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首页> 外文期刊>The Analyst: The Analytical Journal of the Royal Society of Chemistry: A Monthly International Publication Dealing with All Branches of Analytical Chemistry >Electrochemical sensor with substitutional stripping voltammetry for highly sensitive endotoxin assay
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Electrochemical sensor with substitutional stripping voltammetry for highly sensitive endotoxin assay

机译：电化学传感器，带取代溶出伏安法，用于高度灵敏的内毒素测定

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摘要

We have developed a novel method for detection of endotoxin with extra-high sensitivity by using substitutional stripping voltammetry (SSV). In this method, a p-aminophenol (pAP) conjugated peptide (Boc-Leu-Gly-Arg-pAP; LGR-pAP) was used as a substrate for a protease, which is activated at the last step of the endotoxin-induced Limulus amebocyte lysate (LAL) cascade reaction. Extra-highly sensitive detection of pAP liberated by the endotoxin-induced LAL reaction was successfully realized with SSV, based on the accumulation of an amperometric signal owing to exchange of the oxidation current of pAP generated at an electrode in a reaction cell with silver deposition on another electrode in a deposition cell. This reaction is driven by the difference in the redox potential between pAP/ quinoneimine and silver/silver ion. The amount of the deposited silver is quantified by anodic stripping voltammetry (ASV). This SSV-based endotoxin assay was performed with a chip device comprising two cells, each of which was connected via a liquid junction made of Vycor? glass. The reaction cell and the deposition cell contained a standard endotoxin sample with LAL regents containing LGR-pAP and AgNO_3 solution, respectively. After the cells were electrically connected for 60 min, ASV was conducted in the deposition cell to quantify the total electrical charge derived by the oxidation of free pAP in the reaction cell. The ASV signal increased with the increase of the endotoxin concentration in the sample solution in the range of 0.5–1000 EU L~(-1).
机译：我们已经开发出一种新的方法，可通过使用替代溶出伏安法（SSV）来以极高的灵敏度检测内毒素。在此方法中，将对氨基苯酚（pAP）偶联肽（Boc-Leu-Gly-Arg-pAP; LGR-pAP）用作蛋白酶的底物，该蛋白酶在内毒素诱导的induced的最后一步被激活变形细胞裂解液（LAL）级联反应。基于安培信号的积累，SSV成功实现了由内毒素诱导的LAL反应释放的pAP的超高灵敏度检测，这是由于交换了在反应池中电极上产生的pAP的氧化电流而使银沉积在反应池上而产生的沉积池中的另一个电极。该反应是由pAP /醌亚胺和银/银离子之间的氧化还原电势差驱动的。沉积的银量通过阳极溶出伏安法（ASV）进行定量。该基于SSV的内毒素测定法是用包含两个细胞的芯片装置进行的，每个细胞通过由Vycor？制成的液体接头连接。玻璃。反应池和沉积池包含标准内毒素样品，其中的LAL试剂分别含有LGR-pAP和AgNO_3溶液。在将电池电连接60分钟后，在沉积电池中进行ASV，以量化反应电池中游离pAP氧化产生的总电荷。随着样品溶液中内毒素浓度的增加，ASV信号在0.5–1000 EU L〜（-1）范围内增加。

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