Qualitative and quantitative analysis of khellin in AMMI visnaga friuts and pharmaceutical preparations using HPTLC and HPLC

An attempt was made to develop and validate a simple and accurate HPTLC method for the analysis of khellin in Ammi visnaga fruits and some pharmaceutical preparations. HPLC method was developed in order to verify the results of HPTLC analyses. HPTLC method was developed using chloroform-acetone 9:1 (%, v/v) as mobile phase. The analysis was performed on 10 × 20 cm aluminum-backed plates coated with 0.2-mm layers of silica gel 60 F ₂₅₄ (E-Merck, Germany). Camag TLC Scanner III was used for the UV densitometric scanning. RP-HPLC method was developed using a mobile phase composed of methanol-water 60:40 (%, v/v) and C18-ODS-Hypersil column in isocratic mode. Analyses were done in UV absorbance mode at 248 nm for both. The used HPTLC system was found to give sharp, symmetrical, and well-resolved peak at R_f value of 0.29 ± 0.04 and linearity in the range 50-300 ng/spot (r² = 0.9939). On the other hand, the peak of khellin was well-resolved at R_t 4.73 min and linearity in the range of 100-1000 ng/mL (r² = 0.9960) was used in the HPLC method. These results indicated that proposed HPTLC and HPLC methods could be successfully applied for quality control and routine analysis of khellin in commercial formulations.