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一种用于全面测定大鼠血浆中嘌呤途径代谢物的液相色谱-串联质谱法及其在枸杞抗痛风作用中的应用

A liquid chromatography-tandem mass spectrometry method for comprehensive determination of metabolites in the purine pathway of rat plasma and its application in anti-gout effects of Lycium ruthenicum Murr.

机构信息

Analysis and Test Center, State Key Laboratory of Plateau Ecology and Agriculture, Qinghai University, Xining, China.

Shanghai Key Laboratory of Functional Materials Chemistry, School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai, China.

出版信息

J Sep Sci. 2023 Nov;46(21):e2300090. doi: 10.1002/jssc.202300090. Epub 2023 Sep 8.

Abstract

It has been proved that purine metabolites are implicated in various biological syndromes and disorders. Therefore, the realization of panoramic detection of purine metabolites will be of great significance to the pathogenesis of purine metabolic disorders. In the present study, an ultra-high performance liquid chromatography with tandem mass spectrometry method was developed for the comprehensive quantification of purine metabolites in rat plasma. The 17 purine metabolites were separated and quantified in the short running time of 15 min. The proposed method was strictly validated by applying SeraSub solution as a matrix and proved to be linear (R ≥ 0.9944), accurate (the recoveries of all analytes ranged from 85.3% to 103.0%, with relative standard deviation values ≤ 9.3%), and precise (the intra- and inter-day precisions were less than 10.8% and 12.4%, respectively). The method was then successfully applied to the qualification of the endogenous purine metabolites in acute gouty arthritis rats, as well as colchicine and anthocyanin-intervened rats. Results showed that uric acid, xanthine, hypoxanthine, and xanthine were considered the key factors of acute gouty arthritis. The established method and measurement of purines in rat plasma might help the investigation of the action mechanisms between purine disorders and related diseases.

摘要

嘌呤代谢物与多种生物综合征和疾病有关已得到证实。因此,实现嘌呤代谢物的全景检测对嘌呤代谢紊乱的发病机制将具有重要意义。在本研究中,建立了一种超高效液相色谱-串联质谱法,用于大鼠血浆中嘌呤代谢物的全面定量分析。在 15 分钟的短运行时间内分离和定量了 17 种嘌呤代谢物。该方法严格验证了 SeraSub 溶液作为基质的适用性,证明其具有良好的线性(R≥0.9944)、准确性(所有分析物的回收率均在 85.3%至 103.0%之间,相对标准偏差值≤9.3%)和精密度(日内和日间精密度分别小于 10.8%和 12.4%)。该方法随后成功应用于急性痛风性关节炎大鼠以及秋水仙碱和花青素干预大鼠内源性嘌呤代谢物的定性分析。结果表明,尿酸、黄嘌呤、次黄嘌呤和黄嘌呤被认为是急性痛风性关节炎的关键因素。该方法的建立和大鼠血浆中嘌呤的测定可能有助于研究嘌呤紊乱与相关疾病之间的作用机制。

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