RESEARCH ARTICLE Inventi Rapid: Ethnopharmacology Vol. 2, Issue 2 [ISSN 0976-3805] 2011ep331, CCC: $10 © Inventi Journals (P) Ltd Published on Web 05/04/2011, www.inventi.in INTRODUCTION The use of plants as a Source of Remedies for the treatment of many diseases dated back to prehistory and people of all continents have this old tradition. Citrullus colocynthis Linn. (C.CT) Commonly known as bitter apple is an important medicinal herb that traditionally have been used as a strong purgative, antitumor, (Especially of the Abdomen) Antidiabetic, Antioxidant, Antibacterial 2, 4, 8, 3, 10 etc. It is an annual or perennial (in wild) herbaceous, stems are rough and angular, leaves rough, 3 to 7 lobed, 5 to 10 cm Long, flowers monoecious, solitary, peduncled, axillary, corolla 5-lobed, ovary villous, fruit is pepo, nearly globular, 4 to 10 cm in diameter with somewhat elliptical fissures, about size of the small orange, seeds numerous, native to dry areas of north Africa, being common throughout the Sahara, Areas of Egypt, Sudan, Eastwards through Iran to India, Pakistan, Afghanistan & other parts of tropical Asia. The Curative properties of medicinal plants are mainly due to the presence of various complex chemical substances of different composition which occur as secondary metabolites 7&8 . They are grouped as alkaloids, glycosides, flavonoids, saponins, tannins, carbohydrates & essential oils. Plant based constituents can be derived from any part of the plant like bark, leaves, flowers, roots, fruits, seeds 5 etc. The present study deals with the hepatoprotective activity from invivo fruit sample of Citrullus colocynthis Linn. 1 Pest Control & Ayurvedic Drug Research Lab., S.S.L. Jain P.G. College Vidisha, [M.P.], India. E-mail: drarshediqbal@gmail.Com *Corresponding author MATERIALS AND METHODS The plant proposed for the present study belongs to family cucurbitaceae which is found in and around Vidisha district (M.P.) in India. The extraction of shade dried powdered material (40-50 mesh size) of the plant was carried out by soxhlet apparatus using different solvents according to increasing order of polarity. Two polar solvents viz. 70% or 90% Ethanol and Water respectively were used for extraction. Both are polar Solvents which can dissolve the polar compounds present in the cytoplasmic part of the plant material. Since flavonoids are extractable in the most polar solvent only. Hence choice of these two polar solvents was made for extraction purpose. The extraction was carried out for duration of 48 hrs. or till 8 cycles are completed. The crude extract thus obtained was filtered with whatsman filter paper number 1. The filtrate was vacuum evaporated in Rotor vapor (RE- 100) to obtain a semisolid crude material. The vacuum evaporation was done under reduced pressure & at temp. Below 40 0 C. The crude extract gave different percentage of yields in different solvents. The concentrated extract was tested for bioassay and chromatographic separation of the flavonoid compounds. A trace of toluene was added with the crude extract to prevent fungal growth. Extracts were kept in glass vials with stoppers. They were placed in refrigerator for further isolation, purification and bioassay experiments. Chemical Testing Method of Flavonoids • 10 mg of the substance was dissolved in 1ml chloroform. This solution was treated with 2ml bromine-chloroform reagent; an orange coloured precipitate was formed. • 20mg of the substance was dissolved in 1ml ethanol and mixed with 2ml phenyl hydrazine reagent and warmed gently, yellow precipitate of phenyl hydrazine was formed. Experimental Bioassay Methodology Rats were divided into six groups of 6 animals in each group. Group1: (Control) or vehicle were administered a single dose of liquid paraffin (LP) 1ml/kg body weight. Group2: (Diseased) received LP + ccl4 1ml/kg body weight each. Group3, 4 and 5: (Test Groups) were administered orally 50,100 and 200 mg/kg body weight of the plant compounds respectively in the form of aqueous suspension daily once a day. Group6: Received silymerin the known hepatoprotective compound (Sigma chemicals company USA) at a dose of 100mg/kg along with ccl4. After 36 hour of administration of ccl4 blood was collected and serum was separated and used for determination of biochemical parameters. Statistical Method The Results were statistically analyzed by ANOVA and presented as Mean ± SE for 6 animals in each group. Students‘t’ test was used to analyze Statistical significance. P value less than or equal to 0.05 were considered statistically significant. RESULTS AND DISCUSSION Histopathological findings revealed morphological alteration in the Liver of the rats. Administration of 200mg/kg body weight dose of plant fruit extract effectively reduced these pathological damages caused by the ccl4 treatment. It was noticed that liver showed congested central vein, portal inflammation and necrosis in the ccl4 treated rats. On silymarin treatment reduction of inflammation, normal central vein & hepatocytes to the normal stage was seen. The histological changes in the liver reversed when 200mg/kg body weight dose was given to the rats. Thus fruit extract of C.CT. Showed. Hepatoprotective nature against ccl4 induced hepatotoxicity. REFERENCES AND NOTES 1. Akther, T. Ramanathan. An overview of chemical Composition of biomedical potential of Citrullus colocynthis. Asian J.Plant Sci., 9: 394- 401, 1999. 2. Al-Ghaithi F, El-Ridi MR, Adeghate E, Amiri M.H. Biochemical effects of C.CT. in normal and diabetic Rats. Mol. Cell Biochem. 261 (1): 143- 149, 2004. 3. Dallak M, Al-khateeb M, Abbas M, Elessa R, Al-Hashem F, Bashir N, Khalil M. In vivo, Acute, Normohypoglycemic, Antihyperglycemic, insulinotropic Actions of orally Administered Ethanol Extract of Citrullus colocynthis. L. pulp. Am.j. Biochem Biotechnol. 5(3): 119- 126, 2009. Hepatoprotective Activity of Citrullus colocynthis Linn. Arshed Iqbal Dar 1* , Vikas Sharma 1 , R C Saxena 1 , S K Bansal 1 Abstracts: Citrullus colocynthis Linn. is an important medicinal plant belonging to the family of cucurbitaceae. It is used as strong laxative. Seed is rich in fat and proteins. It is supposed to be a traditional plant in Africa. This little known vegetable has potential to improve nutrition and boost up food values. Gurudeeban et al. (2010) have recently overviewed the chemical composition and biomedical potential of this plant. Looking to the wide applicability in Ayurvedic system of medicine, it was proposed to undertake the fruit pulp of this plant for hepatoprotective activity against ccl4 induced hepatotoxicity. Akhter et al (1999) have reported that germination of seeds in dark increases the glycerol content of the plants. Raw et al. (2008) have reported that the ethanolic extract of this plant affect the hair growth in Albino rats. The present paper reports different biochemical parameters in the serum of Albino rats which were examined from the blood of the treated and controlled rats. Liver homogenates were also subjected to various biochemical parameters. It was noticed that total protein concentration was less in group 2 as compared to the normal control and in the group 3 it becomes almost equal to group 1 regarding total lipid, triglycerides and cholesterol; the normal values were 128.46, 8.18 and 62.38 which considerably have got increased in group 2. The different doses between 50mg to 200mg body weight were observed in the present study after the ccl4 induced hepatotoxicity in rats. The other parameters such as AST, ALP, ALT, were also noticed which will be given in full text. On the results it can be concluded that Citrullus colocynthis alcoholic extract reduces the inflammation and protect the hepatocytes. Key Words: Citrullus colocynthis, Hepatoprotective, ccl4 Swiss Albino rats.