黄曲霉毒素（AFT）是一类严重危害食品安全的霉菌毒素，免疫亲和柱-高效液相色谱分析（IAC-HPLC）作为检测AFT的经典方法，存在基质干扰及柱容量实时监测的难题。为探索解决以上弊端，本研究以AFT单抗为亲和配体，从三种噬菌体肽库高通量淘选AFT模拟肽群，经DNA测序并化学合成AFT模拟肽，从中筛选理化特性与AFT相近，但与AFT单抗的亲和力有显著差异的AFT模拟肽组合（高亲和力/低亲和力>10）；在此基础上，采用理化性质、荧光光谱与AFT相同的AMCA-NHS对其进行荧光标记，以荧光标记的高亲和力模拟肽为定量内标，荧光标记的低亲和力模拟肽为柱容量实时监测内标，建立一种可实时监测柱容量的IAC-HPLC内标分析法；并以同位素稀释串联液质法为金标准，开展与常规IAC-HPLC的比较研究，有望解决基质干扰和柱容量实时监测的难题，为提高IAC-HPLC分析方法的准确度提供新思路。

Aflatoxins (AFT) are a series of harmful mycotoxins. The immunoaffinity column-high pressure liquid chromatography (IAC-HPLC) is a typical analysis method. But the method is vulnerable to complex sample matrix interference. Furthermore, the capacity of the IAC cannot be monitored in real time is also a shortcoming. In this study, a group of mimic peptides of aflatoxins would be acquired by panning from three phage random peptide libraries with an anti-AFT monoclonal antibody. After DNA sequencing, those peptides will be chemically synthesized and analyzed for physicochemical characteristic and affinity. Two peptides with similar physicochemical characteristic to AFT and differential affinity (high affinity/low affinity>10) to the antibody would be chosen and conjugated with AMCA-NHS, which bearing the same physicochemical and fluorescent characteristic as AFT. The fluorescence labeled mimic peptide with high affinity would serve as internal standard, and the fluorescence labeled mimic peptide with low affinity would serve as capacity real time monitoring tracer to establish a novel IAC-HPLC method. A comparative methodological study would be done for the new method and the conventional IAC-HPLC method, in which a national standard method (Isotope dilution tandem liquid chromatography mass spectrometry) would be set up for golden standard. The method is hopeful for settle down the matrix interference and real time monitoring problems of IAC. The study could provide novel thought for elevating the precision and accuracy of IAC-HPLC.