The fruits of C.dichotoma were purchased from local market of Hubli and were authenticated by Prof V. S Huddar, Department of Botany, H. S. K Science and S. K Arts College, Hubli. The collected fruits were shade dried, reduced to coarse powder (2 kg) and subjected to repeated exhaustive extraction in batches with ethanol in a soxhlet extractor. After complete extraction the alcoholic extract (68 gm) was then suspended in water and further fractionated using petroleum ether (8 gm), solvent ether (2 gm), ethyl acetate (4 gm) butanol (5.5 gm) and butanon (2.5) in succession. These extracts were vacuum dried and used for anti-ulcer activity. Tween 80 (1%) was used as vehicle to suspend the extracts.

All the extracts were subjected to phytochemical analysis as per standard procedures ₆ to know the nature of phytoconstituents present (TABLE 1). The major active constituents were found in extracts includes flavanoids, alkaloids and saponins. Among all active constituents flavanoids were found to be in higher concentrations.

Figure 1

TABLE 1: Qualitative chemical examination of various extracts obtained by successive solvent fractionation of the . fruits.

The experiments were initiated only after approval Institutional Animal Ethical Committee. Albino mice weighing 25-30 g and albino rats weighing 150-200 gm were obtained from central animal house, K. L. E. S’s College of Pharmacy, Hubli, Karnataka. They were maintained at standard housing conditions and fed with commercial diet (Hindustan Lever Ltd., Bangalore) and water ad libitum during the experiment.

The acute toxicity studies were carried out as per stair case or “Up and down” method ₇ . Accordingly the LD₅₀ of all extracts were found to be 3000 mg/kg bodyweight. One tenth of this dose was selected for the study of aphrodisiac activity.

The Albino rats of Wistar strain are divided into seven groups of six animals each. Animals were fasted 24 hours. First group received normal saline 2ml/kg body weight (negative control), the second group received ranitidine 20mg/kg body weight by oral route (positive group) and reaming group III, IV, V, VI and VII were received extracts of petroleum ether, solvent ether, ethyl acetate, butanol and butanone respectively in the dose of 300mg/kg body weight by oral route.

In this method extracts were administered 30 minutes prior to pyloric ligation, Animals were scarified 4 hour later and the stomach was opened to collect the gastric contents. The total volume of gastric content was measured.

In this method the treated groups received extracts in the dose of 300mg/kg body weight for 8 days p.o. After 8 days of treatment, the animals were fasted for 24 hours. Ulcer was produced by administration of aspirin (a dose of 200 mg/ kg orally) on the day of sacrifice. The animals were sacrificed 4 hour later and stomach was opened to calculate the ulcer index by Kunchandy method ₈ .

Indomethacin was administered orally in a dose of 15 mg/kg ₉ body weight. The animals were sacrificed 6 hours after Indomethacin administration.

Figure 2

Table 2: Effect of fruits extracts and Ranitidine in pyloric ligated in rats

Figure 3

Table 3: Effect of fruits extracts and ranitidine in aspirin induced ulcer in rats

Figure 4

Table 4: Effect of fruits extracts and ranitidine in indomethacin induced ulcer in rats

All values are in mean ± SE. n=6, VGJ = Volume of gastric juice in ml,

TA = Total acidity in mEq/l., FA = Free acidity in mEq/l.

UI = Ulcer index, P <0.001 = *,

The volume of Sodium hydroxide required corresponds to the total acidity. Acidity

(mEq /l/100g) was calculated as; Acidity = {Volume of NaOH X Normality X 100 mEq/l/100g X 0.1}

The stomach was cut open along the greater curvature and washed in Normal saline. The gastric juice was collected in a measuring cylinder. The gastric juices were centrifuged at 1000 rpm for 10 min. 1 ml of the supernatant was diluted with 9 ml of distilled water. The solution was titrated against 0.1 N sodium hydroxide using Topferu reagent till the solution turned to orange colour. Then it was laid flat and the ulcer area was calculated under a dissecting microscope (10xs) with a square grid. Gastric mucosal lesions were seen in the form of haemorrhage or linear breaks.

Ulcer index was calculated using the following method

Ulcer index = 10/x

Where x = Total mucosal area / Total area of mucosal lesions

The effect of extracts of C. dichotoma fruits on pyloric ligation, aspirin and indomethacin induced ulcer model is presented in Table 2, 3 and 4 respectively.

All the results were reported as mean  S.E. Significance of the difference between “control” and” drug treated” were determined by the student “t” test