混合型阴离子交换固相萃取-液相色谱-串联质谱法测定饲料中尼卡巴嗪含量的研究

Determination of Nicarbazin in Feeds Based on Mixed-Anionic Exchange Solid Phase Extraction by High Performance Liquid Chromatography-Tandem Mass Spectrometry

研究旨在建立一种利用混合型阴离子交换固相萃取净化测定饲料中尼卡巴嗪的液相色谱-串联质谱方法。试样(配合饲料、浓缩饲料、预混合饲料)经甲醇:乙腈(1:1)提取,采用混合型阴离子交换固相萃取柱(MAX)净化,用乙腈定容,C18色谱柱(100 mm×2.1 mm,2.6μm)分离,以乙腈和超纯水为流动相进行梯度洗脱,流速为0.3 mL/min,柱温为40℃。待测物采用电喷雾离子源(ESI)负离子多反应监测(MRM)模式检测,外标法定量。不同饲料基质标准溶液中尼卡巴嗪浓度在0.04~50 ng/mL范围内线性关系良好,相关系数(r 2)均在0.99以上,检测限为0.02 mg/kg,定量限为0.05 mg/kg。饲料中尼卡巴嗪含量在0.05~500 mg/kg(预混饲料为0.05~10000 mg/kg)范围内回收率为83.3%~105%,相对标准偏差(RSD)为2.4%~7.6%。该方法灵敏度高、准确性及重现性好,适用于配合饲料、浓缩饲料、预混合饲料中低含量和高含量尼卡巴嗪的分析。

The objective of this study is to develop a method for the determination of nicarbazin in feeds based on mixed-anionic solid phase extraction(SPE)by high performance liquid chromatography–tandem mass spectrometric.The analyzed samples(compound feed,concentrated feed,and premixed feed)were extracted with methanol:acetonitrile(1:1),purified by mixed-anionic solid phase extraction column(MAX),dissolved in acetonitrile,separated on a C18 chromatographic column(100 mm×2.1mm,2.6μm)and performed with acetonitrile and ultrapure water as mobile phases,with the flow rate of 0.3 mL/min and the column temperature of 40℃by gradient elution.The analysis was conducted by using negative-ion electrospray tandem mass spectrometry via multiple reactions monitoring(MRM)scan mode,and the samples were quantified by external standard method.The concentrations of nicarbazin in different feed matrix standard solutions had a good linear relationship in the range of 0.04-50 ng/mL,with correlation coefficients(r 2)above 0.99,the limit of detection(LOD)was 0.02 mg/kg,and the limit of quantitation(LOQ)was 0.05 mg/kg.The recovery rate of nicarbazin with the content of 0.05-500 mg/kg in feed(0.05-10000 mg/kg in premixed feed)was 83.3%-105%,and the relative standard deviation(RSD)was 2.4%-7.6%.The method was sensitive,accurate,repeatable and suitable for the analysis of low and high levels of nicarbazin in compound feed,concentrated feed and premixed feed.