Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

The Korean Society of Food Hygiene and Safety (한국식품위생안전성학회)
권호 표지
DOI QR코드

DOI QR Code

Tetrodotoxin and Its Analogs: A Review of Analysis Methods and Levels in Pufferfish

  • Park, Bong Ki (Department of Food Science and Technology, Chung-Ang University) ;
  • Jang, Woo Jin (Department of Food Science and Technology, Chung-Ang University) ;
  • Park, Ki-Hwan (Department of Food Science and Technology, Chung-Ang University) ;
  • Lee, Hee-Seok (Department of Food Science and Technology, Chung-Ang University) ;
  • Lee, Wan-Ok (Department of Marine Life Science (BK21 FOUR), JeJu National University) ;
  • Choi, Kwang-Sik (Department of Marine Life Science (BK21 FOUR), JeJu National University) ;
  • Lee, Jihyun (Department of Food Science and Technology, Chung-Ang University)
  • Received : 2021.01.20
  • Accepted : 2021.02.25
  • Published : 2021.04.30
Download PDF
⟨ Previous Next ⟩

Abstract

Pufferfish is a major source of tetrodotoxin (TTX), a potent neurotoxin that can lead to death in humans if consumed. The content of TTX and its analogs in different pufferfish species and the distribution in various fish organs are poorly understood. In this study, we compared TTX analysis methods, including mouse bioassay, enzyme-linked immunosorbent assay, high-pressure liquid chromatography (HPLC) coupled with a fluorescence detector, gas chromatography-mass spectrometry, and liquid chromatography-mass spectrometry (LC-MS) methods. Different HPLC columns (e.g., amide, hydrophilic interaction liquid chromatography, and C18 columns) and mobile phases were compared for TTX analysis using LC-MS. Additionally, published extraction methods were compared for LC-MS analysis. The content of TTX and its analogs in different pufferfish species and their distribution in various organs (e.g., liver, muscle, intestine, testis, ovary, and skin) were summarized. This information can help understand the available TTX analysis methods, variation in TTX contents in pufferfish species from different regions, and TTX distribution in various organs.

테트로도톡신(Tetrodotoxin)은 나트륨 통로를 차단하는 신경독소로 가장 오래되고 강력한 해양독소 중 하나이다. 전통적으로 아시아에서 복어 섭취로 인한 테트로도톡신 중독사고가 많이 발생했었다. 최근 아열대 복어 종과 그 잡종 섭취로 인한 테트로도톡신 중독사고가 아시아 외 지역에서도 산발적으로 발생하고 있다. 그러나 복어 종별 테트로도톡신 및 유사체의 함량과 기관 내 분포에 대해서는 잘 알려져 있지 않다. 이 총설에서는 식품공전 복어독(테트로도톡신) 분석법인 쥐를 이용한 생물학적 정량법, 효소면역측정법, 고압 액체 크로마토그래피-형광검출기, 가스크로마토그래피-질량분석기를 이용한 분석법의 장단점을 비교하였다. 또한 최근 주로 사용되는 액체 크로마토그래피-질량분석기를 이용한 테트로도톡신과 테트로도톡신 유사체 분석법을 소개하고 이에 사용된 전처리법, 컬럼 및 이동상을 비교하였다. 또한 복어 종별 테트로도톡신과 그 유사체 함량과 다양한 기관에서의 분포를 정리하였다. 이 총설은 테트로도톡신 분석법과 다양한 복어 종의 테트로도톡신과 유사체 함량 및 기관에서의 분포를 이해하는데 도움을 줄 수 있을 것이다.

Keywords

Acknowledgement

This research was supported by a grant (20163MFDS641) from the Ministry of Food and Drug Safety.

References

  1. Narahashi, T., Pharmacology of tetrodotoxin. J. Toxicol. Toxin Rev., 20, 67-84 (2001). https://doi.org/10.1081/TXR-100102537
  2. Fuhrman, F.A., Tetrodotoxin, tarichatoxin, and chiriquitoxin: Historical perspectives. Ann. N. Y. Acad. Sci., 479, 1-14 (1986). https://doi.org/10.1111/j.1749-6632.1986.tb15556.x
  3. Chowdhury, F.R., Ahasan, H.N., Al Mamun, A., Rashid, A.M., Al Mahboob, A., Puffer fish (Tetrodotoxin) poisoning: An analysis and outcome of six cases. Trop. Doct., 37, 263-264 (2007). https://doi.org/10.1258/004947507782333017
  4. Chua, H.H., Chew, L.P., Puffer fish poisoning: a family affair. Med. J. Malaysia, 64, 181-182 (2009).
  5. How, C.K., Chern, C.H., Huang, Y.C., Wang, L.M., Lee, C.H., Tetrodotoxin poisoning. Am. J. Emerg. Med., 21, 51-54 (2003). https://doi.org/10.1053/ajem.2003.50008
  6. Mahmud, Y., Tanu, M.B., Noguch, T., First occurrence of a food poisoning incident due to ingestion of Takifugu oblongus, along with a toxicological report on three marine puffer species in Bangladesh. J. Food Hyg. Soc. Japan, 40, 473-480 (1999). https://doi.org/10.3358/shokueishi.40.6_473
  7. Acar, C., Ishizaki, S., Nagashima, Y., Toxicity of the Lessepsian pufferfish Lagocephalus sceleratus from eastern Mediterranean coasts of Turkey and species identification by rapid PCR amplification. Eur. Food Res. Technol., 243, 49-57 (2017). https://doi.org/10.1007/s00217-016-2721-1
  8. Katikou, P., Georgantelis, D., Sinouris, N., Petsi, A., Fotaras, T., First report on toxicity assessment of the Lessepsian migrant pufferfish Lagocephalus sceleratus (Gmelin, 1789) from European waters (Aegean Sea, Greece). Toxicon, 54, 50-55 (2009). https://doi.org/10.1016/j.toxicon.2009.03.012
  9. Kosker, A.R., Ozogul, F., Durmus, M., Ucar, Y., Ayas, D., Regenstein, J.M., Ozogul, Y., Tetrodotoxin levels in pufferfish (Lagocephalus sceleratus) caught in the Northeastern Mediterranean Sea. Food Chem., 210, 332-337 (2016). https://doi.org/10.1016/j.foodchem.2016.04.122
  10. Kosker, A.R., Ozogul, F., Durmus, M., Ucar, Y., Ayas, D., Simat, V., Ozogul, Y., First report on TTX levels of the yellow spotted pufferfish (Torquigener flavimaculosus) in the Mediterranean Sea. Toxicon, 148, 101-106 (2018). https://doi.org/10.1016/j.toxicon.2018.04.018
  11. Rambla-Alegre, M., Reverte, L., del Rio, V., de la Iglesia, P., Palacios, O., Flores, C., Caixach, J., Campbell, K., Elliott, C.T., Izquierdo-Munoz, A., Campas, M., Diogene, J., Evaluation of tetrodotoxins in puffer fish caught along the Mediterranean coast of Spain. Toxin profile of Lagocephalus sceleratus. Environ. Res., 158, 1-6 (2017). https://doi.org/10.1016/j.envres.2017.05.031
  12. Campbell, K., Barnes, P., Haughey, S.A., Higgins, C., Kawatsu, K., Vasconcelos, V., Elliott, C.T., Development and single laboratory validation of an optical biosensor assay for tetrodotoxin detection as a tool to combat emerging risks in European seafood. Anal. Bioanal. Chem., 405, 7753-7763 (2013). https://doi.org/10.1007/s00216-013-7106-8
  13. Chen, X.W., Liu, H.X., Jin, Y.B., Li, S.F., Chung, S., Zhang, S.S., Jiang, Y.Y., Separation, identification and quantification of tetrodotoxin and its analogs by LC-MS without calibration of individual analogs. Toxicon, 57, 938-943 (2011). https://doi.org/10.1016/j.toxicon.2011.03.011
  14. Yotsu-Yamashita, M., Chemistry of puffer fish toxin. J. Toxicol. Toxin Rev., 20, 51-66 (2001). https://doi.org/10.1081/TXR-100102536
  15. Jang, J., Yotsu-Yamashita, M., Distribution of tetrodotoxin, saxitoxin, and their analogs among tissues of the puffer fish Fugu pardalis. Toxicon, 48, 980-987 (2006). https://doi.org/10.1016/j.toxicon.2006.07.034
  16. Jang, J.H., Lee, J.S., Yotsu-Yamashita, M., LC/MS analysis of tetrodotoxin and its deoxy analogs in the marine puffer fish Fugu niphobles from the southern coast of Korea, and in the brackishwater puffer fishes Tetraodon nigroviridis and Tetraodon biocellatus from Southeast Asia. Mar. Drugs, 8, 1049-1058 (2010). https://doi.org/10.3390/md8041049
  17. Rodriguez, P., Alfonso, A., Otero, P., Katikou, P., Georgantelis, D., Botana, L.M., Liquid chromatography-mass spectrometry method to detect Tetrodotoxin and Its analogues in the puffer fish Lagocephalus sceleratus (Gmelin, 1789) from European waters. Food Chem., 132, 1103-1111 (2012). https://doi.org/10.1016/j.foodchem.2011.11.081
  18. Nakamura, M., Yasumoto, T., Tetrodotoxin derivatives in puffer fish. Toxicon, 23, 271-276 (1985). https://doi.org/10.1016/0041-0101(85)90149-7
  19. Japan Food Hygiene Association, 1990. Standard methods of analysis in food safety regulation, biology. Japan Food Hygiene Association, Tokyo, Japan, pp. 86-87.
  20. Horie, M., Kobayashi, S., Shimizu, N., Nakazawa, H., Determination of tetrodotoxin in puffer-fish by liquid chromatography-electrospray ionization mass spectrometry. Analyst, 127, 755-759 (2002). https://doi.org/10.1039/b201128j
  21. Bane, V., Lehane, M., Dikshit, M., O'Riordan, A., Furey, A., Tetrodotoxin: Chemistry, toxicity, source, distribution and detection. Toxins, 6, 693-755 (2014). https://doi.org/10.3390/toxins6020693
  22. EFSA Panel on Contaminants in the Food Chain (CONTAM), Knutsen, H.K., Alexander, J., Barregard, L., Bignami, M., Bruschweiler, B., Ceccatelli, S., Cottrill, B., Dinovi, M., Edler, L., Grasl?Kraupp, B., Hogstrand, C., Hoogenboom, L., Nebbia, C.S., Oswald, I.P., Rose, M., Roudot, A.C., Schwerdtle, T., Vleminckx, C., Vollmer, G., Wallace, H., Arnich, N., Benford, D., Botana, L., Viviani, B., Arcella, D., Binaglia, M., Horvath, Z., Steinkellner, H., Manen, M.V., Petersen, A., Risks for public health related to the presence of tetrodotoxin (TTX) and TTX analogues in marine bivalves and gastropods. EFSA Journal, 15, 4752 (2017).
  23. Bane, V., Hutchinson, S., Sheehan, A., Brosnan, B., Barnes, P., Lehane, M., Furey, A., LC-MS/MS method for the determination of tetrodotoxin (TTX) on a triple quadruple mass spectrometer. Food Addit. Contam. A, 33, 1728-1740 (2016). https://doi.org/10.1080/19440049.2016.1235801
  24. Diener, M., Christian, B., Ahmed, M.S., Luckas, B., Determination of tetrodotoxin and its analogs in the puffer fish Takifugu oblongus from Bangladesh by hydrophilic interaction chromatography and mass-spectrometric detection. Anal. Bioanal. Chem., 389, 1997-2002 (2007). https://doi.org/10.1007/s00216-007-1602-7
  25. Shoji, Y., Yotsu-Yamashita, M., Miyazawa, T., Yasumoto, T., Electrospray ionization mass spectrometry of tetrodotoxin and its analogs: liquid chromatography/mass spectrometry, tandem mass spectrometry, and liquid chromatography/tandem mass spectrometry. Anal. Biochem., 290, 10-17 (2001). https://doi.org/10.1006/abio.2000.4953
  26. Kang, Y.W., Lee, Y.S., Park, S.K., Seo, J.H., Kim, M.H., The study on the analysis method of tetrodotoxin in puffer fish. J. Food Hyg. Saf., 27, 37-41 (2012). https://doi.org/10.13103/JFHS.2012.27.1.037
  27. Hwang, D.F., Noguchi, T. Tetrodotoxin poisoning. Adv. Food Nutr. Res, 52, 141-236 (2007). https://doi.org/10.1016/S1043-4526(06)52004-2
  28. Tahara, Y., Uber das tetrodongift. Biochem., Z, 10, 255-275 (1910).
  29. Goto, T., Kishi, Y., Takahashi, S., Hirata, Y., Tetrodotoxin. Tetrahedron, 21, 2059-2088 (1965). https://doi.org/10.1016/S0040-4020(01)98344-9
  30. Tsuda, K., Ikuma, S., Kawamura, M., Tachikawa, R., Sakai, K., Tamura, C., Amakasu, O., Tetrodotoxin. VII. On the structures of tetrodotoxin and its derivatives. Chem. Pharm. Bull., 12, 1357-1374 (1964). https://doi.org/10.1248/cpb.12.1357
  31. Woodward, R.B., Gougoutas, J.Z., The structure of tetrodotoxin. J. Am. Chem. Soc., 86, 5030-5030 (1964). https://doi.org/10.1021/ja01076a076
  32. Chau, R., Kalaitzis, J.A., Neilan, B.A., On the origins and biosynthesis of tetrodotoxin. Aquat. Toxicol., 104, 61-72 (2011). https://doi.org/10.1016/j.aquatox.2011.04.001
  33. Narahashi, T., Tetrodotoxin. Proc. Jpn. Acad., Ser. B, 84, 147-154 (2008). https://doi.org/10.2183/pjab.84.147
  34. Noguchi, T., Ebesu, J.S.M., Puffer poisoning: epidemiology and treatment. J. Toxicol. Toxin Rev., 20, 1-10 (2001). https://doi.org/10.1081/TXR-100103080
  35. Lago, J., Rodriguez, L.P., Blanco, L., Vieites, J.M., Cabado, A.G., Tetrodotoxin, an extremely potent marine neurotoxin: Distribution, toxicity, origin and therapeutical uses. Mar. Drugs, 13, 6384-6406 (2015). https://doi.org/10.3390/md13106384
  36. Pinto, E.P., Rodrigues, S.M., Gouveia, N., Timoteo, V., Costa, P.R., Tetrodotoxin and saxitoxin in two native species of puffer fish, Sphoeroides marmoratus and Lagocephalus lagocephalus, from NE Atlantic Ocean (Madeira Island, Portugal). Mar. Environ. Res., 151, 104780 (2019). https://doi.org/10.1016/j.marenvres.2019.104780
  37. Kudo, Y., Finn, J., Fukushima, K., Sakugawa, S., Cho, Y., Konoki, K., Yotsu-Yamashita, M., Isolation of 6-deoxytetrodotoxin from the pufferfish, Takifugu pardalis, and a comparison of the effects of the C-6 and C-11 hydroxy groups of tetrodotoxin on its activity. J. Nat. Prod., 77, 1000-1004 (2014). https://doi.org/10.1021/np401097n
  38. Wu, B.Q., Yang, L., Kao, C.Y., Levinson, S.R., Yotsu-Yamashita, M., Yasumoto, T., 11-Oxo-tetrodotoxin and a specifically labelled 3H-tetrodotoxin. Toxicon, 34, 407-416 (1996). https://doi.org/10.1016/0041-0101(95)00168-9
  39. Isbister, G.K., Kiernan, M.C., Neurotoxic marine poisoning. Lancet Neurol., 4, 219-228 (2005). https://doi.org/10.1016/S1474-4422(05)70041-7
  40. Arakawa, O., Hwang, D.F., Taniyama, S., Takayani, T., Toxins of pufferfish that cause human intoxications. Coastal Environmental and Ecosystem Issues of the East China Sea, 227-244 (2010).
  41. Joshi, S.K., Mikusa, J.P., Hernandez, G., Baker, S., Shieh, C.C., Neelands, T., Zhang, X.F., Niforatos, W., Kage, K., Han, P., Krafte, D., Faltynek, C., Sullivan, J.P., Jarvis, M.F., Honorea, P., Involvement of the TTX-resistant sodium channel Nav 1.8 in inflammatory and neuropathic, but not postoperative, pain states. Pain, 123, 75-82 (2006). https://doi.org/10.1016/j.pain.2006.02.011
  42. Marcil, J., Walczak, J.S., Guindon, J., Ngoc, A.H., Lu, S., Beaulieu, P., Antinociceptive effects of tetrodotoxin (TTX) in rodents. Br. J. Anaesth., 96, 761-768 (2006). https://doi.org/10.1093/bja/ael096
  43. Hagen, N.A., du Souich, P., Lapointe, B., Ong-Lam, M., Dubuc, B., Walde, D., Love, R., Ngoc, A.H., Canadian Tetrodotoxin Study Group, Tetrodotoxin for moderate to severe cancer pain: a randomized, double blind, parallel design multicenter study. J Pain Symptom Manage., 35, 420-429 (2008). https://doi.org/10.1016/j.jpainsymman.2007.05.011
  44. Biessy, L., Boundy, M.J., Smith, K.F., Harwood, D.T., Hawes, I., Wood, S.A., Tetrodotoxin in marine bivalves and edible gastropods: A mini-review. Chemosphere, 236, 124404 (2019). https://doi.org/10.1016/j.chemosphere.2019.124404
  45. Wang, J., Wang, D., Luo, X., Zhang, J., Cao, M., Preparation and characterization of monoclonal antibodies to tetrodotoxin. J. Hyg. Res., 5, 308-311 (In Chinese) (1996).
  46. Gong, H., Ji, R., Jiang, T., Yang, J., Development of ELISA-kit of quantitative analysis for Tetrodotoxin. Chin. J. Public Health, 21, 1423-1424 (2005). https://doi.org/10.3321/j.issn:1001-0580.2005.12.007
  47. Stokes, A.N., Williams, B.L., French, S.S., An improved competitive inhibition enzymatic immunoassay method for tetrodotoxin quantification. Biol. Proced. Online, 14, 3 (2012). https://doi.org/10.1186/1480-9222-14-3
  48. Narita, H., Noguchi, T., Maruyama, J., Ueda, Y., Hashtmoto, K., Watanabe, Y., Hida, K., Occurrence of tetrodotoxin in a trumpet shell, "boshubora" Charonia sauliae. Bull. Japan. Soc. Sci. Fish, 47, 935-941 (1981). https://doi.org/10.2331/suisan.47.935
  49. Man, C.N., Noor, N.M., Harn, G.L., Lajis, R., Mohamad, S., Screening of tetrodotoxin in puffers using gas chromatography-mass spectrometry. J. Chromatogr. A, 1217, 7455-7459 (2010). https://doi.org/10.1016/j.chroma.2010.09.064
  50. Yasumoto, T., Michishita, T., Fluorometric determination of tetrodotoxin by high performance liquid chromatography. Agric. Biol. Chem., 49, 3077-3080 (1985). https://doi.org/10.1271/bbb1961.49.3077
  51. Yotsu, M., Endo, A., Yasumoto, T., An improved tetrodotoxin analyzer. Agric. Biol. Chem., 53, 893-895 (1989). https://doi.org/10.1271/bbb1961.53.893
  52. Pires Jr, O.R., Sebben, A., Schwartz, E.F., Morales, R.A.V., Bloch Jr, C., Schwartz, C.A., Further report of the occurrence of tetrodotoxin and new analogues in the Anuran family Brachycephalidae. Toxicon, 45, 73-79 (2005). https://doi.org/10.1016/j.toxicon.2004.09.016
  53. Fong, B.M.W., Tam, S., Tsui, S.H., Leung, K.S.Y., Development and validation of a high-throughput double solid phase extraction-liquid chromatography-tandem mass spectrometry method for the determination of tetrodotoxin in human urine and plasma. Talanta, 83, 1030-1036 (2011). https://doi.org/10.1016/j.talanta.2010.11.014
  54. Nzoughet, J.K., Campbell, K., Barnes, P., Cooper, K.M., Chevallier, O.P., Elliott, C.T., Comparison of sample preparation methods, validation of an UPLC-MS/MS procedure for the quantification of tetrodotoxin present in marine gastropods and analysis of pufferfish. Food Chem., 136, 1584-1589 (2013). https://doi.org/10.1016/j.foodchem.2012.01.109
  55. Huang, H.N., Lin, J., Lin, H.L., Identification and quantification of tetrodotoxin in the marine gastropod Nassarius by LC-MS. Toxicon, 51, 774-779 (2008). https://doi.org/10.1016/j.toxicon.2007.12.003
  56. Tsujimura, K., Yamanouchi, K., A rapid method for tetrodotoxin (TTX) determination by LC-MS/MS from small volumes of human serum, and confirmation of pufferfish poisoning by TTX monitoring. Food Addit. Contam. A, 32, 977-983 (2015). https://doi.org/10.1080/19440049.2015.1027879
  57. Nakagawa, T., Jang, J., Yotsu-Yamashita, M., Hydrophilic interaction liquid chromatography-electrospray ionization mass spectrometry of tetrodotoxin and its analogs. Anal. Biochem., 352, 142-144 (2006). https://doi.org/10.1016/j.ab.2006.02.010
  58. Noguchi, T., Arakawa, O., Takatani, T., TTX accumulation in pufferfish. Comp. Biochem. Physiol. Part D Genomics Proteomics, 1, 145-152 (2006). https://doi.org/10.1016/j.cbd.2005.10.006
  59. Noguchi, T., Arakawa, O., Takatani, T., Toxicity of pufferfish Takifugu rubripes cultured in netcages at sea or aquaria on land. Comp. Biochem. Physiol. Part D Genomics Proteomics, 1, 153-157 (2006). https://doi.org/10.1016/j.cbd.2005.11.003
  60. Gao, W., Kanahara, Y., Tatsuno, R., Soyano, K., Nishihara, G.N., Urata, C., Takatani, T., Arakawa, O., Maturation-associated changes in internal distribution and intra-ovarian microdistribution of tetrodotoxin in the pufferfish Takifugu pardalis. Fish. Sci., 84, 723-732 (2018). https://doi.org/10.1007/s12562-018-1209-2
  61. Itoi, S., Ishizuka, K., Mitsuoka, R., Takimoto, N., Yokoyama, N., Detake, A., Takayanagi, C., Yoshikawa, S., Sugita, H., Seasonal changes in the tetrodotoxin content of the pufferfish Takifugu niphobles. Toxicon, 114, 53-58 (2016). https://doi.org/10.1016/j.toxicon.2016.02.020
  62. Yotsu-Yamashita, M., Yamagishi, Y., Yasumoto, T., 5, 6, 11-Trideoxytetrodotoxin from the puffer fish, Fugu poecilonotus. Tetrahedron Lett., 36, 9329-9332 (1995). https://doi.org/10.1016/0040-4039(95)02020-P
  63. Yotsu-Yamashita, M., Mebs, D., Yasumoto, T., Tetrodotoxin and its analogues in extracts from the toad Atelopus oxyrhynchus (family: Bufonidae). Toxicon, 30, 1489-1492 (1992). https://doi.org/10.1016/0041-0101(92)90526-B
  64. Endoa, A., Khora, S.S., Murata, M., Naoki, H., Yasumoto, T., Isolation of 11-nortetrodotoxin-6 (R)-ol and other tetrodotoxin derivatives from the puffer Fugu niphobles. Tetrahedron Lett., 29, 4127-4128 (1988). https://doi.org/10.1016/S0040-4039(00)80434-7
  65. Kodama, M., Ogata, T., Noguchi, T., Maruyama, J., Hashimoto, K., Occurrence of saxitoxin and other toxins in the liver of the pufferfish Takifugu pardalis. Toxicon, 21, 897-900 (1983). https://doi.org/10.1016/0041-0101(83)90081-8
  66. Nakamura, M., Oshima, Y., Yasumoto, T., Occurrence of saxitoxin in puffer fish. Toxicon, 22, 381-385 (1984). https://doi.org/10.1016/0041-0101(84)90082-5
  67. Khora, S.S., Isa, J., Yasumoto, T., Toxicity of puffers from Okinawa, Japan. Nippon Suisan Gakk., 57, 163-167 (1991). https://doi.org/10.2331/suisan.57.163
  68. Mahmud, Y., Yamamori, K., Noguchi, T., Occurrence of TTX in a brackish water puffer "midorifugu", Tetraodon nigroviridis, collected from Thailand. J. Food Hyg. Soc. Japan, 40, 363-367 (1999). https://doi.org/10.3358/shokueishi.40.5_363
  69. Mahmud, Y., Yamamori, K., Noguchi, T., Toxicity and tetrodotoxin as the toxic principle of a Brackish water puffer, Tetraodon steindachneri, collected from Thailand. J. Food Hyg. Soc. Japan, 40, 391-395 (1999). https://doi.org/10.3358/shokueishi.40.5_391
  70. Sato, S., Ogata, T., Borja, V., Gonzales, C., Fukuyo, Y., Kodama, M., Frequent occurrence of paralytic shellfish poisoning toxins as dominant toxins in marine puffer from tropical water. Toxicon, 38, 1101-1109 (2000). https://doi.org/10.1016/S0041-0101(99)00223-8
  71. Deeds, J.R., White, K.D., Etheridge, S.M., Landsberg, J.H., Concentrations of saxitoxin and tetrodotoxin in three species of puffers from the Indian River Lagoon, Florida, the location for multiple cases of saxitoxin puffer poisoning from 2002 to 2004. Trans. Am. Fish. Soc., 137, 1317-1326 (2008). https://doi.org/10.1577/T07-204.1
  72. Azman, A.M.N., Samsur, M., Othman, M., Distribution of tetrodotoxin among tissues of puffer fish from Sabah and Sarawak waters. Sains Malays., 43, 1003-1011 (2014).
  73. Kosker, A.R., Ozogul, F., Ayas, D., Durmus, M., Ucar, Y., Regenstein, J.M., Ozogul, Y., Tetrodotoxin levels of three pufferfish species (Lagocephalus sp.) caught in the North-Eastern Mediterranean sea. Chemosphere, 219, 95-99 (2019). https://doi.org/10.1016/j.chemosphere.2018.12.010