同位素稀释液相色谱-串联质谱法同时测定贝类产品中5种硝基呋喃代谢物

Determination of 5 nitrofuran metabolites in shellfish products by isotope dilution liquid chromatography-tandem mass spectrometry

中文关键词:  液相色谱-串联质谱法  硝基呋喃代谢物  同位素稀释法  贝类

英文关键词:liquid chromatography-tandem mass spectrometry  nitrofuran metabolite  isotope internal standard dilution  shellfish

中文摘要:

      目的 建立贝类产品中呋喃唑酮、呋喃它酮、呋喃西林、呋喃妥因和硝夫索尔代谢物的同位素稀释液相色谱-串联质谱(liquid chromatography-tandem mass spectrometry, LC-MS/MS)检测方法。方法 样品中加入同位素内标, 经盐酸水解及邻硝基苯甲醛衍生, 调节pH为7.2±0.2, 乙酸乙酯提取, 氮吹浓缩, 正己烷去脂净化, 色谱柱分离, 电喷雾电离(electronic spray ion, ESI)正负离子多反应(multiple reaction monitoring, MRM)模式检测, 同位素内标稀释法定量。结果 5种硝基呋喃代谢物的方法定量限为0.1~0.2 μg/kg; 方法的线性范围为0.2~20 μg/kg; 相关系数均大于0.999; 方法平均回收率为88.7%~112.1%, 相对标准偏差为3.9%~13.7%。应用该方法对280份贝类产品进行检测, 其中45份样品中检出了含量较低的氨基脲, 范围值在0.21~1.86 μg/kg。结论 方法灵敏准确, 适合贝类产品中硝基呋喃代谢物的检测。

英文摘要:

      Objective To establish an analytical method for determination of furazolidone, furaltadone, nitrofurazone, nitrofurantoin and nifursol metabolites in shellfish by isotope dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS). Methods Isotopic internal standards were added to the sample, which then were hydrolyzed by hydrochloric acid and derivatized with 2-nitrobenzaldehyde. The solution was neutralized to pH 7.2±0.2, extracted with ethyl acetate, and evaporated to dryness with nitrogen. The residue was reconstituted with mobile phase, and defatted with acetonitrile saturated n-hexane. The analytes were quantified by LC-MS/MS operating in both positive and negative ion multiple monitoring modes (MRM), and quantitatively determined by isotope internal standard dilution method. Results Limits of quantifications for the 5 nitrofuran metabolites were 0.1-0.2 μg/kg. The matrix calibration curves were established with correlation coefficients more than 0.999 in the range of 0.2-20 μg/kg. The average recoveries were ranged from 88.7% to 112.1%, with the relative standard deviations of 3.9%-13.7%. The method was successfully applied for 280 shellfish products, and 45 of them were confirmed with low levels of semicarbazide (SEM) ranging from 0.21 to 1.86 μg/kg. Conclusion The proposed method has high sensitivity and good accuracy, which is suitable for determination of nitrofuran metabolites in shellfish products.