RP-HPLC测定染料木素-4′-葡萄糖苷及其有关物质

 目的建立染料木素-4′-葡萄糖苷的含量测定方法。方法采用反相高效液相色谱法。Diamonsil C₁₈柱(4.6mm×250mm,5μm),流动相为乙腈-水(用磷酸调节pH至2.73)(25∶75),流速1.0mL·min^-1,检测波长261nm,柱温35℃。结果染料木素-4′-葡萄糖苷在0.5055～101.1mg·L^-1内峰面积与浓度呈良好线性关系(r=1.0000,n=6),平均回收率99.2%(RSD=1.1%,n=9),日内精密度RSD<0.5%,日间精密度RSD<0.7%,最低检测限0.2ng。同时对样品中所含染料木素、染料木素-7-葡萄糖苷和染料木素-7,4′-二葡萄糖苷等微量有关物质的分离和检测也获得满意的结果。结论该方法简便、可靠,可用于染料木素及其苷类的质量控制。

Determination of Genistein-4′-Glucoside and Its Related Substances by RP-HPLC

OBJECTIVE To establish a HPLC method for determining the content of genistein-4′-glucoside.METHODS The separation was performed on a Diamonsil C₁₈ column (4.6 mm×250 mm,5 μm). The mobile phase was acetonitrile-water (adjust pH to 2.73 by phosphoric acid) (25∶75). The flow rate was 1.0 mL·min^-1 and the detection wavelength was 261 nm. RESULTS The calibration curve was linear over the range of 0.505 5～101.1 mg·L^-1(r=1.000 0,n=6) for genistein-4′-glucoside.The average recovery was 99.2% (RSD=1.1%,n=9). The RSDs of intra-day and inter-day validatin were below 0.5% and 0.7% respectively. The LOD was about 0.2 ng. The minor substances of genistein, genistein-7-glucoside and genistein-7-4′-diglucoside in the samplealso were isolated and detected satisfactorily. CONCLUSION The method is convenient and reliable for quality control of genistein and its related glycosides.