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A novel CDs-DMIP monolithic capillary column coupled with HPLC-FLD was prepared.The column showed ability of impurities removal and excellent selectivity.Template elution process of CDs-DMIP was easier than that of DMIP.Aflatoxin B1 existed in peanut sample was enriched more than 71 times.The method was prospered to analyze Aflatoxin B1 and LOD achieved 0.118 ng mL−1.In this study, a novel method was proposed to sensitively determine aflatoxin B1 (AFB1) in peanut sample by using a carbon quantum dots-coated dummy molecularly imprinted polymer (CDs-DMIP) monolithic column for pretreatment coupled with high performance liquid chromatography-fluorescence detection (HPLC-FLD). The CDs-DMIP monolithic column was prepared by in-situ polymerization in a water bath using 5,7-dimethoxycoumarin as dummy template molecule. The CDs-DMIP monolithic column was applied to determine AFB1 by HPLC-FLD. Satisfactory linearity was obtained over 0.5–2000 ng mL−1, with a high correlation coefficient of 0.9999. The recoveries of AFB1 in peanut sample ranged from 79.5% to 91.2%, and the intraday and interday relative standard deviation ranged from 1.2% to 4.9%. Limit of detection (S/N = 3) and limit of quantitation (S/N = 10) were 0.118 ng mL−1 and 0.393 ng mL−1, respectively. Under the optimized conditions, the enrichment factor was over 71-fold. AFB1 in peanut sample and even some other samples could be sensitively determined by CDs-DMIP-HPLC-FLD method.