Development of HPTLC method for the simultaneous estimation of quercetin, curcumin, and ascorbic acid in herbal formulations

Antioxidants can scavenge free radicals in the human body and have been associated with a protective effect on the human body against a variety of diseases. As a part of normal cellular function, free radical production occurs continuously within the body at a cellular level. Excess-free radical production from endogenous or exogenous sources may result in a variety of diseases. Vitamins (A, C, and E), carotenoids, polyphenols, and flavonoids are some examples of common natural antioxidants. Among the different antioxidants quercetin, curcumin, and ascorbic acids have been extensively studied, and numerous reports on their antioxidant activity are available in the literature. Antioxidants are regarded as a key player in the current pandemic situation, and they play a key role as a preventive and controlling tool. There are a variety of herbal formulations and immune booster therapies available, many of which claim to contain various antioxidants and are effective against covid-19. Numerous herbal formulations and immune booster therapies are available and claimed to have different antioxidants and other constituents. In the present study simple, rapid, accurate, precise, and sensitive high-performance thin-layer chromatographic analytical method was developed for simultaneous estimation of quercetin, curcumin, and ascorbic acid. The developed method can effectively employ for different polyherbal formulations. The developed method was validated according to ICH guidelines. In the present study, an aluminum plate precoated with 60F 254 silica gel was used as a stationary phase whereas chloroform: ethyl acetate: formic acid (6:6:2.5 v/v/v) was used as a mobile phase. Chromatographic detection was performed with a Camag TLC scanner at 265 nm. The linearity range for the proposed method was found to be 500–1000 ng/band for all three analytes. The antioxidant potentiality of selected antioxidants was evaluated by an optimized UV visible spectroscopic-reducing power method. The reducing power method was optimized for the concentration and volume of reagent, incubation temperature, and time. The results of the study revealed that quercetin is having the best antioxidant potential compared to curcumin and ascorbic acid.