Download Bioassay Techniques for Drug Development by Atta-ur-Rahman, M. Iqbal Choudhary, William J. Thomsen PDF

By Atta-ur-Rahman, M. Iqbal Choudhary, William J. Thomsen

The target of an activity-directed isolation procedure is to isolate bioactive compounds which can offer structural leads of healing value. the conventional means of drug improvement is lengthy and dear, yet easy and quick bioassays can function the place to begin for drug discovery. This booklet provides more than a few "bench most sensible" bioassay recommendations worthwhile for ordinary product and pharmaceutical chemists occupied with drug discovery and pharmacognosy. the vast majority of those bioassays are awarded in a step by step layout, so that they may well also be applied via technical body of workers with little history in microbiology, biochemistry or pharmacology.

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Different concentrations of test sample (1000, 500, 250, 100, 50, 10ppm) are dissolved in distd. water (small bowls) for the test. A minimum of eight replicates are required for each concentration. 6. One-day old larvae* (25 per bowl) are then transferred into bowls containing the test sample. The exposure period is 24 h during which no food is offered to the larvae. 7. The killing effect of the test sample is assessed after 30 min. and 24 hr of exposure. 8. Percent mortality is calculated to represent the larvicidal activity of each sample.

Test sample (plant extract, pure natural product or synthetic compounds) The procedure used for the Hoechst 33258 fluorescence assay is given below: 1. Cells are seeded onto 96-well microtiter plates at a concentration of 5×104−105 ml−1, volume 200µl/well. 2. 5°C in humified CO2 (10%) incubator for 24h. 3. Old medium is removed and fresh medium is added. 4. 10 ul containing various concentrations of test compound is added whereas +ve and −ve control has standard drug(s) and no drug respectively.

11. The bioautogram is sprayed with approx. 5 ml of TTC solution. The plate can be sprayed directly in the polyethylene box. 12. Incubate for 4 h at 37°C. 13. Open lid, spray bioautogram with approx. 5–10 ml EtOH, 70%. 14. Stain control TLC with suitable chromogenic reagent. 15. The bioautogram is evaluted by comparison with the control TLC. Zones of inhibition (indicating the presence of antibacterial compound) on the bioautogram appear as white spots on a pink background. : All manipulations of bacterial materials should be performed in a contained environment (Laminar flow chamber or glove box) with disposable surgical gloves.

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