Chemical Research in Chinese Universities

, Volume 34, Issue 6, pp 905–911 | Cite as

Rapid Determination of Tetrodotoxin in Human Plasma by Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry

  • Weizhu Chen
  • Yiping Zhang
  • Jipeng Sun
  • Quanling Xie
  • Zhuan Hong
  • Ruizao YiEmail author


A sensitive analytical method was developed to determine tetrodotoxin(TTX) in human plasma samples using protein precipitation, followed by ultra performance liquid chromatography(UPLC) analysis coupled with tandem mass spectrometry(MS/MS) using 11-deoxytetrodotoxin(11-deoxyTTX) as an internal standard. The plasma samples were prepared using protein precipitation prior to being analyzed by UPLC-MS/MS to identify TTX over a zwitterionic-hydrophilic interaction liquid chromatography column. The retention time values of TTX and 11-deoxyTTX were 4.12 and 3.67 min, respectively. TTX and 11-deoxyTTX were monitored and quantitated on the basis of their ion transitions for their respective precursor ions to their product ions(i.e., m/z 320.0→162.1 for TTX and m/z 304.0→176.0 for 11-deoxyTTX) in the multiple reaction-monitoring mode. The lower limit of quantification of this method was determined to be 0.0199 ng/mL. This method showed good linearity for plasma samples that contained TTX concentrations in the range of 0.0199—1.99 ng/mL. The specificity, precision, accuracy, matrix effect, and stability characteristics of this method were also examined. The intra-assay precision and accuracy ranged from 1.89% to 6.00% and from 92.21% to 100.00%, whereas the inter-assay precision and accuracy ranged from 0.64% to 7.75% and from 99.38% to 101.26%, respectively. This new method therefore represents a rapid, accurate, reliable, and highly sensitive method for the qualitative and quantitative analyses of a trace amount of TTX in human plasma samples.


Tetrodotoxin(TTX) 11-DeoxyTTX Ultra performance liquid chromatography(UPLC) coupled with tandem mass spectrometry(MS/MS) Plasma Protein precipitation 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. [1]
    Gall B. G., Stokes A. N., Pett J. J., Spivey K. L., French S. S., Brodie E. D. III, Brodie E. D. Jr., Toxicon, 2014, 90(90), 249CrossRefGoogle Scholar
  2. [2]
    Kao C. Y., Pharmacol. Rev., 1996, 18(2), 997Google Scholar
  3. [3]
    Yotsu-Yamashita M., Toxin. Rev., 2001, 20(1), 51Google Scholar
  4. [4]
    Nieto F. R., Cobos E. J., Tejada M. Á., Sánchez-Fernández C., González-Cano R., Cendán C. M., Mar. Drugs, 2012, 10(2), 281CrossRefGoogle Scholar
  5. [5]
    Hagen N. A., Fisher K. M., Lapointe B., Souich P., Chary S., Moulin D., Sellers E., Ngoc A. H., J. Pain Symptom. Manage., 2007, 34(2), 171CrossRefGoogle Scholar
  6. [6]
    Kohane D. S., Smith S. E., Louis D. N., Pain, 2002, 104(1/2), 415Google Scholar
  7. [7]
    Ahasan A. M. N., Mamun A. A., Karim S. R., Bakar M. A., Gazi E. A., Bala C. S., Singapore Med. J., 2004, 45(2), 73Google Scholar
  8. [8]
    Narahashi T., Proc. Jpn. Acad. Ser. B, 2008, 84(5), 147CrossRefGoogle Scholar
  9. [9]
    Alcaraz A., Whipple R. E., Gregg H. R., Andresen B. D., Grant P. M., Forensic Sci. Int., 1999, 99(1), 35CrossRefGoogle Scholar
  10. [10]
    Yotsu-Yamashita M., Goto A., Nakagawa T., Chem. Res. Toxicol., 2005, 18(5), 865CrossRefGoogle Scholar
  11. [11]
    Leung K. S., Fong B. M., Tsoi Y. K., Mar. Drugs, 2011, 9(11), 2291CrossRefGoogle Scholar
  12. [12]
    Cho H. E., Ahn S. Y., Son I. S., In S., Hong R. S., Kim D. W., Woo S. H., Moon D. C., Kim S., Forensic Sci. Int., 2012, 217(1–3), 76CrossRefGoogle Scholar
  13. [13]
    Hagen N., Du S. P., Lapointe B., Ong-Lam M., Dubuc B., Walde D., Love R., Ngoc A., J. Pain Symptom Manag. 2008, 35(4), 420CrossRefGoogle Scholar
  14. [14]
    Kohane D. S., Smith S. E., Louis D. N., Langer R., Pain, 2000, 104(1/2), 415CrossRefGoogle Scholar
  15. [15]
    Kurono S., Hattori H., Suzuki O., Yamada T., Seno H. Anal. Lett., 2001, 34(2), 2439CrossRefGoogle Scholar
  16. [16]
    Fukushima S., Jpn. J. Forensic Toxicol., 1991, 9, 126Google Scholar
  17. [17]
    Kawatsu K., Shibata T., Hamano Y., Toxicon, 1999, 37(2), 325CrossRefGoogle Scholar
  18. [18]
    Islam Q. T., Razzak M. A., Islam M. A., Bari M. I., Basher A., Chowdhury F. R., Sayeduzzaman A. B. M., Ahasan H. A. M., Faiz N. M. A., Arakawa O., Yotsu-Yamashita M., Kuch U., Mebs D., Trans. R. Soc. Trop. Med. Hyg., 2011, 105(2), 74CrossRefGoogle Scholar
  19. [19]
    O’Leary M. A., Schneider J. J., Isbister G. K., Toxicon, 2004, 44(5), 549CrossRefGoogle Scholar
  20. [20]
    Yu C. H., Yu C. F., Tam S., Yu P. H. F., Food Add. Contam.: Part A, 2010, 27(1), 89CrossRefGoogle Scholar
  21. [21]
    Tsai Y. H., Hwang D. F., Cheng C. A., Hwang C. C., Deng J. F., J. Chrmatogr. B, 2006, 832(1), 75CrossRefGoogle Scholar
  22. [22]
    Saito T., Miura N., Namera A., Ota S., Miyazaki S., Inokuchi S., Chromatographia, 2014, 77(9/10), 687CrossRefGoogle Scholar
  23. [23]
    Jena H. C., Lin S. J., Tsa Y. H., Chend C. H., Line Z. C., Hwang D. F., J. Chromatogr. B, 2008, 871(1), 95CrossRefGoogle Scholar
  24. [24]
    Fong B. M., Tam S., Tsui S. H., Leung K. S., Talanta, 2011, 83(3), 1030CrossRefGoogle Scholar
  25. [25]
    Shoji Y., Yotsu-Yamashita M., Miyazawa T., Yasumoto T., Anal. Biochem., 2001, 290(1), 10CrossRefGoogle Scholar
  26. [26]
    Nakagawa T., Jang J., Yotsu-Yamashita M., Anal. Biochem., 2006, 352(1), 142CrossRefGoogle Scholar
  27. [27]
    Matuszewski B. K., Constanzer M. L., Chavez-Eng C. M. Anal. Chem., 2003, 75(13), 3019CrossRefGoogle Scholar
  28. [28]
    Matuszewski B. K., Technol. Biomed. Life Sci., 2006, 830(2), 293CrossRefGoogle Scholar
  29. [29]
    Committee for Medicinal Products for Human Use, Guideline on Validation of Bioanalytical Methods, European Medicines Agency, 2010Google Scholar

Copyright information

© Jilin University, The Editorial Department of Chemical Research in Chinese Universities and Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Weizhu Chen
    • 1
    • 2
    • 3
  • Yiping Zhang
    • 1
    • 2
    • 3
  • Jipeng Sun
    • 1
    • 2
    • 3
  • Quanling Xie
    • 1
    • 2
    • 3
  • Zhuan Hong
    • 1
    • 2
    • 3
  • Ruizao Yi
    • 1
    • 2
    • 3
    Email author
  1. 1.Third Institute of OceanographyState Oceanic AdministrationXiamenP. R. China
  2. 2.Engineering Research Center of Marine Biological Resource Comprehensive UtilizationState Oceanic AdministrationXiamenP. R. China
  3. 3.Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological ResourcesXiamenP. R. China

Personalised recommendations