酶联免疫法快速检测保健食品中吲哚美辛和阿西美辛  

An indirect competitive enzyme-linked immunosorbent assay for determination of indomethacin and acemetacin in health food

作　　者：刘凤银 陆曼芝 梁水源 胡玉璐 黄更生 邹红丽 李攻科 LIU Fengyin;LU Manzhi;LIANG Shuiyuan;HU Yulu;HUAGN Gengsheng;ZOU Hongli;LI Gongke(College of Biology and Food Engineering,Guangdong University of Education,Guangzhou 510303,China;School of Chemistry,Sun Yat-sen University,Guangzhou 510006,China;College of Agriculture,Shanxi Agricultural University,Taigu 030801,China)

机构地区：[1]广东第二师范学院生物与食品工程学院,广东广州510303 [2]中山大学化学学院,广东广州510006 [3]山西农业大学农学院,山西太谷030801

出　　处：《食品与发酵工业》2023年第13期266-271,共6页Food and Fermentation Industries

基　　金：广州市科技计划项目(202102080339);广东省科技创新战略专项资金项目(pdjh2021b0369)。

摘　　要：针对保健食品中吲哚美辛和阿西美辛的非法添加问题,以吲哚美辛和阿西美辛分别作为免疫半抗原和包被半抗原,通过活泼酯法与载体蛋白偶联制备完全抗原,免疫动物筛选获得了可同时识别吲哚美辛和阿西美辛的兔多克隆抗体,并建立了抗风湿类保健食品中吲哚美辛和阿西美辛同时检测的间接竞争酶联免疫分析方法。方法对吲哚美辛的检出限为0.3 ng/mL,半抑制质量浓度(IC 50)为5.6 ng/mL,线性范围为0.9~32.9 ng/mL;对阿西美辛的检出限为1.1 ng/mL,IC 50为7.4 ng/mL,线性范围为2.2~24.3 ng/mL;与多种非甾体抗炎药无交叉反应。片剂、硬胶囊和软胶囊3种剂型的抗风湿类保健食品经甲醇超声提取,通过标准稀释液稀释消除基质效应,吲哚美辛和阿西美辛的添加回收率为80.0%~119.7%,相对标准偏差低于15%;检测结果与液相色谱-串联质谱法检测结果的线性相关系数均大于0.99,结果一致性良好,可用于保健食品中吲哚美辛和阿西美辛的快速筛查。This study focused on the illegal addition of indomethacin and acemetacin in health food.Indomethacin as an immune hapten was coupled to bovine serum albumin to produce immunogen,and acemetacin as a coating hapten was coupled to ovalbumin to produce coating antigen.New Zealand white rabbits were immunized and a polyclonal antibody was obtained which could recognize both indomethacin and acemetacin.Then an indirect competitive enzyme-linked immunosorbent assay(ic-ELISA)for simultaneously detecting indomethacin and acemetacin residues in antirheumatic pain relieving health food samples was developed.The ic-ELISA demonstrated a half inhibition concentration(IC 50)of 5.6 ng/mL for indomethacin and 7.4 ng/mL for acemetacin,respectively.The linear ranges were 0.9-32.9 ng/mL with a limit of detection(LOD)of 0.3 ng/mL for indomethacin,and 2.2-24.3 ng/mL with a LOD of 1.1 ng/mL for acemetacin.No cross-reactivity was observed to all the non-steroidal anti-inflammatory drugs tested.The antirheumatic pain relieving health food in tablets,hard capsules and soft capsules were extracted with methanol and the matrix effect was eliminated by diluting with standard solutions.The average recoveries of indomethacin and acemetacin in spiked samples were 80.0%-119.7%,with relative standard deviation below 15%,respectively.Good correlation between the ic-ELISA and liquid chromatography-tandem mass spectrometry was obtained for spiked health food samples,and the correlation coefficient were greater than 0.99.This proposed ic-ELISA was suitable for rapid screening for indomethacin and acemetacin in health food samples.

关 键 词：吲哚美辛 阿西美辛 抗风湿 保健食品 间接竞争酶联免疫吸附测定 抗体 

分 类 号：TS218[轻工技术与工程—粮食、油脂及植物蛋白工程]