Simultaneous pharmacokinetic assessment of phytopharmaceuticals from Fenugreek extract by liquid chromatography-tandem mass spectrometry in Sprague Dawley rats.

Fenugreek seeds are used in numerous marketed herbal formulations having therapeutic benefits. Some of its bioactive components including 4-hydroxyisoleucine (4-HIL), trigonelline (TG), raffinose (RFO) and pinitol (PT) are reported to have potential therapeutic activity such as antibacterial, antidiabetic, stomach stimulant, anti-invasive, hyperandrogenism and other allied diseases including polycystic ovary syndrome (PCOS). A fully validated, selective and sensitive bioanalytical method for simultaneous rapid quantification of the above mentioned bioactive components has been developed using hyphenated liquid chromatography electrospray tandem mass spectrometry. Separation of the analytes was achieved within 5 minutes by gradient elution in C18 column at 0.5 mL/min flow rate. Plasma protein precipitation technique was employed to isolate the analytes from the samples. Oral pharmacokinetic profile of the four bioactive components in SD rats was then further evaluated using non compartmental analysis using Phoenix WinNonlin software.