基于UHPLC-Q-ExactiveMS分析鉴定隐丹参酮在大鼠体内的代谢产物

Analysis and Identification of Metabolites of Cryptotanshinonein Rats by UHPLC-Q-Exactive Mass Spectrometer

采用超高效液相色谱-四极杆-静电场轨道阱高分辨质谱(UHPLC-Q-ExactiveMS)法对隐丹参酮在大鼠体内的代谢产物进行分析鉴定。大鼠单次灌胃后,收集不同时间点的血浆以及24h内的尿液和粪便,应用固相萃取法处理生物样品,采用AcquityUPLCBEHC18色谱柱(2.1mm×100mm×1.7μm),以0.1%甲酸溶液(A)-乙腈(B)为流动相进行梯度洗脱,在电喷雾离子源正离子模式下对生物样品进行分析。结合精确分子质量、碎片离子信息及相关文献报道,共分析鉴定出45种代谢产物。结果表明,隐丹参酮在大鼠体内的主要代谢途径为还原、羟基化、去甲基化、葡萄糖醛酸化、S-半胱氨酸结合、脱一氧化碳、脱氢、脱乙基以及它们的复合反应等。本研究阐明了隐丹参酮在大鼠体内的代谢情况,可为进一步研究其药效物质基础及质量标准提供依据。

For further study of bioactive components of cryptotanshinone in vivo,the metabolites of cryptotanshinone in rats were analyzed and identified by ultra-high performance liquid chromatography coupled with quadrupole/electrostatic field Obitrap high-resolution mass spectrometry(UHPLC-Q-Exactive MS).Cryptotanshinone was suspended in 0.2%Tween-80 solution.Rats in drug group were given adose of 300 mg/kg body weight orally.0.2%Tween-80 solution was administrated to rats in control group.Plasma at different time points was collected after oral gavage.Urine and feces were collected after single intragastric administration in rats within 24 h.Biological samples(plasma,urine and feces)were separated by solid phase extraction(SPE)to exclude protein and solid residue precipitation.Acquity UPLC BEH C18 column(2.1 mm×100 mm×1.7μm)was used with 0.1%formic acid solution(A)-acetonitrile(B)as the mobile phase for the gradient elution.Then the biological sample was analyzed by quadrupole/electrostatic field Obitrap high-resolution mass spectrometry at positive ion mode.According to accurate molecular weight,primary and secondary mass spectrometry information and literatural reports,a total of 45 metabolites(including cryptotanshinone)were screened and identified.The main metabolic of pathways in rats are reduction,hydroxylation,demethylation,glucuronidation,S-cysteine acid binding reaction,decarbonation,dehydrogenation,deethylation,deacetaldehydeation and their composite reactions.This study not only provides useful data to comprehensively understand the metabolic mechanism and material basis of cryptotanshinone,but also showes that it is reliable for further study of drug-related constituents and quality standard of cryptotanshinone.