摘要
采用液相色谱质谱(LC-MS)方法建立10个不同产地的新鲜当归(FRAS)及干燥当归(RAS)色谱指纹图谱;用乙酰苯肼(APH)和环磷酰胺(CPX)联合复制血虚大鼠模型,观察FRAS和RAS对血虚大鼠生理指标的影响。通过灰色关联度分析(grey relational analysis,GRA)和偏最小二乘(partial least squares,PLS)回归分析法将LC-MS色谱指纹图谱中各共有峰的相对含量和补血药效数据相关联,建立谱效关系。结果显示,FRAS和RAS均有一定的补血作用(P<0.05)。根据PLS回归系数大小确定了FRAS和RAS中各共有峰对补血作用的贡献程度。其中,4个色谱峰对补血作用的贡献较大,分别为P1(未知),P2(未知),P7(阿魏酸),P11(洋川芎内酯A)。该实验研究了FRAS和RAS的LC-MS色谱指纹图谱与补血作用之间的相关性,确定了二者中的补血药效成分,为当归资源的合理应用提供了一定的理论基础。
Ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(LC-MS) was used to establish the chromatography fingerprint for fresh(FRAS) and dry(RAS) roots of Angelica sinensis from 10 different places. The rat model of blood deficiency was established by acetyl-phenyl-hydrazine(APH) and cyclophosphamide(CTX). Then grey relational analysis(GRA) and partial least squares regression(PLS) were used to investigate the spectrum-effect relationship between the relative contents and the data of enriching blood pharmacodynamics efficacy. The results showed that the FRAS and RAS had certain enriching blood activities(P<0.05). The contribution degree of the FRAS and RAS to enriching blood activities of each common peaks were determined by regression coefficient. Among them, 4 common peaks contributed significantly to the effect of enriching blood activities, P1(unknown), P2(unknown), P7(ferulic acid), and P11(senkyunolide A) respectively. This paper investigated the spectrum-effect relationship between enriching blood activities and LC-MS chromatography fingerprint of RAS and FRAS, and determined the effective compositions of RAS and FRAS with enriching blood activities. It lays a theoretical foundation for the comprehensive development and utilization of A. sinensis.
引文
[1] 中国药典.一部[S].2015:133.
[2] 郭玫,李应东,邵晶,等.不同干燥方式对当归质量的影响[J].中成药,2004(1):38.
[3] 李成义,祝永甲,王明伟,等.不同干燥方法对当归质量影响研究[J].甘肃中医学院学报,2013,30(5):21.
[4] 李伟霞,唐于平,尚尔鑫,等.当归、川芎和归芎药对中主要芳香酸类成分在血虚大鼠体内的代谢研究[J].中华中医药杂志,2013,28(5):1212.
[5] 杨英来,崔方,胡芳,等.当归补血、活血作用的谱效关系研究[J].中国中药杂志,2013,38(22):3923.
[6] 李影迪,初杰,范颖,等.当归多糖与黄芪多糖配伍对骨髓造血干细胞增殖分化的影响[J].辽宁中医药大学学报,2016,18(6):31.
[7] 林丽,晋玲,李应东,等.当归干、鲜品中游离阿魏酸和总阿魏酸含量[J].中国实验方剂学杂志,2012,18(22):94.
[8] 马霞.当归不同部位的化学成分与补血作用研究[D].兰州:甘肃中医药大学,2018.
[9] Shen Y W,Liu J J.Application of the gray relevancy method to college teachers research performance assessment of humanities and social sciences[J].Sci Tech Manag Res,2010,22:237.
[10] Shi M J,Zhang Y,Song M M,et al.Screening the marker components in Psoralea corylifolia L.with the aids of spectrum-effect relationship and component knock-out by UPLC-MS2[J].Int J Mol Sci,2018,19(11):3439.
[11] 史旭芹,唐于平,瞿城,等.复合血虚模型中环磷酰胺剂量的探索[J].中药药理与临床,2018,34(5):136.
[12] Wang J M,Peng L N,Shi M J,et al.Spectrum effect relationship and component knock-out in Angelica dahurica radix by high performance liquid chromatography-Q exactive hybrid quadrupole-orbitrap mass spectrometer[J].Molecules,2017,22(7):1231.
[13] 刘洁丽,马霞,张瑛,等.当归地上部分补血作用的谱效关系研究[J].中国中药杂志,2019,44(7):1416.
[14] 宋雁,贾旭东,崔文明,等.不同途径和剂量环磷酰胺建小鼠免疫抑制模型的对比研究[J].中国食品卫生杂志,2013,25(3):218.
[15] 金若敏,宁炼,陈长勋,等.血虚模型动物制备及当归补血汤的作用研究[J].中成药,2001,23(4):268.
[16] 纪鹏.生当归及其不同炮制品干预小鼠血虚模型的代谢组学研究[D].兰州:甘肃农业大学,2015.
[17] 张婕,葛永辉,杨慧,等.LC-MS/MS法同时测定烟草中的16种游离氨基酸和6种Amadori化合物[J].烟草科技,2017,50(11):58.
[18] 牛研,王书芳.LC-Q-TOF-MS和LC-IT-MSn分析当归芍药散中化学成分[J].中草药,2014,45(8):1056.
[19] 朱云祥,陈璐琳,龚婧如,等.LC-Q-TOF-MS及LC-IT-MS分析酸枣仁汤的化学成分[J].中国中药杂志,2014,39(3):457.