作者：李兰英¹, 徐勤¹, 许丽¹, 闻艳丽¹, 武利庆², 任淑贞¹, 刘刚¹

作者单位：1. 上海市计量测试技术研究院, 上海 201203;
2. 中国计量科学研究院, 北京 100029

关键词：铁(III);8-羟基喹啉;三磷酸腺苷(ATP);化学发光分析仪;计量校准

摘要：

化学发光分析仪检测的是微弱光信号,传统的物理标准光源光强度往往超出其检测范围,难以用于计量校准.为此,该文建立并优化两种生物化学模拟发光标准体系.将8-羟基喹啉(8-Ox)引入鲁米诺-过氧化氢-金属离子体系中,实现发光方式由闪光型到辉光型的转变,在100s内发光值的相对标准偏差仅为1.1%;由此建立发光强度稳定的化学发光标准体系,利用此体系考察化学发光分析仪的重复性、交叉污染、线性相关系数.利用三磷酸腺苷(ATP)-荧光素-荧光素酶反应,建立生物发光标准体系,考察化学发光分析仪检测灵敏度.为化学发光分析仪日常的校准工作以及国家计量校准规范的制订提供实验基础.

Research of a biochemical reaction system for the calibration of the microplate luminometer

LI Lanying¹, XU Qin¹, XU Li¹, WEN Yanli¹, WU Liqing², REN Shuzhen¹, LIU Gang¹

1. Shanghai Institute of Measurement and Testing Technology, Shanghai 201203, China;
2. National Institute of Metrology, Beijing 100029, China

Abstract: Microplate Luminometer detects very weak optical signal, which is much lower than the traditional physical standard illuminant. In order to meet the requirement of the calibration for Microplate Luminometer, we establish a stable biochemical luminescence standard system, with proper light intensity, high stability and clear traceability. Firstly, a chemiluminescence standard system of luminol, H₂O₂ and Fe(III) in the presence of 8-hydroxyquinoline has been developed. In this system, 8-hydroxyquinoline inhibited the reaction activity of Fe(III) through complexation, and thus change the flash luminescence to stable glow luminescence. The system was used to research on the metrological properties which including repeatability of measurement, cross contamination, linear correlation coefficient. Secondly, a bioluminescence standard system was established based on the reaction system of ATP, luciferin and luciferase, to research the sensibility of microplate luminometer. Results of this work provide novel experimental support for the estabilishment of the national metrology calibration specification and routine calibration of microplate luminometers.

Keywords: Fe(III);8-hydroxyquinoline;ATP;microplate luminometer;calibration

2015, 41(3): 46-50  收稿日期: 2014-10-11;收到修改稿日期: 2014-12-7

基金项目: 国家质检总局科研计划项目(2013QK132)

作者简介: 李兰英(1984-),女,山东阳谷县人,工程师,硕士,主要从事兽药残留基体标准物质的研制和非标仪器校准方法的研究.

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