Zirconium silicate assisted removal of residual proteins after organic solvent deproteinization of human plasma, enhancing the stability of the LC-ESI-MS response for the bioanalysis of small molecules
Zirconium silicate assisted removal of residual proteins after organic solvent deproteinization of human plasma, enhancing the stability of the LC-ESI-MS response for the bioanalysis of small molecules
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DOI：
10.1016/j.aca.2014.09.014
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发表时间：
2014-12-10
影响因子：
6.2
通讯作者：
Bonn, Guenther K.
中科院分区：
文献类型：
--
作者：
Hussain, Shah;Pezzei, Cornelia;Bonn, Guenther K.
开发了一种有效的血浆净化方法，其中应用甲醇蛋白沉淀，然后用硅酸锆辅助排除残留蛋白。硅酸锆（IV）与蛋白质的强结合使得能够通过快速固相萃取（SPE）程序在溶剂脱蛋白后消除剩余的蛋白质。基质辅助激光解吸/电离飞行时间质谱法（MALDI/TOF MS）用于监测应用于人血浆样品的清洁实践期间的蛋白质。使用二喹啉甲酸（BCA）测定通过比色检测定量蛋白质。所提出的分析策略导致人血浆样品中>99.6%蛋白质的消耗。采用高效液相色谱-二极管阵列-电喷雾质谱联用技术（HPLC-DAD/ESI MS）对人血浆中咖啡酰奎尼酸及其代谢产物进行了定性和定量分析。该方法证明了不同浓度加标标准化合物的高回收率。洋蓟素和绿原酸的回收率分别为81-86%和78- 83%。咖啡酸的提取率超过89- 92%，而阿魏酸和二羟基氢化肉桂酸的回收率分别为87-91%和92- 95%。根据FDA-生物分析方法验证行业指南（2001）对该方法进行了部分验证。该方案提高了甲醇脱蛋白的净化效率，显著降低了基质效应，为从人血浆中分离小分子提供了一个很好的分析工具。(C)© 2014 Elsevier B. V.保留所有权利。
An efficient blood plasma clean-up method was developed, where methanol protein precipitation was applied, followed by zirconium silicate assisted exclusion of residual proteins. A strong binding of zirconium (IV) silicate to the proteins enabled the elimination of remaining proteins after solvent deproteinization through a rapid solid-phase extraction (SPE) procedure. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI/TOF MS) was used for monitoring the proteins during clean-up practice applied to human plasma samples. The proteins were quantified by colorimetric detection using the bicinchoninic acid (BCA) assay. The presented analytical strategy resulted in the depletion of >99.6% proteins from human plasma samples. Furthermore, high-performance liquid chromatography hyphenated to diode-array and electrospray ionization mass spectrometric detection (HPLC-DAD/ESI MS) was applied for qualitative and quantitative analysis of the caffeoylquinic acids (CQAs) and their metabolites in human plasma. The procedure demonstrated high recoveries for the standard compounds spiked at different concentrations. Cynarin and chlorogenic acid were recovered in the range of 81-86% and 78-83%, respectively. Caffeic acid was extracted in the excess of 89-92%, while ferulic acid and dihydroxyhydrocinnamic acid showed a recovery of 87-91% and 92-95%, respectively. The method was partially validated in accordance with FDA-Industry Guidelines for Bioanalytical Method Validation (2001). The presented scheme improves the clean-up efficacy of the methanol deproteinization, significantly reduces the matrix effects and provides a great analytical tool for the isolation of small molecules from human plasma. (C) 2014 Elsevier B.V. All rights reserved.