Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100193, China.
Analyst. 2012 Dec 21;137(24):5845-53. doi: 10.1039/c2an36084e.
An ultrasound-assisted solid-liquid extraction (USLE) coupled to ultra-performance liquid chromatography-evaporative light scattering detection (UPLC-ELSD) method has been developed for the simultaneous extraction and determination of six bile acids (BAs) in natural Calculus bovis and its substitutes, collected from different origins. The USLE conditions, UPLC chromatographic and ELSD conditions for BAs were optimized. Under optimum conditions, the six target analytes were efficiently extracted and baseline separated within 10 min. The limits of detection (LODs) and quantification (LOQs) for six BAs were less than 7 ng and 22 ng, respectively. Average recoveries were within the range of 98.8-100.7% with relative standard deviations (RSDs) <2% for the six analytes. This method, due to its convenience, high selectivity, fast analysis efficiency and good reproducibility can be employed for analyzing the content differences of six BAs in 40 batches of natural C. bovis and its existing substitutes. The differences of the content of each BA in natural C. bovis and its substitutes were significant, and the total contents of six BAs in 13 batches of natural C. bovis were in the range of 7.96-160.17 mg g(-1), in 20 natural C. bovis of 0-245.89 mg g(-1), in 2 artificial cultivated C. bovis of 178.48-194.22 mg g(-1), in 3 cultured C. bovis of 41.01-107.3 mg g(-1), and in 2 counterfeit C. bovis of 144.9-340.25 mg g(-1). The significant differences of multi-component contents reflected the various inherent qualities of these samples, so, the use of these substitutes as the replacers of natural source in clinic should be paid more attention. Some substitutes could not be used as the replacers.
建立了超声辅助固液萃取（USLE）与超高效液相色谱-蒸发光散射检测（UPLC-ELSD）联用测定天然牛黄及其代用品中 6 种胆汁酸的方法。优化了 USLE 条件、UPLC 色谱和 ELSD 条件。在最佳条件下，6 种目标分析物在 10 min 内得到有效提取和基线分离。6 种胆汁酸的检测限（LOD）和定量限（LOQ）均小于 7 ng 和 22 ng。6 种分析物的平均回收率在 98.8%-100.7%范围内，相对标准偏差（RSD）<2%。该方法由于其方便、选择性高、分析效率快、重现性好，可用于分析 40 批天然牛黄及其代用品中 6 种胆汁酸的含量差异。天然牛黄及其代用品中各 BA 含量差异显著，13 批天然牛黄中 6 种 BA 的总含量在 7.96-160.17 mg g(-1)范围内，20 批天然牛黄中 0-245.89 mg g(-1)范围内，2 批人工养殖牛黄中 178.48-194.22 mg g(-1)范围内，3 批培养牛黄中 41.01-107.3 mg g(-1)范围内，2 批假冒牛黄中 144.9-340.25 mg g(-1)范围内。多组分含量的显著差异反映了这些样品的不同内在质量，因此，在临床中应更加注意这些替代品作为天然来源的替代品的使用。一些替代品不能作为替代品使用。
