AbstractBackground: This research focused on HPTLC method for the simultaneous estimation of natural antioxidant markers namely rutin, gallic acid and quercetin from the plant extract of Cucumis melo var agrestis (CME). Aims and Objectives: The main aim of this research is to extract and simultaneously quantitate the three antioxidant markers-rutin, gallic acid and quercetin from CME by using normal phase HPTLC. Materials and Methods: Separation of compounds was performed on TLC aluminium plates precoated with silica 60 F254 after that detection of rutin, gallic acid and quercetin. Camag TLC scanner 3 prepared with win CATS software was used for densitrometric scanning at 254 nm. This method was additional validated in terms of linearity, precision, accuracy and sensitivity as per the ICH guidelines. Results: A good linear relationship was obtained for the calibration plots with R2 = 0.9957, 0.9909, and 0.9920 for rutin, gallic acid and quercetin respectively. Accuracy was checked by recovery study performed at three different stages with the average recovery between 95% and 98% for all the three markers. Conclusion: This developed method of HPTLC can be used for the detection of antioxidant markers in botanicals and herbal formulations.
Keywords: Bioflavonoids; Rutin; Gallic acid; Quercetin; Antioxidant and Developed method of HPTLC.