title: An Efficient Purification Method for Quantitative Determinations of Protodioscin, Dioscin and Diosgenin in Plasma of Fenugreek-Fed Mice.
abstract: 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.
authors.affiliation: Central Research & Development Institute, House Foods Group Inc.
authors.forename: Keiko
authors.initials: K
authors.lastname: Taketani
authors.forename: Shohei
authors.initials: S
authors.lastname: Hoshino
authors.forename: Taku
authors.initials: T
authors.lastname: Uemura
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Goto
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Takahashi
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Tsuge
authors.forename: Teruo
authors.initials: T
authors.lastname: Kawada
mesh_terms: D000818:Animals; D002725:Chloroform; D002851:Chromatography, High Pressure Liquid; D004144:Diosgenin / Q000031:analogs & derivatives / Q000097:blood / Q000302:isolation & purification*; D008297:Male; D000432:Methanol; D008820:Mice, Obese; D010936:Plant Extracts / Q000097:blood / Q000737:chemistry*; D012503:Saponins / Q000097:blood / Q000302:isolation & purification; D052616:Solid Phase Extraction / Q000379:methods*; D053719:Tandem Mass Spectrometry; D029910:Trigonella / Q000737:chemistry*
chemical_list: D010936:Plant Extracts; D012503:Saponins; C019357:dioscin; C056164:fenugreek seed meal; D002725:Chloroform; C459619:protodioscin; D004144:Diosgenin; D000432:Methanol