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基于植物亚硫酸盐氧化酶生物传感器的食品中亚硫酸盐分析的快速灵敏 HPLC 方法。

A fast and sensitive HPLC method for sulfite analysis in food based on a plant sulfite oxidase biosensor.

机构信息

Institute of Nutrition and Food Sciences, University of Bonn, Endenicher Allee 11-13, 53115 Bonn, Germany.

出版信息

Biosens Bioelectron. 2010 Sep 15;26(1):175-81. doi: 10.1016/j.bios.2010.06.009. Epub 2010 Jun 12.

Abstract

A reliable and sensitive analysis of sulfites in food is essential in food monitoring. However, the established methods exhibit deficiencies in the very low concentration ranges (below 10 mg/L SO(2)), especially with more complex food matrices. With a focus on these challenges, an HPLC method with immobilized enzyme reactor (HPLC-IMER) for the analysis of sulfites in food was optimized and compared to a standard method. A modulated sample preparation procedure and the use of a novel sulfite oxidase from Arabidopsis thaliana were explored to make the method applicable for most food samples. The plant sulfite oxidase turned out to be superior to the commercially available animal sulfite oxidase in terms of detection limit (0.01 mg/L SO(2)), linear range (0.04-20 mg/L SO(2)) and stability. In a small scale comparison within our laboratory, as well as in a standardized proficiency testing, the HPLC-IMER was compared to an established distillative method. The enzyme-based method is not only more sensitive and specific, it also yields higher sulfite recoveries in almost all samples while exhibiting better statistic method parameters.

摘要

在食品监测中,可靠且灵敏的亚硫酸盐分析至关重要。然而,已建立的方法在非常低的浓度范围内(低于 10mg/L SO(2))存在缺陷,特别是在更复杂的食品基质中。本研究针对这些挑战,优化了一种基于固定化酶反应器(HPLC-IMER)的高效液相色谱法,用于分析食品中的亚硫酸盐,并与标准方法进行了比较。通过优化调制样品制备程序和使用新型拟南芥亚硫酸盐氧化酶,该方法适用于大多数食品样品。与商业上可用的动物亚硫酸盐氧化酶相比,植物亚硫酸盐氧化酶在检测限(0.01mg/L SO(2))、线性范围(0.04-20mg/L SO(2))和稳定性方面表现出优势。在我们实验室内部的小规模比较以及标准化能力验证中,将 HPLC-IMER 与已建立的蒸馏方法进行了比较。基于酶的方法不仅更灵敏、更特异,而且在几乎所有样品中都能获得更高的亚硫酸盐回收率,同时表现出更好的统计方法参数。

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