利用分子印記磁性聚合物搭配液相層析串聯質譜進行雜環胺類化合物分析

Abstract

過去許多文獻與臨床研究表示雜環胺類化合物具高度致癌性與突變性，希望能開發一種有效分析雜環胺類化合物的方法，分子印記技術是根據抗原-抗體理論所延伸的應用，該技術提供對目標分析物高度的專一性與低廉的成本。本篇研究目的在開發以氧化鐵作磁芯，藉由表位印記合成分子印記磁性聚合物 (Magnetic Molecularly Imprinted Polymers, MMIPs) 並應用於雜環胺類化合物的定量。本篇研究以2-Amino-3-methylimidazo[4,5-f]quinoline (IQ) 為模板分子、Ethylene glycol dimethacrylate (EGDMA)作為交聯劑、2,2′-Azobis(2-methylpropionitrile) (AIBN) 作起始劑並以ACN : MeOH (9:1, v:v) 作為致孔溶劑，以表位印記的方式將分子印記聚合物合成在奈米氧化鐵上形成MMIPs，利用傅立葉紅外光譜儀 (FT-IR)、掃描式電子顯微鏡 (SEM)、X光粉末繞射儀 (XRD) 對MMIPs進行結構解析與性質量測，最後所得MMIP成品結合固相萃取法 (SPE) 搭配高效液相層析串聯質譜儀定量真實樣品中的雜環胺類化合物。在優化了MMIPs-SPE萃取方法後，發現以MAA作官能基單體所合成出的MMIPs對IQ具有高度選擇性，且當模板/官能基單體/交聯劑比例為1:4:20時具有最好的聚合效果，在最終優化條件下，本篇研究所開發的方法提供了5種雜環胺類化合物良好的線性關係 (R >0.995)，定量極限 (LOQ) 達到0.5 ng/mL。本篇研究首次採用MMIPs與固相萃取聯用，分析肉品中的雜環胺類化合物，提供雜環胺類化合物純化的一種新策略。HCAs are highly carcinogenic and mutagenic. Molecularly imprinted technology is an extending application based on the "antigen-antibody binding theory". The study developed the Magnetic molecularly imprinted polymers (MMIPs) for the purification and quantification of HCAs. In this study, novel magnetic molecularly imprinted polymers for 2-Amino-3-methylimidazo[4,5-f]quinoline (IQ) were successfully prepared by surface molecular imprinting method using functionalized Fe particles as the magnetic cores. IQ was selected as a template molecular; Methacrylic acid (MAA), Ethylene glycol dimethyl acrylate, 2,2′-Azobis (2-methylpropionitrile) were served as the functional monomer, crosslinker, and initiator. Acetonitrile-methanol (9:1, v/v) used as the porogen for the preparation process. Fourier transform infrared spectroscopy, scanning electron microscopy, transmission electron microscope and x-ray diffraction were applied to characterize the final product of magnetic molecularly imprinted polymers. The results indicated that excellent selectivity for IQ was obtained using MAA as functional monomer. The template/functional monomer/crosslinking agent at a ratio of 1:4:20 gave better adsorption effect. Furthermore, the molecularly imprinted polymers incorporated with solid-phase extraction and the extraction conditions were optimized. Afterwards, HCAs were detected and analyzed in real samples by MMIPs-SPE and LC-MS/MS. Under the final optimized detection condition, the proposed method offered good linearity (R >0.995) for the five HCAs with acceptable relative standard deviation (RSD), and limit of quantitation (LOQ) of 0.5 ng/mL was successfully achieved.