An Efficient Purification Method for Quantitative Determinations of Protodioscin, Dioscin and Diosgenin in Plasma of Fenugreek-Fed Mice

Abstract

Summary An efficient purification method for simultaneous recovery of polar saponins, protodioscin (PD) and dioscin (DC), and non-polar aglycon, diosgenin (DG), from plasma of mice fed diets containing seed flours of fenugreek (Trigonella foenum-graecum) was established for subsequent quantitative analysis by LC-ESI-MS/MS. Mice plasma samples were first deproteinated by addition of acetonitrile, and the supernatant was applied to a carbon-based solid phase extraction tube. After successive washing with methanol and 35% chroloform/ methanol (v/v), PD, DC and DG were eluted simultaneously with 80% chroloform/methanol (v/v). The eluate was evaporated to dryness, and re-dissolved in 80% methanol (v/v). The filtered sample was analyzed with an LC-ESI-MS/MS system. After the purification procedure, recovery rates between 89.3 to 117.4% were obtained without notable ion suppression or enhancement. The use of internal standards was therefore not necessary. The utility of the method was demonstrated by analyzing plasma of mice from a fenugreek feeding study.

Diosgenin: Recent Highlights on Pharmacology and Analytical Methodology

“…The results showed an increase in the absorption of diosgenin in hyperlipidemic rats when compared with normal rats [134]. Later, Taketani et al [87] developed a purification method for quantitative determination of diosgenin, dioscin, and protodioscin in plasma of fenugreek-fed mice, which consisted in deproteination of plasma samples, SPE with successive washes, and then analysis by LC-ESI-MS/MS. Moreover, the LC-MS technique was used for bioavailability studies of diosgenin in inclusion complexes with cyclodextrins in Caco-2 cell monolayers and rat jejunum.…”

Qualitative and quantitative HPLC-ELSD-ESI-MS analysis of steroidal saponins in fenugreek seed

“…HPLC equipped with a light diffusion detector was applied to fenugreek saponin analysis only by Petit et al (17), though it is the method of choice in steroidal saponin separation in plant matrices (18)(19)(20). Protodioscin was described as a major saponin of fenugreek seeds (21,22). Steroidal saponins are responsible or co-responsible for several pharmacological effects of fenugreek seeds, including antidiabetic, hypolipidemic, anti-inflammatory and anticancer effects (14,23,24).…”

Exploring the impact of a naturally occurring sapogenin diosgenin on underlying mechanisms of Ca2+ movement and cytotoxicity in human prostate cancer cells

“…Several lines of evidence showed the plasma level of diosgenin in in vivo studies. The level of diosgenin may reach ~30 μM . In subjects taking higher doses or with liver or kidney dysfunction the level may go considerably higher.…”