大鼠血浆中槲皮素及其代谢产物测定方法研究进展

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内容来源于Talanta

 

大鼠血浆中槲皮素代谢物具有不同的理化性质，如分子量、亲脂性和酸碱性等。近期，捷克共和国查尔斯大学的OndřejNovák和LucieNováková团队在“Talanta”上发表名为“Simultaneous determination of quercetin and its metabolites in rat plasma by using ultra-high performance liquid chromatography tandem mass spectrometry”论文在槲皮素及其代谢产物测定方法研究方面取得新进展。其团队建立的液质联用定量方法选择性好、灵敏度高，可较好的对酚酸和一对同分异构体（ 柽柳黄素 和异鼠李素）进行分离。另外，本研究对两个样品处理方法，蛋白沉淀法和微量固相萃取方法(MEPS)进行了优化。为了提高槲皮素及其代谢产物的血浆提取效率，其团队在蛋白沉淀过程中对样品进行了酸化，并对微量固相萃取技术中的吸附剂和洗脱溶剂进行了优化。对两种样品处理方法进行了全面验证，结果显示其均具有可接受的精密度、准确度和基质效应。采用蛋白沉淀或微量固相萃取技术，结合液质联用定量方法均可在50ul生物样品中同时测定槲皮素及其代谢物的含量。由于蛋白质沉淀法可更加快速和非选择性的平行制备样品，因此该处理方法最终被选择并应用于血浆药代动力学研究。

 

 

图片来源于Talanta， Volume 185,1 August 2018, Pages 71-79 DOI: 10.1016/j.talanta.2018.03.033

 

原文摘要

Fast, selective, and sensitive ultra-high performance liquid chromatography method with tandem mass spectrometry detection for the determination of quercetin and its metabolites with various physico-chemical properties such as molecular weight, lipophilicity, and acid-base properties has been developed. These compounds included small hydrophilic phenolic acids and more lipophilic metabolites with preserved flavonoid structure in small amount of rat plasma. The developed method enables selective separation of phenolic acids and a pair of isomers tamarixetin and isorhamnetin with satisfactory peak shapes and a high sensitivity using mass spectrometry detection. In addition, two sample preparation procedures including protein precipitation and microextraction in packed sorbent (MEPS) were optimized. The sample acidification included in protein precipitation as well as optimizing of MEPS sorbents and elution solvents improved isolation of quercetin and related compounds from rat plasma. Finally, both methods developed for sample preparation were fully validated to demonstrate sufficient accuracy and precision and acceptable matrix effects. Both sample preparation approaches combined with mass spectrometry-based quantification allowed the simultaneous determination of quercetin and its metabolites from a small amount of biological samples of only 50 μL. Due to the fast and non-selective parallel sample preparation, the protein precipitation was eventually applied to plasma samples derived from pharmacokinetic studies.

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