基于UPLC-Q-Exactive Orbitrap MS/MS分析甘松新酮及其代谢产物在大鼠体内的分布

Distribution of Nardosinone and Its Metabolites in Rats Analyzed by UPLC-Q-Exactive Orbitrap MS/MS

摘要:

目的 采用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱法（UPLC-Q-Exactive Orbitrap MS/MS）分析甘松活性成分甘松新酮在大鼠体内的代谢和分布情况，并推测其代谢途径。方法 大鼠连续3 d灌胃30 mg·kg^-1剂量的甘松新酮混悬液，于设定的时间点收集血浆、尿液、粪便，以及心、肝、脾、肺、肾、脑、胃、肠组织，经处理后进行UPLC-Q-Exactive Orbitrap MS/MS分析，采用Xcalibur 2.2软件分析质谱数据，通过比较给药组和空白组的基峰色谱和提取离子色谱搜索代谢产物，再根据色谱峰相对保留时间（t_R）、准分子离子峰、精确分子质量及二级质谱图碎片离子等信息，利用SciFinder、PubChem等数据库检索元素组成和参考相关文献，从而鉴定可能的代谢产物，并推断代谢途径。结果 共鉴定甘松新酮代谢产物30种，从尿液、粪便、血浆、脑、心、肝、脾、肺、肾、胃、肠中分别鉴定了15、19、12、7、4、11、8、13、13、8、12种代谢产物。甘松新酮在大鼠体内主要通过羟基化、去羟基化、还原、脱氢、水合、脱水、羧基化、葡萄糖醛酸化和去羟基异丙基等途径代谢。结论 甘松新酮可在大鼠体内发生Ⅰ相和Ⅱ相代谢，代谢产物广泛分布在大鼠的主要器官中，该研究结果可为甘松新酮的药效物质基础、作用机制研究，以及临床应用提供依据。

Abstract:

Objective Ultra-performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry（UPLC-Q-Exactive Orbitrap MS/MS） was used to investigate the metabolism and distribution of nardosinone in rats， then metabolic pathways were speculated.Method Rats were administered with 30 mg·kg^-1 of nardosinone suspension by gavage for 3 consecutive days， and plasma， urine， feces， and tissues of heart， liver， spleen， lung， kidney， brain， stomach， and intestine were collected at predetermined time points. After treatment， the samples were processed for UPLC-Q-Exactive Orbitrap MS/MS， and the MS data were analyzed using Xcalibur 2.2 software. The metabolites were searched by comparing the base peak chromatogram and extracted ion chromatogram between the treated group and blank group， and based on the relative retention time（t_R）， quasi-molecular ion peak， precise molecular mass， and fragment ions of MS/MS， the elemental composition were searched using databases such as SciFinder and PubChem， as well as referring to relevant literature， the possible metabolites were identified and the metabolic pathways were inferred.Result A total of 30 metabolites of nardosinone were identified， including 15， 19， 12， 7， 4， 11， 8， 13， 13， 8 and 12 metabolites in urine， feces， plasma， brain， heart， liver， spleen， lung， kidney， stomach and intestine， respectively. The main metabolic pathways of nardosinone in rats were hydroxylation， dehydroxylation， reduction， dehydrogenation， hydration， dehydration， carboxylation， glucuronidation， and dehydroxy-isopropyl.Conclusion Nardosinone can be metabolized by phase Ⅰ and phase Ⅱ metabolism in rats， and the metabolites are widely distributed in the major organs. The results of this study can provide a basis for further research on the pharmacodynamic material basis， pharmacological mechanism and clinical application of nardosinone.