Deepak Kumar Jain.pdf

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Flavonoids by Shinoda test and Alkaline reagent test also 18, 19 tested for Saponins and Tannins. Animals care and handling The experiments were carried out on albino rabbits of 5-8 months of either sex weighing between 1.5 to 1.6 kg and Wistar albino rats of either sex of weight 150 ±20 grams. They were provided from Truba Institute of Pharmacy, Bhopal and were acclimatized to the standard laboratory conditions in well cross ventilated animal house at temperature 25±2°C relative humidity 44 –56% and light and dark cycles of 10 and 14 hours respectively for 1 week before and during the experiments. Rabbits were kept in mesh bottom iron cages to avoid caprophagy habit they fed with cauliflower, cabbage, carrot and water for 10 days before the experiment while Rats were fed with standard diet (Golden Feeds, Delhi) and water ad libitum. The experiments were approved by the Institutional Animal Ethics Committee and a s p e r C P C S E A g u i d e l i n e s ( a p p r o v a l n o . 1196/a/08/CPCSEA). Drugs and chemicals Diclofenac sodium provided by Novartis pharmaceuticals Pvt. Ltd, Paracetabmol provided by Lupin Pharmaceuticals Pvt. Ltd., Typhoid-Paratyphoid A, B vaccine (Sanofi pasture) was purchased and all other chemicals utilized were of analytical grade. Acute toxicity studies The LD 50 was calculated by “Staire case” method. The LD50 was determined in rats and mice by oral and intraperitonea1 route. The initial dosing was 2000 mg/kg p.o. and 800 mg/kg 4 i.p. in both the species . Preparation of extract dose 2% acacia suspension was prepared by suspending 2 gram of accurately weighed acacia powder in 100 ml of 0.9% saline. 20 ml of vehicle was taken separately to which 2 gram of dried extract was added and sonicated, this produce suspension of 100 mg/ml strength. Both TPEE and TPEAE suspension were prepared in such manner. Animal grouping Patel et al Evaluation of Analgesic and Antipyretic Activity of Tridax Procumbens Leaves Extract. For Analgesic activity, rats were divided into six groups and each group contains six animals. Group I (Negative control, vehicle treated), Group II (Standard, Diclofenac sodium) 10mg/kg treated), Group III (TPEE 200mg/kg treated), Group IV (TPEE 300mg/kg treated), Group V (TPEAE200mg/kg treated), Group VI (TPEAE 300mg/kg treated). For Antipyretic activity, rabbits were divided into four groups 227 and each group contains four animals. Group I (Negative control, vehicle treated), Group II (Standard, Paracetamol 100mg/kg treated), Group III (TPEE 300mg/kg treated), Group IV (TPEAE 300mg/kg treated). Screening of Analgesic Activity in Rats Acetic acid induced writhing test Acetic acid 1%v/v 1ml/100gm was injected intraperitonialy to animals of each group. Standard and test groups were treated with diclofenac sodium (I.P.) 15 minute and ethanolic extract, ethyl acetate extract (P.O.) 1 hour prior to the administration of acetic acid respectively while control group was treated with vehicle. Wriths were the stretching of the abdomen with simultaneous stretching of at least one hind limb. The total no. of wriths was recorded for 10 min starting 20, 2 10 min after injection of acetic acid. NaCl induced writhing test Sodium chloride 4%w/v at 1ml/kg concentration was injected intraperitonialy to animals of each group. Standard and test groups were treated with diclofenac sodium (I.P.) 15 minute and ethanolic extract, ethyl acetate extract (P.O.) 1 hour prior to the administration of Nacl respectively while control group was treated with vehicle. Then they were observed for a period of 10 min after 10 min of injection of acetic acid and the number of writhes is recorded for each 20, 21 animal. The percent inhibition of wriths was calculated. Screening of Antipyretic Activity in Rabbits Typhoid-Paratyphoid A, B vaccine 0.1 ml was injected into the marginal ear-vein of rabbits of each group. Standard and test groups were treated with paracetamol (I.V.) 15 minute and ethanolic extract, ethyl acetate extract (P.O.) 1 hour after the administration of Typhoid-Paratyphoid A, B vaccine respectively while control group was treated with vehicle. The rectal temperature each animal was recorded 15 minute before and at the interval of 30 minute after treatment using 22 Telethermometer up to 4 hours Statistical Analysis The results wereexpressed as mean ±SEM. The results of the present study were analyzed using one way ANOVA followed by dunnett's t-test computed statically by using Graph pad prism software (version 5.04). RESULTS Leaves were green, characteristic odour with slight bitter taste. Leaves were of size 3-5 cm in length and 1-1.5 cm width. There fracture was soft and rough texture. (Table 1). The findings of physiochemical analysis were that the total ash value was 6.6%, acid insoluble ash (2.5%), water soluble ash RJPS, Oct - Dec, 2011/ Vol 1/ Issue 3
Table 1: Morphological characteristics of leaves of Tridax procumbens. S.No. Characters Observation 1 Color Green 2 Odour Characteristic 3 Taste Acridic 4 Size 3-5cm lenght,1-1.5cm width 5 Texture Rough 6 Fracture Soft (1.14%), loss on drying was 0.1% and foaming index was less than 20%. These parameters suggest that the plant part contains nil moisture, have sufficient amount of organic content. (Table No.2). On performing several tests for Phyotochemical analysis on the ethanolic and ethyl acetate extract it was observed that both the extract contains appreciable amount of flavonoids and Phytosterols while other Phytoconstituents such as alkaloids, tannins, resins, proteins and carbohydrate were also present while they lack glycosides. (Table No.3). The Analgesic activity of Tridax Table 2: Physiochemical analysis of powdered leaves of Tridax procumbens. S.No. Parameters Observations (%) 1 Loss on drying 0.1 2 Foaming index 20 3 Total ash value 6.6 4 Acid insoluble ash value 2.5 5 Water soluble ash value 1.14 Patel et al Evaluation of Analgesic and Antipyretic Activity of Tridax Procumbens Leaves Extract. procumbens leaves extract was due to the presence of active Phytoconstituents which were determined in qualitative analysis of crude extracts. On administration of extract before I.P. administration of 1% v/v 1ml/100gm acetic acid leads to inhibition of wriths (Table No. 4). The Analgesic activity of Tridax procumbens leaves extract was due to the presence of active Phytoconstituents which were determined in qualitative analysis of crude extracts. On administration of extract before I.P. administration of 4%w/v 1ml/kg sodium chloride leads to inhibition of wriths (Table No. 5). The effect of TPEE and TPEAE after I.V. injection of 0.1 ml of Typhoid-Paratyphoid A, B vaccine into the marginal ear-vein of rabbits leads to decrease in temperature measured by telethermometer (Table No. 6) Fig. 1: Effects of TPEE and TPEAE on Acetic acid induced writhing response in rats CTR-Control, STD-Standard, TPEE-Tridax procumbens ethanolic extract, TPEAE-Tridax procumbens ethyl acetate extract (1-200mg/kg, 2-300mg/kg) Table 3: Phyotochemical Analysis of Tridax procumbens TPEE and TPEAE. S.N Phytoconstituents Test Ethyl Acetate Ethanolic extract(TPEAE) extract (TPEE) 1 Carbohydrates Benedict’s test +ve +ve 2 Proteins Biuret test +ve +ve 3 Flavonoids Shinoda ’ s Test +ve +ve 4 Alkaloids Mayer ’ s test, Dragendorff’s test +ve +ve 5 Glycosides Legal’s test, Baljet’s test -ve -ve 6 Saponins By shaking the extract in test tube +ve +ve 7 Tannins Ferric chloride test +ve +ve 8 Steroids Salkowski test +ve +ve Table 4: Effects of TPEE and TPEAE on Acetic acid induced writhing response in rats. Group No. Drug Dose Wriths % Inhibition of (Mean±SEM) wriths I Vehicle 1 ml/100gm 14.17±0.40 0 II Diclofenac sodium 10 mg/kg 2.67±0.33 ** 81.2 III TP Ethanolic extract 200 mg/kg 10.2±0.93 * 28.0 IV TP Ethanolic extract 300 mg/kg 4.4±0.68 ** 68.2 V TP Ethyl acetate extract 200 mg/kg 12.32±0.86 13.0 IV TP Ethyl acetate extract 300 mg/kg 9.8±1.73 * 30.84 TP-Tridax procumbens, N = 6, Data expressed as mean ± S.E.M, One way ANOVA followed by Dunnett's t-test Group I negative control, II standard, III, IV, V and VI test, All test and standard groups were ompared with group I (negative control) ** P
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Page 5 and 6: as compared to acetic acid which we

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