HPLC-ELSD法同时测定地黄及其不同炮制品中8个糖类成分的含量

Simultaneous determination of eight saccharides contents in Radix Rehmanniae and its different processed products by HPLC-ELSD

目的:建立高效液相色谱-蒸发光散射检测器法同时测定地黄及其不同炮制品中8个糖类成分的含量,分析地黄炮制前后糖类成分的含量变化情况。方法:采用XBridge■BEH Amide色谱柱(250 mm×4.6 mm,5μm);30%乙腈水+0.1%氨水(A)和90%乙腈水+0.1%氨水(B)为流动相,梯度洗脱;柱温40℃;漂移管温度105℃;气体流量2.0 L·min^(-1);增益值为1;进样量为10μL。结果:8个糖类成分在各自浓度范围内线性关系良好(r>0.9990),精密度、稳定性、重复性试验结果的RSD均<3.0%,平均加样回收率为97.7%~101.3%。采用主成分分析和方差分析对上述8个糖类成分的含量进行分析,结果显示地黄及其不同炮制品样品中主要含有糖的种类和含量均存在显著差异。结论:该方法简单、准确、重现性好,能有效分析地黄及其不同炮制品中8个糖类成分的含量,为地黄药材的质量控制提供了参考依据。

Objective:To establish an high performance liquid chromatography evaporative light scattering detection(HPLC-ELSD)method for simultaneous determination of eight saccharides contents in Radix Rehmanniae(RR)and its different processed products and to analyze the changes of saccharides contents before and after RR processing.Methods:Determination was carried out on a Waters XBridge■BEH Amide column(250 mm×4.6 mm,5μm).The mobile phase was composed of 90% acetonitrile containing 0.1%ammonia(B)and 30% acetonitrile containing 0.1% ammonia(A)with gradient elution.The column temperature was 40℃ and the injection volume was 10μL.Furthermore,the drift tube temperature,carrier gas flow rate and gain value of evaporative light scattering detection were set at 105℃,2.0 L·min~(-1)and 1,respectively.Principal component analysis(PCA)and variance analysis(ANOVA)were used to analyze the contents of the above eight saccharides components.Results:Good separation of 8 components was achieved within 64 min and calibration curves of the eight components showed good linear relationship(r>0.9990).The RSDsof precision,stability and repeatability results were all less than 3.00%,and the average recoveries were within 97.7%-101.3%.PCA and ANOVA results showed that the types and contents of the main saccharides in RR and its different processed products were significantly different.Conclusion:The method is simple,accurate and reproducible,and can effectively analyze the contents of eight saccharides in RR and its different processed products,which provides a reference for the quality control of RR.