丹酚酸A甲基结合代谢物的制备与结构鉴定  

Preparation and Chemical Structure Identification of Methylated Metabolites of Salvianolic Acid A

作　　者：许卉[1] 李艳丽[1] 田红翠 翟文婷[1] 

机构地区：[1]烟台大学药学院,烟台264005

出　　处：《分析化学》2014年第1期65-70,共6页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(No.81102780)资助项目~~

摘　　要：利用具有高儿茶酚氧位甲基转移酶(COMT)活力的大鼠肝细胞液体外温孵代谢反应,首次制备了4种丹酚酸A(SAA)的甲基结合代谢物。HPLC分析结果显示,这些体外温孵代谢产物与大鼠经静脉途径给予SAA后在体内生成的代谢产物一致。以ODS为固定相,甲醇-水混合溶剂体系梯度洗脱,经中压柱层析分离,制得纯度在95%以上的代谢产物单体成分。综合应用ESI-MS、MS2,以及1H NMR,13C NMR,HSQC和HMBC等波谱方法,并与母体化合物SAA进行波谱数据的对比分析,鉴定这4种甲基结合代谢物分别为3-甲氧基丹酚酸A(1)、3'-甲氧基丹酚酸A(2)、3,3″-二甲氧基丹酚酸A(3)和3',3″-二甲氧基丹酚酸A(4)。Four methylated metabolites of salvianolic acid A （SAA） were prepared by in vitro enzyme- catalyzed methylation in the incubation system containing rat liver cytosol with high activity of catechol-O- methyltransferase（COMT）. By comparing the chromatographic profiles and retention time in HPLC, these in vitro products were detected to be consistent with those in vivo metabolites found in bile from rat after intravenously administering SAA. The metabolites 1 through 4 with purity more than 95 % were separated by medium-pressure reverse phase column chromatography using ODS and a gradient elution system of methanol and water. The structure characterization was conducted on the basis of spectral methods of MS and NMR, including ESI-MS and MS2 fragmentation, 1H NMR, 13C NMR, heteronuclear single quantum coherence （ HSQC ）, and heteronuclear multiple bond correlation （ HMBC ）. Comparing with the spectral data of parent compound（SAA）, these metabolites were unambiguously identified as 3-methoxy-salvianolic acid A （1）, 3＇-methoxy-salvianolic acid A （ 2 ）, 3,3＂-dimethoxy-salvianolic acid A （ 3 ）, and 3＇, 3＂-dimethoxy-salvianolic acid A （4）, respectively.

关 键 词：丹酚酸A 体外酶促甲基结合代谢 结构鉴定 质谱 核磁共振谱 

分 类 号：R283[医药卫生—中药学]