Method validation using HPLC for the isolation, separation and quantification of some biologically active compounds

4^th International Conference and Exhibition on Materials Science & Engineering

September 14-16, 2015 Orlando, USA

Gouri Sankhar Brahma

The ICFAI Foundation for Higher Education, India

Posters-Accepted Abstracts: J Material Sci Eng

Abstract :

Method validation for the identification, isolation, separation and purification of one typical class of biologically active purine based alkaloid viz. Caffeine was carried out using reverse phase High Performance Liquid Chromatography technique (HPLC) in the isocratic mode. Conventional solvent extraction technique involving differential and incremental solvation procedure by binary solvent mixtures was adopted to extract the total alkaloids present in the samples. Non polar protic constitutive solvent (ethylene glycol), Polar aprotic dissociative solvent (1, 4-dioxane) in aqueous medium at a composition of 20%+70% was used for extraction of purine alkaloids from the samples. UV-visible spectrophotometry was used to ascertain the presence of the alkaloid in the samples. The operational parameters (Caffeine); C-18 column, CH3CN + MeOH+H2O at pH =8.8 (phosphate buffer), u =1ml/min, Pinlet= 130-140 bar,�» = 273 nm (VWD), T = ambient at 27�ºC,�·= 10-3 N sec m-2, D = 10-9 m2 sec-1, �¦= 1000, h = 3 and dP = 5 mm was found to be the optimal parametric combination for the effective analysis of this purine based alkaloid by HPLC. Recovery rate of caffeine from samples are estimated to be: 100%, 97%, 98.5%, 98%, 96% & 98.8% for Lamolate�®, Coffee beans, Tea Leaves, soft drink -1 (Pepsi), soft drink -2 (Coke) & soft drink -3 (Thums up) respectively.

Biography :

Email: gsb_ad@ifheindia.org