目的: 通过建立牛黄清心丸(局方)中牛黄及代用品胆汁酸类成分的特征图谱,达到鉴别牛黄清心丸(局方)中牛黄种类的目的,进而更好地控制牛黄清心丸(局方)中牛黄的质量。方法: Waters SymmetryshieldTM C18色谱柱(250 mm×4.6 mm,5 μm),以0.2%甲酸水溶液(含10 mmol·L-1醋酸铵)(A)-乙腈(B)为流动相,梯度洗脱,流速1.0 mL·min-1;采用负模式电喷雾离子化源(ESI-),以甘氨胆酸(m/z 464.3→m/z 402.4/74.1)、牛磺胆酸(m/z 514.3→m/z 124.1/80.0)、胆酸(m/z 407.4→m/z 343.3/289.1)、猪去氧胆酸(m/z 391.4→m/z 391.4)、鹅去氧胆酸(m/z 391.4→m/z 391.4)和去氧胆酸(m/z 391.4→m/z 343.3/327.2)为检测离子对进行分析,建立牛黄清心丸(局方)中牛黄中胆汁酸类成分的特征图谱,对牛黄清心丸(局方)中牛黄及代用品进行鉴别研究。结果: 所建立的牛黄清心丸(局方)中牛黄及代用品LC-MS/MS特征图谱可以牛磺胆酸、甘氨胆酸和胆酸的峰面积比值对该制剂中牛黄的种类进行区分。同时发现1个批次牛黄清心丸(局方)存在人工牛黄掺伪的问题;其余批次牛黄清心丸中牛黄种类与说明书中一致。结论: LC-MS/MS特征图谱技术可以有效地排除牛黄清心丸(局方)中其他成分的干扰,所建立的牛黄和代用品胆汁酸特征图谱可以用于鉴别牛黄清心丸(局方)中牛黄及代用品种类,以期对含牛黄制剂中牛黄的质量控制提供参考。
Objective: To control the quality of Bovis Calculus in Niuhuang Qingxin pills(prescription of the bureau) by establishing an LC-MS/MS characteristic chromatogram for bile acids. Methods: LC-MS/MS characteristic chromatogram analysis was performed on a Waters SymmetryshieldTM C18(250 mm×4.6 mm, 5μm) with the gradient elution of 0.2% formic acid solution(containing 10 mmol·L-1 of ammonium acetate)(A)-acetonitrile(B) at a flow rate of 1.0 mL·min-1. The electrospray ion (ESI) resource was negative mode and monitoring ion pairs were glycocholic acid(m/z 464.3→m/z 402.4/74.1), taurocholic acid (m/z 514.3→m/z 124.1/80.0), cholic acid(m/z 407.4→m/z 343.3/289.1), hyodeoxycholic acid(m/z 391.4→m/z 391.4), chenodeoxycholic acid(m/z 391.4→m/z 391.4) and deoxycholic acid(m/z 391.4→m/z 343.3/327.2). Results: The LC-MS/MS characteristic chromatogram of bile acids in Bovis Calculus was established in Niuhuang Qingxin pills (prescription of the bureau). The specific value of peaks area of bile acids in Bovis Calculus and its substitutes were different.Bovis Calculus and its substitutes could be identified according to the specific value of peak area of taurocholic acid to cholic acid and whether hyodeoxycholic acid was detected. There was an adulteration use of Bovis Calculus Artifactus in Niuhuang Qingxin pills (prescription of the bureau). The types of Bovis Calculus and its substitutes in other batches were consistent with those in the instructions. Conclusion: LC-MS/MS characteristic chromatogram technology can effectively eliminate the interference of other ingredients in Niuhuang Qingxin pills(prescription of the bureau). The established characteristic chromatogram of bile acids can be used to identify the types of Bovis Calculus and its substitutes in Niuhuang Qingxin pills (prescription of the bureau), which provides reference for the quality control of Bovis Calculus in Niuhuang Qingxin pills (prescription of the bureau).
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