Antihyperglycemic Activity of Various Fractions of Cassia auriculata Linn . in Alloxan Diabetic Rats

Watanabe S, Kayano M, Ishino Y, Miyao K. US Patent No., 3,976, 1. 197. Dollery C, editor. Therapeutic drugs. Edinburgh: Churchill Livingston; 2. 1991. Reynold JE, editor. Martindale, The extra pharmacopoeia. London: The 3. Pharmaceutical Press; 1991. Rowland M, Tozer TN. Clinical pharmacokinetics concepts and 4. applications. 3rd ed. New Delhi: B.I.Waverly Pvt.Ltd; 1996. Satoskar RS, Bhandarkar SD, Ainapure SS. Pharmacology and 5. pharmacotherapeutics. Revised 15th ed. Mumbai: Popular Prakashan; 1997. Rall TW. Drugs used in the treatment of asthma. In: Gillman AG, 6. Rall TW, Nies AS, Taylor P, editors. Goodman and Gillmans the pharmacological basis of therapeutics. 8th ed.New York: Pergamon Press; 1990. p. 618-37. The United States Pharmacopoeia 24. Rockville, MD: The United States 7. Pharmacopoeial Convention; 2000. p. 1942. Indian Pharmacopoeia. 4th ed. Delhi: Controller of publication, Ministry 8. of Health and Family Welfare; 1996. p. A-184. Indian Pharmacopoeia. 4th ed. Delhi: Controller of publication, Ministry 9. of Health and Family Welfare; 1996. p. A-80A-84, 735-36. All the capsules were found to be ß oating for more than 12 h. The drug content in the capsules of each formulation was found within the range of 90-110 percent as specified for theophylline capsules IP8, which confirms to show an excellent drug content uniformity in each formulation (Table 3).

morbidity due to this disease is increasing 2 .The focus has been shifted to treat the various ailments through plant-derived drugs due to their safety, efficacy, cultural acceptability and lesser side effects.
Cassia auriculata Linn., a member of genus Cassia belonging to family Caesalpiniaceae is commonly known as Tanner's Cassia 3 .It is a shrub found throughout southern, western and central India 4 .The various parts of the plant has been reported to posses a number of therapeutic activities to manage disease states like leprosy, asthma, gout, rheumatism 5 and diabetes 6 .It is also used as antipyretic, antiulcer and in the treatment of skin infections 7 .The ß ower has been reported to contain ß avonoids, proanthocyanidins and β-sitosterol [8][9] .
In folk remedies, ß owers of Cassia auriculata Linn.are proposed to have antidiabetic activity.From literature survey, it was evident that the aqueous extract of ß owers has been reported for its antidiabetic activity in streptozotocin-diabetic rats at a dose of 0.45 g/kg body weight 10 .In the present investigation an attempt has been made to find out the potent fraction responsible for the said activity at a dose of 0.20 g/kg body weight.Thus, the present work describes effect of the hydromethanolic extract and its fractions on the blood glucose, total protein, cholesterol and triglyceride level in alloxan-diabetic rats.
The ß owers of Cassia auriculata Linn.were collected from the premises of Shirpur situated in Dhule district of Maharashtra during March 2005.The crude drug was authenticated from the Botany Department, L. K., Dr. P. R. Ghogrey Science College, Dhule (MS).Herbarium specimen was deposited in Botany Department for future reference.The flowers were dried under shade and crushed into coarse powder.
Coarse powder was extracted by cold maceration method using methanol:water (1:1).The extract thus obtained was concentrated in rotary flash vacuum evaporator and further dried in vacuum oven.The hydromethanolic extract was subjected to fractionation with ethyl acetate and n-butanol.The fractions were dried in vacuum oven and utilized for animal studies.
In house bred Wistar male rats weighing between 180-220 g were utilized for study.Animals were housed in polypropylene cages and given standard pellet diet and water ad libitum.The ethical clearance was obtained from the Institutional Animal Ethical Committee (Approval no.152/02/C/CPCSEA).Diabetes was induced in rats by intraperitoneal injection of alloxan monohydrate 11 (S.D. Fine Chemicals, Mumbai) in ice-cold citrate buffer, pH 4.5 at a dose of 0.12 g/kg.The diabetic state was conÞ rmed 48 h after alloxan injection by weight loss and hyperglycemia 12 .Rats with blood sugar levels 200-350 mg/dl were selected for the study.
The rats were divided into six groups each carrying six animals.Group I served as diabetic control and received 0.3% w/v CMC, orally and alloxan.Group II served as positive control and received standard drug phenformin (0.04g/kg).Group III received hydromethanolic extract of ß ower (0.20g/kg).Group IV received n-butanol fraction of hydromethanolic extract (0.20g/kg).Group V received ethyl acetate fraction of hydromethanolic extract (0.20g/kg).Group VI served as normal untreated group.The treatment was continued for 8 d by administering the respective fractions/drug in 0.3% w/v CMC.On ninth day of the treatment, blood samples were collected by puncturing the retro-orbital plexus under mild ether anesthesia and kept aside for ½ h for clotting.Serum was separated by centrifuging the samples at 6000 rpm for 20 mins and stored in the refrigerator until analyzed.
The serum was analyzed for blood glucose level (GOD-PAP method) 13 , total protein (Biuret method) 14 , cholesterol (CHOD-PAP method) 15 and triglycerides (enzymatic method) 16 according to the kit manual.The quantitative measurements were made on six animals in each group and the values of biochemical estimations are expressed as mean±SE.The data obtained were subjected to one-way ANOVA followed by Bonferroni multiple comparison test 17 .
Alloxan-induced β-cell cytotoxicity is well established animal model for investigating antidiabetic activity.
From the results, it is evident that hydromethanolic, n-butanol and ethyl acetate fraction shown signiÞ cant (P<0.001)reduction in blood glucose level.However n-butanol fraction was highly effective and results are comparable with that of reference drug, phenformin (Table 1

TABLE 1 : EFFECT OF EXTRACT AND FRACTIONS OF CASSIA AURICULATA FLOWERS ON BODY WEIGHT AND BLOOD SUGAR LEVEL
The observations are given as mean±SEM, n= 6, *p<0.01,compared with diabetic control group

Table 2
).In comparison to other extracts, n-butanol fraction was found to be more potent in normalizing the blood lipids and protein level.Proximate chemical analysis reveals the presence of steroids, ß avonoids, tannins and carbohydrates in hydromethanolic extract

TABLE 2 : EFFECT OF EXTRACT AND FRACTIONS OF CASSIA AURICULATA FLOWERS ON BIOCHEMICAL PARAMETERS
The observations are given as mean±SEM, n= 6, *P<0.01 compared with diabetic control group