Validated RP-UHPLC-QTOF-MS and HPTLC-Densitometry Methods For Simultaneous Quantification Of Hypoxoside And Other Polar Secondary Metabolites In Hypoxis hemerocallidea (African Potato)

Hypoxoside, the perceived active ingredient in Hypoxis raw materials and products, has never been simultaneously quantified with other polar secondary metabolites present in Hypoxis. Our validated high resolution reversed-phase ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry and low-cost high-performance thin-layer chromatography-densitometry methods detected and simultaneously quantified hypoxoside, bis-dehydroxyhypoxoside, hemerocalloside, and galpinoside with good linearity (R² = 0.999–0.963), limit of detection (0.035–9.80 ng mL⁻¹), limit of quantitation (0.11–59.04 ng mL⁻¹), percentage recoveries (88–115), and precision (%RSD 0.36–3.50) from Hypoxis raw materials. The methods also separately quantified geraniol glycoside with good validation parameters.