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Simultaneous identification and quantification of tetrodotoxin in fresh pufferfish and pufferfish-based products using immunoaffinity columns and liquid chromatography/quadrupole-linear ion trap mass spectrometry

  • Ecology
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Chinese Journal of Oceanology and Limnology Aims and scope Submit manuscript

Abstract

In this study, we established a comprehensive method for simultaneous identification and quantification of tetrodotoxin (TTX) in fresh pufferfish tissues and pufferfish-based products using liquid chromatography/quadrupole-linear ion trap mass spectrometry (LC-QqLIT-MS). TTX was extracted by 1% acetic acid-methanol, and most of the lipids were then removed by freezing lipid precipitation, followed by purification and concentration using immunoaffinity columns (IACs). Matrix effects were substantially reduced due to the high specificity of the IACs, and thus, background interference was avoided. Quantitation analysis was therefore performed using an external calibration curve with standards prepared in mobile phase. The method was evaluated by fortifying samples at 1, 10, and 100 ng/g, respectively, and the recoveries ranged from 75.8%–107%, with a relative standard deviation of less than 15%. The TTX calibration curves were linear over the range of 1–1 000 μg/L, with a detection limit of 0.3 ng/g and a quantification limit of 1 ng/g. Using this method, samples can be further analyzed using an information-dependent acquisition (IDA) experiment, in the positive mode, from a single liquid chromatography-tandem mass spectrometry injection, which can provide an extra level of confirmation by matching the full product ion spectra acquired for a standard sample with those from an enhanced product ion (EPI) library. The scheduled multiple reaction monitoring method enabled TTX to be screened for, and TTX was positively identified using the IDA and EPI spectra. This method was successfully applied to analyze a total of 206 samples of fresh pufferfish tissues and pufferfish-based products. The results from this study show that the proposed method can be used to quantify and identify TTX in a single run with excellent sensitivity and reproducibility, and is suitable for the analysis of complex matrix pufferfish samples.

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Authors and Affiliations

  1. Key Laboratory of Testing and Evaluation for Aquatic Product Safety and Quality, Ministry of Agriculture, Chinese Academy of Fishery Sciences, Qingdao, 266071, China

    Mengmeng Guo  (郭萌萌), Haiyan Wu  (吴海燕), Zhijun Tan  (谭志军), Chunxia Zhao  (赵春霞), Guanchao Zheng  (郑关超), Zhaoxin Li  (李兆新) & Yuxiu Zhai  (翟毓秀)

  2. Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, 266071, China

    Mengmeng Guo  (郭萌萌), Haiyan Wu  (吴海燕), Zhijun Tan  (谭志军), Chunxia Zhao  (赵春霞), Guanchao Zheng  (郑关超), Zhaoxin Li  (李兆新) & Yuxiu Zhai  (翟毓秀)

  3. National Center for Quality Supervision and Test of Aquatic Products, Qingdao, 266071, China

    Mengmeng Guo  (郭萌萌), Haiyan Wu  (吴海燕), Zhijun Tan  (谭志军), Chunxia Zhao  (赵春霞), Guanchao Zheng  (郑关超), Zhaoxin Li  (李兆新) & Yuxiu Zhai  (翟毓秀)

  4. Key Laboratory of Food Safety Risk Assessment, Ministry of Health, China National Center for Food Safety Risk Assessment, Beijing, 100021, China

    Tao Jiang  (江涛)

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  1. Mengmeng Guo  (郭萌萌)
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  2. Haiyan Wu  (吴海燕)
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  3. Tao Jiang  (江涛)
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  4. Zhijun Tan  (谭志军)
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  6. Guanchao Zheng  (郑关超)
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  7. Zhaoxin Li  (李兆新)
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  8. Yuxiu Zhai  (翟毓秀)
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Correspondence to Zhijun Tan  (谭志军).

Additional information

Supported by the National Natural Science Foundation of China (No. 41106109) and the China National Food Safety Standards Development Project (No. ZHENGHE-2015-356)

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Guo, M., Wu, H., Jiang, T. et al. Simultaneous identification and quantification of tetrodotoxin in fresh pufferfish and pufferfish-based products using immunoaffinity columns and liquid chromatography/quadrupole-linear ion trap mass spectrometry. Chin. J. Ocean. Limnol. 35, 883–893 (2017). https://doi.org/10.1007/s00343-017-6097-2

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