CAJ | 학술논문

目的：建立一种用于检测牛奶中雌二醇的酶热传感技术。方法羧基化雌二醇经1-乙基-(3-二甲基氨基丙基)碳酰二亚胺盐酸盐(EDC)与β-内酰胺酶偶联。酶热传感器中装SPG酶柱,通过ELISA原理使游离雌二醇与结合雌二醇竞争结合雌二醇抗体,从而检测基质中的雌二醇浓度。优化酶热传感检测体系中的反应底物、系统流速、抗原抗体稀释比例等实验因素。结果磷酸盐缓冲液(PBS)基质中测定IC50值为10.2 ng/mL,线性检测范围：5.1~19.4 ng/mL,最低检测限2.8 ng/mL,变异系数为3.4%,与多种雌激素类结构类似物之间均无交叉反应,特异性良好。牛奶样品中测定IC50值：0.45 ng/mL,线性检测范围：0.24~0.79 ng/mL,最低检测限0.12 ng/mL。结论酶热传感技术可以快速检测牛奶中的雌二醇残留,操作便捷、样品前处理简单,是一种高效、经济的适用于食品安全工作现场检测的方法。
목적：건립일충용우검측우내중자이순적매열전감기술。방법최기화자이순경1-을기-(3-이갑기안기병기)탄선이아알염산염(EDC)여β-내선알매우련。매열전감기중장SPG매주,통과ELISA원리사유리자이순여결합자이순경쟁결합자이순항체,종이검측기질중적자이순농도。우화매열전감검측체계중적반응저물、계통류속、항원항체희석비례등실험인소。결과린산염완충액(PBS)기질중측정IC50치위10.2 ng/mL,선성검측범위：5.1~19.4 ng/mL,최저검측한2.8 ng/mL,변이계수위3.4%,여다충자격소류결구유사물지간균무교차반응,특이성량호。우내양품중측정IC50치：0.45 ng/mL,선성검측범위：0.24~0.79 ng/mL,최저검측한0.12 ng/mL。결론매열전감기술가이쾌속검측우내중적자이순잔류,조작편첩、양품전처리간단,시일충고효、경제적괄용우식품안전공작현장검측적방법。
Objective To establish an enzyme thermistor technique for determination of estradiol in milk. Methods Carboxyl transformation and coupled estradiol withβ-lactamase through 1-ethyl-(3-dimethyl amino propyl) carbonyl imine hydrochloride (EDC). SPG was installed in thermal sensor as enzyme column, and direct competition ELISA principle was used to detect estradiol concentrations in matrix. Experimental factors such as the reaction substrate, the flow rate of the system, and the antigen-antibody dilution ratio were optimized.Results The linerity range was 5.1~19.4 ng/mL, IC50 was 10.2 ng/mL and the detection limit was 2.8 ng/mL in PBS sample. The result was stable, reliable and specific, no cross-reaction could be demonstrated with a variety of estrogen analogues and coefficient variation (CV) was 3.4%. The linerity range was 0.24~0.79 ng/mL, IC50was 0.45 ng/mL, and the detection limit was 0.12 ng/mL in milk sample.Conclusion This method is successfully applied to detect estradiol in milk, it is simple, efficient, economic, and suitable for the on-site use.