Quantification of Two Biomarker Compounds by a Validated High-Performance Thin-Layer Chromatographic Method from Different Extracts of Pluchea dioscoridis Growing in Saudi Arabia

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 × 10 cm glass-backed silica gel 60 F₂₅₄) 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 λ_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 μg 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.