High Performance Thin Layer Chromatographic Method for Simultaneous Estimation of Vitamin C, Thymoquinone and Thymol in Plant Extracts

Background: Lack of an effective HPTLC simultaneous estimation method for vitamin C, thymoquinone and thymol in plant extracts. Aim: The present study involves the development of an accurate, precise, specific, and specific HPTLC method for the identification and quantification of three phytomarkers thymol and thymoquinone in Nigella sativa (kali jiri) seed extract and vitamin C in Hylocereus polyrhizus (dragon fruit) extract. Methods: Using an aluminum plate pre-coated with silica gel 60 F254 and methanol-n hexane-ammonia (15%) (8.5:1.5:0.2 v/v/v) as the mobile phase, thin layer chromatographic development was performed. Results: For each of the three markers, densitometric quantification was carried out at the isobestic point of 271 nm. Vitamin C, thymol, and thymoquinone bands were separated chromatographically at Rf values of 0.66, 0.35, and 0.19, respectively, by using developed mobile phase. For thymol, thymoquinone, and vitamin C, linearity range was 2000-8000 ng/band. The three markers showed 99.39%–99.91% recovery for thymol, 99.22%–99.89% recovery for thymoquinone, and 99.19%–99.69% recovery for vitamin C. Conclusion: The optimized method was used to quantify three thymol and thymoquinone in N. sativa (kali jiri) seed extract and vitamin C in H. polyrhizus (dragon fruit) extract.