色谱

色谱

• 研究报告 • 上一篇    下一篇

高效液相色谱-荧光检测法测定牛奶中氯霉素的残留量

潘莹宇,许茜,康学军,张建新

  

  1. Department of Nutrition and Food Hygiene, School of Public Health, Southeast University, Nanjing 210009, China
  • 收稿日期:2005-06-13 修回日期:2005-08-31 出版日期:2005-11-30 发布日期:1987-12-25
  • 通讯作者: 许茜

Determination of Chloramphenicol Residues in Milk by Reversed-Phase High Performance Liquid Chromatography with Fluorescence Detection

PAN Yingyu, XU Qian, KANG Xuejun, ZHANG Jianxin   

  1. Department of Nutrition and Food Hygiene, School of Public Health, Southeast University, Nanjing 210009, China
  • Received:2005-06-13 Revised:2005-08-31 Online:2005-11-30 Published:1987-12-25

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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: 残留, 高效液相色谱-荧光检测法, 氯霉素, 牛奶 , 荧光胺, 柱前衍生化

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×4.6 mm i.d., 4.0 μm) 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 ℃. 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 μg/L to 800 μg/L. The limit of detection (LOD) was 0.2 μg/L, and the limit of quantitation (LOQ) was 0.4 μg/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.

Key words: chloramphenicol, fluorescamine, milk , 　pre-column derivatization, residues, high performance liquid chromatography-fluorescence detection