Sensitive detection of α-amylase based on host-guest inclusion system of γ-cyclodextrin and dansyl-derived diphenylalanine.

Yu Liu, Yutian Jiao, Longjun Xiong, Gongli Wei, Baocai Xu, Guiju Zhang, Ce Wang, Li Zhao
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Abstract

A highly sensitive detection system for α-amylase was developed via host-guest complexation between γ-cyclodextrin and dansyl-modified diphenylalanine (FF-Dns). The host-guest inclusion of FF-Dns into the cavity of γ-CD in a HEPES buffer solution (10 mM, pH 7.4) significantly enhanced the fluorescence intensity, and the emission wavelength gradually shifted from 558 to 535 nm. The hydrolysis of γ-CD by the addition of α-amylase released FF-Dns, leading to the recovery of the fluorescence emission characteristics. Therefore, the FF-Dns/γ-CD host-guest complexation system can serve as a platform for the sensitive detection of α-amylase with good selectivity against potential interference. The limit of detection (LOD) of the system was 0.004 U/mL, with a linear working range of 0-6 U/mL. The detection assay was successfully applied in 0.1 % serum, achieving an LOD of 0.017 U/mL and a linear working range of 0-10 U/mL.

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基于γ-环糊精和丹酰衍生二苯丙氨酸的主-客包合系统的α-淀粉酶灵敏检测。
通过γ-环糊精和丹酰改性二苯基丙氨酸(FF-Dns)之间的主-客复合物,开发了一种高灵敏度的α-淀粉酶检测系统。在 HEPES 缓冲溶液(10 mM,pH 7.4)中,将 FF-Dns 加入到 γ-CD 的空腔中,可显著增强荧光强度,发射波长从 558 nm 逐渐转移到 535 nm。加入α-淀粉酶后,γ-CD被水解,释放出FF-Dns,从而恢复了荧光发射特性。因此,FF-Dns/γ-CD 主-客复合物系统可作为灵敏检测α-淀粉酶的平台,对潜在干扰具有良好的选择性。该系统的检测限(LOD)为 0.004 U/mL,线性工作范围为 0-6 U/mL。该检测方法成功地应用于 0.1 % 血清中,检测限为 0.017 U/mL,线性工作范围为 0-10 U/mL。
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