HIGH PERFORMANCE THIN LAYER CHROMATOGRAPHIC (HPTLC) METHOD FOR SIMULTANEOUS QUANTITATION OF LUTEOLIN AND QUERCITIN FROM DRIED WHOLE PLANT POWDERS OF GMELINA ARBOREA LINN. AND KALANCHOE PINNATA (LAM.) PERS

Objectives: To develop and validate High Performance Thin Layer Chromatographic (HPTLC) method for simultaneous quantitation of luteolin and quercitin from dried whole plant powders of Gmelina arborea Linn. and Kalanchoe pinnata (Lam.) Pers.. Methods: Chromatography was performed using methanolic extracts of dried whole plant powder of Gmelina arborea Linn. and Kalanchoe pinnata (Lam.) Pers.. Separation of luteolin and quercitin from methanolic extracts of both the plant materials was achieved on Silica gel 60F254 TLC plates using chloroform : methanol in the volume ratio of 96.0 : 4: 0 as the mobile phase. Detection and quantitation of luteolin and quercitin was done by densitometric scanning at λ=254 nm. The developed HPTLC method has been validated using International Conference on Harmonization (ICH) guidelines. Results: The validated HPTLC method was used for simultaneous quantitation of luteolin and quercitin from methanolic extracts of dried whole plant powders of Gmelina arborea Linn. and Kalanchoe pinnata (Lam.) Pers. using their respective calibration curves. Amounts of luteolin and quercitin present in dried plant powder of Gmelina arborea Linn. are 0.9236mg/g and 0.0836 mg/g respectively. Amounts of luteolin and quercitin present in dried plant powder of Kalanchoe pinnata (Lam.) Pers are 0.0529mg/g and 0.2331mg/g. Conclusion: The developed method is simple, precise and accurate and can also be used for routine quality control analysis and for the quantitation of luteolin and quercitin in herbal raw materials as well as in their formulations. Keywords: Luteolin, quercitin, Gmelina arborea Linn. and Kalanchoe pinnata (Lam.) Pers., High Performance Thin Layer Chromatography, Method validation