Determination of Bioactive Compounds of Fenugreek (Trigonella foenum-graecum) Seeds Using LC-MS Techniques.

Trigonella foenum-graecum (fenugreek), a legume has been used as spice throughout the world to enhance the sensory quality of foods, is considered to have a high nutraceutical value. The antidiabetic actions of Trigonella seeds have been speculated due to the presence of steroid saponins, alkaloids, and fiber content. Five bioactive compounds, namely, trigonelline, isoorientin, orientin, vitexin, and isovitexin, were detected and quantified by HPLC system. Further rapid detection of bioactive compounds was achieved using high-performance liquid chromatography hybrid electrospray quadrupole time-of-flight mass spectrometric (HPLC-ESI-QTOF-MS/MS) method. Twenty-five compounds were tentatively identified based on their retention time and measured accurate mass. The fragmentation patterns of known compounds were utilized to elucidate their structures. Six of these identified compounds, that is, 4-hydroxyisoleucine, trigonelline, isoorientin, isovitexin, pinitol, and sarsasapogenin, were simultaneously quantified by UHPLC-ESI-MS/MS method under the multiple reactions monitoring (MRM) mode using ultrahigh-performance liquid chromatography hybrid electrospray triple quadrupole linear ion trap mass spectrometer. The analytical method was validated and successfully applied for simultaneous determination of compounds in Fenugreek seeds, which indicated the suitability of the validated method for its quality control.