微波辅助衍生-离子液体分散液液微萃取-高效液相色谱法检测牛奶中氨基糖苷类抗生素残留

Determination of Aminoglycosides Residues in Milk by Microwave-assisted Derivatization and Ionic Liquid-based Dispersive Liquid-liquid Microextraction Coupled with HPLC

摘要:

将微波辅助衍生法与离子液体分散液液微萃取法相结合，建立了牛奶中链霉素、庆大霉素、卡那霉素、妥布霉、小诺霉素、阿米卡星和新霉素等7种氨基糖苷类抗生素（AGs）的快速萃取、衍生方法。本研究以氯甲酸芴甲酯（FMOC-Cl）为衍生试剂，1-辛基-3-甲基咪唑六氟磷酸盐（Omim]PF6）为提取剂，甲醇为分散剂，将微波辅助衍生和离子液体分散液液微萃取相结合，使目标化合物被衍生的同时被萃取和富集到离子液体中，并通过高效液相色谱荧光检测器进行定量检测。实验结果表明，各化合物在线性范围内具有较好的线性关系（r>0.9989），检出限为0.11~0.57 μg/L。加标样品回收率在91.95~106.33%之间，其RSD为1.39~5.92%。本法操作简单，灵敏度高，结果可靠，可广泛应用于牛奶样品中氨基糖苷类抗生素的检测。

Abstract:

A new method based on simultaneous microwave-assisted derivatization and ionic liquid-based dispersive liquid-liquid microextraction was developed for the derivatization and extraction of aminoglycosides (AGs) including streptomycin (STR), gentamicin (GEN), kanamycin (KAN), tobramycin (TOB), micronomicin (MIC), amikacin (AMI) and neomycin (NEO) in milk samples. In the experiment, 9-fluorenylmethyl chloroformate (FMOC-Cl), 1-octyl-3- methylimidazolium hexafluorophosphate ([Omim]PF6) and methanol were used as derivatization reagent, extraction solvent and disperser, respectively. Aminoglycosides were simultaneously derivatized and extracted into ionic liquid phase by this method in a single step. Derivatives of the aminoglycosides were then determined by high-performance liquid chromatography coupled with fluorescence detection. The calibration curve had good linear relationship (r＞0.9989). The limits of detection (LODs) of analytes were in range of 0.11~0.57 μg/L. The recoveries were between 91.95% and 106.33% with the relative standard deviations of 1.39~5.92%. The proposed method has the advantages of simple operation, high sensitivity and reliable results and should be extensively applied for determination of aminoglycosides in milk samples.