丹酚酸B在大鼠体内的代谢产物及代谢途径研究

Study on Metabolites and Metabolic Pathway of Salvianolic Acid B in Rats

关键词:  丹酚酸B  代谢产物  代谢途径  液相色谱-串联质谱法

Key words:  salvianolic acid B  metabolites  metabolic pathway  LC-MS/MS

摘要:

目的 建立大鼠血浆、胆汁、尿液与粪便中丹酚酸B代谢产物的测定方法，并考察其代谢途径。方法 取SD大鼠18只，平分为灌胃和静注2组，每组再分成3个小组，分别采集血浆、胆汁、尿液及粪便，每小组2只大鼠分别单剂量灌胃（500 mg·kg^-1）和尾静脉注射（16.5 mg·kg^-1）给予丹酚酸B，另一只分别灌胃水和尾静脉注射生理盐水作为空白对照。采用高效液相色谱串联四极杆飞行时间质谱（HPLC-Q-TOF-MS/MS）联用法测定血浆、胆汁、尿液与粪便中的丹酚酸B代谢产物，同时推断代谢途径。结果 根据一级质谱分子离子信息和二级质谱碎裂离子信息，在大鼠体内共发现8个丹酚酸B的代谢产物，其中胆汁、粪便中均含有这8个代谢产物，尿液中发现3个代谢产物，血浆中发现2个代谢产物。由丹酚酸B及其代谢产物结构推断丹酚酸B体内主要通过甲基化反应、酯键水解反应代谢。结论 建立的分析方法准确、灵敏、快速，符合生物样品分析要求，适用于大鼠血浆、胆汁、尿液与粪便中丹酚酸B代谢产物的分析，并分析鉴定了8个代谢产物。代谢实验结果表明，胆汁排泄是丹酚酸B最主要的排泄方式。

Abstract:

OBJECTIVE To establish methods for the determination of salvianolic acid B metabolites in rats' plasma, bile, urine and feces, and investigate their metabolic pathways. METHODS Eighteen SD rats were equally divided into two groups:intragastric administration and intravenous injection. Each group was divided into three groups to collect plasma, bile, urine and feces, respectively. Two rats of each group were given salvianolic acid B single-dose by intragastric administration(500 mg·kg^-1) and tail vein injection(16.5 mg·kg^-1) respectively, and the other one was administered gavage water and saline intravenously as a blank control. Determine the metabolites of salvianolic acid B in plasma, bile, urine and feces by HPLC-Q-TOF-MS/MS, and infer the metabolic pathways. RESULTS According to information of MS and MS², eight metabolites of salvianolic acid B were found in rats, including eight metabolites in bile and feces, three metabolites in urine and two metabolites in plasma. Based on the structure of salvianolic acid B and its metabolites, the in vivo metabolism of salvianolic acid B was mainly metabolized by methylation reaction, ester bond hydrolysis reaction. CONCLUSION The established analytical method is accurate, sensitive and rapid, and meets the requirements of biological sample analysis. It is applicable to the analysis of the metabolites of salvianolic acid B in rat plasma, bile, urine and feces, and 8 metabolites were identified. Metabolic experiments show that bile excretion is the most important excretion of salvianolic acid B.