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用于酶法测定特定食品和饮料中蔗糖和D-葡萄糖的Enzytec™液体组合蔗糖/D-葡萄糖试剂的验证:2024年5月首次行动

Validation of Enzytec™ Liquid Combi Sucrose/D-Glucose for Enzymatic Determination of Sucrose and D-Glucose in Selected Foods and Beverages: First Action 2024.05.

作者信息

Lacorn Markus, Hektor Thomas

机构信息

R-Biopharm AG, R&D Food & Feed, An der Neuen Bergstr. 17, Germany.

出版信息

J AOAC Int. 2025 Mar 1;108(2):151-172. doi: 10.1093/jaoacint/qsae100.

Abstract

BACKGROUND

Sucrose is produced in the greatest quantity of all industrially produced organic substances and can be used in almost all processed foods. Glucose is the most abundant monosaccharide and contained in many foodstuffs naturally or added as an ingredient.

OBJECTIVE

To validate the performance of the Enzytec™ Liquid Combi Sucrose/D-Glucose test kit for the determination of sucrose and D-glucose in juices, chocolate, breakfast cereals, ice cream, sweetened condensed milk, wine, beer, and soft drinks.

METHODS

The kit contains all reagents in a ready-to-use format and is suitable for automation. Sucrose is cleaved by β-fructosidase. The resulting D-glucose from sucrose and glucose originally in the sample are phosphorylated and react in a NADH-generating reaction afterwards. β-Fructosidase is not present in the glucose system. The amount of NADH produced is equivalent to the amounts of sucrose and D-glucose and is measured at 340 nm within 30 min.

RESULTS

The linear measurement range for a 100 µL test volume is from 17 to 2500 mg/L sucrose and 4 to 2000 mg/L D-glucose. Trueness was checked by using four Certified Reference Materials (CRM) and resulted in recoveries from 96 to 105%. Spiking of juices, ice cream, sweetened condensed milk, and shandy resulted in recoveries between 98 and 106% for both systems. Intermediate precision was 3% or lower for sucrose and glucose. For automation, three applications with different test volumes were validated. Linearity is given from 3.8 to 9500 mg/L for sucrose and 2.4 up to 10000 mg/L for glucose.

CONCLUSION

The method is robust and accurate for manual and automated applications. The method was approved as an AOAC Official Method of Analysis℠.

HIGHLIGHTS

The ready-to-use components of the test kit have a shelf of at least 29 months.

摘要

背景

蔗糖是所有工业生产的有机物质中产量最大的,几乎可用于所有加工食品。葡萄糖是最丰富的单糖,天然存在于许多食品中或作为一种成分添加。

目的

验证Enzytec™ 液体组合蔗糖/D-葡萄糖检测试剂盒在测定果汁、巧克力、早餐谷物、冰淇淋、甜炼乳、葡萄酒、啤酒和软饮料中蔗糖和D-葡萄糖的性能。

方法

该试剂盒包含所有即用型试剂,适用于自动化检测。蔗糖被β-果糖苷酶裂解。蔗糖产生的D-葡萄糖和样品中原有的葡萄糖被磷酸化,随后在产生NADH的反应中发生反应。葡萄糖检测体系中不存在β-果糖苷酶。产生的NADH量与蔗糖和D-葡萄糖的量相当,并在30分钟内在340nm处进行测量。

结果

100µL检测体积的线性测量范围为蔗糖17至2500mg/L,D-葡萄糖4至2000mg/L。通过使用四种有证标准物质(CRM)检查了准确性,回收率为96%至105%。对果汁、冰淇淋、甜炼乳和混合啤酒进行加标,两个检测体系的回收率均在98%至106%之间。蔗糖和葡萄糖的中间精密度为3%或更低。对于自动化检测,验证了三种不同检测体积的应用。蔗糖的线性范围为3.8至9500mg/L,葡萄糖的线性范围为2.4至10000mg/L。

结论

该方法对于手动和自动化应用均稳健且准确。该方法已被批准为AOAC官方分析方法℠。

要点

检测试剂盒的即用型组件保质期至少为29个月。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eb1c/11879208/72b10c0ca2cc/qsae100f1.jpg

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