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酶修饰毛细管作为平台用于同时荧光检测 d-葡萄糖和 l-乳酸。

An enzyme-modified capillary as a platform for simultaneous fluorometric detection of d-glucose and l- lactate.

机构信息

Department of Chemistry, College of Humanities and Sciences, Nihon University, Sakurajousui, Setagaya, Tokyo, 156-8550, Japan; School of Pharmacy, Tokyo University of Pharmacy and Life Science, 1432-1 Horinouchi, Hachioji, Tokyo, 192-0392, Japan.

Department of Chemistry, College of Humanities and Sciences, Nihon University, Sakurajousui, Setagaya, Tokyo, 156-8550, Japan.

出版信息

J Pharm Biomed Anal. 2019 Jan 30;163:1-8. doi: 10.1016/j.jpba.2018.09.028. Epub 2018 Sep 19.

Abstract

The preparation of a glass capillary pattered with lipid layers on which lactate dehydrogenase (LDH) and glucose dehydrogenase (GDH) were regionally adsorbed and its application for simultaneous detection of d-glucose and l-lactate in human serum is described. A lipid layer was formed on the surface of BSA-unabsorbed octadecyltrichlorosilane (OTS) inner wall of a glass capillary. The electrostatic charge of the lipid layer was a key factor for adsorbing the enzymes on the lipid layer. The fluorescence intensities were observed at each enzyme site in the presence of diaphorase (DIA), β-nicotinamide-adenine dinucleotide oxidized (NAD), resazurin, d-glucose and l-lactate. The fluorescence intensities at each enzyme site increased with an increase in the concentration of d-glucose and l-lactate=with the detection limits of 32 μM and 4.9 μM, respectively.

摘要

描述了一种玻璃毛细管的制备方法,该毛细管的表面有脂质层图案,其中乳酸脱氢酶(LDH)和葡萄糖脱氢酶(GDH)被区域吸附,可用于同时检测人血清中的 d-葡萄糖和 l-乳酸。在玻璃毛细管的 BSA 未吸附的十八烷基三氯硅烷(OTS)内壁上形成一层脂质层。脂质层的静电荷是将酶吸附到脂质层上的关键因素。在存在二氢还蛋白(DIA)、β-烟酰胺腺嘌呤二核苷酸氧化型(NAD)、resazurin、d-葡萄糖和 l-乳酸的情况下,观察到每个酶位的荧光强度。随着 d-葡萄糖和 l-乳酸浓度的增加,每个酶位的荧光强度增加,检测限分别为 32μM 和 4.9μM。

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