Precise and sophisticated HPTLC and HPLC methods were developed for determination of caffeine from coffee bean extract. HPTLC analysis was performed on silica gel 60 F254 plate using ethyl acetate: methanol: water in the proportion of 10: 1.4: 1 (v/v/v) as mobile phase. A clear band of caffeine was observed at Rf value 0.55 (+0.1). Caffeine shows linearity within the concentration range of 200-600 ppm with regression coefficient of 0.9987, at 254 nm. HPLC analysis was performed by using mobile phase of methanol and water (40:60) with Zorbax SB C-18 column. Quantification was carried out by using PDA detector at 272nm. The method was validated in terms of specificity, precision, linearity, robustness, accuracy, limit of detection (LOD) and limit of quantification (LOQ) as per ICH Q2 (R1) guidelines. The % RSD for validation parameters including accuracy, method precision, intermediate precision, robustness was found to be less than 2.0 %. In the present study, accurate and sophisticated HPTLC and HPLC methods were validated for determination of caffeine in coffee bean extract. Also qualitative analysis was performed for the estimation of caffeine from prepared capsule formulation using HPTLC method.
Loading....