**Abbreviations**

GC: gas chromatography; GLC: gas liquid chromatography; MS: mass spectrometry; HPLC: high performance liquid chromatography; MTBSTFA: N-methyl-N-(t-butyldimethylsilyl) trifluoroacetamide; tBDMS: *tert*-butyldimethylsilyl; TMS: trimethylsilyl; PTC: phenylthiocarbamoyl; PITC: phenyl isothiocyanate; SPME: solid-phase microextraction.

Ala: alanine; Gly : glycine; Val: valine; Leu: leucine; Ile: isoleucine; Pro: proline; Ser: serine; Thr: threonine; Phe: phenylalanine;Asp: aspartate; Glu: glutamate;Orn: ornithine; Asn:

asparagine;Gln: glutamine; Arg: arginine;Tyr: tyrosine; Lys: lysine; Cys: cysteine; Met: methionine, His: histidine; Hcy: homocysteine.

Chromatographic Analysis of Nitrogen Utilization and Transport in Arbuscular Mycorrhizal Fungal Symbiosis 243

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## **7. References**


Chromatographic Analysis of Nitrogen Utilization and Transport in Arbuscular Mycorrhizal Fungal Symbiosis 243

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**Chapter 10** 

© 2012 Asakawa et al., licensee InTech. This is an open access chapter distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

© 2012 Asakawa et al., licensee InTech. This is a paper distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

**Instrumental Analysis of Tetrodotoxin** 

Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/48668

**1. Introduction** 

Manabu Asakawa, Yasuo Shida, Keisuke Miyazawa and Tamao Noguchi

Tetrodotoxin (TTX), a pufferfish ("fugu" in Japanese) toxin named after its order name Tetraodontiformes, is the toxic principle of puffer fish poisoning. This toxin (C11H17N3O8; a molecular weight of 319) is one of the most potent nonproteinaceous toxins as well as the best-known marine natural toxins (Figure 1). In Japan, pufferfish have been a traditional food for many years, and since people have become accustomed to eating them, cases of TTX poisoning are frequent. It poses a serious hazard to public health. These cases have occurred on a regular basis not only in Japan but also in Asia for a number of years, sporadically resulting in severe poisoning or even death. On the other hand, the Japanese are aware of the its toxicity and have devised methods to reduce TTX levels especially in the liver. However, TTX poisoning incidents continue to occur in Japan. Since there is no antidote for the toxin, patient mortality is very high. Judging from statistics provided by the Japanese Ministry of Health, Labour and Welfare, the number of deaths due to puffer poisoning has steadily declined, from more than 10 cases every year between 1960 and 1981 to less than 10 cases with low mortality every year since 1982, generally with low mortality. This decline is probably due to not only strict adherence to government regulations but also an increase in cultured puffer rather than a decrease in wild puffer. The toxicosis is characterized by the onset of symptoms in the victim. Treatment of the illness is mainly based on the symptoms of the patient. More fruitful treatment can be provided if the causative toxin is identified. In 1950, TTX was isolated for the first time as a crystalline prism from toxic pufferfish ovaries by Yokoo [1]. Its structure was elucidated by three groups in 1964 [2-4]. TTX is a powerful and specific sodium channel blocker [5]. When ingested by humans, it acts to block the sodium channels in the nerve cells and skeletal muscles, and to thereby block excitatory conduction, resulting in the occurrence of typical symptoms and signs such as respiratory paralysis and even death in severe cases. The lethal potency is 5000 to 6000 MU/mg. One MU (mouse unit) is defined as the amount of toxin required to kill a 20g male mouse within 30 min after intraperitoneal administration, and the minimum lethal dose (MLD) for humans is estimated to be approximately 10,000 MU

