Background: A simple, accurate, and reliable high-performance thin-layer chromatography (HPTLC) method was developed for separation and detection of ferulic acid and vanillin. Methods: Separation of ferulic acid and vanillin was carried out on 20 × 10 cm thin layer chromatography (TLC) plates using mobile phase containing toluene/1, 4-dioxan/acetic acid in the ratio 9:2.5:0.4 (v/v). The FA and vanillin were scanned at 320 and 312 nm, respectively. Method was validated for linearity, accuracy, precision, robustness, limit of detection, limit of quantification, and specificity.