应用液质联用技术测定水产品中孔雀石绿及无色孔雀石绿残留量.pdfVIP

应用液质联用技术测定水产品中孔雀石绿及无色孔雀石绿残留量 *陆柏益 蔡增轩任一平 浙江省疾病预防控制中心 杭州 310009 浙江大学生物系统工程与食品科学学院 杭州 310029 摘 要:目的:建立一种应用 LC-MS/MS快速测定水产品中孔雀石绿及其代谢产物含量的方 法。方法:采用液相色谱-电喷雾串联四极杆质谱仪,用多离子反应监测(MRM)定量法。 通过对试样净化预处理、色谱条件、质谱参数等的优化选择和方法回收率、精密度等的验证, 建立了定量测定水产品中孔雀石绿及其代谢产物的理想方法。结果:孔雀石绿和代谢产物- 无色孔雀石绿的定量限LOD分别为 4pg/mL和 5pg/mL,在 0.001-1ng/ kg的线性范围内,相 关系数 r均为 0.99,回收率分别为 78.5%、65.0%。结论:采用液相色谱-电喷雾串联四极杆 质谱仪,可快速测定水产品中孔雀石绿和无色孔雀石绿含量,结果准确可靠,重复性好,灵 敏度高。 关键词:液相色谱-电喷雾串联四极杆质谱,固相萃取,孔雀石绿,无色孔雀石绿 Determination of malachite green and leucomalachite green residues in aquatic products by HPLC-MS/MS Lu BaiyiCai Zengxuan Ren Yiping (College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310029) 2 (Zhejiang Provincial Center for Disease Prevention and Control,Hangzhou 310009) ABSTRACT A fast determination method for malachite green and its metabolite leucomalachite green residues in aquatic products was established. A liquid chromatography coupled electrospray ionization quadrupole tandem mass spectrometry was applied in this method. The residues were quantified with multiple reaction monitoring MRM mode. The analytical method in the present study was well validated and good results were obtained with respect to precision, repeatability and spiked recovery. The limit of detection of residues were 4 pg/ml and 5 pg/ml for malachite green and leucomalachite green respectively, and with linear range of 0.001 to 1 ng/ml. The recover ratios were 78.5% and 65.0% for malachite green and leucomalachite green respectivelyThe method can be applied for the determination of residues of malachite green and leucomalachite green in aquatic products for its good repeatability and high sensitivity KEYWORDS: HPLC-MS/MS , SPE, Malachite Green, Leucomalachite Green 绪 言 孔雀石绿(Malachite green)又名碱性绿,商品名有草酸盐和氯化盐两种形式,为翠绿 色有光泽的结晶,属三苯甲烷类染料[1]。化学名称为四甲基代二氨基三苯甲烷,分子式为 C H CIN,结构式见图 1,易溶于水、乙醇和甲醇,水溶液呈兰绿色。其在水生生物体中 23 25 2 的主要代谢产物为无色孔雀石绿结构式见图 2,无色孔雀石绿由于不溶于水,残留毒性比 孔雀石绿更强。孔雀石绿在细胞分裂时阻碍蛋白肽的形成,使细胞内的氨基酸无法转化为蛋白肽,细胞分