日本沼虾中氨基脲含量检测方法的优化

Optimization of determination method of semicarbazide (SEM) content in Macrobrachium nipponensis

中文关键词:  氨基脲  日本沼虾  高效液相色谱-串联质谱法

英文关键词:semicarbazide  Macrobrachium nipponensis  high performance liquid chromatography-tandem mass spectrometry

中文摘要:

      目的 以农业部783 号公告-1–2006 方法为基础, 优化高效液相色谱-串联质谱法及样品前处理方法,使其更适用于日本沼虾中不同组织中氨基脲含量的检测。方法 样品经过HCl 水解, 2-硝基甲苯衍生, 乙酸乙酯萃取后, 采用高效液相色谱-串联质谱法检测氨基脲含量, 内标法定量。为考察优化后方法在日本沼虾不同组织测定结果的准确性和稳定性, 对不同基质样品进行标准添加实验。结果 氨基脲的添加水平为1.0~120.0 μg/kg 时平均回收率为74.3%~113.5%, 精密度小于6.3%, 方法检出限为0.25 μg/kg(S/N≥3), 定量限为0.5 μg/kg(S/N≥10)。结论 虾壳中氨基脲含量最高, 肌肉中含量最低; 肌肉中氨基脲主要以结合态形式存在,壳中氨基脲主要以游离态形式存在。

英文摘要:

      Objective Based on 783 announcement-1-2006 of the Ministry of Agriculture, to optimize high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) and sample pre-treatment methods,so as to make it more suitable for the detection of semicarbazide content in different tissues of Macrobrachium nipponensis. Methods Samples were hydrolyzed in HCl solution, derived with 2-nitrobenzaldehyde, and then samples were extracted by ethyl acetate, finally it were determined and analyzed by HPLC-MS/MS with internal standard. In order to investigate the accuracy and stability of the method in different tissues of Macrobrachium nipponensis, the standard addition experiment was conducted for different matrix samples. Results The average recovery rates were 74.3%-113.5% when the additions of semicarbazide were 1.0-120.0 μg/kg with precisions were less than 6.3%. The limit of detection was 0.25 μg/kg (S/N≥3), and the limit of quantitation was 0.5 μg/kg (S/N≥10). Conclusion The concentration of total semicarbazide in shells is the highest, and in muscles is the lowest, and the semicarbazide in muscle is mainly in the binding form, and in shell is mainly in the free form.