基于1H-NMR结合LC-MS技术分析熊胆粉及常见胆类中药中的胆酸类成分

目的 建立基于核磁共振氢谱技术（¹H-NMR）结合液质联用技术（LC-MS）的分析方法，阐明熊胆粉中胆酸类成分组成，找出熊胆粉与其他常见胆类中药中的差异性成分。方法 利用¹H-NMR非靶标分析熊胆粉及其他常见胆类中药的化学成分组，通过信号归属表征熊胆粉中胆酸类成分组成，以¹H-NMR图谱分段积分后峰面积为变量进行多元统计分析。利用LC-MS对熊胆粉及其他常见胆类中药中10个胆酸类成分（胆酸、熊去氧胆酸、猪去氧胆酸、鹅去氧胆酸、牛磺熊去氧胆酸、牛磺鹅去氧胆酸、甘氨熊去氧胆酸、甘氨猪去氧胆酸、牛磺胆酸、甘氨胆酸）进行含量测定，并以含量为变量进行多元统计分析。结果 基于¹H-NMR图谱及多元统计分析发现胆酸类成分为胆类中药的主要成分，也是熊胆粉与其他常见胆类中药的主要差异性成分。结合文献及对照品比对，从熊胆粉中初步鉴定出7个胆酸类成分。进一步结合LC-MS含量测定结果和多元统计分析验证熊胆粉与其他常见胆类中药的主要差异性胆酸类成分为熊去氧胆酸、鹅去氧胆酸、牛磺熊去氧胆酸和牛磺鹅去氧胆酸。结论 熊胆粉与其他常见胆类中药的胆酸类成分存在显著差异。与其他胆类中药相比，熊胆粉中熊去氧胆酸、鹅去氧胆酸及其牛磺型结合胆酸含量较高。¹H-NMR技术具有对化学成分组全面、快速分析的优势，可与LC-MS检测相互补充，共同应用于中药复杂体系成分分析和代谢组学的深入研究。

Analysis of bile acid profiles in bear bile powder and other medicinal bile using 1H-NMR and LC-MS

Objective To establish an analysis method based on ¹H-NMR combined with LC-MS technology to clarify the bile acid profiles in bear bile powder, and find out the differences between bear bile powder and other medicinal bile. Methods The ¹H-NMR spectra of each medicinal bile was recorded in parallel, the bile acid profiles of bear bile powder were characterized by signal attribution, and the datasets of piecewise integration were introduced for multiple statistical analysis to find the potential biomarkers. Then, the content of 10 bile acidscholic acid (CA), ursodeoxycholic acid (UDCA), hyodeoxycholic acid (HDCA), chenodeoxycholic acid (CDCA), tauroursodeoxycholic acid (TUDCA), taurochenodeoxycholic acid (TCDCA), glycyrsodeoxycholic acid (GUDCA), glycohyodeoxycholic acid (GHDCA), taurocholic acid (TCA), glycocholic acid (GCA)] were determined using LC-MS, and then the content was used as variables for multivariate statistical analysis. Results Based on ¹H-NMR spectrum and multivariate statistical analysis results, it was found that bile acid profiles were the main components of medicinal bile and the main differences between bear bile powder and other medicinal bile. With the assistance of literature and reference compounds, seven bile acids were tentatively identified from the representative spectrum of bear bile powder. Combined with LC-MS results, it was found that UDCA, CDCA, TUDCA and TCDCA contributed to primary differences. Conclusion The bile acid profiles in bear bile powder is significantly different from that in other medicinal bile, UDCA, CDCA, TUDCA and TCDCA have the potential to be the biomarkers. Furthermore, ¹H-NMR spectroscopy can provide comprehensively qualitative and quantitative information for TCMs, and it should be an eligible tool as well as LC-MS for the quality evaluation of TCMs.