Novel Reverse-Phase High-Performance Liquid Chromatography (RPHPLC) Method for the Quantification of Apigenin in Ocimum Basilicum Linn Seeds (Tukmaria)

Background: Ocimum basilicum L. seeds, is commonly also known as Takhmaria in Gujarat. It is a common constituent present in fruits, plant-derived beverages, vegetables, wheat, sprouts, and some seasonings. Objective: A simple, specific, precise, accurate, and sensitive method for the quantification of apigenin by reverse-phase high-performance liquid chromatography (RP-HPLC) was developed and validated. Methods: Analysis was carried out on Inertsil ODS-3V-C18 column (250 mm × 4.6 mm i.d, 5 μm) as stationary phase and methanol-acetonitrile (55:45 v/v) as a mobile phase at a flow rate of 1.0 mL/ min. Detection was carried out at 340 nm. The retention time of apigenin was found to be 8.30 min. The proposed method was validated according to ICH Guidelines, Q2 (R1). Results: The developed method showed good linearity in the range of 10-50 μg/mL with a correlation coefficient (R2= 0.9998). The LOD and LOQ were found to be 1.23 μg/mL and 4.05 μg/mL, respectively. The percentage recovery for apigenin was found to be 97.75-100.5%. All validation parameters were found within acceptable limits and demonstrated good reliability in the quantification of apigenin. Conclusion: Thus, the newly developed and validated method can be successfully applied for the quantification of apigenin from seeds of O. basilicum L. and can also be applied for the standardization of polyherbal formulation containing O. basilicum seeds. Assay results showed good recovery when statistically compared with the high-performance thin-layer chromatography (HPTLC) method.