超高效液相色谱-荧光法测定蜂蜜中百里香酚的残留量

Determination of Thymol in Honey by Ultra Performance Liquid Chromatography with Fluorescence Detector

建立了超高效液相色谱-荧光检测法测定蜂蜜中百里香的残留量的检验方法。用40%乙腈-水溶液提取蜂蜜中的百里香酚,经离心、上清液过滤后,以0.1%甲酸水溶液-乙腈为流动相进行梯度洗脱,采用Accucore aQ色谱柱(150 mm×2.1 mm,2.6μm)分离,荧光检测器检测(激发波长274 nm,发射波长297 nm),外标法定量。结果表明,百里香酚在0.01~3.00μg/mL范围内线性关系良好,决定系数(R 2)为0.9999,在蜂蜜基质中的平均加标回收率在87.4%~106.9%之间,变异系数在0.7%~7.3%之间,定量限为0.10 mg/kg。利用该方法测定了48份市售蜂蜜样品,仅有1份样品存在百里香酚残留。本研究建立的方法简单、迅速、可靠,适用于蜂蜜中百里香酚残留的批量筛查和定量测定。

The presented method was developed to determine the concentration of thymol residues in honey by ultra performance liquid chromatography with a fluorescence detector.The thymol in honey was extracted by 40%acetonitrile-water solution.After centrifugation and filtration,the separation of thymol was performed on an Accucore aQ liquid chromatography column(150 mm×2.1 mm,2.6μm)with the gradient elution of acetonitrile and water(containing 0.1%formic acid)as mobile phases,and quantitated by external standard method with fluoresce detector(excitation wavelength 274 nm,emission wavelength 297 nm).A good linear relationship(R 2=0.9999)was observed with concentration ranging from 0.01μg/mL to 3.00μg/mL.The average recovery of thymol was from 87.4%to 106.9%,the coefficient of variation was between 0.7%and 7.3%,and the limit of quantification was 0.10 mg/kg.The developed method was applied to determine residual thymol in 48 commercial honey samples,and the thylmol was only detected in one sample.The presented method is simple,quick,reliable,and suitable to be used for batch screening and quantitative determination of residual thymol in honey.