Sains Malaysiana 43(7)(2014): 1003–1011

 

Distribution of Tetrodotoxin among Tissues of Pufferfish fromSabah and Sarawak Waters

(Taburan Tetrodotoksin antara Tisu di dalam Ikan Buntal dari Perairan Sabah dan Sarawak)

 

 

A.  MOHD NOR AZMAN2*, M. SAMSUR1 & M. OTHMAN2

 

1Faculty of Resource Science and Technology, Universiti Malaysia Sarawak

94300 Kota Samarahan, Sarawak, Malaysia

 

2Fisheries Research Institute, 11960 Batu Maung, Penang, Malaysia

 

Diserahkan: 10 Oktober 2012/Diterima: 16 September 2013

 

ABSTRACT

Puffer fish, mainly from Tetraodontidae family known to possess a neurotoxin or tetrodotoxin (TTX) which can cause a puffer fish poisoning and adverse effect to human health. In current study, the tetrodotoxin (TTX) concentration in different tissues (liver, skin, muscle) of 14 species of puffer fish from Sabah and Sarawak waters were analysed and determined by application of liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). Overall, extracted toxin for all specimens were shown to be toxic with result more than 0.2 μg/g as calculated based on TTX standard curve. Among the tissues, liver were found to be highest in TTX concentration (91.0 μg/g), followed by muscle (51.1 μg/g) and skin (6.87 μg/g). Moreover, TTX concentrations among puffer fish species were significantly differences (p<0.05) with Arothron immaculatus (275 μg/g) showed highest mean value, while the lowest value was detected in Lagocephalus lunaris (4.92 μg/g). From this finding, LC-MS/MS application could be a potential tools to determine the TTX and advisedly used as a procedure in screening of seafood for monitoring program. Furthermore, baseline data of TTX levels in selected puffer fish from the study could be important information and used as guideline in order to mitigate puffer fish poisoning cases especially in East Malaysia waters.

 

Keywords: East Malaysia; LC-MS/MS; puffer fish; Tetrodotoxin; tissues

 

 

ABSTRAK

Ikan buntal, terutamanya daripada famili Tetraodontidae diketahui mengandungi neurotoksin atau tetrodotoxin (TTX) yang boleh menyebabkan keracunan ikan buntal dan memberi kesan buruk kepada kesihatan manusia. Dalam kajian ini, kepekatan tetrodotoxin (TTX) dalam tisu yang berbeza (hati, kulit, otot) dalam 14 spesies ikan buntal dari perairan Sabah dan Sarawak telah dianalisis dan ditentukan menggunakan spektrometri jisim kromatografi cecair/spektrometri jisim (LC-MS/MS). Secara keseluruhannya, toksin yang diekstrak daripada semua spesimen adalah toksik dengan keputusan melebihi daripada 0.2 μg/g berdasarkan pengiraan daripada keluk piawai TTX. Antara tisu yang dikaji, kepekatan TTX yang tertinggi didapati dalam hati (91.0 μg/g), diikuti oleh otot (51.1 μg/g) dan kulit (6.87 μg/g). Tambahan lagi, terdapat perbezaan ketara (p<0.05) dalam kepekatan TTX antara spesies ikan buntal dengan Arothron immaculatus (275 μg/g) menunjukkan nilai min tertinggi, manakala nilai terendah dikesan pada Lagocephalus lunaris (4.92 μg/g). Daripada penemuan ini, penggunaan LC-MS/MS boleh menjadi alat yang berpotensi untuk menentukan TTX dan boleh digunakan sebagai prosedur dalam pemeriksaan makanan laut bagi program pemantauan. Tambahan pula, data asas tahap TTX dalam ikan buntal yang dipilih daripada kajian boleh menjadi maklumat penting dan digunakan sebagai garis panduan dalam usaha untuk mengurangkan kes keracunan ikan buntal terutamanya di perairan Malaysia Timur.

 

Kata kunci: Ikan buntal; LCMS/MS; Malaysia Timur; Tetrodotoksin; tisu

RUJUKAN

Ahasan, H.A.M.N., Mamun, A.A., Karim, S.R., Bakar, M.A., Gazi, E.A. & Bala, C.S. 2004. Paralytic complications of puffer fish (Tetrodotoxin) poisoning. Singapore Medical Journal 45(2): 73-74.

Arakawa, O., Hwang, D.F., Taniyama, S. & Takatani, T. 2010. Toxins of pufferfish that cause human intoxications. In Coastal Environmental and Ecosystem Issues of the East China Sea, edited by Ishimatsu, A. & Lie, H.J. pp. 227-244.

Brillantes, S., Samosorn, W., Faknoi, S. & Oshima, Y. 2003. Toxicity of puffers landed and marketed in Thailand. Fisheries Science 69: 1224-1230.

Berry, P.Y. & Hassan, A.A. 1973. Comparative lethality of tissue extracts from the Malaysian puffer fishes Lagocephalus lunaris lunaris, L. l. spadiceus and Arothron stellatus. Toxicon 11: 249-254.

Che Nin, M., Norjuliana, M.N., Gan, L.H., Razak, L. & Samsur, M. 2010. Screening of tetrodotoxin in puffers using gas chromatography-mass spectrometry. Journal of Chromatography A 1217(47): 7455-7459.

Chew, S.K., Goh, C.H., Wang, K.W., Mah, P.K. & Tan, B.Y. 1983. Puffer fish (Tetrodotoxin) poisoning: Clinical report and role of anti-cholinesterase drugs in therapy. Singapore Medical Journal 24: 168-171.

Chou, S.S., Tsai, Y.H., Chang, P.C. & Hwang, D.F. 1994. Tetrodotoxin-associated food poisoning due to ingesting fish. Journal Food and Drug Analysis 2(1): 77.

Chua, H.H. & Chew, L.P. 2009. Puffer fish poisoning: A family affair. Medical Journal of Malaysia 64(2): 181-182.

Chulanetra, M., Soonkrung, N., Srimanote, P., Indrawattana, N., Thanongsaksrikul, J., Sakolvaree, Y., Chongsa-Nguan, M., Kurazono, H. & Chaicumpa, W. 2011. Toxic marine puffer fish in Thailand seas and tetrodotoxin they contained. Toxins 3: 1249-1262.

Diener, M., Christian, B., Ahmed, M.S. & Luckas, B. 2007. Determination of tetrodotoxin and its analogs in the puffer fish Takifugu oblongus from Bangladesh by hydrophilic interaction chromatography and mass-spectrometric detection. Analytical Bioanalytical Chemistry 389: 1997- 2002.

EC (European Commission). 2004. Corrigendum to Regulation (EC) No 853/2004 of the European Parliament and of the Council of 29 April 2004 laying down specific hygiene rules for food of animal origin. Official Journal of the European Union L 139. pp L226/68 (25/6/2004).

El-Sayed, M., Yacout, G.A., El-Samra, M., Ali, A. & Kotb, S.M. 2003. Toxicity of the Red Sea pufferfish Pleuranacanthus sceleratus “El-Karad”. Ecotoxicology and Environmental Safety 56(3): 367-372.

Fisheries Research Institute (FRI). 2004. Field Guide to Selected Commercial Marine Fishes of Malaysian Waters. Pulau Pinang, Department of Fisheries Malaysia.

Froese, R. & Pauly, D. 2011. FishBase. www.fishbase.org. Accessed on August 2011.

Helbig, T. & Luckas, B. 2010. Determination of tetrodotoxins (TTXs) and paralytic shellfish poisoning (PSP) toxins in puffer fish from Malaysia and Japan by application of a ZIC-HILIC column and mass spectrometric detection. Current Development in Oceanography 1(2): 69-83.

Isbister, G.K., Son, J., Wang, F., Maclean, C.J., Lin, C.S., Ujma, J., Balit, C.R., Smith, B., Milder, D.G. & Kiernan, M.C. 2002. Puffer fish poisoning: A potentially life-threatening condition. Medical Journal of Australia 177(11-12): 650-653.

Jang, J.H., Lee, J.S. & Yotsu-Yamashita, M. 2010. 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 South East Asia. Marine Drugs 8: 1049-1058.

Japan Food Hygiene Association. 2005. Puffer toxin. In Environmental Health Bureau, Ministry of Health and Welfare, edited by Shokuhin Eisei Kensa Shishin. Manual for Methods for Food Sanitation Testing, Tokyo. pp. 661-673.

Jen, H.C., Lin, S.J., Tsai, Y.H., Chen, C.H., Lin, Z.C. & Hwang, D.F. 2008. Tetrodotoxin poisoning evidenced by solid-phase extraction combining with liquid chromatography-tandem mass spectrometry. Journal of Chromatography B 871: 95-100.

Kan, S.K., Chan, M.K. & David, P. 1987. Nine fatal cases of puffer fish poisoning in Sabah, Malaysia. Medicine Medical Journal of Malaysia 42(3): 199-200.

Katikou, P., Georgantelis, D., Sinouris, N., Petsi, A. & Fotaras, T. 2009. First report on toxicity assessment of the Lessepsian migrant pufferfish Lagocephalus sceleratus (Gmelin, 1789) from European waters (Aegean Sea, Greece). Toxicon 54: 50-55.

Kungsuwan, A. 1994. Survey on poisonous pufferfish in Andaman Seas. Proceeding of the Seminar on Fisheries 1993, Department of Fisheries, Bangkok, Thailand, 15-17 September 1993.

Kungsuwan, S., Arakawa, O., Promdet, M. & Onoue, Y. 1997. Occurrence of paralytic shellfish poisons in Thai freshwater puffers. Toxicon 35: 1341-1346.

Laobhripatr, S., Limpakarnjanarat, K., Sangwonloy, O., Sudhasaneya, S., Anuchatvorakul, B., Leelasitorn, S. & Saitanu, K. 1990. Food poisoning due to consumption of the freshwater puffer Tetraodon fangi in Thailand. Toxicon 28(11): 1372-1375.

Lin, S.J. & Hwang, D.F. 2001. Possible source of tetrodotoxin in the starfish Astropecten scoparius. Toxicon 39: 573-579.

Lyn, P.C.W. 1985. Puffer fish poisoning: Four case reports. Medical Journal of Malaysia 40(1): 31-34.

Mahmud, Y., Tanu, M.B., Takatani, T., Asayama, E., Arakawa, O. & Noguchi, T. 2001. Chelonodon patoca, a highly toxic marine puffer in Japan. Journal of Natural Toxins 10: 69-74.

Matsui, T., Taketsugu, S., Sato, H., Yamamori, K., Kodama, K., Ishi, A., Hirose, H. & Shimizu, C. 1990. Toxification of cultured puffer fish by the administration of tetrodotoxin producing bacteria. Nippon Suisan Gakkaishi 56(4): 705.

Miyazawa, K. & Noguchi, T. 2001. Distribution and origin of tetrodotoxin. Journal of Toxicology Toxin Reviews 20: 11-33.

Mohamad, S., Lim, P.T., Bojo, O., Nyanti, L., Ismail, N., Rahim, K.A. & M. Long, S. 2008. Toxicity of freshwater puffer fish Xenopterus naritus (Tetraodontidae) collected from East Malaysia. IOC-WESTPAC Scientific Symposium. 21st- 25th May 2008.

Monaliza, M.D. & Mohamad, S. 2011. Toxicity and toxin properties study of pufferfish collected from Sabah waters. Health and the Environment Journal 2(1): 14-17.

Mosher, H.S. & Fuhrman, F.A. 1984. Occurrence and origin of tetrodotoxin. In Seafood Toxins, edited by Edward, P.R. Washington, D.C.: American Chemical Society. pp. 333-344.

Nagashima, Y., Matsumoto, T., Kadoyama, K., Ishizaki, S. & Terayama, M. 2011. Toxicity and molecular identification of Green Toadfish Lagocephalus lunaris collected from Kyushu Coast, Japan. Journal of Toxicology 2011: 801285.

Nakagawa, T., Junho, J. & Yotsu-Yamashita, M. 2006. Hydrophilic interaction liquid chromatography-electrospray ionization mass spectrometry of tetrodotoxin and its analogs. Analytical Biochemistry 352: 142-144.

Nakamura, M. & Yasumoto, T. 1985. Tetrodotoxin derivatives in puffer fish. Toxicon 23: 271-276.

Ngy, L., Yu, C.F., Taniyama, S., Takatani, T. & Arakawa, O. 2009. Co-occurence of tetrodotoxin and saxitoxin in Cambodian marine pufferfish Takifugu oblongus. African Journal of Marine Science 31(3): 349-354.

Noguchi, T. & Arakawa, O. 2008. Tetrodotoxin – distribution and accumulation in aquatic organisms and cases of human intoxication. Marine Drugs 6: 220-242.

Noguchi, T., Arakawa, O. & Takatani, T. 2006. TTX accumulation in pufferfish. Comparative Biochemistry and Physiology, Part D 1: 145-152.

Nunez-Vazquez, E.J., Yotsu-Yamashita, M., Sierra-Beltran, A.P., Yasumoto, T. & Ochoa, J.L. 2000. Toxicities and distribution of tetrodotoxin in the tissues of puffer fish found in the coast of the Baja California Peninsula, Mexico. Toxicon 38: 729-734.

Oliveira, J.S., Fernandes, S.C.R., Schwartz, C.A., Bloch Jr., C., Melo, J.A.T., Pires Jr., O.R. & Freitas, J.C. 2006. Toxicity and toxin identification in Colomesus asellus, an Amazonian (Brazil) freshwater puffer fish. Toxicon 48: 55-63.

Othman, B., Khairul Adha, A.R., Samsur, M., Shabdin, M.L., Lee, N., Norhadi, I. & Lim, P.T. 2006. Toxicity of the yellow puffer fish Xenopterus naritus from Sungai Saribas Sarawak. Research Bulletin, FRST 1 (June). pp 5-6.

Pires, O.R., Sebben, A., Schwartz, E.F., Largura, S.W.R., Bloch, C., Morales, R.A.V. & Schwartz, C.A. 2002. Occurrence of tetrodotoxin and its analogues in the Brazilian frog Brachycephalus ephippium (Anura: Brachycephalidae). Toxicon 40(6): 61-766.

Rodriquez, P., Alfonso, A., Otero, P., Katikou, P., Georgantelis, D. & Botana, L.M. 2012. Liquid chromatography-mass spectrometry method to detect Tetrodotoxin and its analogues in the puffer fish Lagocephalus sceleratus (Gmelin, 1789) from European waters. Food Chemistry 132: 1103-1111.

Sato, S., Kodama, M., Ogata, T., Saitanu, K., Furuya, M., Hirayama, K. & Kakinuma, K. 1997. Saxitoxin as a toxic principle of a freshwater puffer, Tetraodon Fangi, in Thailand. Toxicon 35: 137-140.

Shoji, Y., Yotsu-Yamashita, M., Miyazawa, T. & Yasumoto, T. 2001. Electrospray ionization mass spectrometry of tetrodotoxin and its analogs: Liquid chromatography/ mass spectrometry, tandem mass spectrometry and liquid chromatography/tandem mass spectrometry. Analytical Biochemistry 290: 10-17.

Simon, K.D., Mazlan, A.G. & Usup, G. 2009. Toxicity of puffer fishes (Lagocephalus wheeleri Abe, Tabeta and Kitahama, 1984 and Lagocephalus sceleratus Gmelin, 1789) from the east coast waters of Peninsular Malaysia. Journal of Biological Sciences 9(5): 482-487.

Tani, I. 1945. Studies on Japanese Pufferfish in Association with Poisoning due to Ingestion of Them. Tokyo: Teikoku, Tosho. p. 103.

Tsai, Y.H., Chao, S.M., Lin, G.T., Noguchi, T. & Hwang, D.F. 2004. Tetrodotoxins of the starfish Astropecten vappa collected from western Taiwan. Fisheries Science 70: 930- 932.

Yasumoto, T. & Michishita, T. 1985. Fluorometric determination of tetrodotoxin by high performance liquid chromatography. Agricultural and Biological Chemistry 49: 3077-3080.

Yasumoto, T., Yasumura, D., Yotsu, M., Michishita, T., Endo, A. & Kotaki, Y. 1986. Bacterial production of tetrodotoxin and anhydrotetrodotoxin. Agricultural and Biological Chemistry 50(3): 793-795.

Yotsu-Yamashita, M., Jang, J.H., Cho, Y. & Konoki, K. 2011. Optimization for simultaneous analysis of tetrodotoxin, 4-epitetrodotoxin, 4,9-anhydrotetrodotoxin and 5,6,11-trideoxytetrodotoxin by hydrophilic interaction liquid chromatography-tandem mass spectrometry. Forensic Toxicology 29: 61-64.

Yu, C.F. & Yu, P.H.F. 2002. Are puffer fish more toxic in their spawning seasons? Marine Biology 140: 1053-1057.

 

 

*Pengarang untuk surat-menyurat; email: mohayo01@yahoo.com

 

 

sebelumnya