Chromatographic Fingerprinting of Ajwain (Trachyspermum ammi) Fruits for Quality Control and Standardization
Ayman Kirmani, Rasikh Javaid Nengroo, Abdul Wahab, Aisha Siddiqui, M. Mujeeb AhmedAbstract
Background:
Objective:
To establish thin-layer chromatography (TLC) and high-performance thin-layer chromatography (HPTLC) fingerprint profiles of aqueous and hydroalcoholic extracts of
Materials and Methods:
Authenticated dried fruits were extracted using aqueous and hydroalcoholic solvents. Chromatographic analysis was performed on silica gel 60 F254 plates using toluene: ethyl acetate: formic acid (7:2:1) as the mobile phase. Plates were visualized at 254 nm and 366 nm. Densitometric scanning was conducted to determine Rf values and peak areas. Gallic acid and thymol were used as reference standards.
Results:
TLC analysis showed well-resolved bands corresponding to phenolic and aromatic constituents. HPTLC fingerprinting revealed multiple peaks (9–11 peaks) in both extracts within an Rf range of 0.02–0.80, indicating complex phytochemical composition. The hydroalcoholic extract exhibited a major peak at Rf 0.17 with 38.85% area, while the aqueous extract showed a prominent peak at the same Rf with 34.16% area. A characteristic peak corresponding to gallic acid was observed at Rf 0.80. Thymol was identified qualitatively based on its fluorescence behavior at 366 nm, whereas gallic acid was detected quantitatively in densitometric analysis. Fluorescent bands observed at 366 nm suggested the presence of thymol-related compounds. The hydroalcoholic extract demonstrated a broader distribution of phytoconstituents compared to the aqueous extract.
Conclusion:
The developed chromatographic fingerprints provide reliable reference standards for quality control and pharmacopoeial standardization of