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

Abstract

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.

Keywords

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

References

[1] Gall B. G., Stokes A. N., Pett J. J., Spivey K. L., French S. S., Brodie E. D. I., Brodie E. D. Jr. Toxicon, 2014, 90(90): 249.

[2] Kao C. Y. Pharmacol. Rev., 1996, 18(2): 997.

[3] Yotsu-Yamashita M. Toxin. Rev., 2001, 20(1): 51.

[4] Nieto F. R., Cobos E. J., Tejada M. Á, Sánchez-Fernández C. González-Cano R., Cendán C. M., Mar. Drugs, 2012, 10(2): 281.

[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): 171.

[6] Kohane D. S., Smith S. E., Louis D. N. Pain, 2002.

[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): 73.

[8] Narahashi T. Proc. Jpn. Acad. Ser. B, 2008, 84(5): 147.

[9] Alcaraz A., Whipple R. E., Gregg H. R., Andresen B. D., Grant P. M. Forensic Sci. Int., 1999, 99(1): 35.

[10] Yotsu-Yamashita M., Goto A., Nakagawa T. Chem. Res. Toxicol., 2005, 18(5): 865.

[11] Leung K. S., Fong B. M., Tsoi Y. K. Mar. Drugs, 2011, 9(11): 2291.

[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): 76.

[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): 420.

[14] Kohane D. S., Smith S. E., Louis D. N., Langer R. Pain, 2000, 104: 415.

[15] Kurono S., Hattori H., Suzuki O., Yamada T. Seno H. Anal. Lett., 2001, 34(2): 2439.

[16] Fukushima S. Jpn. J. Forensic Toxicol., 1991, 9: 126.

[17] Kawatsu K., Shibata T., Hamano Y. Toxicon, 1999, 37(2): 325.

[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): 74.

[19] O’Leary M. A., Schneider J. J., Isbister G. K. Toxicon, 2004, 44(5): 549.

[20] Yu C. H., Yu C. F., Tam S., Yu P. H. F. Food Add. Contam.: Part A, 2010, 27(1): 89.

[21] Tsai Y. H., Hwang D. F., Cheng C. A., Hwang C. C., Deng J. F. J. Chrmatogr. B, 2006, 832(1): 75.

[22] Saito T., Miura N., Namera A., Ota S., Miyazaki S., Inokuchi S. Chromatographia, 2014, 77(9): 687.

[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): 95.

[24] Fong B. M., Tam S., Tsui S. H., Leung K. S. Talanta, 2011, 83(3): 1030.

[25] Shoji Y., Yotsu-Yamashita M., Miyazawa T., Yasumoto T. Anal. Biochem., 2001, 290(1): 10.

[26] Nakagawa T., Jang J., Yotsu-Yamashita M. Anal. Biochem., 2006, 352(1): 142.

[27] Matuszewski B. K., Constanzer M. L., Chavez-Eng C. M. Anal. Chem., 2003, 75(13): 3019.

[28] Matuszewski B. K. Technol. Biomed. Life Sci., 2006, 830(2): 293.

[29] Committee for Medicinal Products for Human Use, Guideline on Validation of Bioanalytical Methods, European Medicines Agency, 2010