目的 基于丹荷颗粒中15个特征性成分（丹酚酸B、虎杖苷、大黄素、大黄素-8-O-β-D-葡萄糖苷、白藜芦醇、金丝桃苷、异槲皮苷、紫云英苷、橙皮苷、川陈皮素、荷叶碱、O-去甲基荷叶碱、N-去甲基荷叶碱、去氢荷叶碱、乌药碱）在大鼠血浆暴露量的测定，筛选丹荷颗粒中适于作为质量标志物（quality marker，Q-Marker）的成分。方法 以EC-C₁₈柱为预柱、ZORBAX RRHD Eclipse Plus C₁₈柱为色谱柱，以0.1%甲酸水溶液-乙腈为流动相，采用动态多反应监测扫描质谱，建立高效液相色谱-串联质谱（HPLC-MS/MS）方法；大鼠ig丹荷颗粒，采集不同时间点的血浆样品，并测定血浆样品中各特征性成分的质量浓度，计算其血浆暴露量；确定体内高暴露的成分，对其进一步进行活性及制剂含量分析，依据体内暴露量、高暴露成分活性、制剂相应成分含量确定Q-Marker。结果 丹酚酸B、虎杖苷、大黄素、大黄素-8-O-β-D-葡萄糖苷、白藜芦醇、金丝桃苷、异槲皮苷、紫云英苷、橙皮苷、川陈皮素、荷叶碱、O-去甲基荷叶碱、N-去甲基荷叶碱、去氢荷叶碱、乌药碱在大鼠血浆中暴露量分别为（1340.45±167.64）、（232.49±37.05）、（677.51±71.61）、（33.99±20.74）、（281.53±32.32）、（25.01±4.90）、（16.85±2.88）、（11.89±2.69）、（113.71±23.05）、（11.13±1.94）、（168.69±42.68）、（2595.00±314.78）、（9.55±1.36）、（1.29±0.11）、（13.96±0.88）ng·h/mL，其中丹酚酸B、虎杖苷、大黄素、金丝桃苷、紫云英苷、橙皮苷、荷叶碱具有调血脂活性且在制剂中质量分数均大于0.1 mg/g。结论 丹酚酸B、虎杖苷、大黄素、槲皮素、橙皮苷、柚皮苷和荷叶碱可作为丹荷颗粒的Q-Marker。

Objective To screen Danhe Granules for quality marker (Q-Marker) based on determination of plasma exposure of 15 characteristic components (salvianolic acid B, polydatin, emodin, emodin-8-O-β-D-glucoside, resveratrol, hyperin, isoquercitrin, astragalin, hesperidin, nobiletin, nuciferine, O-nornuciferine, N-nornuciferine, dehydronuciferin, and coclaurine) in Danhe Granules in rats. Methods EC-C₁₈ column was used as pre-column, ZORBAX RRHD Eclipse Plus C₁₈ column was used as column, 0.1% formic acid aqueous solution-acetonitrile was used as mobile phase, dynamic multiple response monitoring scanning mode was used for mass spectrometry and HPLC-MS/MS method was established. Rats were ig given Danhe Granules, plasma of rats at different time points were collected, concentration of each component in plasma samples of rats were determined, plasma exposure in vivo were calculated. Components with high exposure in vivo were determined, of which activity and preparation content were further analyzed. Q-Marker were determined according to amount of exposure in vivo, activity of high exposure ingredients, corresponding composition and content of the preparation. Results Plasma exposure levels of salvianolic acid B, polydatin, emodin, emodin-8-O-β-D-glucoside, hyperin, astragalin, astragalin, hesperidin, nobiletin, nuciferine, O-nornuciferine, N-nornuciferine, dehydronuciferin, coclaurine were (1340.45 ±167.64), (232.49 ±37.05), (677.51 ±71.61), (33.99 ±20.74), (281.53 ±32.32), (25.01 ±4.90), (16.85 ±2.88), (11.89 ±2.69), (113.71 ±23.05), (11.13 ±1.94), (168.69 ±42.68), (2595.00 ±314.78), (9.55 ±1.36), (1.29 ±0.11), and (13.96 ±0.88) ng∙h/mL, salvianolic acid B, polydatin, emodin, hyperin, astragalin, hesperidin, and nuciferine showed lipid-lowering activity and their contents in preparations were all greater than 0.1 mg/g. Conclusion Salvianolic acid B, polydatin, emodin, quercetin, hesperidin, naringin, and nuciferine can be used as Q-Marker of Danhe Granules.

R285.5;R285.61

国家重大新药创制科技重大专项资助项目（2019ZX09201004-001-022）