Tang Xi-Yang, Gao Meng-Xue, Xiao Hui-Hui, Yun Wei-Jing, Dai Yi, Yao Zhi-Hong, Wong Man-Sau, Yao Xin-Sheng
Institute of Traditional Chinese Medicine and Natural Products, College of Pharmacy, Jinan University, West Huangpu Avenue No.601, Guangzhou 510632, China.
State Key Laboratory of Chinese Medicine and Molecular Pharmacology (Incubation), The Hong Kong Polytechnic University Shenzhen Research Institute, Nanshan District, Shenzhen 518057, China.
J Chromatogr Sci. 2020 Jun 5;58(6):494-503. doi: 10.1093/chromsci/bmaa012.
Xian-Ling-Gu-Bao (XLGB), a famous traditional Chinese medicine prescription consisted of six herbal medicines, was used for prevention and treatment of osteoporosis in China. As an oral formulation, the multiple components contained in XLGB were inevitably biotransformed by the intestinal microflora before absorption via the gastrointestinal tract. However, the dynamic profiles of biotransformation products of XLGB remain unknown. In this paper, a rapid and sensitive ultra-performance liquid chromatography tandem triple quadrupole mass spectrometry method was developed for the simultaneous quantitative analysis of multiple biotransformation products of XLGB with rat intestinal microflora. For 10 selected quantitative compounds, all calibration curves revealed good linearity (r2 > 0.99) within the sampling ranges considered. The whole intra- and inter-day precisions (as relative standard deviation) of all analytes were <13.5%, and the accuracies (as relative error) were in the range from -11.3 to 11.2%. The lower limits of quantification were 20, 10, 5, 20, 2, 2, 2, 5, 2 and 2 ng/mL for sweroside, timosaponin BII, epimedin C, asperosaponin VI, psoralen, isobavachin, icariside II, timosaponin AIII, isobavachalcone and icaritin, respectively. The matrix effects, extraction recoveries and stabilities were all satisfactory. Meanwhile, dynamic profiles of 21 additional biotransformation products were also monitored by their area-time curves. The analytical method was successfully applied to describe dynamic profiles of 31 biotransformation products of XLGB and the recipes with removal of a definite composed herbal medicine (Anemarrhenae Rhizoma or Rehmanniae Radix).
仙灵骨葆（XLGB）是一种由六种草药组成的著名中药方剂，在中国用于预防和治疗骨质疏松症。作为一种口服制剂，XLGB中含有的多种成分在通过胃肠道吸收之前不可避免地会被肠道微生物群进行生物转化。然而，XLGB生物转化产物的动态变化情况仍然未知。本文建立了一种快速灵敏的超高效液相色谱串联三重四极杆质谱法，用于同时定量分析XLGB与大鼠肠道微生物群的多种生物转化产物。对于10种选定的定量化合物，所有校准曲线在所考虑的采样范围内均显示出良好的线性（r2>0.99）。所有分析物的日内和日间精密度（以相对标准偏差计）均<13.5%，准确度（以相对误差计）在-11.3%至11.2%范围内。獐牙菜苷、知母皂苷BII、朝藿定C、asperosaponin VI、补骨脂素、异补骨脂素、淫羊藿苷II、知母皂苷AIII、异补骨脂查尔酮和淫羊藿苷的定量下限分别为20、10、5、20、2、2、2、5、2和2 ng/mL。基质效应、提取回收率和稳定性均令人满意。同时，还通过其面积-时间曲线监测了另外21种生物转化产物的动态变化情况。该分析方法成功应用于描述XLGB以及去除一种特定组成草药（知母或地黄）的配方的31种生物转化产物的动态变化情况。
