inc of china.
The rat hyaluronic acid HAand lamininLN
ELISA kits are manufactured by Market inc of
U.S.A.
Methods of animal experiment
Rats are randomly divided into 6 groups
(6/group), group A, B, C,D, E and F respectively.Rats
at group BCDE and F received a single oral dose
of 30% CCl4 (0.2ml/100g) in sesame oil by gavage
for 8 weeks.After 2 weeksrats at group C received a
single oral doses of 75mgkg-1d-1 malotilate
suspensions in water by gavage for 6 weeksand
group A, D, E and F received a single oral doses of 0,
0.15, 0.6, 2.4 mgkg-1d-1 polyhydroxystilbene
fluidextract fluidextract suspensions in water/tween
80 by gavage for 6 weeks.Rats fasted 16 h and then
sacrificed after the last dose.At the same timesblood
and liver sample were also collected.
Spciepus yagara Ohwi
In this study, the effects of Polyhydroxystilbene extraction from Spciepus yagara on lever
fibrosis used an induced lever fibrosis model by
carbon tetrachloride in rats. The antioxidant effect to
be one of the cardinal mechanisms in the therapeutic
effect to hepatic fibrosis was employed to examine.
Assay methods for serum biochemiscal

values
The serum biochemistry values including
alanine
aminotransferase
(ALT),
aspartate
aminotrasferase(AST), total protein(TP), albumin
64
NADPH/Vit-C[14] and cysteine/FeSO4 reactive

system[16]. The concentration of MDA in rats liver
microsomes suggesting peroxidized levels of liver
microsomes, were measured with thiobarbituric acid
method (TBAcolorimetry).
(ALB) and the values of superoxide dismutase

(SOD)hydroxyproline (Hyp) in liver were assayed
using automatic biochemistry analyzer with the
methods offered by biochemistry assay kits[16]. The
serum values of HA LN were assayed using
ELIASA with the methods offered by ELISA
kits.The concentration of MDA in liver was measured
with thiobarbituric acid (TBA) method (colorimetry).
Statistical analysis
All data were analyzed by SPSS for windows
statistical package. Measurement data were analyzed
by ANOVA, ranked data were analyzed by
Kruskal-Wallis test. P values were less than 0.05
were considered to be statistically significant.
Pathological examination
The pathological change of hepatic slice by HE
dyeingwere observed under the light microscope.
The classification and statistical analysis of hepatic
fibrosis refer to Consensus on evaluation of the
diagnosis and efficacy of hepatic fibrosis[11].
Results
Animal experiment
Effect on liver functions At the end of
treatment period5/12 animals died in group B and D,
3/12 animals in group C, E and F died respectively
while no died in group A. Decrease in mortality was
observed in group C, E and F.
Methods of anti-peroxidized experiment in

vitro
In vitro,hepatic microsomes of rats were
prepared by centrifugation[12,13]. The production of
MDA in rats liver microsomes were induced with
Table 1. Effect of polyhydroxystibene fluidextract on serum AST, ALT, TP and ALB values of rats (Mean SD)
Animal
ALT
amountn
u/L
AST
u/L
**
TP
ALB
g/L
g/L
**
74.15.5
33.83.5*
Group A
31.89.2
115.634.7
Group B
183.959.0
346.8110.7
70.36.2
29.92.9
Group C
87.631.2
**
**
71.54.4
30.12.2
Group D
152.751.3
265.664.9
72.25.5
29.32.9
Group E
176.593.8
328.6131.1
69.64.9
29.72.9
Group F
109.762.0*
195.853.2*
71.04.3
30.42.6
167.463.2
Significant relative to group B* p <0.05, ** p <0.01

Table 2. Effect of polyhydroxystibene fluidextract on SOD, MDA, Hyp levels in liver tissue and
HA,LN content in serum of rats (Mean SD)
Animal
SOD
MDA
-1
Hyp
HA
-1
LN
-1
ngml-1
amountn
umg
(OD)
umg
ngml
Group A
26.67.8
0.1170.021
0.5610.086
49.66.3**
29.54.2**
Group B
22.01.9
0.1370.019
0.9110.097
181.372.0
75.010.6
25.52.4
**
105.624.5
42.717.8**
0.8270.157
129.037.0
59.924.4
0.1090.029
0.7000.178
89.922.0
53.214.0*
0.1030.017**
0.7650.227
74.318.1**
48.421.8*
Group C
Group D
25.03.6
0.1280.029
Group E
24.72.3
Group F
31.13.2**
0.1150.013
Significant relative to group B* P <0.05,** P<0.01
65
0.6110.039
**
in the MDA values in rats liver.As shown in Table 2

the serum HA and LN values are increased
significant in group B (vs group A, P<0.01).
Compared with group B, the content of serum HA in
group F decreased significantly (P<0.01), and the
content of serum LN in group E and F decreased
significantly (P<0.05).
As shown in Table 1, the ALT, AST values in

group B is significant higher than that in group A
(P<0.01)and TP and ALB values in group B is
lower than that in group A.Rats treated with
malotilatethe ALTAST values of its is significant
lower than that in group B (P<0.01). Compared with
group Bdecrease in the values of ALT, AST is
observed in treatment groups of polyhydroxystibene
fluidextract. The decreasing tendency of the ALT and
AST values is observed in group F (P<0.05).
The SOD values in rats liver in group B are
obvious lower than that in group A. Malotilate and
polyhydroxystibene
fluidextract
produced
dose-dependent increase in the SOD levels in rats
liver.Compared with group Bpolyhydroxystibene
fluidextract at 2.4 mgkg-1d-1 and Malotilate at 75
mgkg-1d-1 produced significant increase in the SOD
values in rats liverand produced significant decrease
Pathomorphological examination
As shown in pathomorphological examination
hepatocyte necrosis, inflammatory cellular infiltrated,
hyperplasia and hepatocirrhosis were observed at the
liver slice in group B.Compared with group B
pathomorphological changes are slight in group C
and F.After the analysis of Kruskal-Wallis testresult
is shown in Table 3.
Table 3. The statistical result of pathomorphological examination on rat hepatocirrhosis

Graded to hepatocirrhosisanimal amount
0
Group A
9/9
Group B
1/7
2/7
4/7
Group C*
2/9
4/9
3/9
Group D
1/7
5/7
1/7
Group E
1/8
1/8
5/8
1/8
Group F*
1/8
2/8
5/8
Significant relative to group B* P <0.05.
Fig 2. Hepatic pathological photos of rats in group A (normal control group).

The structure hepatocyte and hepatic lobulesis are normality
The In intro experiment

The basal MDA value OD of lipid
peroxidation was 0.830.51 in reaction system. After
stimulated by Vit-C and NADPH, the MDA value
was 0.900.03 in reaction system. As shown in Table

4, PHS fluidextract completely inhibited the lipid
peroxidation induced by Vit-C and NADPH in
microsomes at the concentration of 15 gml-1 (Fig 2).
66
At the concentrations of 75gml-1, and 375ml-1, the

lipid peroxidation value was signaficantly lower than
that of control group (p <0.01).
Fig 3. Photos of group B. denaturation and necrosis are obversed in hepatocyte .Inflammatory cellular infiltratedhepatic
plate arrayed turbulenthyperplasia and hepatocirrhosis were obversved in above photos
Fig 4. Photos of group C. Compared with B, pathomorphological changes is slight.
Fig 5. Photos of group E. Compared with Bpathomorphological changes is slight
Photos of group B. denaturation and necrosis are

obversed in hepatocyte .Inflammatory cellular
infiltrated hepatic plate arrayed turbulent,
hyperplasia and hepatocirrhosis were obversved in
above photos (Fig 3). The pathomorphological
changes of group C and E compared with B as shown

in Fig 4 and 5.
As shown in Table 5, the basal value of lipid
peroxidation was 0.890.05 (MDA, OD). After
stimulated by cysteine and FeSO4the MDA value
67
was 1.040.05 in reaction system. Polyhydroxy-
group (P<0.01). The fine liner relationship was
stilbene fluidextract completely inhibited the lipid
obversed between the dose and the effect (Fig 6).
peroxidation induce by cysteine and FeSO4 in the
Effects of polyhydroxystibene fluidextract on lipid
-1
system at the concentration of 5gml . At the

-1
perxidation induced by cysteine/FeSO4 in liver
-1
concentrations of 5gml and 10 gml , the lipid
microsomes of rats are shown in Fig 7.
peroxidation value was lower than that of control

Table 4. Effect of polyhydroxystibene fluidextract on lipid peroxidation induced by
Vit-C and NADPH in rats liver microsomes (n=5, Mean SD)
Group
Concentration (gml-1)
Control
Induced model
PHS fluidextract
PHS fluidextract
PHS fluidextract
PHS fluidextract
3
15
75
375
MDA (OD)
0.830.51*
0.900.03
0.870.03
0.720.03**
0.590.01**
0.540.02**
Significant relative to Induced model group** P <0.01.
Fig 6. Effects of polyhydroxystibene fluidextract on lipid peroxidtion induced by

Vit-c and NADPH in rats liver microsomes
Table 5. Effects of polyhydroxystibene fluidextract on lipid perxidation induced by cysteine/FeSO4

in rat liver microsomes (n=5, MeanSD)
Group
Concentration (g/ml)
MDA(OD)
Control
0.890.05**
Induced model
1.040.05
PHS fluidextract
2.5
0.940.03**
PHS fluidextract
0.890.03**
PHS fluidextract
10
0.800.03**
Significant relative to Induced model group** P<0.01.
68
Discussion
of its traditional effect, such as effect of activating

blood circulation and releasing pains.From a
literature survey ,it appears that none of the previous
investigators is concerned with the anti-hepatic
fibrosis effect of Spcirpus yagara Ohwi. According
to the Chinese tridetional medical theories, stagnation
and block of the blood vessel was the pathogenesis
on hepatic fibrosis patients.Ameliorated the blood
sludging very avails to hepatic fibrosis patients.
Spcirpus yagara Ohwi can meliorate economy
microcirculation, and can meliorate blood providing
of liver.
Stigma of blood stasis always appears in hepatic

fibrosis patients. For its effect of activating blood
circulation to dissipate blood stasis, Spcirpus yagara
Ohwi always occurs in herbalist doctor`s
prescriptions for treatment of the hepatic fibrosis.
Pharmacological studies in animal models indicated,
the pharmacological effect of Spcirpus yagara Ohwi
is in accord with the traditional efficacy. In fact, it is
a question that the pharmacological research of
Spcirpus yagara Ohwi still stagnated in the research
Fig 7. Effects of polyhydroxystibene fluidextract on lipid perxidation induced

by cysteine/FeSO4 in liver microsomes of rats
by Vit-c and NADPH in microsomes at the

concentration of 5mgml-1 and completely inhibited
the lipid peroxidation induced by cysteine and
FeSO4 in the system at the concentration of
1.6mgml-1.
In conclusion, polyhydroxystibene fluid extract
is an effective substantiate in treating hepatic
fibrosis. Its mechanism might be probably related to
the effect of anti-lipid peroxidation.
Thereforethe therapeutic effect of Spcirpus

yagara Ohwi on hepatic fibrosis was studied with the
harmacological method by us in our laboratory, at the
same time, we hope to reveal the mechanism of its.
Experimental
observation
shown,
polyhydroxystibene fluidextract which has certain
effect on hepatic fibrosis of rats model, could
significantly meliorate the hepatic function of model
rats,protect hepatocyte and abate hepatocyte
necrosis.Rats striked by CCl4shown obvious stigma
of injured by lipid peroxidationthe SOD values in its
liver decreased significantlythe content of MDA
increased.It is generally accepted that lipid
peroxidation is the bridge between hepatic fibrosis
and continuate hepatitis (Liu, 2002). Thereforby
employing reactional system of NADPH/Vit-C or
cysteine/FeSO4 we focused attention on the
mechanism of therapeutic hepatic fibrosis effect of
Spcirpus yagara Ohwi.
As the result in above, PHS fluidextract can
resist liver microsame injury of lipid peroxides
completely inhibited the lipid peroxidation induced
Acknowledgments
Authors are very grateful to
Professor Liu CX for his helps. This work is completed

partly by Guo CM, Wang JJ, Cheng XX, Hu LZhang JX
and Wang H. This study was supported by National Basis
Research Plan (973 Plan) No 2007CB516807.
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Nie XP et al. Asian Journal of Pharmacodynamics and Pharmacokinetics 2009; 9(1):71-76
Asian Journal of
Pharmacokinetics
ISSN
1608-2281

Changes of the adenosine content in single and mixed decoctions

of Gualou-xiebai-baijiu decoction
Xiao-Pu Nie1, Wen-Yuan Gao1*, Pei-Gen Xiao2
1
School of Pharmaceutical Science and Technology, Tianjin University, Tianjin 300072, China
Institute of Medicinal Plant, Chinese Academy of Medical Sciences and Peking Union Medical College,
Beijing 100094, China
Abstract
Aim To investigate the adenosine content in single and mixed decoctions of

Gualou-xiebai-baijiu decoction, we developed a simple and rapid High-performance liquid
chromatographic (HPLC) method for the determination of adenosine, an active constitute to
restrain platelet aggregation in Gualou-xiebai-baijiu decoction. Methods The decoctions were
separated on a HiQsilC18 column4.6mm250mm5mand detected by ultraviolet detection
at a wavelength of 260nm and at the temperature of 30 C. The use of methanol-water (10:90,
v/v) as the mobile phase at a flow rate of 1.0 mLmin-1 enabled the baseline separation of the
drugs free from interferences with isocratic elution. Results The content of adenosine in the
mixed decoction is much higher than those in the single decoction which was extracted with
20% ethanol. But it is much less than in the single decoction which was extracted with 10% and
30% ethanol. The content of adenosine in single and mixed decoctions have exceeding
significant difference (P<0.01). The linearity of the calibration curve was obtained (r = 0. 9997)
over the analytical range from 1.984 to 19.84g mL1. The average recovery was 100.0 %
(R.S.D. = 1.6 %, n=6). Conclusion The adenosine content in the single and mixed decoctions
has exceeding significant difference. The dissolution of adenosine is related with both the
ethanol concentration and decoction.
Key words
Gualou-xiebai-baijiu decoction; adenosine; single

High-performance liquid chromatographic method.
Article history
Received 9 July, 2008;
Publication data
Pages: 7 Tables: 1
Professor Wen-Yuan Gao, School of Pharmaceutical Science and Technology, Tianjin
Accepted
decoction;
mixed
decoction;
25 December 2008
Figures: 3 References: 15
Paper ID 1608-2281-2009-09010071-06
University, Tianjin 300072, China. E-mail: pharmgao@tju.edu.cn
adenosine receptors, four of which have been cloned

and pharmacologically characterized: A1, A2A, A2B,
and A3[1]. Among the four subtypes, A1and A 2A are
associated with behavioral effects that are exhibited
in animals treated with adenosine analogs[2].
Introduction
Adenosine is an endogenous nucleoside that
generally acts as a neuromodulator in the central
nervous system (CNS). Its actions are mediated by
71
Numerous studies have shown substantial evidence

that adenosine modulates cardiovascular control at
the level of the nucleus tractus solitarius (NTS) [3-6].
In the last few years, there are more and more
researches on the differences between the single
decoction and the mixed decoction [7].To our
knowledge, however, there is no report on
Gualou-xiebai-baijiu decoction in this aspect.
Adenosine (Fig. 1) is an active constitute to inhibit
platelet aggregation and has been separated from both
Fructus trichosanthisi and Bulbus alliimacrostemi [8,9].
Moreover, Fructus trichosanthisi and Bulbus
alliimacrostemi are often paired a basic prescription
in many prescriptions to treat cardiovascular disease
in traditional Chinese medicine[10].
Nowadays, coronary heart disease CHD is
severely harming human health for its high
incidence and death rate. CHD belongs to
arthromyodynia category according to traditional
Chinese medicine. The traditional Chinese medicine
plays more and more important role in the treatment
of CHD. Gualou-xiebai-baijiu decoction is a
well-known classic preparation in traditional
Chinese medicine. It originates from Jinkui Yaolue,
a famous pharmaceutical book written by Zhang
Zhong-Jing in the Han dynasty. The prescription
consists of Fructus trichosanthisi and Bulbus
alliimacrostemi and is extracted with wine [11]. It has
been used to treat CHD and angina pectoris for
several hundred years and has achieved good results
in modern clinical practice [12].
NH
of Gualou-xiebai-baijiu decoction.

Chemicals and reagents
The crude herb of Fructus trichosanthis and
Bulbus allii macrostem were purchased from Anguo
Meiwei Pharmaceutical Co. (Hebei, China).
Adenosine was obtained from National Engineering
Research Center of technique of manufacture of the
traditional Chinese medicine solid preparation
(Jiangxi, China).
Methanol (chromatographic
grade) was purchased from Concord Technological
Company (Tianjin, China). Water was twice
distilled water. Other reagents used were at least
analytical grade.
Equipment
The HPLC system consists of an Agilent1100
serials pump, solvent degasser, column oven, UV
detector. This system was operated with a Chem
Station software (Version: A.09.01) (Agilent,
USA).The HPLC consists of a 5m C18 column
(4.6250mm) (HiQ silC18, Japan) and a safeguard
column of the same material.
Chromatographic conditions
The column temperature was kept constant at
30 and the mobile phase flow rate was
1.0mLmin-1. All of injection volume was 10L.
For quantitative assay, the mobile phase was
methanol -water (10:90, v/v) and the UV detector
measured absorbance at 260 nm.
N
N
Standard solutions
Stock solution of adenosine of 19.84gmL-1
was prepared in 90% methanol. Appropriate
dilutions (v/v) of the stock solutions were made
with 90% methanol to obtain working solutions
from 1.984 to 19.84gmL-1 of adenosine. These
solutions were used for the study of linearity,
accuracy, repeatability and recovery.
The linearity of the method was established by
using 5 standard solutions for adenosine
(concentration range of 1.98419.84gmL-1),
assayed in triplicate on the experiment.
N
O
HO
H
OH
OH
Fig 1. Chemical structure of adenosine
In this paper, we established a simple and rapid

High-performance liquid chromatographic (HPLC)
method for the determination of adenosine in
Gualou-xiebai-baijiu-tang and then investigated the
content of adenosine in single and mixed decoction
72
injected into HPLC.
Sample preparation
Mixed decoction preparation The Fructus
Trichosanthis and Bulbus Allii Macrostemi 5.0g
(the proportion of Fructus Trichosanthis and Bulbus
Allii Macrostemi was 3:2), were soaked by 20%
ethanol (40ml) for 0.5h. Then they were extracted in
a regurgitate bath for 1 h. The decoction was
filtered through absorbent gauze. The remainder
slags were continually extracted in a regurgitate
bath for 1 h by 20% ethanol (30ml). The decoction
was filtered through absorbent gauze and merged
with the first decoction. For determination of
adenosine, the solution was filtered through a
membrane (0.22m) and then injected into HPLC.
Data analysis
Statistical analysis was performed using the
repeated-measures ANOVA test, followed by t test, to
compare different group.
Results and discussion

Selection of chromatographic conditions
A good separation condition should satisfy the
need that the analyzed peaks have baseline
separation with adjacent peaks within a short
analysis time as far as possible.
To obtain the
chromatograms with good separation, we tried
various mobile phase. According to literatures [13, 14],
buffer salt solutions which were prepared
complicatedly were used and damage the
chromatographic column. So we tried the mixtures
of acetonitrile and water (18:92, 4:96, v/v) as
mobile phase but separation was not satisfactory.
When the mixtures of methanol and water (15:85,
13:87 and 12:88, v/v) were used as mobile phase,
the analyzed peaks did not have baseline separation
with adjacent peaks. Only when using methanol and
water (10:90, v/v) satisfactory separation result can
be obtained.
Single decoction preparation The Fructus

Trichosanthis 3.0g was soaked by 20% ethanol
(24ml) for 0.5h. Then they were extracted in a
regurgitate bath for 1 h. The decoction was filtered
through absorbent gauze. The remainder slags were
continually extracted in a regurgitate bath for 1 h by
20% ethanol (18ml). The decoction was filtered
through absorbent gauze and merged with the first
decoction.
The Bulbus Allii Macrostemi 2.0g was
extracted by the same method as Fructus
Trichosanthis. Finally, the decoction of Fructus
Trichosanthis and Bulbus Allii Macrostemi was
filtered through a membrane (0.22m) and then
Sample
Table 1. Recovery of adenosine in Gualou-xiebai-baijiu decoction

Initial content (g) Amount added (g) Recovery (%) Average value of recovery (%)
100.5985
100.192
99.4
108.3180
108.128
102.0
99.6212
103.168
99.7
98.3459
98.208
98.5
102.5576
102.176
101.8
100.1929
100.192
98.5
100.0
R.S.D. (%)
1.6
resulted in straight lines over the concentration

range. The linearity equations were calculated by
using linear regression analysis, and typical
calibration curves were defined by the following
equations: y = 30.206x-5.3233, r=0.9997.
It is possible to conclude that, for adenosine,
the functions are linear in the working range.
Method validation
Linearity The linear range for adenosine
(1.98419.84gmL-1) was evaluated. In linearity
range was used 5 different standard solutions of the
lower and upper limit concentration. Calibration
curves were obtained by plotting the peak area
versus adenosine concentrations (gmL-1) and
73
19.047
VWD1 A, Wavelength=260 nm (NIE\GX000037.D)

mAU
4
-2
-4
10
15
20
25
mi
A
mAU
mAU
100
100
80
80
60
60
40
18.970
18.968
40
20
20
0
-20
0
10
15
20
25
mi
10
15
20
25
mi
mAU
mAU
80
80
60
60
40
18.968
18.940
40
20
20
0
0
-20
0
10
15
20
25
mi
10
15
20
25
mi
mAU
mAU
100
80
80
60
60
40
40
18.944
18.845
20
20
0
-20
0
10
15
20
25
mi
F
Fig 2.
10
15
20
25
mi
G
HPLC chromatograms of chemical reference substance and sample
Accuracy The accuracy of this method was

determined by comparing the calculated
concentration using calibration curves to the known
concentrationR.S.D.=0.41%,n=6.
Stability The stability study shows that the

decoction containing adenosine is stable at room
temperature for 12hR.S.D.=1.5%,n=6.
Repeatability The content of adenosine in 5
74
different decoction styles are uniform on the whole.

A chromatogram of a adenosine standard
solution sample; B chromatogram of a mixed
decoction sample by 10% ethanol; C chromatogram
of a single decoction sample by 10% ethanol; D
chromatogram of a mixed decoction sample by 20%
ethanol; E chromatogram of a single decoction
sample by 20% ethanol; F chromatogram of a
mixed decoction sample by 30% ethanol; G
chromatogram of a single decoction sample by 30%
ethanol.
samples was prepared according to the method in

2.5.1. The obtained R.S.D. values were 1.55%.The
method is precise when working under the same
operating conditions (e.g. same operator and
equipment).
Recovery The recoveries of six samples were
investigated. Average recovery of adenosine was
generally above 98%.The results are shown in Table
1.
Sample analysis
Chromatogram Typical chromatograms of
samples are shown in Fig.2. All samples were
completely separated. The results presented here
show that a HPLC method for the measurement of
adenosine has been developed. The categories of
compounds in Gualou-xiebai-baijiu decoction
extracted from different solution have changed. The
categories of compounds in Gualou-xiebai-baijiu
decoction which extracted by same solution but
Analysis mixed decoction samples by

different ethanol concentrations Samples were
prepared according to the method in 2.5.1, by 10%
ethanol, 20% ethanol and 30% ethanol, respectively.
Then the contents of adenosine in them were
investigated. The results are shown in Fig.3.
Fig 3. Results of adenosine content in Gualou-xiebai-baijiu decoction

single decoction compared to mixed decoction which were extracted from the same solution.( **P<0.01)
was extracted with 20% ethanol. But it is much less

than the single decoction which was extracted with
10% and 30% ethanol.
The dissolution of adenosine is related with
both the ethanol concentration and decoction.
Single decoction conduces to the dissolution of
adenosine at 10% ethanol and 30% ethanol.
However, the dissolution of adenosine in mixed
decoction is better than in single one. Mixed
decoction, Gualou-xiebai-baijiu decoction, at 20%
Analysis single decoction samples by

different ethanol concentrations Samples were
prepared according the method in 2.5.2, by 10%
ethanol, 20% ethanol and 30% ethanol, respectively.
Then the contents of adenosine in them were
investigated. The results are shown in Fig.3.
The contents of adenosine in single and mixed
decoction has very significant difference (P<0.01).
The content of adenosine in the mixed decoction is
much higher than that in the single decoction which
75
ethanol had the best protection effect to the rats

which had undergone a thirty-minute ligature of
coronary left anterior descending branch and used
as a model of ischemic reperfusion[15]. It is possible
that the content of adenosine in the decoction is
higher at 20% ethanol than in other concentration
ethanol solutions (10% and 30%). The traditional
Chinese medicine prescriptions often produce a
joint action through many ways and targets. More
active constitutes and pharmacology researches on
the traditional Chinese medicine prescriptions
should be done in more aspects.
Nerv Syst 1996; 59: 113124.

4.
Barraco RA, Phillis JW. Subtypes of adenosine receptors in

the brainstem mediate opposite blood pressure responses.
Neuropharmacology 1991; 30: 403407.
5.
Mosqueda-Garcia R, Tseng CJ, Appalsamy M, et al.

Modulatory effects of adenosine on baroreflex activation in
the brainstem of normotensive rats. Eur J Pharmacol 1989;
174: 119122.
6.
Scislo TJ, OLeary DS: Mechanisms mediating regional

sympathoactivatory responses to stimulation of NTS A1
adenosine receptors. Am J Physiol Heart Circ Physiol 2002;
283: H1588H1599.
7.
Deng Y Y, Gao W Y, Chen H X. et al. Advances in studies

on variability of single and mixed decoction of the
traditional
Chinese
medicine
prescription.
Chinese
Traditional and Herbal Drugs 2005; 36(12):1909-1911.
Conclusion
8.
Liu D L, Qu G X, Wang N L. et al. Antiplatelet aggregation

constituents
A simple and rapid High-performance liquid

chromatographic (HPLC) method for the
determination of adenosine is developed. This
method is achieved by methanol and reversed phase
HPLC with ultraviolet detection. This assay method
developed the quality control of the preparations
including Fructus Trichosanthis and Bulbus Allii
Macrostemi. The adenosine content in the single
and mixed decoctions has very significant
difference. The dissolution of adenosine isrelated
with both the ethanol concentration and decoction.
Active constitutes and pharmacology researches on
the traditional Chinese medicine prescriptions,the
influencing action of decoction style should be
considered.
from
Trichosanthes
kirilowii.
Chinese
Traditional and Herbal Drugs 2004; 35(12):1334-1336.

9.
Peng J P. Chemical constituents in Allium macrostemon

Bunge and Allium chinense GDon and their inhibitory
effects on human platelet aggregation. ShenyangShenyang
College of Pharmacy 1993.
10.
Chao
M,
He
B.
Overviews
in
studies
on
Gualou-xiebai-tang. Chin J Exp Tradit Med Form 1999;

5(1):57-60.
11.
Xu P Y. Pharmacology of traditional Chinese medical

formulae. Beijing: Peoples Medical Publishing House, 1995;
371-373.
12.
Wu B, Cheng X J, Wang M W. The Selected Effective

Prescription of Gualou-xiebai-baijiu-tang on Myocardial
Anoxia and Ischemia in Rodents.Chinese Traditional and
Herbal Drugs 2000; 31(11):844-845.
13.
Jin H. Determination of Adenosine in Chongcaoyifei

Capsule by HPLC. Chinese Traditional Patent Medicine,
2006; 28(1):147-148.
14.
Capsule by HPLC.Chin J Pharm Anal 2005; 25(1):101-103.
References
15.
1.
Yang F M. Determination of Adenosine in Jinhe Tiangen

Nie X P, Man S L, Zhang Y J et al. The extracts isolated by
Fredholm BB, Abbracchio MP, Burnstock G, et al.
different ethanol concentrations from Fructus trichosanthis
Nomenclature
and Bulbus allii macrostemonis influencing the experimental
and
classification
of
purinoceptors.
Pharmacol Rev 1994; 46: 143156.
myocardial
2.
Fredholm BB: Purinoceptors in the nervous system.
Metabonomics
Pharmacol Toxicol 1995; 76: 228239.
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Barraco RA, OLeary DS, Ergene E, et al. Activation of
2007.
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elicits specific regional vascular response patterns. J Auton
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ischemia.
in
Biopharmaceutical
Research
of
Traditional
Forum
Chinese
Successful the 2nd AsianPacific Regional ISSX Meeting

International Society for Study of
Xenebiotics (ISSX) held the 2nd AsianPacifi c
Regional Meeting at the School of Pharmacy,
Fudan University in Shanghai, China, from May
1113. The meeting, co-chaired by Honghao
Zhou, Yuichi Sugiyama and Zhuohan Hu,
attracted a total of 517 meeting participants and
additional accompanying persons. The four short
courses held on Monday, May 11 had a
combined enrollment of 398. The regional
breakdown was 79% Asian Pacific,19% North
American, 2% European. Eighteen countries
were represented (Table 1).
than 10 abstracts presented from 6 institutions in

China (Table 2).
Table 1 Asian-Pacific Regional ISSX Meeting

attendees by countries
Country/ Regional
Attendees
China
USA
Japan
South
Taiwan, China
UK
Australia
239
93
83
42
20
8
7
Fig 1. News of this meeting
340 abstracts were accepted by this meeting.

297 abstracts were accepted as posters and
presented during the poster sessions held on
Monday, May 12 and Tuesday, May 13. All
accepted abstracts have been published in a
special supplemental issue of Drug Metabolism
Reviews. ISSX is very appreciative of the
extraordinary efforts of the meetings able
organizing committee, led by Honghao Zhou,
Yuichi Sugiyama, and Zhuohan Hu, as well as
the strong support and involvement of the
members of the ISSX Meeting
Organizing Committee. The ISSX Web site
will feature a full report of the scientific sessions
of the 2nd AsianPacific Regional Meeting.
News of this meeting was published in
ISSX Newsletters 2008;28(2):6 (Fig 1).
ISSX published a special issue of Drug
Metabolism Reviews (IF=5.59) (Fig 2). 340
abstracts were published in the issue. From the
publication, we found that drug metabolism
research is an active study filed in China. More
Fig 2. Abstracts publication of this Meeting

Table 2 More than 10 abstracts presented
from 6 institutions
No.
1
2
3
4
5
6
77
Institution
Shanghai Research Center of Drug
Metabolism
Shanghai Institute of liver Disease
Tianjin Institute of Pharmaceutical
Research
Center South University
China Pharmaceutical University
Second Military Medical University
Paper
number
33
20
15
12
11
11
Asian Journal of
Pharmacokinetics
ISSN
1608-2281

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function in hot environment. Amsterdam: Elsevier; 1998;

129-39.
4.
Wnag LCK. Current drug safety testing in USA.

Proccedings of International Symposium on Traditional
Medicines and Modren Pharmacology; 1987 May 2-4;
Beijing, China. Beijing: Chinese Pharmacological Society;
1987; 257-61.
5.
World Health Organization. Good manufacturing practices

for pharmaceutical products. Annex 1. Thirty-second
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for pharmaceutical preparation. WHO Technical Report
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6.
N.823.
Geneva,
Switzerland:
World
Health
Guo FK, Li YL, Wu SG. Antisense IRAK-2 oligonucleotide

inhibits interleukin-1-induced nuclear factor-kB activiation
in vitro. Acta Pharmacol Sin 2000; 21: in press.
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Sample references
1.
Abraham BK, Adithan C, Usha Kiran P, Asad M,

Koumaravelou K. Genetic polymorphism of CYP2D6 in
kamataka and andhra pradesh population in india. Acta
Pharmacol Sin 2000; 21: 494-8.
2.
Liu
CX.
Studies
on
Drug
Metabolism
and
pharmacokinetics in China. ISSX Newsletter 1990; 9(2):

1-2.
3.
Milton AS. Prostaglandins and fever. In: Sharma HS,

Westman J, editors. Progress in brain research; v 115. Brain
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A Lanata

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Official Journal of International Quarterly Publication

Chang-Xiao Liu and Yuichi Sugiyama

Hong Kong Medical Publisher

Asian Journal of Pharmacodynamics and Pharmacokinetics

Assistant Academic Editor-in-Chief

Members of Editorial Committee

Terry D. Lindstrom (Indiana, USA)

Chang-Koo Shim (Seoul, KOREA)

Chang-Xiao Liu (Tianjin, CHINA)

Guang-Ji Wang (Nanjing, CHINA)

Zheng-Min Chen (Tianjin, CHINA)

Xiao-Dong Liu (Nanjing, CHINA)

Hui Wang (Wuhan, CHINA)

Moses SS Chow (Hong Kong, CHINA)

Ke-Xin Liu (Dalian, CHINA)

Ming-Wei Wang (Shanghai, CHINA)

Guang-Hua Du (Beijing, CHINA)

Jian-Shi Lou (Tianjin, CHINA)

Zhi-Min Wang (Beijing, CHINA)

Paul Fawcett (Otago, NEW ZEALAND)

Takeo Murakawa (Osaka, JAPAN)

Yi-Tao Wang (Macau, CHINA)

Jack Gao (Carborough, CANADA)

Terumichi Nakagawa (Kyoto, JAPAN)

Guang-Li Wei (Tianjin, CHINA)

Mei-Yu Geng (QingdaoCHINA

Charles H. Nightingale (Hartford, USA)

Patrick M. Gennissel (Paris, FRANCE)

Inotsume Nobuo (Hokkaido, JAPAN)

Kun Han (Chungbuk, KOREA)

Richard ZM Qian (Hong Kong, CHINA)

Fu-Ming Xie ( Indiana, USA)

Guo-Zhu Han (Dalian, CHINA)

Madhubala Rentala (New Delhi,

Guo-Wang Xu (Dalian, CHINA)

Pei Hu (Beijing, CHINA)

(New Jersey, USA)

Ming Xue (Beijing, CHINA)

Zhuo-Han Hu (Shanghai, CHINA)

Jin-Xiu Ruan (Beijing, CHINA)

Bao-Feng Yang (HarbinCHINA)

Xi Huang (Chengdu, CHINA)

Guo-Wei Sang (Beijing, CHINA)

Zong-Hui Yuan (Wuhan, CHINA)

Jun-Yan Hong (New Jersey, USA)

Xiang-Guo Shi (Boston, USA)

Fan-Dian Zeng (Wuhan, CHINA)

Li-Ya Ju (Charenton le Pont, FRANCE)

Duan-Yun Si (Tianjin, CHINA)

Su Zeng (Hangzhou, CHINA)

Devarakonda R. Krishna (Kakatiya,

Chang Koo Shim (Seoul, KOREA)

Qiang Zhang (Beijing, CHINA)

Yuichi Sugiyama (Tokyo, JAPAN)

Jiang Zheng (SeattleUSA)

June Lee (NIH, USA)

Ding-Feng Su (Shanghai, CHINA)

Da-Fang Zhong (Shanghai, CHINA)

Nikolaus Sucher (Hong Kong, CHINA)

Hong-Hao Zhou (Changsha, CHINA)

Lin-Lin Li (Xinjiang, CHINA)

Yong-Da Sun (Bradford, UK)

Yao-Wei Zhu (PA, USA)