超高效液相色谱-串联质谱法定量检测日本沼虾不同组织中氨基脲含量

Quantitative determination of semicarbazide content in different tissues of Macrobrachium nipponense based on ultra performance liquid chromatography-tandem mass spectrometry

中文关键词:  液相色谱串联质谱联用仪  日本沼虾  氨基脲

英文关键词:ultra performance liquid chromatography-tandem mass spectrometry  Macrobrachium nipponense  semicarbazide

中文摘要:

      目的 建立超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)测定日本沼虾(Macrobrachium nipponense)不同组织部位中不同形态氨基脲(semicarbazide, SEM)的检测方法。方法 样品用66.7%甲醇溶液洗脱后, 经盐酸酸化水解, 2-硝基苯甲醛过夜衍生, 调pH至7.3~7.4后, 用乙酸乙酯提取, 分析物采用UPLC-MS/MS定性检测, 稳定同位素内标法进行定量测定。结果 SEM在0.25~20.00 μg/kg范围内线性关系良好, 相关系数大于0.999, 肌肉、头壳、背壳、头足胸、眼柄和鳃6个组织的方法检出限为0.25 μg/kg (S/N≥3), 定量限为0.50 μg/kg (S/N≥10), 肝胰腺的方法检出限为0.5 μg/kg, 定量限为1.0 μg/kg, 相对标准偏差不大于5.20%。虾壳中SEM含量最高, 肌肉中含量最低, 肌肉中SEM主要以游离态形式存在, 虾壳中SEM主要以结合态形式存在。结论 本方法灵敏度高、重现性好, 可用于日本沼虾不同组织中不同形态SEM的定量测定。日本沼虾不同组织中SEM的存在形态与含量均存在巨大差异。

英文摘要:

      Objective To establish an ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the determination of different forms of semicarbazide (SEM) in different tissue parts of Macrobrachium nipponense. Methods After washing with 66.7% methanol solution, the samples underwent hydrochloric acid acidification and hydrolysis, followed by overnight derivatization of 2-nitrobenzaldehyde. After adjusting the pH value to 7.3?7.4, it was extracted with ethyl acetate. The analyte was qualitatively detected using UPLC-MS/MS and quantitatively determined using stable isotope internal standard method. Results The SEM exhibited a good linear relationship within the range of 0.25?20.00 μg/kg, with a correlation coefficient exceeding 0.999. For 6 tissues (muscle, head shell, back shell, head foot chest, eye handle, and gills), the limit of detection was 0.25 μg/kg (S/N≥3) and the limit of quantitation was 0.50 μg/kg (S/N≥10). The limit of detection and limit of quantitation for liver and pancreas was 0.5 μg/kg and 1.0 μg/kg, respectively, with relative standard deviations not more than 5.20%. The shrimp shell possessed the highest SEM content, while the muscle exhibited the lowest content. SEM in muscle primarily existed in the free form, whereas SEM in the shrimp shell predominantly existed in the bound form. Conclusion This method has high sensitivity and good reproducibility, and can be used for quantitative determination of SEM in different tissues of Macrobrachium nipponense. There are significant differences in the morphology and content of SEM in different tissues of Macrobrachium nipponense.