High-Performance Thin Layer Chromatography Method Development and Validation for Simultaneous Determination of Phenolic Acids in Selected Indian Bamboo Species
Background: Indian bamboo species like Bambusa arundinaceae Retz, Bambusa vulgaris, Schard and Dendrocalamus strictus Roxb is sucessively proved in various pharmacological activities like antihyperglycemic, anti-diabetic, anti-cancer, anti-inflamatory, anti-obesity, anti-fatigue, anti-lipidemic and cardiovascular diseases. Hence, there is a need to optimize selective and sensitive methodologies using sophisticated analytical equipment to accurately quantify the levels of bioactive compounds such as phenolic acids in Indian bamboo species. Material and Method: In the present study to decide the robustness of HPTLC analytical method by the factorial design; analyze the four factors (developed distance, saturation time, mobile phase ratio, band length) in terms of in order which associated to the main factors and interaction effects; determine method development various parmeters (precision, accuracy, lineraty, limit of detetion, limit of quantification, etc). Results: Phenolic acids chromatographed on top of silica gel 60 F254 TLC plates using toluene: ethyl acetate: formic acid: methanol (3:3:0.6:0.8 v/v/v/v) as optimized mobile phase. A prominent spot for chlorogenic acid (ChA), gallic acid (GA), caffeic acid (CA) and ferulic acid (FA) was simultaneously observed with retardation factor (Rf) 0.15 ± 0.02, 0.57 ± 0.03, 0.78 ± 0.01, 0.87 ± 0.01 when the densitometric scanning was implemented at 280nm.The linearity for the calibration plots showed r2 = 0.999 with concentration from (100-700 ng/band) for ChA and (50-350 ng/band) for GA, CA, FA simultaneously. Conclusion: Developed method successfully employed for the simultaneously quantification of selected Indian Bamboo- species.