直接竞争酶联免疫吸附-高效液相色谱法测定烯效唑

引用本文: 张良, 王泉振, 王宁, 刘曙照. 直接竞争酶联免疫吸附-高效液相色谱法测定烯效唑[J]. 分析化学, 2012, 40(11): 1730-1734. doi: 10.3724/SP.J.1096.2012.20123 shu

Citation: ZHANG Liang, WANG Quan-Zhen, WANG Ning, LIU Shu-Zhao. Determination of Uniconazole Residue in Apple by Direct Competitive Enzyme-linked Immunosorbent Assay and High Performance Liquid Chromatography[J]. Chinese Journal of Analytical Chemistry, 2012, 40(11): 1730-1734. doi: 10.3724/SP.J.1096.2012.20123 shu

直接竞争酶联免疫吸附-高效液相色谱法测定烯效唑

张良 ^1, ,
王泉振 ^1, ,
王宁 ^1, ,
刘曙照 ^{2,
,}

通讯作者: 刘曙照

基金项目:
本文系863专题课题(No.2009AA03Z331) (No.2009AA03Z331)

国家自然科学基金(No.20777064)资助项目 (No.20777064)

摘要: 应用本实验室制备的对烯效唑具特异性亲和力的多克隆抗体,采用包被抗体直接竞争模式建立烯效唑的酶联免疫吸附分析法。在优化条件下,对烯效唑标样检测的线性范围为1-10^-4 mg/L,线性中浓度为1.31 μg/L,5次重复测定的相对标准偏差(RSD)为8.4%; 检出限为0.022 μg/L。在苹果中分别添加烯效唑标样1.0, 0.1和0.01 mg/kg; 直接竞争ELISA法测定的回收率分别为84.6%-101.0%, 99.6%-117.0%和80.8%-92.8%; RSD(n=5)分别为8.6%, 6.9%和6.7%。ELISA法对苹果中烯效唑的检出限为0.024 μg/kg。在同样前处理条件下,高效液相色谱-紫外检测(HPLC-UVD)法对苹果中烯效唑的检出限为0.25 mg/kg,苹果中添加1.00 mg/kg烯效唑,HPLC-UVD法测定的回收率为92.6%。

关键词:

English

Determination of Uniconazole Residue in Apple by Direct Competitive Enzyme-linked Immunosorbent Assay and High Performance Liquid Chromatography

1.
¹ College of Horiculture and Plant Protection, Yangzhou University, Yangzhou 225009, China;

Corresponding author: 刘曙照

Received Date: 15 February 2012
Fund Project:
本文系863专题课题(No.2009AA03Z331)

国家自然科学基金(No.20777064)资助项目

Abstract: With the high specific affinitive antibody to uniconazole prepared in our lab, the method of antibody-immobilized direct competitive ELISA was employed to determine uniconazole residue in apple. Under the optimized conditions the antibody of 4 mg/L dissolved in 0.02 mol/L pH7.6 PB was immobilized on microplate,the peroxidase-hapten conjugate was 105 times diluted in the reaction medium of 0.02 mol/L pH 6.8 PBS containing 0.15 mol/L NaCl, the liner concentrations ranged from 1 mg/L to 10^-4 mg/L, the half-maximal inhibition (IC₅₀) was 1.31 μg/L with relative standard deviation (RSD) of 8.4%, the IC₂₀ was 0.022 μg/L. The recoveries were 84.6%-101%, 99.6%-117% and 80.8%-92.8% with the RSD of 8.6%, 6.9% and 6.7% at the spiked level of 1.0, 0.1, 0.01 mg/kg in apple respectively. The limit of detection for ELISA was 0.024 μg/kg, and the limit of detection for HPLC-ultraviolet visible detector was 0.25 mg/kg.

Key words:

Uniconazole
/ Apple
/ Direct competitive enzyme-link immunosorbent assay
/ High-performance liquid chromatography

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1.
沈阳化工大学材料科学与工程学院沈阳 110142

直接竞争酶联免疫吸附-高效液相色谱法测定烯效唑

通讯作者: 刘曙照

English

Determination of Uniconazole Residue in Apple by Direct Competitive Enzyme-linked Immunosorbent Assay and High Performance Liquid Chromatography

Corresponding author: 刘曙照

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

直接竞争酶联免疫吸附-高效液相色谱法测定烯效唑

通讯作者: 刘曙照

English

Determination of Uniconazole Residue in Apple by Direct Competitive Enzyme-linked Immunosorbent Assay and High Performance Liquid Chromatography

Corresponding author: 刘曙照

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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