CAJ | 학술논문

丙烯酰胺由于分子量小、结构简单,制备其特异性抗体难以实现。本研究将丙烯酰胺与对巯基苯乙酸衍生合成半抗原,偶联载体蛋白并免疫动物制备针对丙烯酰胺衍生物的特异性抗体,并进行辣根过氧化物酶标记,进而建立通过检测丙烯酰胺衍生物实现对丙烯酰胺定量分析的直接竞争酶联免疫分析方法。本方法对丙烯酰胺的检出限为3.0μg/L,线性范围为9.2~195μg/L,对饼干、薯片及咖啡样品中丙烯酰胺的平均添加回收率为83.6%~112.7%,结果与标准检测方法HPLC-MS/MS符合。本方法可用于食品样品中丙烯酰胺的快速检测。
병희선알유우분자량소、결구간단,제비기특이성항체난이실현。본연구장병희선알여대구기분을산연생합성반항원,우련재체단백병면역동물제비침대병희선알연생물적특이성항체,병진행랄근과양화물매표기,진이건립통과검측병희선알연생물실현대병희선알정량분석적직접경쟁매련면역분석방법。본방법대병희선알적검출한위3.0μg/L,선성범위위9.2~195μg/L,대병간、서편급가배양품중병희선알적평균첨가회수솔위83.6%~112.7%,결과여표준검측방법HPLC-MS/MS부합。본방법가용우식품양품중병희선알적쾌속검측。
Due to the low molecular weight and simple structure, the production of specific antibodies against acrylamide is unavailable. In this study, a novel hapten was synthesized through the derivatization of acrylamide and 4-mercaptophenylacetic acid. The hapten was then coupled to carrier protein and used to immunize New Zealand rabbits. Polyclonal antibody which showed specific binding to the acrylamide derivative ( hapten) was obtained. The antibody was labeled with horseradish peroxidase ( HRP) and used to develop a direct competitive enzyme-linked immunosorbent assay ( dc-ELISA) . The dc-ELISA was used to determine the content of acrylamide derivative, and then transferred to the content of acrylamide. The assay showed an IC50 value of 45. 49 μg/L, a limit of detection of 3. 0 μg/L and the linear range of 9. 2-195 μg/L for acrylamide. The recovery of acrylamide from spiked food sample was determined ranging from 83 . 6% to 112 . 7%. Good correlations between the results of dc-ELISA and standard HPLC-MS/MS were obtained. The proposed dc-ELISA is suitable for the determination of acrylamide in food samples.