CAJ | 학술논문

对呋喃妥因代谢物1－氨基乙内酰脲（ AHD）进行半抗原改造，采用活化酯法将半抗原与卵清蛋白（ OVA）偶联为包被原，与标准品竞争AHD单克隆抗体的抗原结合位点，加入辣根过氧化物酶标记羊抗鼠二抗，建立了AHD间接竞争化学发光酶联免疫（ CLEIA）检测法，并对化学发光液、包被原与抗体最优稀释度、包被条件、封闭液和竞争时间五项参数进行优化。结果表明：该方法具有良好的特异性， IC50为0．753 ng／mL，在鸡肉组织中的检测限为0．028μg／kg，添加回收率在80．54％～102．64％之间，变异系数均小于10％。与国标中液相色谱－串联质谱法（ LC－MS／MS）和我国出入境行业标准中酶联免疫检测法（ ELISA）相比，不仅降低了检测限，而且操作简便，为动物源性食品中呋喃妥因代谢物的残留提供了准确、便捷的分析检测手段。
대부남타인대사물1－안기을내선뇨（ AHD）진행반항원개조，채용활화지법장반항원여란청단백（ OVA）우련위포피원，여표준품경쟁AHD단극륭항체적항원결합위점，가입랄근과양화물매표기양항서이항，건립료AHD간접경쟁화학발광매련면역（ CLEIA）검측법，병대화학발광액、포피원여항체최우희석도、포피조건、봉폐액화경쟁시간오항삼수진행우화。결과표명：해방법구유량호적특이성， IC50위0．753 ng／mL，재계육조직중적검측한위0．028μg／kg，첨가회수솔재80．54％～102．64％지간，변이계수균소우10％。여국표중액상색보－천련질보법（ LC－MS／MS）화아국출입경행업표준중매련면역검측법（ ELISA）상비，불부강저료검측한，이차조작간편，위동물원성식품중부남타인대사물적잔류제공료준학、편첩적분석검측수단。
The haptene of nitrofurantoin metabolite ( AHD) was modified by hapten, then made it conjugate with ovalbmin ( OVA ) by using N-hyduoxysuccinimide ester method to synthesize envelope antigen. Then the compound was coupled to AHD monoclonal antibody competitively with standard reference. After the HRP labeled goat-anti-mouse secondary antibody coupled to the monoclonal antibody which conjugated with envelope antigen, an indirect competitive chemiluminescent enzyme immunoassay ( CLEIA) for AHD was established. Then five test parameters, such as chemiluminescent solution, envelope antigen concentration and antibody concentration, enveloped condition, sealed liquid and competitive reaction time were optimized. The results indicated that the AHD CLEIA had good specificity for AHD.We obtained an IC50 value of 0.753 ng/mL.We also obtained the limit of detection of 0.028μg/kg and recoveries rates of AHD ranged from 80.54% to 102.64% in spiked chicken samples. The coefficient of variation was less than 10%. Compared with LC-MS/MS in national standards and ELISA in industry standard for CIQ, this method not only improved the detection limit, but also operated easily. It could provide a convenient and accurate way for detection of AHD, which will have great value for the evaluation of food safety.