雌马醇对映体的高效液相色谱分离

Enantiomeric separation of equol by HPLC

目的:考察色谱条件对 equol 对映体手性分离的影响,建立反相条件下手性流动相添加剂法色谱分离方法。方法:采用高效液相色谱手性流动相法,固定相为 ODS 柱(250mm×4.6mm,5μm),流动相为环糊精水溶液与有机改性剂(甲醇、乙醇、异丙醇、乙腈)的混合溶液,流速1.0mL·min^(-1),检测波长254nm,柱温为室温。结果:首次用β-环糊精作为手性流动相添加剂成功拆分了 equol 对映体,并确立最佳分离条件:30mmol·L^(-1)β-CD 水溶液(pH 6.4)-乙腈(95:5),分离因子(α)可达1.21。结论:本方法简便易行,操作成本低,可直接放大到制备分离。

Objective：To develop and optimize a reversed - phase chromatographic method for separation of equol enantiomers by studying the effect of chromatographic conditions on retention and separation factors. Method：A reversed -phase chromatography with a chiral mobile phase additive （CMPA） was adapted for separation of equol enantiomers. The column used was a Base Line C18 （250mm×4.6mm,5μm） ,and the mobile phases were composed of aqueous solution of β - cyclodextrin and different organic modifiers such as methanol, ethanol, isopropyl alcohol and acetonitrile. The flow rate was 1.0 mL ·min^-1 and the detection wavelength was 254 nm. The column temperature was ambient. Result：The enantiomers of equol were separated on a ODS column with β - CD as CMPA with separation factor up to 1.21 under optimum conditions：30 mmol ·L^-1 β -CD aqueous（ pH 6. 4） -acetonitrile（95 ： 5 ）. Conclusion： Being simple, rapid and economic, the method is suitable for separation and determination of enantiomers of equol.