Abstract
20-Hydroxyecdysone (20E), an imperative phytoecdysteroid, regulates several biochemical and physiological processes during the different developmental stages in insects. The aim of the present work was to validate an analytical method for the detection and identification of the 20E content in Sesuvium portulacastrum using high-performance thin-layer chromatography (HPTLC) combined with image analysis and using high-resolution mass spectrometry (HRMS). Using HPTLC, the better separation of 20E was achieved on TLC plate prewashed with ethanol—ethyl acetate—water (8:2:0.5) and using the solvent system consisting of chloroform—methanol—benzene (12.5:2.5:1.5) at RF value of 0.30. The bands were documented using a TLC visualizer and scanned at 254 nm in absorption mode. The method was validated for specificity, linearity, precision, robustness, stability, accuracy, limit of detection (LOD), and limit of quantification (LOQ) as per the International Conference on Harmonization (ICH) guidelines. The linear calibr...
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