Peng Can, Tian Jixin, Lv Mengying, Huang Yin, Tian Yuan, Zhang Zunjian
1Key Laboratory of Drug Quality Control and Pharmacovigilance (China Pharmaceutical University), Ministry of Education, Nanjing 210009, China.
J Chromatogr Sci. 2014 Feb;52(2):128-36. doi: 10.1093/chromsci/bms256. Epub 2013 Jan 11.
Artificial Calculus Bovis is a major substitute in clinical treatment for Niuhuang, a widely used, efficacious but rare traditional Chinese medicine. However, its chemical structures and the physicochemical properties of its components are complicated, which causes difficulty in establishing a set of effective and comprehensive methods for its identification and quality control. In this study, a simple, sensitive and reliable liquid chromatography-tandem mass spectrometry method was successfully developed and validated for the simultaneous determination of bilirubin, taurine and major bile acids (including six unconjugated bile acids, two glycine-conjugated bile acids and three taurine-conjugated bile acids) in artificial Calculus Bovis using a Zorbax SB-C18 column with a gradient elution of methanol and 10 mmol/L ammonium acetate in aqueous solution (adjusted to pH 3.0 with formic acid). The mass spectra were obtained in the negative ion mode using dehydrocholic acid as the internal standard. The content of each analyte in artificial Calculus Bovis was determined by monitoring specific ion pairs in the selected reaction monitoring mode. All analytes demonstrated perfect linearity (r(2) > 0.994) in a wide dynamic range, and 10 batches of samples from different sources were further analyzed. This study provided a comprehensive method for the quality control of artificial Calculus Bovis.
人工牛黄是临床治疗中广泛应用、疗效显著但来源稀缺的牛黄的主要替代品。然而，其化学结构和成分的理化性质复杂，导致难以建立一套有效且全面的鉴别和质量控制方法。本研究成功开发并验证了一种简单、灵敏且可靠的液相色谱 - 串联质谱法，用于同时测定人工牛黄中的胆红素、牛磺酸和主要胆汁酸（包括六种未结合胆汁酸、两种甘氨酸结合胆汁酸和三种牛磺酸结合胆汁酸）。采用Zorbax SB - C18柱，以甲醇和10 mmol/L醋酸铵水溶液（用甲酸调至pH 3.0）进行梯度洗脱。以脱氢胆酸为内标，在负离子模式下获得质谱图。通过在选择反应监测模式下监测特定离子对来测定人工牛黄中各分析物的含量。所有分析物在较宽的动态范围内均表现出良好的线性关系（r(2) > 0.994），并进一步分析了10批不同来源的样品。本研究为人工牛黄的质量控制提供了一种全面的方法。
