RP-HPLC法同时测定猪胆粉药材中3种牛磺结合型胆酸的含量

目的: 建立同时测定猪胆粉药材中牛磺猪胆酸、牛磺猪去氧胆酸和牛磺鹅去氧胆酸含量的RP-HPLC方法。 方法: 采用 Welchrom C₁₈色谱柱 (250 mm×4.6 mm,5 μm),流动相为甲醇-磷酸二氢钠溶液(0.03 mol·L^-1)(70∶30),用磷酸调节pH为4.4,流速为1.0 mL·min^-1,检测波长200 nm,柱温25 ℃。 结果: 牛磺猪胆酸、牛磺猪去氧胆酸和牛磺鹅去氧胆酸进样量分别在1.65~16.5 μg (r=0.9998),2.03~20.3 μg (r=0.9998),1.09~10.9 μg (r=0.9996)范围内线性关系良好;检测限分别为26.4,7.6,38.2 ng;平均加样回收率(n=9)分别为101.2% (RSD=1.9%),99.1% (RSD=1.7%),100.6% (RSD=2.1%)。 结论: 该方法简便、准确,重复性好,在同一色谱条件下实现多指标成分的同时测定,为猪胆粉的全面质量评价提供参考。

Objective: To develop an RP-HPLC method for simultaneous determination of taurohyocholic acid,taurohyodeoxycholic acid and taurochenodeoxycholic acid in the dried bile of Sus scrofa domestica Brisson (Pulvis Fellis Suis). Methods: Welchrom C₁₈ (250 mm×4.6 mm,5 μm) column was used,with mixture of methanol and monobasic sodium phosphate solution (0.03 mol·L^-1) (70∶30) as the mobile phase,and adding phosphate acid to adjust to pH 4.4,the flow rate was 1.0 mL·min^-1,the detection wavelength was at 200 nm and the column temperature was at 25 ℃. Results: The linear ranges of taurohyocholic acid,taurohyodeoxycholic acid and taurochenodeoxycholic acid were 1.65-16.5 μg (r=0.9998),2.03-20.3 μg (r=0.9998),1.09-10.9 μg (r=0.9996),respectively;The limits of detection for taurohyocholic acid,taurohyodeoxycholic acid and taurochenodeoxycholic acid were 26.4 ng,7.6 ng,38.2 ng,respectively;The average recoveries(n=9) of taurohyocholic acid,taurohyodeoxycholic acid and taurochenodeoxycholic acid were 101.2% (RSD=1.9%),99.1% (RSD=1.7%),100.6% (RSD=2.1%),respectively. Conclusion: This method is simple and accurate with good reproducibility for simultaneous determination of multiple index components under the same chromatographic condition,and provides a reference for comprehensive quality evaluation of Pulvis Fellis Suis.