A novel UPLC–MS/MS method for simultaneous quantification of trigonelline, 4‐hydroxyisoleucine, and diosgenin from Trigonella foenum‐graecum extract: Application to pharmacokinetic study in healthy and type 2 diabetic rats

Abstract

Trigonelline (TR), 4‐hydroxyisoleucine (4‐HI), and diosgenin (DG) are the main bioactives of the purified standardized extract of the popular plant Trigonella foenum‐graecum L. (TFG), and it has been proven effective for the treatment of various diseases. However, to the best of our knowledge, no study has investigated the pharmacokinetic parameters of purified standardized T. foenum‐graecum extract in normal and diabetic Wistar rats. The present study has developed and validated a rapid, reliable, and sensitive simultaneous ultra‐performance liquid chromatography MS method to estimate these bioactives. The chromatographic separation was achieved using methanol, acetonitrile, and 0.1% formic acid with the ideal gradient flow system on a BEH Shield RP 18 column. A positive electrospray ionization mode was selected to estimate m/z values of TR (138.14 > 94.63), 4‐HI (148.19 > 74.08), and DG (415.54 > 271.33). The method was robust and reproducible over the linearity range of 60–5000, 6–5000, and 15–5000 ng/mL for TR, 4‐HI, and DG, respectively. Using this novel validated method, we investigated the pharmacokinetic parameters of bioactives using Phoenix WinNonlin version 8.0 (Certera) in normal and diabetic rats. The assay was successfully applied for the estimation of pharmacokinetic parameters using noncompartmental analysis. This investigation shows that the absorption rate increased, whereas distribution and elimination processes slowed down in diabetic rats compared with normal rats.

The Flavonoid Rich Black Currant (Ribes nigrum) Ethanolic Gemmotherapy Extract Elicits Neuroprotective Effect by Preventing Microglial Body Swelling in Hippocampus and Reduces Serum TNF-α Level: Pilot Study

“…Surprisingly, no anthocyanins could be revealed in the BC-GTE, though others have already reported their presence in BC fruits. The trigonelline and 4-hydroxy isoleucine were detected in the BC-GTE, while their incidence in the fenugreek (Trigonella foenum-graecum) was invoked to explain the associated antidiabetic properties [15]. Based on this similarity, it seems logical to predict an antidiabetic property for the BC-GTE, whereas this has already been confirmed for the fruits of BC [9].…”

Integration of molecular docking, molecular dynamics and network pharmacology to explore the multi‐target pharmacology of fenugreek against diabetes

“…In addition, extractions derived from fenugreek have been used for the treatment of diabetes, with good results. The therapeutic effects of fenugreek manifest mainly as improvements to insulin sensitivity and resistance as well as reduced fasting blood glucose and HbA1c levels 33 . These pharmacodynamic activities may be related to the flavonoids, alkaloids, volatile oils and unsaturated fatty acids derived from fenugreek 14 .…”

“…The therapeutic effects of fenugreek manifest mainly as improvements to insulin sensitivity and resistance as well as reduced fasting blood glucose and HbA1c levels. 33 These pharmacodynamic activities may be related to the flavonoids, alkaloids, volatile oils and unsaturated fatty acids derived from fenugreek. 14 However, which compositions are most effective, the potential target genes and the mechanisms of action are not fully understood, which limits the application of fenugreek.…”

Simultaneous pharmacokinetic assessment of phytopharmaceuticals in fenugreek extract using LC–MS/MS in Sprague–Dawley rats

“…A few individual bioanalytical methods for the quantification of 4-HIL (Wadhwa et al, 2020), TG (Wadhwa et al, 2021), and PT (Qiu et al, 2021), excluding RFO, have been reported earlier in literature, but they were time consuming and expensive. A few molecules isolated from TFG, including isoxanthochymol, xanthochymol, garcinol, camboginol, and diosgenin, have been analyzed using LC-MS/MS techniques (Faeste et al, 2010;Wadhwa et al, 2022). The current bioanalytical method has been developed and validated for its selectivity, sensitivity, and rapid and simultaneous measurement of 4-HIL, TG, RFO, and PT in rat plasma.…”