Mrázková Michaela, Caslavska Jitka, Thormann Wolfgang, Krivánková Ludmila
Institute of Analytical Chemistry, Academy of Sciences of the Czech Republic, Brno, Czech Republic.
Electrophoresis. 2006 Dec;27(23):4772-8. doi: 10.1002/elps.200600148.
The analysis of ethyl glucuronide (EtG), a marker of recent alcohol consumption, in serum with an optimized CZE assay is reported. The method uses a 0.1-mm id fused-silica capillary of 50 cm effective length that is coated with linear polyacrylamide, a pH 4.4 nicotinic acid/epsilon-aminocaproic acid (EACA) BGE, reversed polarity and indirect analyte detection. The assay is based on a 1:1 dilution of serum with deionized water and has LODs for EtG, lactate and acetate of 3.8 x 10(-7) M, 2.60 x 10(-6 )M and 2.18 x 10(-6 )M, respectively. Separation of EtG from endogenous macro- and microcomponents (anionic serum components of high and low concentration, respectively) and its quantification are shown to be possible for a wide range of lactate (stacker) and acetate (destacker) concentrations, macrocomponents that have an impact on the CZE behavior of EtG and that change after intake of ethanol. The assay has been successfully applied to the analysis of EtG, lactate and acetate in (i) sera of volunteers that ingested known amounts of alcohol and (ii) samples of patients that were classified (teetotalers and social drinkers vs. alcohol abusers) via analysis of carbohydrate-deficient transferrin.
报道了采用优化的毛细管区带电泳(CZE)法分析血清中近期酒精摄入标志物——葡萄糖醛酸乙酯(EtG)。该方法使用内径0.1 mm、有效长度50 cm的熔融石英毛细管,内壁涂有线性聚丙烯酰胺,以pH 4.4的烟酸/ε-氨基己酸(EACA)为背景电解质(BGE),采用反相极性和间接分析物检测。该分析方法基于血清与去离子水1:1稀释,EtG、乳酸和乙酸的检测限分别为3.8×10⁻⁷ M、2.60×10⁻⁶ M和2.18×10⁻⁶ M。结果表明,对于广泛的乳酸(堆积剂)和乙酸(去堆积剂)浓度,以及摄入乙醇后会对EtG的CZE行为产生影响并发生变化的大量内源性宏量和微量成分(分别为高浓度和低浓度的阴离子血清成分),都可以实现EtG与它们的分离及其定量分析。该分析方法已成功应用于以下样本中EtG、乳酸和乙酸的分析:(i)摄入已知量酒精的志愿者血清;(ii)通过分析缺糖转铁蛋白分类的患者样本(禁酒者和社交饮酒者与酗酒者)。