目的 建立HPLC-MS/MS法同时测定体外培育牛黄Calculus Bovis Sativus（CBS）和天然牛黄Calculus Bovis（CB）中26种胆汁酸成分。方法 Symmetry C₁₈色谱柱（150 mm×2.1 mm，3.5μm），流动相为水（0.1%甲酸+10 mmol/L乙酸铵）-甲醇（0.1%甲酸+10 mmol/L乙酸铵），体积流量0.15 mL/min，梯度洗脱，进样量为10 μL。质谱采用电喷雾离子源，负离子模式监测，喷射电压-4 500 V，离子源温度350℃，采用多反应监测模式（MRM）。结果 牛黄中26种胆汁酸在线性范围内峰面积与摩尔浓度均呈良好的线性关系（r ≥ 0.991 4），加样回收率为98.2%~102.3%，精密度RSD值小于0.95%，CBS和CB的胆汁酸类成分在种类和含量上存在一定差异。结论 该法灵敏度高、专属性好、结果可靠，适用于牛黄中胆汁酸成分的测定。

Objective To develop an HPLC-MS/MS method for the simultaneous determination of 26 bile acids in Calculus Bovis Sativus (CBS) and Calculus Bovis (CB).Methods Chromatographic separation was performed on a Symmetry C₁₈ column (150 mm×2.1 mm, 3.5 μm) with linear gradient elution of water (0.1% formic acid + 10 mmol/L ammonium acetate) and methanol (0.1% formic acid + 10 mmol/L ammonium acetate) at a flow rate of 0.15 mL/min, and the injection volume was 10 μL. Switching electrospray ion source polarity in negative mode. The ion spray voltage was set at-4 500 V, the turbo spray temperature was maintained at 350℃, and multiple-reaction monitoring (MRM) was employed.Results The 26 bile aicds showed good linearity within the linear ranges (r ≥ 0.991 4). The average recoveries of the bile acids ranged from 98.2% to 102.3%. The precisions (RSD) for the investigated components were less than 0.95%. There are some differences in variety and chemical components between CBS and CB.Conclusion The proposed method is reliable and has high sensitivity and good specificity, which allows the determination of various bile acids in CB.

国家自然科学基金资助项目（81573788，81503225）