Development of a Magnetic Molecularly Imprinted Microsphere-Based Signal Amplified Semi-Homogeneous Method for Multidetection of Five Progestins in Milk

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.