基于序贯代谢的五味子入血及脑脊液成分分析

Analysis on Components Absorbed into Blood and Cerebrospinal Fluid of Schisandrae Chinensis Fructus Based on Sequential Metabolism

目的:基于序贯代谢方法,结合液质联用技术鉴定五味子入血和入脑脊液原型成分和代谢产物。方法:雄性SD大鼠在灌胃给药和在体肠灌流给药后收集其综合代谢、肠代谢、肝代谢血浆和脑脊液样品,利用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱法(UPLC Q-Exactive Orbitrap MS)分析并比对五味子提取液、空白血浆、含药血浆、空白脑脊液、含药脑脊液样品谱图差异,采用ACQUITY UPLC BEH Shield RP18色谱柱(2.1 mm×100 mm,1.7μm),流动相乙腈(A)-0.1%甲酸水溶液(B)梯度洗脱(0~7 min,95%B;7~12 min,95%~35%B;12~17 min,35%~15%B;17~20 min,15%~12%B;20~22 min,12%~5%B;22~23 min,5%B;23~25 min,5%~95%B;25~28 min,95%B),加热电喷雾离子化源(HESI),正、负离子检测模式,扫描范围m/z 100~1500。根据化合物的保留时间、特征碎片、分子式及对照品信息鉴定五味子入血及脑脊液的原型成分和代谢产物。结果:从五味子提取物中共鉴定出42个化学成分,包含木脂素、黄酮、氨基酸、鞣质等化合物,其中以木脂素类化合物为主。从不同部位获取的血浆样品中共鉴定出27种原型入血成分与14种代谢产物;在脑脊液中共鉴定出15种原型成分和9种代谢产物;代谢产物生成过程中涉及的代谢反应主要有去甲基化、甲基化、去甲氧基化和羟基化等。结论:通过对五味子在大鼠体内的原型成分与代谢产物的系统鉴定,为后续更好地挖掘五味子中治疗中枢神经系统疾病的药效物质基础提供了数据支撑。

Objective:To identify the prototypical components and metabolites absorbed into blood and cerebrospinal fluid of Schisandrae Chinensis Fructus(SCF)based on sequential metabolism combined with liquid chromatography-mass spectrometry.Method:Blood and cerebrospinal fluid samples of integrated metabolism,intestinal metabolism and hepatic metabolism were collected from male SD rats after gavage and in situ intestinal perfusion administration,and ultra-performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry(UPLC Q-Exactive Orbitrap MS)was used to analyze and compare the differences in the spectra of SCF extract,blank plasma,administered plasma,blank cerebrospinal fluid and administered cerebrospinal fluid with ACQUITY UPLC BEH Shield RP18 column(2.1 mm×100 mm,1.7μm),the mobile phase was acetonitrile(A)-0.1%formic acid aqueous solution(B)for gradient elution(0-7 min,95%B;7-12 min,95%-35%B;12-17 min,35%-15%B;17-20 min,15%-12%B;20-22 min,12%-5%B;22-23 min,5%B;23-25 min,5%-95%B;25-28 min,95%B).And heated electrospray ionization(HESI)was used with positive and negative ion modes,the scanning range was m/z 100-1500.The prototypical constituents and their metabolites absorbed into blood and cerebrospinal fluid of SCF were identified according to the retention time,characteristic fragments,molecular formulae and the information of reference substances.Result:A total of 42 chemical components were identified in the extract of SCF,including lignans,flavonoids,amino acids,tannins,and others,of which lignans were the main ones.A total of 27 prototypical components and 14 metabolites were identified in plasma samples from different sites.A total of 15 prototypical components and 9 metabolites were identified in cerebrospinal fluid.The main metabolic reactions involved in the formation of metabolites were mainly demethylation,methylation,demethoxylation and hydroxylation.Conclusion:Through the systematic identification of the prototypical components and metabolites of SCF in rats,it provides data support for further better exploring the material basis of SCF in the treatment of central nervous system diseases.