DiscoveryDSC-18净化填料-超高效液相色谱串联质谱法测定蜂蜜中氟苯尼考和氟苯尼考胺

Determination of florfenicol and florfenicol amine in honey by ultra performance liquid chromatography tandem mass spectrometry with DiscoveryDSC-18 purification packing

优化了蜂蜜中的氟苯尼考和氟苯尼考胺的测定方法.蜂蜜样品加入d5-氯霉素内标、d3-氟苯尼考胺内标,其中氟苯尼考和氟苯尼考胺用乙腈提取,提取液经DiscoveryDSC-18净化填料进行净化,净化液经过稀释后,在微弱的氮气流下吹干,复溶后,用乙腈与水(体积比10:90)梯度洗脱,通过超高效液相色谱串联质谱仪进行测定,内标法进行定量.结果显示:氟苯尼考质量浓度在0.1~5.0 ng/mL范围内与相应峰面积之间呈现良好的线性关系,线性回归方程为y=0.267x+0.00125,线性相关系数r^(2)=0.9998;氟苯尼考胺质量浓度在1.0~50.0 ng/mL范围内与相应峰面积之间呈现良好的线性关系,线性回归方程为y=0.139x+0.0256,线性相关系数r^(2)=0.9994;氟苯尼考、氟苯尼考胺检出限分别为0.05、0.5μg/kg,加标回收率为91.2%~101.0%,相对标准偏差为0.83%~5.22%.该方法改进了前处理方法,提高了回收率,增加了可靠性,可作为蜂蜜中氟苯尼考和氟苯尼考胺的改进检测方法使用.

The determination method of florfenicol and florfenicol amine in honey was optimized.The honey sample was added with d5-chloramphenicol internal standard and d3-florfenicol amine internal standard,in which florfenicol and florfenicol amine were extracted with acetonitrile,and the extracted solution was purified with DiscoveryDSC-18 purification packing.After dilution,the purification solution was dried under a weak nitrogen flow.After resolution,gradient elution was carried out by the volume ratio of acetonitrile and water was 10∶90,and the determination was carried out by UPLC-MS,and the quantification was carried out by internal standard method.The results showed that there was a good linear relationship between the florfenicol mass concentration in the range of 0.1~5.0 ng/mL and the corresponding peak area.The linear regression equation was y=0.267 x+0.00125,and the correlation coefficient r^(2)=0.9998.There was a good linear relationship between the concentration of florfenicol amine and the corresponding peak area in the range of 1.0~50.0 ng/mL,the linear range was y=0.139 x+0.0256,the correlation coefficient r^(2)=0.9994,the detection limits were 0.05,0.5μg/kg,respectively.The recoveries were 91.2%~101.0%,and the relative standard deviations were 0.83%~5.22%.The method improved the pretreatment method,increased the recovery rate,and increased the reliability.It could be used as an improved method for the determination of florfenicol and florfenicol amine in honey.