体外培育牛黄中主要胆酸类成分的测定

目的: 建立准确度高、重复性好的测定体外培育牛黄中主要胆酸类成分的方法。 方法: 采用高效液相色谱仪与蒸发光散射检测器联用技术对体外培育牛黄进行测定。色谱条件:Wondasil C₁₈色谱柱(4.6 mm×250 mm,5 μm),柱温25 ℃;流动相为0.2%甲酸溶液(A)-乙腈(B),梯度洗脱(0~7 min,5%B;7~15 min,5%B→80%B;15~20 min,80%B→90%B;20~23 min,90%B→96%B),流速1.0 mL·min^-1;ELSD检测器氮气流速2.3 L·min^-1,漂移管温度110 ℃。 结果: 同时测定了体外培育牛黄中胆酸和去氧胆酸含量,胆酸和去氧胆酸进样量分别在0.48~2.42 μg和0.16~0.83 μg范围内线性关系良好,平均回收率(n=6)分别为102.1%(RSD=1.1%)和100.7%(RSD=1.3%)。 结论: 该法可同时测定体外培育牛黄中主要胆酸类成分胆酸和去氧胆酸的含量,适用于体外培育牛黄的质量评价与控制。

Objective: To establish a method for the determination of main cholic acids from Bovis Calculus Sativus. Methods: Chromatography was performed on a Wondasil C₁₈ column(4.6 mm×250 mm,5 μm)with a column temperature of 25 ℃.The mobile phase composition was 0.2% aqueous formic acid(A)and acetonitrile(B)with a flow rate of 1.0 mL·min^-1 using a gradient elution of 0-7 min,5%B;7-15 min,5%B→80%B;15-20 min,80%B→90%B;20-23 min,90%B→96%B.An evaporate light-scattering detector(ELSD)was used as the detector with a nitrogen flow rate of 2.3 L·min^-1,at a drift tube temperature of 110 ℃. Results: Cholic acid and deoxycholic acid were determined simultaneously.The linear ranges of cholic acid and deoxycholic acid were 0.48-2.42 μg and 0.16-0.83 μg,and the average recoveries (n=6)were 102.1%(RSD=1.1%)and 100.7%(RSD=1.3%). Conclusion: The method can simultaneously determine the main cholic acids (cholic acid and deoxycholic acid)in Bovis Calculus Sativus and it can be used for the evaluation and quality control of Bovis Calculus Sativus.