Chemometrics guided UHPLC-RI method for simultaneous quantification of food bioactive components in fenugreek herbal preparations.

Abstract Trigonella foenum graecum (TFG) also known as Fenugreek is a popular herb commonly used as a spice, flavoring, and medicinal agent. It is enriched with bioactive constituents namely pinitol (PT), 4-hydroxy isoleucine (4-HIL), and raffinose (RFO) which are proven to be responsible for TFGs medicinal and health benefits. The current study focuses chemometric guided simultaneous RP-UHPLC method development and validation of three phytocomponents using a universal refractive index detection (RID) technique for quantifying the bioactive components present in the herbal nutraceutical preparations using Design Expert. The nucleosil ® 5 NH 2 column maintained at 35°C was used for chromatographic separation against optimum mobile phase composition of acetonitrile: water in the ratio of 70:30, v/v at a flow rate of 0.4 mL min − 1 . Finally, this novel approach was used to measure bioactive component concentrations in commercial herbal products intended for human use.