Characterization and chemometric based optimization of bioactive metabolites in Hypoxis hemerocallidea with the aid of UPLC-QqQ-MS/MS

Abstract

β-sitosterol and solasodine are major bioactive ingredients in Hypoxis hemerocallidea (H. hemerocallidea) with significant pharmacological properties. As a result, developing a simple and efficient extraction method for simultaneous extraction of both analytes is critical. The purpose of this study was to identify and separate β-sitosterol and solasodine from ethanolic extracts of H. hemerocallidea using a modified QuEChERS method and subsequent analysis via UPLC triple quadrupole mass spectrometry. Response surface methodology was carried out, which included numerical parameters such as ultrasonication time, centrifugation time, and ultrasonication power. The categorical factors included the type of salt used to facilitate extraction, which was (NH4)2SO4 and Na2SO4. Fitting the response surface model to the experimental data produced a quadratic model with a good fit (R2 = 0.9966 for solasodine and R2 = 0.9857 for β-sitosterol). The optimum conditions for extraction of β-sitosterol and solasodine were an ultrasonication time of 30 min, ultrasonication power of 300 W and centrifugation time of 12 min. The generally higher concentrations of analytes obtained for (NH4)2SO4 indicated that it had a superior salting-out ability compared to Na2SO4. In conclusion, for the first time, β-sitosterol and solasodine were simultaneously extracted using modified QuEChERS with good yields through the salting-out action of (NH4)2SO4 in the presence of environmentally friendly solvents, ethanol and water. This modified QuEChERS technique can potentially be applied on a large scale as a sustainable and quick method for enrichment of therapeutic compounds from natural products.