Nováková Michaela, Krivánková Ludmila
Institute of Analytical Chemistry, Academy of Sciences of the Czech Republic, Brno, Czech Republic.
Electrophoresis. 2008 Apr;29(8):1694-700. doi: 10.1002/elps.200700663.
The determination of ethyl glucuronide (EtG), a marker of recent alcohol consumption, in human serum by hyphenation of capillary ITP (CITP) and CZE is reported. For CITP step, 1 x 10(-2) M hydrochloric acid adjusted with epsilon-aminocaproic acid (EACA) to pH 4.4 was used as the leading electrolyte, and 1 x 10(-2) M nicotinic acid with EACA, pH 4.4, was used as the terminating electrolyte (TE). All electrolytes contained 0.2% hydroxypropylcellulose to suppress electroosmosis. In CITP, EtG was separated from fast serum macrocomponents chloride, phosphate, lactate, and acetate. Zones of microcomponents including EtG that migrated between acetate and nicotinate were forwarded to the second capillary filled with a BGE identical with the TE. Conductivity detection was used in the CITP step. Sensitive detection in the CZE step was performed using indirect spectrophotometric detection at 254 nm. The assay is based on a 1:5 dilution of serum with deionized water and has a concentration LOD for EtG in diluted sample of 9.8 x 10(-9) M. The method was used for the determination of EtG in sera of volunteers consuming alcohol.
报道了通过毛细管等速电泳(CITP)和毛细管区带电泳(CZE)联用技术测定人血清中近期酒精摄入标志物——葡萄糖醛酸乙酯(EtG)的方法。在CITP步骤中,用ε-氨基己酸(EACA)将1×10⁻² M盐酸调节至pH 4.4作为前导电解质,用含EACA、pH 4.4的1×10⁻² M烟酸作为终止电解质(TE)。所有电解质均含有0.2%羟丙基纤维素以抑制电渗。在CITP中,EtG与血清中快速移动的大分子成分氯离子、磷酸根、乳酸根和醋酸根分离。包括EtG在内的在醋酸根和烟酸根之间迁移的微量成分区带被转移至填充有与TE相同背景电解质(BGE)的第二根毛细管中。CITP步骤采用电导检测。CZE步骤采用254 nm间接分光光度检测进行灵敏检测。该测定方法基于血清用去离子水1:5稀释,稀释样品中EtG的浓度检测限为9.8×10⁻⁹ M。该方法用于测定饮酒志愿者血清中的EtG。