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高效液相色谱-荧光检测法测定牛奶中氯霉素的残留量
引用本文:潘莹宇,许茜,康学军,张建新.高效液相色谱-荧光检测法测定牛奶中氯霉素的残留量[J].色谱,2005,23(6):577-580.
作者姓名:潘莹宇  许茜  康学军  张建新
作者单位:东南大学公共卫生学院营养与食品卫生学系,江苏,南京,210009
摘    要: 建立了对牛奶中氯霉素的残留量进行检测的高效液相色谱-荧光检测方法。氯霉素还原后在温和条件下与荧光胺发生衍生化反应,采用十八烷基键合硅胶固定相,以乙腈/四氢呋喃/0.02 mol/L醋酸钠-醋酸缓冲液(pH 6.0)(体积比为16∶8∶76)为流动相,流速1.0 mL/min,柱温40 ℃,荧光检测激发波长为410 nm,发射波长为508 nm。在上述实验条件下,氯霉素检测的线性范围为0.4~800 μg/L (r2=0.9999),检出限为0.2 μg/L。当空白样品中氯霉素添加水平为2~40 μg/L时,该方法的回收率为66.6%~92.8%,相对标准偏差为4.5%~9.4%。该方法适用于牛奶中氯霉素痕量残留的监测,具有干扰小、选择性好、灵敏度高等优点。

关 键 词:高效液相色谱-荧光检测法  柱前衍生化  荧光胺  氯霉素  残留  牛奶
文章编号:1000-8713(2005)06-0577-04
收稿时间:2005-06-13
修稿时间:2005年6月13日

Determination of Chloramphenicol Residues in Milk by Reversed-Phase High Performance Liquid Chromatography with Fluorescence Detection
PAN Yingyu,XU Qian,KANG Xuejun,ZHANG Jianxin.Determination of Chloramphenicol Residues in Milk by Reversed-Phase High Performance Liquid Chromatography with Fluorescence Detection[J].Chinese Journal of Chromatography,2005,23(6):577-580.
Authors:PAN Yingyu  XU Qian  KANG Xuejun  ZHANG Jianxin
Institution:Department of Nutrition and Food Hygiene, School of Public Health, Southeast University, Nanjing 210009, China
Abstract:A high performance liquid chromatographic method with fluorescence detection for the determination of chloramphenicol (CAP) residues in milk was developed. Although CAP itself is non-fluorescent the aromatic nitro group of CAP could be reduced to aromatic primary amino group and fluorescamine can be used as a selective reagent for primary amines. Therefore, the CAP was derivatized with fluorescamine prior to injection in this project. The HPLC method was performed on a Diamond C18 column (250 mm x 4.6 mm i. d., 4.0 microm) with the mobile phase composed of sodium acetate buffer (0.02 mol/L, pH 6.0) -acetonitrile-tetrahydrofuran (76: 16: 8, v/v/v). The flow rate of the mobile phase was set at 1.0 mL/min, and the column was maintained at 40 degrees C. Analytes were detected by a fluorescence detector at 410 nm excitation and 508 nm emission wavelength. The standard curve was linear in the range from 0.4 microg/L to 800 microg/L. The limit of detection (LOD) was 0.2 microg/L, and the limit of quantitation (LOQ) was 0.4 microg/L. Overall recoveries were between 66.6% and 92.8% with relative standard deviations between 4.5% and 9.4%. The procedure provides a rapid, reliable and sensitive method for the determination of CAP in milk.
Keywords:high performance liquid chromatography-fluorescence detection  pre-column deri-vatization  fluorescamine  chloramphenicol  residues  milk
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