Ricinus Communis

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Transcript Ricinus Communis

“ANTIUROLITHIC ACTIVITY OF
EXTRACTS OF ROOTS OF RICINUS
COMMUNIS”
PRESENTED
BY
JUHI TIWARI
1. Introduction

Herbal medicine : Medicinal plants are used by all the cultures across the world .
According to W.H.O estimates around 80% of population in developing countries use
herbal drugs for some aspects of primary health care. Herbs are the staple of medical
treatment in many developing countries.

The urinary system: The urinary system consists of two kidneys, two ureters, the urinary
bladder, and the urethra.
Kidney: kidneys are the principal regulators of the internal environment of the body. The
composition of all body fluids is either directly or indirectly regulated by the kidneys as
they form urine from blood plasma.
The kidneys which are two in number, lies on the posterior abdominal wall, one on
each side of vertebral column, behind the peritoneal cavity and below the diaphragm.
Kidneys are bean-shaped organs, about 11 cm long, 6 cm wide, 3 cm thick and
weigh 150 g. They are embedded in, and held in position by, a mass of fat (Scanlon et al.,
2007).

Introduction to Urolithiasis
It is also called Nephrolithiasis or kidney stone.
 Urolithiasis is the presence of calculi in the urinary tract.
 Eighty percent of calculi are composed of calcium (either oxalate or phosphate).
 Others composed of struvite, uric acid, xanthine or cystine.
Urolithiasis
Martin L.J.A(2002)

Urolithiasis may be defined as the solid accumulation of the material that
form in the urinary system which may be in the kidney, ureter, urinary
bladder or other parts of system.

This process is also known as super saturation of the urine i.e greater
number of solute in a solvent.
Epidemiology Joual, A. (1997)

Urolithiasis is the third most common cause of urinary tract disease ,
predominantly in male gender in proportion of approximately 2:1 and is
characterized by a recurrence rate of 50% and reaching 70% within 10
years.
 Incidence of upper urinary tract stone was 2.4 fold greater in men than
in women.
Process of stone formation (Atmania et al., 2004)
Saturation
Supersaturation
Nucleation
Stone formation
Etiology of Urolithiasis
(Pfien et al., 1990)
 Family History of Renal Stone Disease
 Occupational or Situational Risk Factors
 Systemic Diseases
 Medications
 Dietary Factors
 Bacterial infection
Symptoms of Kidney Stones (Parmar et al., 2004).
 Renal pain
 Hematuria
 Pyuria
 Dysuria
 Oligouria
Abdominal distension
 Nausea/vomiting
 Fever and chills
 Postrenal azotemia
Crystal aggregation
Crystal growth
Classification of Kidney Stone Coe FL (2005)
 Calcium containing Stone
 Calcium Oxalate Stone (60%)
 Calcium Phosphate Stone (20%)
 Struvite Stone (8%)
 Uric Acid Stone (10%)
 Cystine Stone(1.5%)
 2,8-di hydroxy adenine stone(0.5%)
Causes for kidney stone
Stones were formed when there is a decrease in urine volume and or
excess in stone forming substance in urine. Dehydration , different
medical condition and also low level of dietary calcium intake may
alter calcium oxalate balance thus resulting in increased excretion of
oxalate and propensity to form oxalate stone.
Need of study
 There are so many research has been already done in respect to
urolithiasis but there is still need to research a safe and effective drug
for the treatment of urolithiasis because there is no satisfactory drug in
modern medicine which can dissolve stone therefore physician still
remain to be depend on alternate system of medicine for their better
relief , which is economically costly and produces major side effects .
Objective of study
To conduct systematic phytochemical investigation of roots of
Ricinus Communis .
To conduct acute toxicity .
To develop urolithiasis model in rats .
To evaluate anti urolithiatic activity of extract of roots of Ricinus
Communis .
To evaluate antimicrobial activity of roots of Ricinus Communis .
Plant profile :
RICINUS COMMUNIS :
Ricinus Communis Tewari, D.D. (2012)
Ricinus Communis : Castor Oil Plant .
Herbaceous plant or semi woody large , dicot , shrub or
small tree belonging to the family Euphorbiaceae .
Reported activities
Anti asthmatic , anti inflammatory, anti microbial ,
anti fungal , anti histaminic activities, Anti hepato
toxicity, Antinociceptive, Antimicrobial activity,
Antifungal activity, Antibacterial activity.
• Animals And Experiments.
Sprague Dawley rats , weighing 100+5 gm from the animal house of
Pinnacle Biomedical Research Institute, Bhopal.
Animal maintenance and handling were in accordance to CPCSEA.
• Extraction Pradnya Onkar et.al (2012)
The powdered roots of Ricinus communis was extracted by
means of maceration process . powder was subjected to cold
maceration with pet ether initially and afterward marc was
treated with hydro-alcoholic solution in a jar, for about 7 days
at room temperature.
•Authentication
Botansit : Dr. Pradeep Tiwari and Herbarium No. Bot./Her./B/1123
was deposited in the herbarium of the Hari Singh Gaur Vishwa
Vidyalaya, Sagar (Madhya Pradesh).
cont.
PRELIMINARY PHYTOCHEMICAL TESTING Khandelwal
K.R., (2003)
1)
2)
3)
4)
5)
6)
The crude extract of Ricinus Communis was
screened for the presence of different phyto chemical
substance
Test for alkaloids : Dragendorff’s test, Mayer’s test,
Hager’s test, Wagner’s test.
Test for saponins
Test for Glycosides : Legal’s test, Baljet’s test, Keller
killani test and Borntrager’s test.
Test for carbohydrates : Molisch test, Fehling’s test,
Benedict’s test
Test for tannins and phenolic compound
Test for flavonoids
Acute oral toxicity
OECD 2001- guideline
 Acute oral toxicity study of hydro alcoholic and petroleum ether extract was
carried out according to OECD 423 guidelines.
Anti microbial activity
 Well Diffusion Method (El-Mahmood et al., 2008 & Bauer et
al.,1966): Zone of inhibition was measured.
 Anti oxidant activity
 DPPH Activity (Patil et al, 2009) : percentage of inhibition was
measured
 Hydrogen Peroxide Scavanging Assay (Patil et al, 2009) :
percentage of inhibition was measured
 Reducing Power Assay (Jayanthi and Lalitha 2011) : Increase in
absorbance of the reaction mixture indicates the reducing power
of the Samples.
 Total Phenolic Content ( Maurya S , Singh D, 2010) : Line of
regression from Gallic acid was used for estimation .
 Total flavonoid Content ( Maurya S , Singh D, 2010) : Line of
regression from rutin was used for estimation
Procedure of dpph and hydrogen peroxide
assay
 Preparation of Standard Ascorbic acid solutions
 Preparation of Test solutions
 Preparation of Control solution
 Percentage of antioxidant activity of plant extract
and Ascorbic acid was calculated by using formula:
Procedure of reducing power assay
 Preparation of Standard Ascorbic acid solutions
 Preparation of Test solutions.
 Increase in absorbance of the reaction mixture
indicates the reducing power of the Samples.
• Procedure of total phenolic content
 Different concentration of gallic acid and plant
extract was prepared in methanol.
 0.5ml of each sample was introduced into test tube
and mixed with 2.5ml of a 10 fold dilute folin
Ciocalteu reagent and 2ml of 7.5% sodium carbonate
was prepared in methanol.
Calculation of tpc

Line of regression from Gallic acid was used for estimation of unknown
phenol content.From standard curve of gallic acid line of regression was
found to be :
y = 0.005x + 0.065 and
R2 = 0.976
Thus the goodness of fit was found to be good for selected standard curve.
By putting the absorbance of test sample (y = absorbance) in line of
regression of above mentioned GA.
Total flavonoid content : Total flavonoids were measured by a
colorimetric method . standard solution of rutin was added to a 75 μl of
NaNO2 solution, and mixed for 6 min, before adding 0.15 mL AlCl3 (100
g/L). After 5 min, 0.5 mL of NaOH was added. The final volume was
adjusted to 2.5 ml with distilled water and thoroughly mixed.
Absorbance of the mixture was determined at 510 nm against the same
mixture, without the sample, as a blank. Total flavonoid content was
expressed as mg rutin/g dry weight (mg rutine/g DW), through the
calibration curve of Rutin. All samples were analysed in three
replications.
Calculation of tfc
Line of regression from rutin was used for estimation of unknown
flavonoid content. From standard curve of rutin, line of
regression was found to be :
y = 0.001x - 0.118 and
R2 = 0.985
Thus the goodness of fit was found to be good for selected
standard curve. By putting the absorbance of test sample (y =
absorbance) in line of regression of above mentioned rutin.
InVitro Assesement of Anti urolithic Activity
• Calcium Oxalate crystallization (Beghalia M., Ghalem S.,Allali H.,
Belouatek A., Marouf A.2008 ,2009) : By determining the crystal size.
• Procedure
 Preparation of synthetic urine.
 Simulation of the sedimentary crystal formation
 The crystal size development by microscope was carried out at
different time intervals of formation of crystals . Calculated
percentage of Inhibition was based on the formula:
I% = [(TSI- TAI) / TSI] * 100
In vivo assesement of anti urolithic
activity Anil T. Pawar (2012)
 Ethylene glycol and ammonium chloride induced urolithiasis
model was used to asses the antiurolithiatic activity in rats.
 Cystone was taken to serve as a standard for anti urolithic
activity.
•
Preparation of doses : All the doses were prepared in hydro
alcoholic and DMSO solvent . In all cases, the concentration was
prepared according to body weight of animal.
• Evaluation of Antiurolithic activity : Animals were divided
into 7 groups containing six animals in each group and kept in a
metabolic cages individually for entire duration of experiment.
All animals had free access to regular rat chow and drinking
water ad libitum .
Experimental design for Antiurolithiatic
activity







Group I- Control group
Group II- Lithiatic Control
Group III- Preventive Regimen
Group IV- Preventive Regimen
Group V- Preventive Regimen
Group VI- Preventive Regimen
Group VII- Cystone treated
• Description of groups
 Group I- Control group: Normal saline was given to the rats.
 Group II- Control group: Lithiatic induction was done by ethylene
glycol and ammonium chloride.This group serve as control for gp.
III to gp. VI.
 Group III- Hydroalcoholic extract of Ricinus Communis 200 mg / kg
bwt.was given with inducing agent to determine the antiurolithic
ability of extract.
Cont…
 Group IV- Hydroalcoholic extract of Ricinus Communis 400 mg
/ kg bwt.was given with inducing agent to determine the
antiurolithic ability of extract.
 Group V- Petroleum ether extract of Ricinus Communis 200
mg / kg bwt.was given with inducing agent to determine the
antiurolithic ability of extract.
 Group VI- Petroleum ether extract of Ricinus Communis 400
mg / kg bwt.was given with inducing agent to determine the
antiurolithic ability of extract.
 Group VII- Animals was treated with standard drug cystone
750 mg/kg bwt. To compare the result of extracts.
All rats were housed in metabolic cages
individually for entire duration of the experiment. On 22nd day
all the groups was sacrificed.
After the termination of experiment blood was
collected by cardiac puncture and serum was separated by
centrifugation at 15,000 rpm for 10 min.
Histopathology
Divakar K et .al
 The abdomen was cut open to remove either kidney from each animal.
Isolated kidneys were cleaned off extraneous tissue and preserved in 10%
formalin. One of the isolated kidneys was then embedded in paraffin using
conventional method and cut into 5 µm thick sections and stained using
hematoxylin-eosin dye and finally mounted in diphenyl xylene. Then the
sections were observed under microscope for histopathological changes in
kidney architecture and their photomicrographs was taken.
Measurement of Biochemical Marker

Creatinine , BUN , Urea , Uric Acid , Phosphate , Calcium was determined
by the method given in the protocol of Erba Diagnostic Ltd..
 Creatinine : Creatinine react with alkaline picrate to produce an orangeyellow color. The absorbance of the orange-yellow color formed is directly
proportional to creatinine concentration . Creatinine was calculated by
determining the ratio of change in absorbance of test and standard
multiplied by concentration of standard.
Cont….
 Blood urea nitrogen and urea
The estimation of urea in serum
involves the enzyme catalysed reactions in which the urea is react with water
to form ammonia, which in turn reacts with α-ketoglutarate and NADH to
form glutamate. The rate of decrease in absorbance is monitored at 340 nm
and is directly proportional to urea concentration in the sample.
:
 Uric Acid : Estimation is done with a modified Trinder peroxidase method
using TBHB. Estimation of uric acid requires reaction with water and oxygen
forming hydrogen peroxide which reacts with 2, 4, 6 Tribromo 3- hydroxyl
benzoic acid and 4 Aminoantipyrine to form Quinoneimine. This is proportion
to uric acid concentration.
 Calcium
Calcium has numerous function within the body , not only as a
structural factor in bones and teeth , but also in normal neuro muscular
function and the clotting of the blood .OCPC reacts with calcium in alkaline
solution to form a purple coloured complex . The intensity of the purple colour
formed is proportional to the calcium concentration .
:
 Phosphorus The principle of the method is based on the reaction in which
:
phosphorus reacts with ammonium molybdate to form phosphomolybdate.
This unreduced Complex (phospho molybdate) is directly proportional to
inorganic phosphorus present in sample.
Kidney Homogenate Analysis
Spakal VD (2008)
 Another isolated left kidney was then homogenized. The
homogenate was centrifuged at 2,000 rpm for 10 min and
supernatant separated. The enzymatic estimation of kidney
homogenate was determined by SOD , GSH and LPO.
Preparation of homogenate
• Organ was rinsed with ice cold normal saline followed by 0.15
M tris HCl (pH 7.4).
• Then the organ was divide and 10% w/v tissue was
homogenize with 0.15 M tris HCl and 0.1 M Phosphate buffer .
Then this homogenate was centrifuged at 15,000 rpm ,15 min ,
4 °C, and take supernatant as sample. Supernatant was
preserved in deep freeze at 2 °C.
Analytical Parameter of Kidney homogenate
 Super oxide Dismutase, (Spakal VD 2008) : Percentage of
inhibition was measured.
 Lipid Peroxidase, (Spakal VD 2008) : Obtained result are
equivalent to MDA.
 Glutathione (Spakal VD 2008) : 5,5’-dithio bis-2-nitrobenzoic
acid is reduced in presence of GSH to produce a yellow
compound. The reduced chromogen is directly proportional to
GSH conc.
 SUPEROXIDE DISMUTASE
:
Chemicals Used sodium pyrophosphate buffer, phenazine
methosulphate ,Nitroblutetrazolium , NADH.
Take OD at 560 nm (take butanol as blank)
 LIPID PEROXIDE :
Chemicals Used : SDS , acetic acid ,TBA, butanol , pyridine ,
Take OD at 532 nm (blank – butanol : pyridine / 15:1) (this
absorbance will be of total MDA formed).
 Glutathione :
Chemicals Used : TCA, EDTA , Ellman’s reagent, sodium citrate
solution. Take OD at 412 nm (water as blank)
Result
 Physical Characters of Extracts :
Hydroalcoholic
Petroleum Ether
 Color Brown
Greenish Brown
 Odour Sweet
Aromatic
 Appearance Sticky
Sticky
 %age yeild 6.82%
1.45%
 Phytochemical testing : Carbohydrate , reducing
sugar, flavonoids, glycosides, tannin and phenolic
compound are present in hydroalcoholic extract
whereas tannin and phenolic and triterpenoids and
steroids are present in petroleum ether extract.
Graphs showing antioxidant activity
DPPH Assay
20
% Inhibition
% Inhibition
25
15
10
5
y = 0.1894x + 4.233
R² = 0.9886
0
0
50
100
y = 0.4648x + 3.938
R² = 0.9963
60
50
40
30
20
10
0
0
20
40
150
60
80
100
120
Concentration (µg/ml)
Concentration (µg/ml)
% Inhibition of DPPH by HA Extract at
different concentration
% Inhibition of DPPH by PE Extract at
different concentration
% Inhibition
35
30
25
20
15
10
5
0
% Inhibition
Hydrogen peroxide scavenging assay
y = 0.3392x - 0.597
R² = 0.9586
40
35
30
25
20
15
10
5
0
y = 0.3368x + 3.823
R² = 0.9799
0
0
20
40
60
80
Concentration (µg/ml)
100
120
Inhibition of Hydrogen peroxide by PE Extract
at different concentration
50
100
150
Concentration (µg/ml)
Inhibition of Hydrogen peroxide by HA
Extract at different concentration
Graphs showing Antimicrobial activity
S.aureus
E.Coli
B.Subtillus
Kleibsiella
50
45
PE Extract (50mg/ml)
PE Extract (100mg/ml)
PE Extract (200mg/ml)
0floxacin (1mg/ml)
25
40
20
30
Zone of inhibition (mm)
Zone of inhibition (mm)
35
25
20
15
15
10
10
5
5
0
50 mg/ml
100 mg/ml
200 mg/ml
OFLOXACIN (1
mg/ml)
0
S.aureus
E.Coli
B.Subtillus
K. pneumoneae
Concentration (mg/ml)
Concentration (mg/ml)
Zone of inhibition due to Ricinus communis Zone of inhibition due to Ricinus communis
HA extract in different microorganism
PEEextract in different microorganism
Graph showing invitroanti urolithic
activty
25%
50%
75%
100%
25%
96
50%
75%
100%
6000%
94
5000%
92
4000%
% Inhibition
% Inhibition
90
88
3000%
86
84
2000%
82
1000%
80
0%
78
5
5 min
10 min
15 min
20 min
25 min
30 min
10
15
20
25
30
Time
Time
% inhibition of calcium oxalate crystal % inhibition of calcium oxalate crystal
at different time interval due to PEE
at different time interval due to HAE
Acute Oral Toxicity
 No mortality was observed up to 2000 mg/kg, hence 2000
mg/kg was considered as NOAEL.1/10th and 1/5th of NOAEL,
i.e.200 mg/kg and 400 mg/kg were selected as dose for further
in vivo investigation.
InVivo Antiurolithic Activity (Serological Analysis)
CREATININE
BUN
2.5
35
2.064
28.285
30
2
23.877
25
20
1.5
27.682
28.888
21.81
18.318
15
1
10
0.5
0.281
0.062
0.323
5
0.304
0.151
0.163
0
0
VEHICLE VEHICLE HAE (200 HAE (400 PEE(200 PEE(400 CYSTONE
+ EG
MG/KG) MG/KG) MG/KG) MG/KG)
Graph showing serum creatinine
concentration in different groups.
Graph showing blood urea nitrogen
concentration in different groups.
19.802
URIC ACID
4.5
4
3.5
3
2.5
2
1.5
1
0.5
0
UREA
4.122
4.048
70
3.957
60
3.057
50
2.18
1.835
60.53
2.125
40
59.239
51.096
61.821
46.673
42.376
39.201
30
20
10
0
VEHICLE VEHICLE HAE (200 HAE (400 PEE(200 PEE(400 CYSTONE
+ EG
MG/KG) MG/KG) MG/KG) MG/KG)
Graph showing urea concentration
in different groups.
Graph showing uric acid concentration
in different groups.
8
7
6
5
4
3
2
1
0
6.765
6.257
4.817
2.96
CALCIUM
PHOSPHATE
5.623
6
6.132
5
3.002
3
5.702
3.944
4
3.945
5.407
2.712
2.503
2.528
2
1
0
VEHICLE VEHICLE HAE (200 HAE (400 PEE(200 PEE(400 CYSTONE
+ EG
MG/KG) MG/KG) MG/KG) MG/KG)
Graph showing phosphate concentration
in different groups.
Graph showing serum calcium
concentration in different groups.
Graph showing effect on enzyme involved in
oxidative stress
SUPEROXIDE DISMUTASE
LIPID PEROXIDASE
0.06
0.05
0.048
0.04
0.03
0.032
0.02
0.017
0.02
0.019
0.02
0.013
0.01
0
0.472
0.5
0.45
0.4
0.35
0.3
0.25
0.2
0.15
0.1
0.05
0
0.39
0.246
0.095
0.093
0.121
VEHICLE VEHICLE + HAE (200 HAE (400 PEE(200
EG
MG/KG) MG/KG) MG/KG)
Graph showing effect on LPOenzyme due
to both extract in various groups.
46.708
50
40
0.36
PEE(400 CYSTONE
MG/KG)
Graph showing effect on SOD enzyme
due to both extract in various groups
GLUTATHIONE
35.815
35.085
26.714
26.183
30
39.803
16.79
20
10
0
VEHICLE
VEHICLE + EG
HAE (200
MG/KG)
HAE (400
MG/KG)
PEE(200 MG/KG)PEE(400 MG/KG)
CYSTONE
Graph showing effect on glutathione enzyme due to both extract
in various groups
Conclusion
 Recently, there is increasing evidence that many healthy natural food and
medicinal herbal and supplements have the potential to become valuable
complementary therapy in the treatment of various renal disorders. In spite
of tremendous advances in the field of medicine, there is no truly satisfactory
drug for the treatment of urolithiasis.
 In the present study, dried powder of Ricinus Communis was subjected to
extraction using petrolium ether and hydro-alcohol (70% Dis.water+30%
Methanol). Some part of both extracts was reserved for preliminary
phytochemical investigation and rest was utilized for pharmacological
screening.
 The preliminary phytochemical investigation of H A extract showed presence
of carbohydrate , reducing sugar, flavanoids , glycoside , tannin and phenolic
compound and in PE extract tannin and phenolic compounds and
triterpenoids and steroids are present.
 The present study indicates that the administration of Ricinus Communis
extract to rats with ethylene glycol (0.75 %) and ammonium chloride (1 %)
induced urolithiasis, reduced and prevented the growth of kidney stones
and renal impairment. The hydroalcoholic extract was having significant
protective effect against ethylene glycol with ammonium chloride induced
urolithiasis than petrolium ether extract. Accordingly, it can be concluded
that the supplementation of Ricinus Communis has a beneficial effect on
urolithiasis induced by ethylene glycol with ammonium chloride solution
and may be also by other chemical factors.
 Whenever ,a patient is suffering from urolithiasis the chances of microbial
infection in the kidney or urinary tract is much higher. This infection may
be due to any particular pathogenesis or due to the histological damage
that can occur due to sharp edges of the crystals. Some time the urinary
tract itself gets injury and further microbial infection due to movement of
small stone through the tract. The Ricinus Communis showed significant
antimicrobial potential against S.aureus, Kleibsiella , B. subtillus and E.Coli.
These microorganisms are common for urinary tract infection. Some time
these microorganism themselves create an environmental condition in the
kidney or urinary tract that favours the formation of kidney stone. Thus
being antimicrobial against these microorganisms the Ricinus Communis
extract will decrease the chances of microbial infection as well as the
chances of favourism for the condition in support of crystal growth.
This study also includes the in vitro antiurolithic study and it was observed that
extract significantly inhibited formation of crystals. Hence extract was having
some component that is avoiding formation of calcium oxalate crystals.
To ascertain safety of any component intended to used inside the
body, it needs to be confirmed. In present study extracts acute oral toxicity
was confirmed on the basis of OECD 423 guidelines. It was observed that
extract was not toxic upto the dose of 2000 mg/kg and hence it was
considered as Not Observed Adverse Effect Limit (NOAEL).
As mentioned above for assessment of antiurolithic activity of HA
and PE extract urolithiasis was induced by using ethylene glycol and
ammonium chloride. Ist group was not having any treatment and hence was
called as vehicle treated. IInd group was having only induced with vehicle
and rest groups were having treatments including standard drug Cystone in
last group.
Assessment was done on three bases, first on the basis of kidney
function test (including Creatinine and BUN), estimation of factor involved in
stone formation (Uric acid, Urea, Phosphate, and calcium), and effect of
enzymes involved in oxidative stress (LPO, SOD, and GSH).

 It was observed that in induced group (group 2) level of Creatinine, BUN was
significantly higher as compared to vehicle treated animals. In chronic renal
failure and uremia, an eventual reduction occurs in the excretion of creatinine.
The most frequently determined clinical indices for estimating renal function
depends upon concentration of urea in the serum. It is useful in differential
diagnosis of acute renal failure and pre renal condition where BUN–creatinine
ratio is increased.This confirmed that by administration of EG and AC
malfunctioning in kidney occurred. In extract treated animals with HA extract
has significant decrease in Creatinine and BUN level as compared to inducer
group . Effect of PEextract was not significant.
 Uric acid, Urea , Phosphate , and calcium level in kidney were also elevated in
inducer group as compared to vehicle treated group. In extract treated
animals with HA extract significant decrease in Uric acid, Urea, Phosphate,
and calcium level as compared to inducer group . Effect of PE extract was not
significant. Level of SOD and Glutathione was significantly less in inducer
treated group as compared to vehicle treated animals and LPO level was
significantly higher in induced treated animals this confirmed that in inducer
group oxidative stress was much higher. In extract treated animals level of
GSH and SOD was significantly higher as compared to inducer group and LPO
was significantly less as compared to inducer group.
 Thus from present investigation it can be concluded that hydroalcoholic
extract of roots of Ricinus communis possess significant antiurolithic activity.
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