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Biology Articles » Botany » Anti-Inflammatory Activity of Piper nigrum, Linn and Ricinus communis, Linn. » Materials and Methods
Materials and Methods
- Anti-Inflammatory Activity of Piper nigrum, Linn and Ricinus communis, Linn.
Plant: Thefruits of Piper nigrum (Family: Piperiaceae) were collected from local market of Kalbadevi, Mumbai and roots of Ricinus communis (Family: Euphorbeaceae) were collected from S.N.D.T. University campus, Santacruz (W), Mumbai and both the samples were authenticated by Zandu Pharmaceuticals Pvt. Limited, Dadar, Mumbai. Preparation of Extract: The collected drugs were cleaned, air dried and powdered. The dried drugs were exhaustively extracted in the soxhlet apparatus (18 hrs of extraction for each batch) using analytical grade solvent. All the extracts were concentrated in vacuo to a syrupy consistency. Preliminary Phytochemical investigation: Various chemical tests were performed on the extracts of fruits of Piper nigrum and roots of Ricinus communis to determine the presence of carbohydrates, amino acids, alkaloids, proteins, glycosides, flavoniods, phenolic compounds, fats, oils, steroids and volatile oil 6. TLC and HPTLC Study: TLC studies were carried out on various extracts of Piper nigrum and Ricinus communis. The stationary phase used was precoated silica gel GF-254 (E. Merk) aluminum plate (Thickness 200µm). Various analytical grade solvents and their combinations were used as mobile phase. The system which gave best resolution of the spots was selected for chromatographic studies. HPTLC studies were carried out to obtain the fingerprint of each extract, which gave better quantitative resolution 7. Animals:Healthy male and female Wistar Albino rats with body weight 150-250g were used for study. They were fed with standard chaw diet and water ad libitum. They were housed in polypropylene cage maintained under standard conditions (12 hrs light / 12 hrs dark cycles, 25 ± 3°, 35-60% humidity). The experimental protocol was subjected to the scrutiny of the Institutional Animal Ethics Committee and was cleared by the same before starting. Acute Toxicity Study: Healthy adult albino rats of either sex were starved overnight and divided into five groups, each containing six animals. Animals were fed per os (p.o.) with an increasing dose of 5, 50, 300, 2000 mg/kg body weight of aqueous and non aqueous extract of Ricinus communis and 5, 50, 300, 1000 mg/kg body weight of aqueous and non aqueous extract of Piper nigrum. After oral administration the animals were observed for signs of toxicity, gross behavioural changes, and mortality up to 14 days. Evaluation of Anti- inflammatory Activity: All the extracts were evaluated for their anti- inflammatory activity by the carrageenan induced rat paw edema method. Healthy adult albino rats of either sex were divided into twelve groups of six animals each. First group received normal saline, second group received aspirin and remaining group received p.o.150 mg/kg body weight of each extract. Food was withdrawn overnight, but adequate supply of water was given to the rats before the experiment. The drugs were given orally with the help of an oral catheter. After one hour a sub plantar injection of 0.1 ml of 1% freshly prepared carrageenan was given to the left hind paw to all the animals. The paw volume was measured with help of Plethysmometer immediately after injection. The paw volume was measured after 1,2,3 and eventually after four hrs. The average fourth hour paw volume of the extract treated rats was compared with the control group and the standard drug (aspirin) group 8,9,10. Statistical analysis: Results were expressed as mean ± SEM and evaluated by Dunnett multiple comparison test. Values of P<0.001 were considered statistically significant.
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