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O R I G I N A L R E S E A R C H

Analysis of Glucosamine and


Chondroitin Sulfate Content in Marketed
Products and the Caco-2 Permeability of
Chondroitin Sulfate Raw Materials
Abimbola O. Adebowale, PhD, Donna S. Cox, MS, Zhongming Liang, MS, Natalie D. Eddington, PhD,*
Pharmacokinetics-Biopharmaceutics Laboratory, Department of Pharmaceutical Sciences,
School of Pharmacy, University of Maryland, Baltimore, Maryland

ABSTRACT
Objective: The purpose of this report is to evaluate to one dollar per 1200 mg of chondroitin sulfate were found
and present the results of analysis of actual contents of sev- to be seriously deficient in meeting label claim (less than
eral products in the marketplace containing glucosamine 10% of label claim). The permeability of the different mol-
and/or chondroitin sulfate and to determine if they signifi- ecular weight chondroitin sulfates was found to be signifi-
cantly deviate from label claim. In addition, the study cantly different (p<0.05), with the permeability coefficient
examined the intestinal transport of several marketed increasing with decreasing molecular weight. This suggests
sources of chondroitin sulfate. that molecular weight of chondroitin sulfate could be a pos-
Methods: A total of fourteen products containing glu- sible predictor of permeability.
cosamine hydrochloride or sulfate and eleven products con-
taining chondroitin sulfate were evaluated using a UV- INTRODUCTION
HPLC method. In addition, a total of 32 products contain-
Dietary supplements containing glucosamine and/or
ing chondroitin sulfate were tested using a titration method.
chondroitin sulfate are numerous and popular. Since the
The permeability of various marketed sources of raw mate-
Dietary Supplement Health and Education Act (DSHEA)
rials of chondroitin sulfate across Caco-2 cell monolayers
does not demand the same rigorous requirements for quali-
were assessed. This analysis was an attempt to evaluate
ty manufacturing as pharmaceuticals, many of the dietary
whether different suppliers of chondroitin sulfate use dif-
supplements marketed may not provide high quality mater-
ferent grades of material.
ial or meet labeled quantities.
Results and conclusions: The amounts of glu-
Over the last five years there has been a significant
cosamine and chondroitin found after analysis were signif-
increase in the number of dietary supplements that have
icantly different from the label claim in some products, with
been introduced into the market. The glucosamine and chon-
deviations from label claims ranging from as low as 0% to
droitin market has been estimated to be over five hundred
over 115%. Products with a retail price of less than or equal
million in retail sales between July 1998 and May 1999.1
Unfortunately the quality of dietary supplements remains in
* Correspondence: question due to the lack of regulatory provisions that direct-
Natalie D. Eddington, PhD ly evaluate the actual content of active ingredients.2 Hence
Department of Pharmaceutical Sciences consumers have limited ways to judge the quality of the
School of Pharmacy products they are purchasing. Independent analysis on many
University of Maryland supplements, including calcium, St. Johns Wort, carnitine,
20 North Pine Street Ginkgo biloba, and super oxide dismutase, showed that
Baltimore, Maryland 21201-1180 many marketed brands contained subpotent content, includ-
Phone: 410-706-6710 Fax: 410-706-6580 ing some with zero active ingredients.3,4
Email: neddingt@rx.umaryland.edu

Spring 2000 Vol. 3, No. 1 JANA 37


Dietary supplements aimed at the management of have been evaluated by meta-analysis in the United
osteoarthritis, (OA) a common disease that affects more States.34,35 Studies completed in the United States have
than 40 million Americans,5 have also proliferated. These used the same pure low molecular weight chondroitin sul-
supplements generally contain glucosamine, chondroitin fate in combination with the amino sugar glucosamine with
sulfate, or a combination thereof. As with other dietary sup- good results.22-27 It has been noted that results of clinical tri-
plements, many products containing either glucosamine or als are directly related to the quality of material used.7
chondroitin sulfate have been reported to not meet label Pure low molecular weight chondroitin sulfate has been
claims.6,7 shown to have up to a three month efficacy carry-over
OA is a disease process associated with alteration in effect.32 This is an interesting point as individuals who take
the structure and function of synovial joints,8 resulting from a quality product and switch to an impure product may have
a loss of balance between synthesis and degradation of the three-months of results before symptoms reoccur. This cou-
macromolecules needed to provide joint tissue with its bio- pled with the standard 20-30% placebo effect in all oral
mechanical and functional properties.9 OA usually occurs arthritis treatments may lead to a false sense of security in the
insidiously, apparently as part of the aging process and consumer or prescribing physician. When the name of the
without obvious initiating cause (primary or idiopathic product or label makes a claim such as arthritis cure or
OA). The disease progresses in the majority of patients.10 pain formula this could possibly potentate the placebo
The typical clinical symptoms are pain, stiffness, and limi- effect.
tation of motion. These factors are reflected in difficulties Since chondroitin sulfate is a large molecule, bioavail-
in performing activities of daily living, eventually altering ability has been questioned.36,37 However, recent research
the patient's quality of life.11 has supported absorption after oral administration.31,38 One
Glucosamine, a bioavailable amino sugar when admin- factor that may affect absorption is the actual chain length
istered as a sulfate or hydrochloride salt, has been shown to of the molecule. It has been shown that low molecular
help relieve the symptoms of osteoarthritis.12-17 European weight chondroitin sulfate has a superior kinetic profile
research has shown glucosamine sulfate to be effective in than high molecular weight.39 The molecular weight as well
clinical trials.12-14 However the radiolabeled pharmacoki- as the molecular composition of chondroitin sulfate are
netic studies on glucosamine sulfate radiolabeled glu- dependent upon the species and/or tissue of origin and may
cosamine HCL, not sulfate.17,18 This research also states be affected by the extraction method.39
that the salts of glucosamine sulfate are a prodrug for glu- Referenced studies highlight the current problems with
cosamine.19 Glucosamine base is the active component not quality control of dietary supplements in general, and
the salt. Glucosamine is also completely ionized in the specifically products containing glucosamine and/or chon-
stomach, so any salt is likely to be cleaved off, leaving the droitin sulfate. The purpose of this report is to evaluate and
glucosamine base.17 Comparative cell culture studies have present the results of an analysis of actual content of sever-
also shown that glucosamine base, glucosamine HCL, as al products containing glucosamine and/or chondroitin sul-
well as glucosamine sulfate, are equally active.20,21 The fate and to determine if they significantly deviate from label
active moiety of any form is the glucosamine molecule, not claim. In addition, as an index of bioavailability, perme-
the salt carrier. To date all the controlled, blinded, pub- ability studies were performed using Caco-2 cell monolay-
lished studies in North America with any statistically sig- ers to examine the intestinal transport of several marketed
nificant positive outcomes have used glucosamine HCL sources of chondroitin sulfate.
with the most convincing results in combination with chon-
Caco-2 cell monolayers are recognized as excellent
droitin sulfate.15,22-27 The only randomized, placebo-con-
models of intestinal transport. Caco-2 cells are derived
trolled US trial evaluating glucosamine sulfate alone as
from human colon adenocarcinoma, and have morphologi-
treatment for osteoarthritis of the knee found that it was no
cal features similar to intestinal epithelia.40 When grown on
better than placebo in reducing pain.28 A recent review arti-
semipermeable filters, Caco-2 cells spontaneously differen-
cle from a well known rheumatologist has also stated that
tiate in culture to form a confluent monolayer which both
a higher dosage of the sulfate salt rather than the
structurally and functionally resembles the small intestinal
hydrochloride salt is required to provide a 1500 mg dose of
epithelium. The Caco-2 cell line exhibits a well-differenti-
glucosamine.7
ated brush border on the apical surface and tight junctions.
Chondroitin sulfate, a much larger molecule than glu- The monolayer also has bipolar properties of an apical sur-
cosamine, is a glycosaminoglycan made up of glucuronic face as well as a basolateral surface with differentiated and
acid and galactosamine. When given orally in the pure form different transport properties. The pharmaceutical industry
of a molecular weight of aproximately 16,900 Dalton, it has is increasingly using Caco-2 cells as a model to screen
been shown to be bioavailable and efficacious in decreasing compounds for intestinal absorption and to predict trans-
pain and slowing the progression of osteoarthritis in port routes and even rates of flux before compounds go to
humans.29-33 The efficacy studies, completed in Europe, preclinical testing. A citation study done through PubMed

38 JANA Vol. 3, No. 1 Spring 2000


showed that since 1979 over 1490 publications dealt with Preparation of Samples of Glucosamine Products
Caco-2 cells, and 527 of those publications have appeared for Quality Control Testing: The contents of each capsule
in the last two years, attesting to the rapidly growing use or tablet were weighed and transferred quantitatively to a
and interest in this model system. 250 ml volumetric flask and about 150 ml of water added.
The mixture was sonicated for 20 minutes and brought to
volume with water. A portion of each solution was filtered
MATERIALS AND METHODS and 20 l of the filtered solution was derivatized with
A total of 14 products containing glucosamine PITC. Simultaneously, a standard solution of glucosamine
hydrochloride or sulfate and 11 products containing chon- hydrochloride was treated in a similar manner. The result-
droitin sulfate were evaluated using the UV-HPLC method ing solutions were assayed by a validated HPLC method.16
described below.16-41 In addition, a total of 32 products con- Permeability of Chondroitin Sulfate Across Caco-2
taining chondroitin sulfate were tested using the titration Cell Monolayers: The growth, maturation, and seeding of
method as described in the assay methods section. The
Caco-2 cells have been previously described.40 Caco-2
products were randomly gathered from the marketplace.
cells with a passage number between 45 and 55 were used
Chondroitin Sulfate Assay Materials: 95% chon- in an effort to derive Caco-2 monolayers with consistent
droitin 4-sulfate (from bovine trachea) was purchased from morphological and biochemical properties. Caco-2 cells
Bioiberica (Poligono Industrial, Barcelona, Spain). Sodium were grown in T-150 flasks at 37C in an atmosphere of 5%
phosphate monobasic was purchased from Fisher Scientific CO2 using Dulbeccos modified Eagle medium (DMEM)
(Pittsburgh, PA). Sodium phosphate dibasic was purchased supplemented with 2% L-glutamine, 1% nonessential
from Sigma Chemical Co. (St. Louis, MO). Deionized amino acids, 1% penicillin-streptomycin, and 10% fetal
water was prepared by ultrapure water system Pyrosystem bovine serum.
Plus* (Hydro, Research Triangle Park, NC).
Caco-2 cells grown for 21 days on the transwell filters
Glucosamine Assay Materials: D(+) glucosamine (2- were washed free of medium and allowed to incubate at
amino-2-deoxy-D-glucose) hydrochloride was purchased 37C in 1.5 ml Dulbeccos phosphate buffered saline (PBS)
from Sigma Chemical Co. Methanol, phenyl isothio- in the insert (apical chamber) and 2.5 ml in the basolateral
cyanate (PITC), sodium dibasic phosphate, and glacial chamber. Bi-directional studies (apical to basolateral and
acetic acid were purchased from J.T. Baker Chemical Co. basolateral to apical) were performed, with 1.5 ml (apical)
(Phillipsburg, NJ). All chemicals and solvents were ACS or 2.5 ml (basolateral) fresh PBS containing chondroitin
analytical grade or HPLC grade. sulfate (0.2 -0.8 x 10-3 M). At 5, 10, 15, 20, 30, 45, 60, 75,
Caco-2 Cell Culture Materials: The Caco-2 cell line 90, and 120 minutes the insert was transferred to fresh solu-
was obtained from American Type Culture Collection tions (2.6 ml) of PBS in the basolateral chamber. At the end
(Rockville, MD). Dulbeccos modified eagle medium of the experiment (120 minutes), a sample was withdrawn
(DMEM), Dulbeccos modified phosphate-buffered saline from the donor chamber to evaluate for metabolism and
with and without Ca2+ and Mg2+ (PBS), nonessential amino cumulative transport. The incubations were maintained at
acids (NEAA), fetal bovine serum (FBS), L-glutamate, 37C and the monolayers were agitated orbitally at 50-60
trypsin (0.25%)-EDTA (1mM), and penicillin G-strepto- rpm during the course of the permeability study. The per-
mycin sulfate antibiotic mixture were purchased from centage amounts of compound appearing in the apical or
Gibco Laboratories (Lenexa, KS). T-flasks were obtained basolateral chamber after each sampling interval were
from Becton Dickinson Labware (Franklin Lakes, NJ); added to obtain the cumulative transport. The molecular
Transwell clusters, 24 mm in diameter (surface area 4.71 weights of the raw materials of chondroitin sulfate were
cm2) and 0.4-m pores were from Corning Costar determined by using a linear calibration method and the
(Cambridge, MA). All other chemicals were of analytical samples were analyzed by a validated HPLC method for
grade from Sigma Chemical Co. and Fisher Scientific (Fair chondroitin sulfate.40
Lawn, NJ). To determine drug transport from the apical to basolater-
Preparation of Chondroitin Sulfate Products for al chambers, effective permeability coefficients, Peff (cm s-1),
Quality Control Testing: The contents of each capsule or were calculated from the following relationships:
tablet were weighed, transferred to a 100 ml volumetric
flask, and made up to volume with 0.1 M phosphate buffer
pH 7.0. The resulting solutions (raw materials) or suspen- Peff =
( )
dC
VR . dt
ss
sions (capsule and powder) were sonicated for 20-30 min- A .C

utes and subsequently filtered through a 0.45 m membrane
filter. Ten l of the final solution was assayed by a validat-
Where VR = volume of the basolateral or apical (receiver)
ed HPLC method.41
chamber (cm3), A = cell monolayer surface area (ie, 4.71

Spring 2000 Vol. 3, No. 1 JANA 39


cm2), Co = initial donor concentration of solute (mMol ml-1), Data Analysis: The percent of the label claim for the
and (dC/dt)ss = initial linear portion of a plot of the cumula- capsules was calculated as follows:
tive receiver concentration of permeant with time, i.e., flux Assayed amount (mg)
across the monolayer at steady state (mMol ml-1s-1).
% Label claim = ------------------------------------------- * 100
Chondroitin Sulfate Assay Method from University
Labeled amount (mg)
of Maryland (HPSEC): A simple, specific, rugged, and
precise high-performance size exclusion chromatographic RESULTS
(HPSEC) method was used to quantify chondroitin sulfate Figure 1 illustrates the percent label claim of 14 products
in dosage forms and cell culture samples using a Polysep- that contained glucosamine hydrochloride or sulfate, some
GFC-P linear column.41 The samples were solubilized in with chondroitin sulfate. As can be seen from the figure,
0.1M-phosphate buffer and the resulting solutions filtered the amount found after analysis was significantly different
through a 0.45 membrane filter. The mobile phase was from the label claim in some products, with deviations from
phosphate buffer (0.1M; pH 7.0) and detection was by ultra- label claims ranging from as low as 25% to over 115%.
violet absorbance at a wavelength of 207 nm. The intraday These results highlight the inconsistencies between label
and interday precision as indicated by the relative standard
deviation was less than 2.83% and 3.53%, respectively.
Phototrode Titration Method (Bioiberica): Thirty-
two products were also tested by titration (Phototrode)
(Bioiberica). This method uses potentiometric titration with
photometric indication by titrating with 0.1% solution w/v
of N-cetylpyridinium chloride to quantify chondroitin sul-
fate. The sample preparation and procedure is as follows:
weigh accurately 100 mg of sample dried for 2 h at 105C
into a graduated flask. Dissolve in about 30 ml of water, add
10 ml of pH 7.0-phosphate buffer and dilute to the mark with
water. Take 5 ml aliquots, bringing these to 30 ml with
water and start titration. Adjust the initial transmittance to
70% in the phototrode, adjusting the wavelength at 420 nm.
When titration is complete, the sodium chondroitin sulfate Figure 1. Percent label claim of products containing glu-
percentage is determined. The amount of chondroitin sul- cosamine hydrochloride or sulfate as determined by a vali-
dated UV-HPLC method with pre-column derivatization
fate content is determined by the following equation: (University of Maryland Analysis).45 The bars indicate the
V x F x 2000 standard error of the mean.
Chondroitin Sulfate Sodium Content%= P(mg)
Where: V = mL of N-cetylpyridinium chloride used, P
= sample weight in mg, 2000 = dilution factor introduced in
the titrator as constant CO2, and F = N-cetylpyridinium
chloride factor against sodium chloride sulfate standard,
calculated as sodium chondroitin sulfate assay in mg for 1
ml of N-ceytlpyridinium chloride.
Glucosamine Hydrochloride Assay Method: A spe-
cific high performance liquid chromatography method was
developed to quantitate glucosamine hydrochloride.16
Reverse phase chromatography using pre-column derivati-
zation with phenyl isothiocyanate, and ultraviolet detection
(=254nm) was used to quantitate the eluate. The mobile
phase consisted of MeOH:H2O:CH3COOH (10:89.6:0.04)
and was pumped at a flow rate of 1.2 ml/min. The precision
of the dosage form assay, expressed as the % relative stan-
dard deviation (RSD), was <5% at all concentrations. The
intraday and interday accuracy, as indicated by the relative Figure 2. Percent label claim of products containing chon-
error (RE), ranged from -2.54 to 2.70% for glucosamine droitin sulfate as determined by a validated UV-HPLC
method using size exclusion chromatography (University
hydrochloride. of Maryland Analysis).45 The bars indicate the standard
error of the mean.

40 JANA Vol. 3, No. 1 Spring 2000


Price in retail dollars per daily dose of 1200 mg chondroitin sulfate

Figure 3. Relationship between products label claim and their standard retail price (SP)

claims of dietary supplements and actual content found in Our initial hypothesis was that the quality of product
the product. Eleven chondroitin sulfate containing products, may be a function of retail price. In a separate analysis of
some also containing glucosamine, were analyzed for con- the data contained in Figure 3, the supplement retail pur-
tent with a validated HPLC method. The results of the label chased prices, not including any consumer rebates, were
claim testing are presented in Figure 2. The percentage transformed to reflect a standard retail price (SP) per daily
label claims ranged from as low as 33% to as much as dose of 1200 mg of chondroitin sulfate as reflected on the
110%. It should be noted that some products (eg, R and Y) label. The content-analyzed products and their percentages
displayed significantly large relative standard deviations. relative to each products label claim (PLC) were calculat-
This suggests that in addition to these products having per- ed. Figure 3 is a scatter plot showing that products with a
centage label claims of less than 40%, the variability in the standard retail price of less than or equal to one dollar are
amount of chondroitin sulfate found in each capsule also seriously deficient in meeting label claim (less than 10%
varies significantly. PLC). When the retail price exceeds one dollar, the figure
Figure 3 presents the percent label claim and adjusted indicates that there are two different clusters of products,
retail price of chondroitin sulfate in 32 chondroitin sulfate with the majority having less than acceptable label claim.
containing products purchased from pharmacies and health Only a few products (5 out of 32) satisfied the industry
food stores during the period of September 1998 through accepted 10% variation in label claim in levels of chon-
November 1999 and analyzed by the titration method. A droitin sulfate. Variation greater than 10% suggests poor
few of these products were labeled to contain chondroitin quality of raw material or poor manufacturing processes
sulfate alone but the majority was combined with glu- and lack of quality control.
cosamine. Twenty-six out of 32 products were found to con- The results of these studies demonstrate that in some
tain less than 90% of the chondroitin sulfate stated on the instances the amount of the chondroitin sulfate or glu-
label with 17 products containing less than 40% of label cosamine found to be present in the sample product varies
claim. Only five out of 32 products contained the labeled significantly from the amount reported on the label. The
amount of chondroitin sulfate in the product. This would greatest inconsistencies were obtained in products contain-
suggest that in many cases, 84% of brands that were tested ing chondroitin sulfate, especially when the daily 1200 mg
are inferior products. dose price was less than one dollar. The implications of
these results are significant and support the need for regu-

Spring 2000 Vol. 3, No. 1 JANA 41


latory intervention of dietary supplements.
Raw Material Permeability Coefficient
In addition to examining the label claims of chon- (x 10-6) (n=3) (cm/sec)
droitin sulfate products, we also examined the permeability
of various marketed sources of raw materials of chondroitin
sulfate across Caco-2 cell monolayers. This analysis was an
A1 10.1 (0.6)
attempt to evaluate whether different suppliers of chon-
droitin sulfate use different grades of material. Studies have
B 8.73 (9.07)
shown a very good correlation between drug permeability
in the Caco-2 model and intestinal drug absorption in
humans.38 Drugs administered orally have displayed C 7.94 ( 7.35)
absorption between 50 to 100% when the Papp value has
been in the range of 0.20 to 54.5 x 10-6 cm/sec. However, D 0.00 ( 0.00)
it should be noted that the molecular weights evaluated
were all less than 1,000 Dalton. In order to show that the E 3.63 ( 2.07)
Caco-2 cells model is acceptable for assessing permeability
of polymers, three different MW chondroitin sulfate mate- F 1.03 ( 1.78)
rials were extracted and purified by the same manufacturer
(Bioiberica) to be used specifically for calibration. The per- 1mol Wt = 16,900 dalton (95% Bioiberica)
meability of the different molecular weight chondroitin sul-
fates was found to be significant (p < 0.05), with the per- Table 1. Permeability coefficient for transport across Caco-
meability coefficient increasing (10.1 x 10-6 cm/sec, 12.5 x 2 cells of chondroitin sulfate from different manufacturers
10-6 cm/sec, 16.2 x 10-6 cm/sec) with decreasing molecular
weight (16.9, 8.0, and 4.0 x 103 Dalton). This suggests that weight, Papp = 10.1 x 10-6 cm/sec) should be absorbed after
the molecular weight of chondroitin sulfate could be a pre- oral administration. Any differences observed experimental-
dictor of permeability. ly (eg, after clinical or animal bioavailability studies) may
A comparison of the permeability of chondroitin sulfate be due to molecular weight influences or first pass metabo-
from different manufacturers with permeability of known lism prior to systemic circulation. In addition, characteris-
molecular weight chondroitin sulfate, (ie, raw material A) tics other than molecular weight such as flexibility of struc-
(Bioiberica 95% mol wt = 16,900 Dalton) is presented in ture, degree of sulfation, and method of manufacture may
Table 1. This reference standard of low molecular weight also be important for oral absorption.
material has been shown to be efficacious and bioavailable
in European and US trials.22-25, 29-33 These results indicate a
DISCUSSION
low permeability for raw materials F (Papp = 1.03 x 10-6
cm/sec), E (3.63 x 10-6cm/sec), C (7.94 x 10-6cm/sec), and Glucosamine and chondroitin sulfate when adminis-
B (8.73 x 10-6cm/sec) raw materials. The data suggests that tered in a pure bioavailable form have been shown to be
the molecular weights of these raw materials are probably > safe and efficacious.22,23,34,35 Our results suggest that there
than 16,900. The proposed higher molecular weight chon- is a significant deviation between the content of the active
droitin sulfates (ie, with exceedingly low permeability) sug- ingredients (glucosamine or chondroitin sulfate) and what
gest that their intestinal permeability and hence absorption is stated by the manufacturers on the label. It would appear
would be expected to be significantly less than the low mol- that this deviation is greater for those products containing
ecular weight chondroitin sulfate (eg,16,900 dalton). Similar chondroitin sulfate. In light of these findings, an interest-
results have been observed in studies examining the perme- ing question emerges: what can the consumer or health care
ability of polyethyleneglycol (PEG) of various molecular provider do to obtain a quality product containing pure
weights. The permeability of PEG was found to be inverse- chondroitin sulfate or glucosamine? A recent review by
ly correlated with molecular weight.40 The Arthritis Foundation provides the following sugges-
tions: (1) consumers should consult with their physician or
In summary, it would appear that the molecular weight
health care provider concerning these two supplements; (2)
of chondroitin sulfate has a direct influence on its perme-
healthcare professionals should become knowledgeable
ability across the gastrointestinal tract, where higher perme-
about glucosamine and chondroitin sulfate products; (3)
ability is observed for chondroitin sulfate with lower molec-
consumers should not purchase through the mail or internet
ular weight. Further, when comparing Papp values from our
unless they know the vendor; and (4) consumers should buy
chondroitin sulfate studies with values reported in the litera-
from companies that use USP material (neither chondroitin
ture with agents such as dexamethasone (Papp = 12.5 X10-6
sulfate nor glucosamine have a monograph as of yet).6
cm/sec) and salicylic acid (Papp = 11.9 x10-6 cm/sec), it
would appear that chondroitin sulfate (of low molecular Prior to obtaining any supplement containing chon-

42 JANA Vol. 3, No. 1 Spring 2000


droitin sulfate or glucosamine, the consumer should animal tissues; therefore impurities may cause allergic or
become informed about the manufacturer and the product. other side effects that raise serious concerns.44 It is
The most useful technique recommended by the Arthritis unknown if less than pure, non-researched sources have a
Foundation is to ask the manufacturer for research showing good safety profile.
that their brand has been scientifically proven or studied. It The HPLC and titration assay methods used in our
is important for manufacturers to evaluate their product, study will not detect other compounds that might be present
and especially the source of chondroitin sulfate used since in the product along with chondroitin sulfate or glu-
not all chondroitin sulfates are identical. Furthermore, con- cosamine. It is possible that some suppliers may dilute
sumers should shy away from products that are backed chondroitin sulfate with materials that can cause analytical
only by testimonials and not scientific research. Products methods to overestimate contents. These compounds could
that make overt claims such as regenerates cartilage, include sugars such as maltose, or other glycosaminogly-
renews cartilage, rebuilds cartilage, cures arthritis, or free- cans such as dermatan sulfate, keratan sulfate, heparin, or
dom from pain, should be looked at with skepticism as hyaluronic acid. None of these glycosaminoglycans have
these statements seldom mean that the product has been efficacy in osteoarthritis when given orally.
researched to make these claims.42 Topical and liquid glu-
cosamine/chondroitin products are also promoted, claiming In summary, this report highlights the problems that
a higher bioavailability than capsules or tablet dosage consumers encounter when attempting to purchase quality
forms. To the authors knowledge, there is no published dietary supplements containing glucosamine and chon-
research to substantiate this claim. droitin sulfate. Despite terms like quality tested appear-
ing on labels, consumers and healthcare providers have no
Based on a 1200 mg/day chondroitin sulfate dosage, basis to compare one product against another or to judge
products that retailed in the pharmacy or health food store the quality of the products they are purchasing or recom-
for less than $1.00 per day contained a sub-potent level of mending. Over fifty products were tested and a substantial
CS and did not meet label claims. However, it must be noted number could not be classified as a quality product based
that some of the more costly products also did not meet on the general agreement of their content with label claim.
label claims. Therefore, proposed recommendations for From our data it is obvious that certain manufacturers of
consumers are to be wary of inexpensive products and com- dietary supplements are unwilling to self-regulate their
pare the calculated daily price based on 1200 mg of CS/day. manufacturing practices.
These findings verify the scientific communitys skep-
ticism towards nutraceuticals based on the lack of quality
control by some manufacturers. These substances are not ACKNOWLEDGEMENT
pharmaceuticals; there is no requirement for pharmaceuti- Partial funding for this study was provided as an unrestrict-
cal Good Manufacturing Practices to guarantee high quali- ed educational grant from Nutramax Laboratories Inc.,
ty, batch-to-batch consistency. The use of validated analyt- Edgewood, Maryland.
ical methods for the raw materials and finished products is
the only mechanism to verify purity. It is not surprising to
REFERENCES
know that the Arthritis Foundation has recently recom-
mended that when a supplement has been studied with 1. Hartman Groups Natural Products Census Supplement Report,
1999, Bellevue, Wash.
good results, find out which brand was used in the study,
2. Foley CM, Kratz AM. Nutraceuticalschallenges and opportuni-
and buy that.6 ties for the new millennium that affect consumers and healthcare
It is worthy to note that there is great variability in the professionals who use and recommend nutraceuticals. JANA
1999; 2(2): 6-10.
permeability of the chondroitin sulfate raw materials.
3. Moore T. Messing with Mother Nature. The Washingtonian July
Permeability could have a direct effect on efficacy. 1999; 58-115.
Therefore, caution is warranted to not extrapolate the 4. Beal B. Evaluation of active enzyme activity in six oral super-
results of previously reported experimental and clinical oxide dismutase products. Paper presented at the 25th annual
studies to all forms of chondroitin. Some products that test- conference of the Veterinary Orthopedic Society, Snowmass,
Colorado, 1998;65.
ed satisfactory on content could be lacking efficacy
because of poor permeability. 5. Lawrence RC, Helmick CG, Arnett FC, Deyo RA, Felson DT,
Giannini EH, Heyse SP, Hirsch R, Hochberg MC, Hunder GG,
In addition to concerns on the content, bioavailability, Liang MH, Pillemer SR, Steen VD, Wolfe F. Estimates of the
prevalence of arthritis and selected musculoskeletal disorders in
and effectiveness of dietary supplements, an often-over- the United States. Arthritis Rheum 1998; 41, 778-799.
looked area is safety. Pure chondroitin sulfate and glu- 6. Horstman J. The Arthritis Foundations Guide to Alternative
cosamine evaluated in published studies is shown to be safe Therapies. The Arthritis Foundation. Atlanta, Georgia:1999;179-
including hemostatic, hematological, and biochemical 180.
parameters with minimal side effects.15,22,23,30,32,33,43 7. Deal CL, Moskowitz RW. Nutraceuticals as therapeutic agents in
Chondroitin sulfate and glucosamine are extracted from osteoarthritis: the role of glucosamine, chondroitin sulfate, and
collagen hydrolysate. Rheum Dis Clin North Am May 1999; 25,
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Spring 2000 Vol. 3, No. 1 JANA 43


8. Dieppe P, Brandt KD, Lohmander S, Felson DT. Detecting and 27. Lippiello L, Hammad T. Dose response and synergistic effect of
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44 JANA Vol. 3, No. 1 Spring 2000