凝胶色谱/超高效液相色谱-串联质谱法测定饲料中16种脂溶性染料的研究

Simultaneous Determination of 16 Kinds of Oil-soluble Synthetic Dyes in Feeds by Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry Coupled with Gel Permeation Chromatography Clean-up Procedure

应用凝胶色谱/超高效液相色谱-串联三重四极杆质谱建立了饲料中16种脂溶性染料测定的新方法。样品经20 m L乙酸乙酯-环己烷（1∶1）提取,凝胶色谱系统净化后,以0.1%甲酸溶液-甲醇为流动相进行梯度洗脱,各脂溶性染料经C18柱得到良好分离。16种染料在2-100μg/L范围内线性关系良好,相关系数（r2）不小于0.99。其检出限为25μg/kg,定量下限为50μg/kg。饲料样品在低、中、高3个加标浓度下,16种脂溶性染料的回收率为66.4%-108.5%,结果表明方法适用于饲料中16种脂溶性染料的检测。该方法操作简单、准确、杂质干扰少。

A method based on ultra performance liquid chromatography -tandem mass specitrometry （UPLC -MS/MS）with gel permeation chromatography was developed for the quantitative analysis of 16 kinds of oil-soluble synthetic dyes in feeds. The conditions of sample preparation and analysis were optimized in this thesis. The dyes were firstly extracted with 20 mL of ethyl acetate - cyclohex- ane（ 1 ： 1 ）, and 5 mL of the extract was injected into the gel permeation chromatography（GPC） cleanup system. The elution was performed with a mixture of ethyl acetate - cyclohexane （ 1 ： 1 ） at a flow rate of 5 mL/min. The elution fraction in 15.8 - 48.3 min was collected. A UPLC separation was carried on a BEH Cls column（ 1.0 mm × 100 mm, 1.7 μm）by gradient elution using methanol -0. 1% formic acid as mobile phase at a flow rate of 0. 3 mL/min. 16 kinds of dyes were detected un- der positive electrospray ionization（ESI）. The method was quantified by the external standard method and the linear ranges for 16 kinds of dyes were in the range of 2 - 100 μg/L with correlation coeffi- cients greater than 0. 99. The average recoveries for 16 kinds of dyes in feeds ranged from 66.4% to 108.5% at three different levels. The limits of detection were 25 μg/kg and the limits of quantitation were 50 μg/kg. The results indicated that the method developed was highly sensitive and selective in determination of 16 kinds of oil-soluble synthetic dyes in feeds.