PhIP化学模型体系中前处理及UPLC-MS检测方法的研究

Study on the Sample Pretreatment Procedure and Determination by UPLC-MS in PhIP Chemical System

本文以PhIP化学模型体系为对象,建立了利用冻干浓缩萃取进行样品前处理,用UPLC-MS进行PhIP含量检测的分析方法。本研究简化了样品前处理步骤,将反应体系冻干浓缩后采用碱性乙酸乙酯体系提取,用超高效液相色谱-三重四级杆串联方式(UPLC-MS)进行PhIP检测,采用Waters BEH phenyl色谱柱,以乙腈和0.1%甲酸溶液为流动相进行梯度洗脱。结果表明,本方法检测PhIP的线性范围为0.5~15 ng/mL,决定性系数为0.999,检出限为0.110 ng/mL,定量限为0.332 ng/mL,在加标水平为10、15、30 ng/mL时的平均收率分别为78.16%、81.22%、82.03%,相对标准偏差(RSD)分别为6.32%、5.40%、4.28%。本方法操作简便、可重复性高,可作为测定化学模型中PhIP的有效方法。

The sample preparation procedure and analytical method were developed for the determination of PhIP in chemical model system. The reaction system was freeze-dried by a lyophilizer,then extracted with alkaline ethyl acetate. Finally,the solvent was removed by nitrogen blower. The extract was separated on a BEH phenyl column,with 0.1% formic acid in acetonitrile-water as mobile phases in gradient elution. The qualitative and quantitative operation of PhIP is based on ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS). The results showed that linear range of PhIP was 0.5~15 ng/mL,the coefficient of determination was 0.999. The limits of detection and limits of quantitation for the PhIP was 0.110 and 0.332 ng/mL respectively. The average recoveries of all the compounds spiked in wine samples at three levels of 10,15,30 ng/mL were in the ranges of 78.16%,81.22%,82.03%,with the corresponding relative standard deviations(RSD)of 6.32%,5.40%,4.28%,respectively. The experimental results indicated that the method is simple,sensitive,accurate and can meet the requirement for the determination of PhIP in chemical model system.