High-performance thin layer chromatography HPTLC analysis

HPTLC fingerprinting analysis was performed with CAMAG LINOMAT 5, where 2 µL of sample was applied by using a Hamilton syringe on a 60F₂₅₄ TLC plate as a band length of 5 mm. Later, it was kept in a TLC developing chamber, which was saturated with solvent vapor (mobile phase) of toluene: ethyl acetate: methanol (7:3:1). The plate was then dried with hot air, placed in a photo documentation chamber, and scanned at 254 nm, 366 nm, and 550 nm, following derivatization with anisaldehyde-sulfuric acid reagent.¹⁴