Toxicology Mechanisms and Methods, 2011; 21(3): 246250

RESEARCH ARTICLE

Toxicological assessment of Ricinus communis Linn root

extracts
Raju Ilavarasan1, Moni Mallika2, and Subramanian Venkataraman3
1
Department of Pharmacology, C. L. Baid Metha College of Pharmacy, Old Mahabalipuram Road, Jyothi Nagar,
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Thorapakkam, Chennai 600096, India, 2Department of Microbiology, Sri Ramachandra Medical College and Research
Institute (Deemed University), Porur, Chennai 600116, India, and 3Department of Pharmacology and Environmental
Toxicology, Dr. A. L. Mudaliar P. G. Institute of Basic Medical Sciences, University of Madras, Taramani, Chennai 600113,
India

Abstract
Ricinus communis Linn (Euphorbiaceae) plant parts are claimed to be used as carminative, asthma, bronchitis,
leprosy, anti-inflammatory, cathartic, and aphrodisiac. The toxicological study was carried out in the root part
of the plant. The collected root was extracted with methanol and water. The extracts were vacuum-dried to
yield the respective aqueous (AE) and methanol (ME) extracts. Toxicological assessment sought to determine
the safety of Ricinus communis root extracts. The extracts were evaluated in the acute toxicity study (OECD-423
guidelines) and 90 days repeated dose toxicological assessment in Wistar albino rats. The acute oral toxicity
For personal use only.

of the aqueous (AE) and methanol (ME) extracts did not produce any toxic symptoms or mortality at the dose
level of 2000mg/kg in rats. In the 90 days (sub-chronic toxicity) repeated dose toxicity study the extracts (AE
and ME) were administered 1000mg/kg daily through oral route. The sub-chronic toxicity study demonstrated
no significant changes in body weight, food, and water intake. Hematology parameters RBC, WBC, DLC, Hb,
blood clotting time, and the biochemical parameters glucose, blood urea nitrogen, creatinine, total cholesterol,
total protein, total bilirubin AST, ALT, and ALP were estimated. Histopathology observation of the major vital
organs (liver, kidney, heart, spleen, lungs, ovary, testis, and brain) were tested. The hematology, biochemical
and histopathology evaluations did not show any adverse effects in any of the organs tested. These results
demonstrate the non-toxic nature of the root extracts AE and ME can be used for long-term usage in clinical
practice.

Keywords: Ricinus communis; acute toxicity; 90 days repeated dose toxicity

Introduction 1994), analgesic (Gupta etal. 1982), antibacterial (Misas etal.

Ricinus communis Linn (Euphorbiaceae) is a soft wooded
small tree which is widespread throughout tropics and
warm temperature regions of the world (Ivan 1998). The leaf
parts are claimed to be used as carminative for asthma and
bronchitis, and cathartic for leprosy, and the root parts are
claimed as anti-inflammatory for diseases of the liver, spleen,
and pile disorders (Kirtikar and Basu 1991). Hepatoprotective
(Yanfg etal. 1987; Visen etal. 1992), anti-fungal (Verpoorte
and Dihal 1987), and hypoglycemic (Dhar etal. 1968) activi-
ties were reported. Diuretic (Abraham etal. 1986; Tanira etal.
1989), anti-convulsant (Adesina 1982), laxative (Capasso etal.
1979; Chhabra etal. 1991), and anti-fertility (Sandhyakumary
etal. 2003) activities were reported. Commercially-available
preparations, for example syrup Jaundex and Arthnex,
Osteoguard, En.Liv tablets, included Ricinus communis root
extracts as one of the ingredients. However, the toxicological
assessment of Ricinus communis root has not been studied.
It is important to evaluate the safety of the plant extracts in-
vivo animal models. Therefore, we sought to determine the
safety profile of aqueous (AE) and methanol (ME) extracts
of Ricinus communis root, including acute oral toxicity and
90-day sub-chronic toxicity in Wistar albino rats.

Address for Correspondence: Raju Ilavarasan, Captain Srinivasa Murti Research Institute for Ayurveda & Siddha Drug Development (CCRAS, Dept of AYUSH, Ministry
of Health and Family Welfare, Government of India), Arumbakkam, Chennai600106, India. Tel: 91-44-26214823. Email: arilavarasan@yahoo.co.in.

(Received 14 July 2010; revised 21 October 2010; accepted 24 October 2010)

ISSN 1537-6516 print/ISSN 1537-6524 online 2011 Informa Healthcare USA, Inc.
DOI: 10.3109/15376516.2010.538752 http://www.informahealthcare.com/txm

Materials and methods
Plant material
The fresh root of the Ricinus communis was collected from
the Erode District (Tamil Nadu, India) in the month of July
to August and the plant was identified and authenticated by
the Research Officer (Pharmacognosy), Central Research
Institute (Siddha), Ministry of Health and Family welfare,
Governement of India. The voucher specimen of the plant
(01/2000) has been kept in the Department of Pharmacology,
C.L. Baid Metha College of Pharmacy (Chennai, India) for
future references.
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Preparation of aqueous and methanol extracts
The freshly collected root of this plant was chopped, shade
dried, and coarsely powdered. The powder was defatted with
petroleum ether (6080C), then successively extracted with
methanol and distilled water using soxhlet extractor. The
aqueous and methanol extracts were dried under reduced
pressure using a rotary flash evaporator. The percentage yield
was 5% w/w for aqueous extract (AE) and 6% w/w for methanol
(ME). The extracts were kept in a refrigerator for further use.
Animals
Adult Wistar albino rats (170200g) of either sex were used
For personal use only.
for the evaluation of toxicological studies. Female rats were
used for acute toxicity study, and male and female rats were
used for repeated dose toxicity. They were kept in polypro-
pylene cages at 252C, relative humidity 4555% under 12h blood urea nitrogen (Natelson etal. 1951), creatinine (Slot
light:dark cycles. All the animals were acclimatized for 1 week
before use. They were fed with standard laboratory animal
feed (Poultry Research Station Tamil Nadu Veterinary and
Animal Sciences University, Chennai, India.) and tap water
ad libitum. The test extracts were administered in the form
of a suspension using 1% Sodium carboxymethyl cellulose
(SCMC). The experimental protocols were approved by the
IAEC (Institutional Animal Ethics Committee, Sanction no:
IAEC/10/01/CLBMCP/20032004, dated 19 April 2004).
Chemicals
All the fine chemicals where purchased from Sigma (Chennai,
India) and other chemicals from SRL (Chennai, India),
Aldrich (Chennai, India), CDH (Chennai, India), Qualigens,
and Hi-media.
Acute toxicity study
An acute oral toxicity (Ecobichon 1997) study was performed
as per OECD-423 guidelines (acute toxic class method).
Wistar rats (n=3) of female sex, selected by random sam-
pling technique, were used for the study. The animals were
kept fasting overnight, providing only water, after which the
extracts were administered orally at the dose level of 5mg/kg
body weight by intragastric tube and the animals observed
for 14 days. If mortality was observed in two-to-three ani-
mals, then the dose administered was assigned as toxic. If
mortality was observed in one animal, then the same dose
was repeated again to confirm the toxic dose. If mortality was
Ricinus communis acute toxicity study 247
not observed, the procedure was repeated for further higher
doses such as 50, 300, and 2000mg/kg body weight.
90 days repeated oral dose toxicity study
In the 90 days repeated oral dose study the OECD test
guidelines-408 specified a test at one dose level equivalent
to at least 1000mg/kg/day (OECD 1998), and based on the
OECD-408 guideline we selected the dose. The animals were
divided into three groups of six animals each, three males and
three females. Group I served as control to receive 1% SCMC
10ml/kg for 90 days. The group II and III animals received
aqueous and methanol extract of Ricinus communis root
1000mg/kg, respectively, for 90 days. Body weight, food, and
water intake were recorded at weekly intervals with simul-
taneous observation for toxic manifestation and mortality
if any. At the end of the 90 days treatment, all the animals
were sacrificed by cervical decapitation after the collection
of blood sample through retro-orbital puncture.
The collected blood sample was used for the evaluation
of hematological and biochemical studies. Liver and kidney
were dissected out and homogenated in 0.1 m Tris buffer,
and the homogenate was used for biochemical investigations.
Sections of the liver, kidney, lung, heart, brain, ovary, testis,
and spleen were dissected out and kept in 10% formalin for
histopathological studies. The hematological parameters
like RBC, WBC, DLC, and Hb (Ghai 1993) were estimated.
The blood clotting time (Pal and Pal 2001) was estimated.
The biochemical parameters of glucose (Sasaki etal. 1972),
1965), total cholesterol (Parekh and Jung 1970), total protein
(Lowry etal. 1951), total bilirubin (Malloy and Evelyn 1937),
AST (King 1965b), and ALP (King 1965a) were estimated.
Statistical analysis
The data were expressed as mean SEM. Results were ana-
lyzed statistically by One-Way ANOVA followed by Tukeys
multiple comparison using SPSS software students version.
The difference was considered significant if p<0.05.
Results
Acute toxicity study (OECD423)
Ricinus communis root (AE, ME) extracts did not produce
any toxic symptoms or mortality up to the maximum dose
of 2000mg/kg/po in rats and, hence, the drugs were consid-
ered to be safe for pharmacological study. According to acute
toxic class method (OECD423 guidelines), the LD 50 dose
of 2000mg/kg and above is categorized as X unclassified
(OECD 1996). Based on these results and under the condi-
tions of this study, the median lethal dose (LD50) of AE and
ME extracts after single oral administration in female Wistar
rats was found to be more than the maximum dose level of
2000mg/kg body weight.
90 days repeated oral dose toxicity study
In 90 days repeated dose study of AE and ME extracts at
the dose of 1000mg/kg/po did not produce any significant
 248 R. Ilavarasan etal.

changes in the body weight, food, and water intake shown
in Table 1. During the study period the AE and ME extracts
treated animals did not show any abnormal behavioral
changes and mortality or morbidity status. The hemato-
logical parameters white blood cells (WBC), red blood cells
(RBC), hemoglobin, clotting time, and differential leukocyte
count did not produce any significant changes when com-
pared with control animals (Table 2). Biochemical param-
eters total serum protein, alkaline phosphates, blood urea
nitrogen, creatinine, AST, ALT, cholesterol, glucose, and total
bilirubin parameters were determined in each sample. No
significant differences were observed in AE and ME extracts
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ME. In the present acute oral toxicity study, AE and ME did
not cause any mortality, abnormal body weight changes,
or gross necropsy findings in female rats at an oral dose of
2000mg/kg. These results demonstrate that the LD50 of AE
and ME is greater than 2000mg/kg, when administered once
orally via gastric intubation to female Wistar rats. In 90 days
repeated dose toxicity study the extracts AE and ME did not
cause any mortality, morbidity, or adverse changes in the
general behaviors. Generally changes in body weight have
been used to access the course of the disease and response to
the therapy of drugs (Winder etal. 1969), and also indicate the
adverse effects of drugs (Teo etal. 2002). The blood glucose

treated animals when compared to the control animals level, which remained constant in extract treated animals,
(Tables 3 and 4). shows the normoglycemic activity in the extracts. The blood
The histopathological observation (Figures 14) of the urea nitrogen and creatinine observation shows that the AE
major vital organs like liver, kidney, heart, spleen, lungs, and ME extract did not produce any renal dysfunction, as
ovary, testis, and brain showed normal architecture suggest- increase in the above parameters reveals the impaired renal
ing no detrimental changes, and no morphological distur- function or acute renal failure (Varley 1964). AST, ALT, ALP,
bances were caused by the daily oral administration of the
AE and ME extracts compared to control animals.
Table 3. Biochemical effect of serum in 90 days repeated dose treatment
of AE and ME extracts in Wistar rats.
Discussion Aqueous extract Methanol extract
Parameters Control (1000mg/kg) (1000mg/kg)
The toxicity studies were focused to determine and demon-
Blood glucose 122.331.65 123.162.28 1162.77
strate the toxicity and safety aspects of the extracts AE and Blood urea 17.50.69 18.160.86 18.200.81
For personal use only.

nitrogen
Serum creatinine 0.330.02 0.340.04 0.320.03
Table 1. 90 days repeated dose treatment of AE and ME extracts on food Cholesterol 75.207.89 70.754.54 67.598.91
intake, water intake, and body weight in Wistar rats.
AST 113.3314.70 13814.89 124.6611.65
Aqueous extract Methanol extract
ALT 58.665.67 61.6616.41 54.668.14
Parameters Control (1000mg/kg) (1000mg/kg)
Total protein 7.200.39 7.220.69 6.600.27
Water intake 16.830.68 16.660.76 17.160.64
(ml/day) ALP 96.263.93 95.6713.29 82.7711.42
Food intake 10.160.89 9.50.87 9.160.64 Total bilirubin 0.470.12 0.350.06 0.440.06
(g/day) Data represents mean SEM of six animals (one-way ANOVA).
Body weight (g) The values were non-significant compared to control.
Units: AST/ALT, IU/L; ALP, KA units; Blood glucose, BUN, Creatinine,
Day 1 147.331.59 146.831.47 147.331.75
Cholesterol, Total protein, Total bilirubin, mg/dL.
4th week 183.832.21 185.162.03 1802.63
8th week 221.661.98 2201.66 224.162.74
13th week 250.662.15 254.163.80 2533.22 Table 4. Biochemical effect of liver and kidney tissue in 90 days repeated
Data represents mean SEM of six animals (one-way ANOVA). dose treatment with AE and ME extracts in Wistar rats.
The values were non-significant compared to control. Aqueous extract Methanol extract
Parameters Control (1000mg/kg) (1000mg/kg)
Liver tissue
Table 2. Hematological effect of 90 days repeated dose treatment of AE Cholesterol 23.851.76 22.453.71 24.443.05
and ME extracts in Wistar rats. AST 16015.05 161.6616.41 15020.33
Aqueous extract Methanol extract ALT 9633.56 86.3311.15 97.669.75
Parameters Control (1000mg/kg) (1000mg/kg) Total protein 2.430.14 2.230.149 2.410.04
RBC ( 106/mm3) 6.450.34 6.430.77 6.150.15 ALP 2.500.41 2.790.87 2.261.07
WBC ( 103/mm3) 8.70.68 8.450.78 8.751.55 Kidney tissue
Hb (g/dL) 130.37 13.631.15 14.160.69 Cholesterol 44.551.53 36.134.62 35.784.44
Blood clotting 2.50.22 2.660.21 2.430.19 AST 136.3317.94 13822.96 148.3322.92
(min)
ALT 87.339.08 7115.37 70.338.01
Neutrophils (%) 29.830.75 27.51.76 29.331.67
Total protein 1.630.25 1.380.22 1.380.12
Lymphocytes (%) 70.53.04 70.662.98 70.53.03
ALP 7.432.17 6.172.23 8.631.22
Eosinophils (%) 4.50.43 4.161.01 4.661.52
Data represents mean SEM of six animals (0ne-way ANOVA). The values
Monocytes (%) 2.330.80 2.330.76 2.50.76 were non-significant compared to control.
Basophils (%) 0.50.22 0.330.21 0.50.22 Units: AST/ALT, moles of pyruvate liberated/mg protein/min; ALP,
Data represents mean SEM of six animals (one-way ANOVA). The values moles of phenol liberated/mg protein/min; Total protein, mg/gm tis-
were non-significant compared to control. sue; Cholesterol, mg/dL.
 Ricinus communis acute toxicity study 249

LIVER KIDNEY HEART LUNGS

Group I Group I Group I Group I

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Group II Group II
Group II Group II

Group III Group III

Group III Group III
Figure 1. Histopathology of liver and kidney showing normal control
(Group I), aqueous extract (Group II), and methanol extract (Group III). Figure 2. Histopathology of heart and lungs showing normal control
Histopathology section of the tissues stained with alum-hemotoxylin and (Group I), aqueous extract (Group II), and methanol extract (Group III).
eosin. Magnification of the figures 1010=100. Histopathology section of the tissues stained with alum-hemotoxylin and
eosin. Magnification of the figures 1010=100.
For personal use only.

TESTIS OVARY
SPLEEN BRAIN

Group I Group I Group I Group I

Group II Group II Group II Group II

Group III Group III Group III Group III

Figure 3. Histopathology of spleen and brain showing normal control Figure 4. Histopathology of testis and ovary treated with normal control
(Group I), aqueous extract (Group II), and methanol extract (Group III). (Group I), aqueous extract (Group II), and methanol extract (Group III).
Histopathology section of the tissues stained with alum-hemotoxylin and Histopathology section of the tissues stained with alum-hemotoxylin and
eosin. Magnification of the figures 1010=100. eosin. Magnification of the figures 1010=100.

and total bilirubin are good indices of liver function (Rodwell to the liver and kidney. This was further confirmed by the
1993). The extracts AE and ME treatment did not show any histopathological assessment of these organs. In conclusion,
significant changes in the enzyme levels in serum, liver, and the AE and ME extracts were considered to be safe for long-
kidney tissues. Hence, the extracts did not induce any toxicity term treatment.
 250 R. Ilavarasan etal.
Acknowledgement
The authors are highly thankful to the Secretary and the
Principal, C.L. Baid Metha College of Pharmacy, Chennai-600
096, for providing the facilities to carry out this work.
Declaration of interest
The authors report no conflicts of interest. The authors alone
are responsible for the content and writing of the paper.
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