By Atta-ur-Rahman, M. Iqbal Choudhary, William J. Thomsen
The aim of an activity-directed isolation approach is to isolate bioactive compounds that can supply structural leads of healing value. the conventional strategy of drug improvement is lengthy and dear, yet uncomplicated and quick bioassays can function the start line for drug discovery. This publication provides various "bench best" bioassay strategies precious for traditional product and pharmaceutical chemists eager about drug discovery and pharmacognosy. the vast majority of those bioassays are provided in a step by step structure, in order that they may well also be carried out by way of technical group of workers with little history in microbiology, biochemistry or pharmacology.
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Extra info for Bioassay Techniques for Drug Development
Saline solution 22. Liquid nitrogen 23. Biological freezer unit 24. Water bath 25. E. histolytica is cultured in Falcon flasks; it should be subcultured twice a week. The culture is examined daily through an inverted microscope in order to monitor the growth and detect the presence of contamination. 2. Subculturing is generally required when amoebae grow very rapidly and attach to the wall. 3. For subculturing the Falcon flasks are cooled in ice bath for 30 min. in order to detach amoebae from the walls of the flasks.
9. Incubate overnight at 37°C. 10. Aqueous solution of INT (20 mg/ml) is prepared and filtered through the membrane filter into the glass TLC sprayer. 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.
TLC plates (silica gel) 2. Inorganic stock solution. The following inorganic salts are dissolved in 1 liter of dist. 7H2O 1g NaCl lg 3. UV lamp 4. )*. 5. Organic solvents 6. TLC tanks 7. Autoclave 8. Glucose 9. Incubator 10. Hot air blower 11. Test sample (crude extract, pure natural product or synthetic compound) *Caution ! Pathogenic. A. Bioassay techniques for drug development 22 Here are the steps which involved in direct bioautography method: 1. The TLC chromatogram is developed in a TLC tank by using a suitable solvent system.