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首页> 外文期刊>RSC Advances >Rapid discovery and identification of multiple absorbed chemical constituents and metabolites in rat cerebrospinal fluid after oral administration of Jitai tablets by a LC-MS based metabolomics approach
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Rapid discovery and identification of multiple absorbed chemical constituents and metabolites in rat cerebrospinal fluid after oral administration of Jitai tablets by a LC-MS based metabolomics approach

机译：基于LC-MS的代谢组学方法快速发现和鉴定吉泰片口服后大鼠脑脊液中多种吸收的化学成分和代谢产物

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摘要

An integrative strategy using LC-Q/TOF-MS and LC-QqQ-MS/MS coupled with multi-variate statistical analysis of principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) was developed to screen and identify the constituents transported into the rat cerebrospinal fluid (CSF) after oral administration of Jitai tablets (JTTs). CSF samples were pretreated with cold methanol prior to liquid chromatography, and the separation was carried out on a HSS T3 column with a linear gradient elution. Mass spectra were acquired in positive ion mode for analyte identification and targeted MS/MS mode for quantification. In the S-plot of OPLS-DA, twenty-six interested ions were extracted, among which, sixteen absorbed prototype components of JTT and seven metabolites were identified in vivo. Cysteine conjugation, demethylation and glucuronidation were the major metabolic reaction types of the identified constituents. An LC-MS/MS method with targeted multiple-reaction monitoring of scopolamine, tetrahydrocoptisine, tetrahydroberineper, protopine, tetrahydropulmatine and corydaline in rat CSF was developed to validate the identification results. The concentration of the six compounds in rat CSF was in the range of 0.014-1.678 ng mL(-1). In conclusion, a LC-MS based metabolomics approach can provide a rapid and sensitive method for characterizing bioactive components of JTT at micro concentration on pathological biopsy, which benefits further pharmacology and mechanism research of JTT.
机译：开发了一种使用LC-Q / TOF-MS和LC-QqQ-MS / MS的集成策略，以及主成分分析（PCA）和正交偏最小二乘判别分析（OPLS-DA）的多元统计分析，以筛选和筛选确定口服吉泰片（JTTs）后运输到大鼠脑脊液（CSF）中的成分。在液相色谱分析之前，先用冷甲醇对CSF样品进行预处理，然后在具有线性梯度洗脱的HSS T3色谱柱上进行分离。以正离子模式采集质谱用于分析物鉴定，以目标MS / MS模式采集以进行定量。在OPLS-DA的S图中，提取了26种感兴趣的离子，其中，在体内鉴定了16种JTT吸收的原型组分和7种代谢物。半胱氨酸的结合，去甲基化和葡糖醛酸化是确定成分的主要代谢反应类型。建立了针对大鼠脑脊液中东pol碱，四氢吗啡碱，四氢小ine碱，原松，四氢普马汀和羟乙基萘的靶向多反应监测的LC-MS / MS方法，以验证鉴定结果。大鼠脑脊液中六种化合物的浓度在0.014-1.678 ng mL（-1）的范围内。总之，基于LC-MS的代谢组学方法可为病理活检中微量浓度的JTT的生物活性成分表征提供快速，灵敏的方法，这有利于JTT的进一步药理和机理研究。

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