Abstract
The aim of this study was to develop a validated high-performance thin-layer chromatography (HPTLC) procedure for resolution of chemical constituents and identification and quantification of two selected natural anticancer compounds, stigmasterol (PD-1) and cinnamic acid (PD-2), in Pluchea dioscoridis chloroform fraction (PDCF). The chromatographic estimations were conducted on normal HPTLC (20 cm x 10 cm glass-backed silica gel 60 F-254) plates with chloroform-methanol-acetic acid (93: 5: 2, V/V) used as the mobile phase. para-Anisaldehyde was used for the derivatization of the developed plate, and compact spots were scanned at lambda(max) = 513 nm. A well resolved, compact, and intense peaks of PD-1 and PD-2 were recorded at R-F = 0.57 and 0.19, respectively. The proposed analytical method for both biomarker compounds was found to be handy, simple, precise, linear (% RSD = 1.03-1.45), accurate (98.91-99.14%), reliable, and sensitive for the analysis of both biomarkers. The LOD/LOQ (ng) for PD-1 and PD-2 were recorded as 38.73/117.37 and 42.58/129.04, respectively, in the linearity range of 200-1400 ng per spot. The obtained result showed maximum quantities of PD-1 and PD-2 (5.36 and 16.98 mu g mg(-1), respectively). The developed HPTLC was found to be suitable for the routine analysis of these 2 biomarkers in the chloroform fraction of Pluchea dioscoridis and can be further employed in the process quality control of herbal formulations containing the said biomarkers.