Taketani Keiko, Hoshino Shohei, Uemura Taku, Goto Tsuyoshi, Takahashi Nobuyuki, Tsuge Nobuaki, Kawada Teruo
Central Research & Development Institute, House Foods Group Inc.
J Nutr Sci Vitaminol (Tokyo). 2015;61(6):465-70. doi: 10.3177/jnsv.61.465.
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.
建立了一种高效纯化方法，用于从喂食含有胡芦巴（Trigonella foenum - graecum）种子粉日粮的小鼠血浆中同时回收极性皂苷、原薯蓣皂苷（PD）和薯蓣皂苷（DC）以及非极性苷元薯蓣皂苷元（DG），以便随后通过液相色谱 - 电喷雾串联质谱（LC - ESI - MS/MS）进行定量分析。首先通过加入乙腈使小鼠血浆样品脱蛋白，然后将上清液应用于碳基固相萃取管。依次用甲醇和35%氯仿/甲醇（v/v）洗涤后，用80%氯仿/甲醇（v/v）同时洗脱PD、DC和DG。将洗脱液蒸发至干，再溶解于80%甲醇（v/v）中。过滤后的样品用LC - ESI - MS/MS系统进行分析。经过纯化过程，回收率在89.3%至117.4%之间，且无明显的离子抑制或增强。因此无需使用内标。通过分析来自胡芦巴喂养研究的小鼠血浆证明了该方法的实用性。
