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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >On-line coupling of size exclusion and capillary zone electrophoresis via a reversed-phase C18 trapping column for the analysis of structurally related enkephalins in cerebrospinal fluid
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On-line coupling of size exclusion and capillary zone electrophoresis via a reversed-phase C18 trapping column for the analysis of structurally related enkephalins in cerebrospinal fluid

机译：通过反相C18捕集柱在线耦合尺寸排阻和毛细管区带电泳，以分析脑脊液中结构相关的脑啡肽

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摘要

On-line coupled analytical techniques can be advantageous in the assay of smaller peptides in complex biological matrices such as plasma, cerebrospinal fluid (CSF) and tissues. The present study shows the feasibility of a recently developed system, consisting of a size-exclusion chromatographic (SEC) separation followed by a trapping procedure on an RP18 microcolumn with subsequent elution of the trapped fraction and separation by capillary zone electrophoresis (CZE) for the quantification of structural-related peptides in biological matrices, as demonstrated for a number of enkephalins in CSF. After SEC separation of the enkephalins from large proteins present in CSF a heart-cut of 200 muL, containing the enkephalin peak, is taken, concentrated on the RP18 microcolumn and, after elution of the enkephalins with 80% acetonitrile, a fraction of the eluate is electrokinetically injected into the CZE system, where stacking and separation is achieved. The degradation of the peptides, caused by endogenous peptidases in the matrix, is sufficiently inhibited with imipramine HCl. The assay has a satisfactory linearity and intraday (9.70-16.3%) precision considering the complexity of this multidimensional separation system. The sensitivity of the method, with a concentration limit of quantification of 2.5 mug/mL, is comparable with other CZE assays for peptides and sufficient for the quantification of peptide drugs in biological matrices. [References: 32]
机译：在线耦合分析技术在复杂的生物基质（例如血浆，脑脊液（CSF）和组织）中较小肽的测定中可能是有利的。本研究显示了最近开发的系统的可行性，该系统包括尺寸排阻色谱（SEC）分离，然后在RP18微柱上进行捕获，然后洗脱所捕获的馏分，并通过毛细管区带电泳（CZE）进行分离。定量分析生物基质中与结构相关的肽，如脑脊液中许多脑啡肽所证明的。从脑脊液中存在的大蛋白中对脑啡肽进行SEC分离后，将包含脑啡肽峰的200μL心切下来，浓缩在RP18微柱上，用80％乙腈洗脱脑啡肽后，洗脱液的一部分电动注入CZE系统，实现堆叠和分离。由丙咪嗪盐酸盐充分抑制由基质中的内源肽酶引起的肽降解。考虑到这种多维分离系统的复杂性，该测定具有令人满意的线性和日内（9.70-16.3％）精度。该方法的灵敏度为2.5马克/毫升的定量限，可与其他CZE肽分析方法相媲美，并且足以定量生物基质中的肽药物。 [参考：32]

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