HPLC法测定苦杏仁苷两种差向异构体的含量

Determination of The Two Epimers of Amygdalin by HPLC

目的分离纯化L-苦杏仁苷对照品,建立苦杏仁药材中D-、L-苦杏仁苷的含量测定方法。方法采用YMC-Pack Ph高效液相色谱柱(250 mm×4.6 mm,5μm);乙腈-水(5∶95)为流动相;流速:1.00 mL·min^(-1);柱温:30℃;检测波长:207 nm。结果本实验分离纯化的L-苦杏仁苷对照品纯度为96.3%,D-、L-苦杏仁苷分别在12.118 0～363.540 0μg·mL^(-1)和1.521 6～45.648 0μg·mL^(-1)范围内具有良好的线性关系,相关系数(r)分别为0.999 8和0.999 9,平均加样回收率分别为98.86%(RSD=0.64%)和97.33%(RSD=0.87%)。结论本实验分离纯化的L-苦杏仁苷对照品纯度高,建立的高效液相色谱法(HPLC)简单可行,结果准确可靠,对D-、L-苦杏仁苷具有良好的分离效果,可用于苦杏仁药材中苦杏仁苷的质量控制,为中成药中苦杏仁苷的质量控制提供参考。

Objective To separate and purify L-amygdalin reference substance,and to establish a HPLC methodfor the content determination of D-amygdalin and L-amygdalin. Methods The YMC-Pack Ph chromatographiccolumn(250 mm×4.6 mm,5 μm) was used. The 5% acetonitrile solution was used as the mobile phase. The flowrate was 1.00 mL · min^-1. The column oven was set at 30 ℃, and the detection wavelength was set at 207 nm.Results The purity of the L-amygdalin reference substance was 96.3%. Under the above chromatographicconditions,good linear relationships were shown between peak area and the concentration of D-amygdalin and L-amygdalin in the ranges of 12.118 0-363.540 0 μg · mL^-1and 1.5216-45.6480 μg · mL^-1, respectively. Thecorrelation coefficients were 0.999 8 and 0.999 9,respectively. The average recoveries were 98.86%(RSD=0.64%)and 97.33%(RSD=0.87%),respectively. Conclusion The purity of the L-amygdalin reference substance obtainedin this experiment was high. The method is simple, feasible, and accurate with high specificity. It has goodseparation of D-and L-amygdalin,which can be used for the quality control of amygdalin in Armeniacae semenand provide reference for the quality control of amygdalin in Chinese patent medicines.