Su Yan, Liu Gelin, Hou Haozhe, Peng Yaojia, Wang Jianping
College of Veterinary Medicine, Hebei Agricultural University, Baoding 071000, China.
Foods. 2023 Jul 25;12(15):2818. doi: 10.3390/foods12152818.
The residues of progestins in milk are significant risk factors for teenage acne and may cause hormone-dependent cancers in consumers, so the determination of these residues in milk is very important. However, an immunoassay or immunoassay-like method capable of determining multiple progestins in milk has not been reported so far. The present study, for the first time, synthesized a type of magnetic molecularly imprinted microsphere that was capable of simultaneously recognizing five progestins. At the same time, an enzyme labeled conjugate was synthesized by coupling progesterone 3-(o-carboxymethyl)oxime with streptavidinated horseradish peroxidase. The above two reagents were used to develop a semi-homogeneous method for the simultaneous detection of the residues of the five progestins in milk. During the experiments, biotinylated horseradish peroxidase was used to amplify the signal, so the sensitivity to the five drugs (limits of detection 0.04-0.1 pg/mL) was increased 44-75-fold. In addition, the magnetic molecularly imprinted microsphere could be regenerated four times by using simple elution. Through general comparison of its detection spectrum, sensitivity, simplicity, and reusability, the present method exhibited better performance than the previous immunoassays for the detection of progestins, and so it could be used as a routine tool for the screening of progestins residues in milk.
牛奶中孕激素残留是青少年痤疮的重要风险因素，且可能导致消费者患激素依赖性癌症，因此测定牛奶中的这些残留非常重要。然而，目前尚未见有能够测定牛奶中多种孕激素的免疫分析或类似免疫分析的方法报道。本研究首次合成了一种能够同时识别五种孕激素的磁性分子印迹微球。同时，通过将孕酮3-(邻羧甲基)肟与链霉亲和素化辣根过氧化物酶偶联，合成了一种酶标记物。利用上述两种试剂建立了一种半均相法，用于同时检测牛奶中五种孕激素的残留。实验过程中，使用生物素化辣根过氧化物酶放大信号，因此对五种药物的灵敏度（检测限为0.04 - 0.1 pg/mL）提高了44 - 75倍。此外，磁性分子印迹微球通过简单洗脱可重复使用四次。通过对其检测谱、灵敏度、简便性和可重复使用性的综合比较，本方法在检测孕激素方面比以往的免疫分析方法表现更优，因此可作为筛查牛奶中孕激素残留的常规工具。
