HPLC-CAD法测定蛇胆汁中牛磺胆酸钠的含量

目的：建立测定蛇胆汁中牛磺胆酸钠含量的高效液相色谱-电雾式检测器（HPLC-CAD）法，同时对紫外检测器、蒸发光检测器和电雾式检测器3种不同检测器的灵敏度进行比较。方法：采用 ZORBAX SB C₁₈色谱柱（4.6 mm×250 mm，5 μm），流动相为乙腈-0.1%三氟乙酸水（30：70），等度洗脱，流速为1.0 mL·min^-1，柱温35 ℃，CAD雾化温度35 ℃，气压4.206×105 Pa。结果：蛇胆汁进样量在0.174 2~3.484 0 μg 范围内与峰面积线性关系良好（r=0.999 5），平均回收率为99.0%，RSD 为0.76%（n=6），阴性对照无干扰。本实验测得牛磺胆酸钠含量范围眼镜蛇科为5.69~28.85 mg·g^-1；游蛇科为23.37~29.65 mg·g^-1；蝰科为6.54~24.00 mg·g^-1。结论：该方法经方法学验证，可作为蛇胆汁质量控制的方法。

Objective: To establish an HPLC-CAD method for determination of Sodium taurocholate in snake bile, and to compare the sensitivity of three different detectors of ultraviolet detector(UV), evaporative light scattering detector(ELSD)and charged aerosol detector(CAD). Methods: ZORBAX SB C₁₈ chromatographic column(4.6 mm×250 mm, 5 μm)was used;mobile phase was acetonitrile: 0.1% trifluoroacetic acid(30:70)at a flow rate of 1.0 mL·min^-1;the column temperature was 35 ℃;the detector was CAD(the spray temperature was 35 ℃), at gas pressure of 4.206×105 Pa. Results: The snake bile showed good linear rang with 0.174 2-3.484 μg (r=0.999 5);the average recovery was 99.0%;RSD was 0.76%(n=6);and the negative control had no interference. The measured contents of sodium taurocholate were 5.69-28.85 mg·g^-1 for Elapidaerang, 23.37-29.65 mg·g^-1 for Colubridaeas, and 6.54-24.00 mg·g^-1 for Viperidaeas. Conclusion: The method can be used for the quality control of snake bile.