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首页> 外文期刊>Analytica chimica acta >Analysis of urinary neurotransmitters by capillary electrophoresis: Sensitivity enhancement using field-amplified sample injection and molecular imprinted polymer solid phase extraction
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Analysis of urinary neurotransmitters by capillary electrophoresis: Sensitivity enhancement using field-amplified sample injection and molecular imprinted polymer solid phase extraction

机译：毛细管电泳分析尿中神经递质：使用现场放大的样品进样和分子印迹聚合物固相萃取提高灵敏度

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Capillary electrophoresis (CE) has been investigated for the analysis of some neurotransmitters, dopamine (DA), 3-methoxytyramine (3-MT) and serotonin (5-hydroxytryptamine, 5-HT) at nanomolar concentrations in urine. Field-amplified sample injection (FASI) has been used to improve the sensitivity through the online pre-concentration samples. The cationic analytes were stacked at the capillary inlet between a zone of low conductivity - sample and pre-injection plug - and a zone of high conductivity - running buffer. Several FASI parameters have been optimized (ionic strength of the running buffer, concentration of the sample protonation agent, composition of the sample solvent and nature of the pre-injection plug). Best results were obtained using H3PO4-LiOH (pH 4, ionic strength of 80 mmol L~(-1)) as running buffer, 100 μmol L~(-1) of H3PO4 in methanol-water 90/10 (v/v) as sample solvent and 100 μmol L~(-1) of H3PO4 in water for the pre-injection plug. In these conditions, the linearity was verified in the 50-300 nmol L~(-1) concentration range for DA, 3-MT and 5-HT with a determination coefficient (r~2) higher than 0.99. The limits of quantification (10 nmol L~(-1) for DA and 3-MT, 5.9 nmol L~(-1) for 5-HT) were 500 times lower than those obtained with hydrodynamic injection. However, if this method is applied to the analysis of neurotransmitters in urine, the presence of salts in the matrix greatly reduces the sensitivity of the FASI/CE-UV method/Therefore, a solid phase extraction (SPE) on a dedicated imprinted polymer (MIP) was developed to extract specific neurotransmitters, catecholamines, metanephrines and indolamines, from urine. Matrix salts were thus discarded after sample extraction on AFFINIMIP? Catecholamine & Metanephrine (100 mg) cartridge. Therefore, lower limits of quantification were determined in artificial urine (46 nmol L~(-1) for DA, 11 nmol L~(-1) for 3-MT and 6 nmol L~(-1) for 5-HT).The application of this protocol M1P-SPE/FASI-CE-UV analysis of neurotransmitters in human urine gave rise to electropherograms with a very good base line and signal to noise ratios above 15.
机译：已经研究了毛细管电泳（CE），用于分析尿液中纳摩尔浓度的一些神经递质，多巴胺（DA），3-甲氧基酪胺（3-MT）和5-羟色胺（5-羟色胺，5-HT）。现场放大样品进样（FASI）已用于通过在线预浓缩样品提高灵敏度。阳离子分析物堆积在毛细管入口处，该区域位于低电导率区域（样品和预进样塞）和高电导率区域之间（运行缓冲液之间）。已优化了几个FASI参数（运行缓冲液的离子强度，样品质子化剂的浓度，样品溶剂的组成和预进样塞的性质）。使用H3PO4-LiOH（pH 4，离子强度为80 mmol L〜（-1））作为运行缓冲液，100μmolL〜（-1）的H3PO4在甲醇/水90/10（v / v）中获得最佳结果作为样品溶剂和100μmolL〜（-1）的H3PO4在水中作为预进样塞。在这些条件下，在50-300 nmol L〜（-1）浓度范围内对DA，3-MT和5-HT的线性进行了验证，测定系数（r〜2）大于0.99。定量限（DA和3-MT的定量限为10 nmol L〜（-1），5-HT的定量限为5.9 nmol L〜（-1））比水力注射法低500倍。但是，如果将此方法应用于尿液中神经递质的分析，则基质中盐的存在会大大降低FASI / CE-UV方法的灵敏度，因此，需要在专用的印迹聚合物上进行固相萃取（SPE）（开发了MIP以从尿液中提取特定的神经递质，儿茶酚胺，间肾上腺素和吲哚胺。因此，在AFFINIMIP？上提取样品后，将基质盐丢弃。儿茶酚胺和肾上腺素（100毫克）药筒。因此，在人造尿液中确定了较低的定量限（DA的定量值为46 nmol L〜（-1），3-MT的定量值为11 nmol L〜（-1），5-HT的定量值为6 nmol L〜（-1））。该协议的应用M1P-SPE / FASI-CE-UV对人尿液中神经递质的分析产生了具有非常好的基线和高于15的信噪比的电泳图。

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