Preparation of molecularly imprinted polymer nanoparticles for the extraction and purification of progesterone combined with UPLC-ESI-MS mass spectrometry detection in royal jelly

This study aimed to develop molecularly imprinted polymer (MIP) nanoparticles specifically for the selective extraction and enrichment of progesterone (P) from royal jelly (RJ), and quantitatively analyzed them by ultra-performance-liquid chromatography electrospray ionization mass spectrometry (UPLC-ESI-MS). Gaussian software-based theoretical calculations identified methacrylic acid (MAA) as the optimal functional monomer for its strong binding affinity to P. MIP was synthesized by precipitation polymerization, and the preparation process of MIP was optimized by one-way variance design and response surface methodology. The optimal formulation was determined to be 0.26 mmol of P, 0.77 mmol of MAA, and 3.0 mmol of trimethylolpropane trimethacrylate (TRIM). The synthesized MIPs demonstrated a maximum adsorption capacity of 34.41 mg/g. The detection limit of P by mass spectrometry was 1 ng/ml. In RJ samples, P content ranged from 0.02 mg/g to 1.53 mg/g, with an average content of 0.266 mg/g. The study successfully developed a stable and reliable MIP, providing an effective method for the selective enrichment and detection of P in complex samples.