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在线受限分子印迹固相萃取与液相色谱-质谱联用选择性测定血清胆汁酸。

Online restricted access molecularly imprinted solid-phase extraction coupled with liquid chromatography-mass spectrometry for the selective determination of serum bile acids.

机构信息

Laboratory of Toxicant and Drug Analyses, Gabriel Monteiro da Silva St. 700, Federal University of Alfenas - Unifal-MG, 37130-001, Alfenas, MG, Brazil.

出版信息

Analyst. 2022 Jun 13;147(12):2779-2792. doi: 10.1039/d2an00446a.

Abstract

A rapid and sensitive online restricted access molecularly imprinted solid-phase extraction method coupled to a liquid chromatography-mass spectrometry (LC-MS) system for simultaneous determination of serum bile acids as well as their taurine and glycine conjugates was developed. Reversible liver damage of workers exposed to volatile organic solvents can be investigated based on the level of the analyzed molecules. A restricted access molecularly imprinted polymer coated with bovine serum albumin (RAMIP-BSA) was synthesized and used as the extraction phase. The column switching liquid chromatography system was able to exclude about 100% of the macromolecules and extract/separate nine bile acids from blood human serum samples, in a total time of 40 minutes. The developed method was validated based on the Food and Drug Administration (FDA) guidelines, being linear for all the analytes in their respective analytical ranges (coefficients of determination higher than 0.99), with limits of detection (LOD) and quantification (LOQ) ranging from 2.0 to 5.7 μg L and from 10.0 to 25.0 μg L, respectively. Suitable results for precision (relative standard deviation ranged from 3.2 to 14.5% and 0.7 to 14.8%, respectively for intra and inter-assay) and accuracy (relative error ranged from -14.8 to 14.2%; -13.8 to 14.3%) were obtained. The validated analytical method was applied to determine bile acids in serum samples of five workers occupationally exposed to volatile organic solvent, demonstrating its applicability in the assessment of these toxicants.

摘要

建立了一种快速灵敏的在线受限-access 分子印迹固相萃取-液相色谱-质谱(LC-MS)联用方法,用于同时测定血清胆汁酸及其牛磺酸和甘氨酸缀合物。基于分析分子的水平,可以研究接触挥发性有机溶剂的工人的可逆性肝损伤。合成了涂有牛血清白蛋白(RAMIP-BSA)的受限-access 分子印迹聚合物,并用作萃取相。该柱切换液相色谱系统能够排除约 100%的大分子,并从人血清样品中提取/分离出 9 种胆汁酸,总时间为 40 分钟。该方法基于美国食品和药物管理局(FDA)指南进行了验证,对于所有分析物在各自的分析范围内均具有线性(决定系数均高于 0.99),检测限(LOD)和定量限(LOQ)范围分别为 2.0 至 5.7 μg/L 和 10.0 至 25.0 μg/L。精密度(日内和日间的相对标准偏差分别为 3.2%至 14.5%和 0.7%至 14.8%)和准确度(相对误差分别为-14.8%至 14.2%和-13.8%至 14.3%)的结果均合适。该经过验证的分析方法已应用于测定 5 名职业接触挥发性有机溶剂工人的血清胆汁酸,证明其适用于评估这些毒物。

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