• Korean Journal of Fisheries and Aquatic Sciences
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• v.27 no.6
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• pp.682-689
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• 1994

Ninety-six specimens of nine puffer fishes landed at a fish market of Pusan, Korea were assayed for anatomical distribution of toxicity. The puffers, Lagocephalus gloveri ('geommilbog'), Fugu rubripes rubripes ('jajubog'), Fugu rubripes chinesis ('geomjajubog'), Lagocephalus wheeleri ('hinmilbog') and Arothron firmamentum ('byeolbog'), were shown to be nontoxic in muscle and skin. However, toxicities in skins of four specimens of Lagocephalus gloveri and a specimen of Arothron firmamentum were more than 10MU/g In the puffer, Fugu xanthopterus ('ggachibog'), the skin showed to be weakly toxic as far as nine specimens out of 27 ones are concerned, and the muscle was nontoxic in all specimens. Meanwhile, it was noted that the puffer, Fugu vermicularis radiatus was found to be a toxic species, differing from the toxicity known so far. Their highest toxicities were 3,880, 1,191, 1,115, 219, 289, 5,620 and 753MU/g for liver, intestine, skin, muscle, testis, ovary and bile, respectively.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.5
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• pp.612-617
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• 2008

Tetrodotoxin (TTX) analogues were first determined from the liver extracts of puffer fish, Takifugu niphobles, by LC/MS with Hydrophilic Interaction Liquid Chromatography (HILIC). In total, 7 TTX analogues were detected within 20 minutes as follows; 5,6,11-trideoxyTTX (34.0%, 1,029.6 nmol/g), 6,11-dideoxyTTX (29.3%, 887.6 nmol/g), TTX (22.1%, 667.8 nmol/g), 4,9-anhydro-TTX (11.2%, 339.3 nmol/g), 11-deoxyTTX＋5-deoxyTTX (2.6%, 78.6 nmol/g), and 4-epiTTX (0.8%, 23.6 nmol/g). Mouse toxicity of diluted liver extracts showed the highest toxicity at pH 3 (8.7 MU/mL) and decreased, as increasing pH, to 1.4 MU/mL at pH 10. At acidic (pH 5) and neutral conditions (pH 7), mouse toxicity of liver extracts (79 MU/mL) decreased slowly, as increasing temperature from $80^{\circ}C$ to $115^{\circ}C$, and time until 1 hour; in contrast, at the akaline condition (pH 9), the toxicity decreased rapidly to the more than half within 10 minutes. Individual toxicity of the fillet of T. niphobles were between $43.2{\sim}106.7$ MU, and $64{\sim}78%$ of its toxicity was eluted to soup when boiled with 3 volumes of water during 10 minutes.

• Journal of Food Hygiene and Safety
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• v.27 no.1
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• pp.37-41
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• 2012

The current standard for testing tetrodotoxin (TTX) in foodstuffs is the mouse bioassay (MBA) in Korea as in many other countries. However, this test suffers from potential ethical concerns over the use of live animals. In addition, the mouse bioassay does not test for a specific toxin thus a sample resulting in mouse incapacitation would need further confirmatory testing to determine the exact source toxin (e.g., TTX, STX, brevotoxin, etc.). Furthermore, though the time of death is proportional to toxicity in this assay, the dynamic range for this proportional relationship is small thus many samples must be diluted and new mice be injected to yield a result that falls within the quantitative dynamic range. Therefore, in recent years, there have been many efforts in this field to develop alternative assays. High performance liquid chromatography (HPLC) coupled with mass spectrometry (MS) has been emerged as one of the most promising options. A LC-MS-MS method involves solid-phase extraction (SPE) and followed by analysis using an electrospray in the positive ionization mode and multiple reactions monitoring (MRM). To adopt LC-MS-MS method as alternative standard for testing TTX, we performed a validation study for the quantification of TTX in puffer fish. This LC-MS-MS method showed good sensitivity as limits of detection (LOD) of $0.03{\sim}0.08{\mu}g/g$ and limits of quantification (LOQ) of $0.10{\sim}0.25{\mu}g/g$. The linearity ($r^2$) of tetrodotoxin were 0.9986~0.9997, the recovery were 80.9~103.0% and the relative standard deviations (RSD) were 4.3~13.0%. The correlation coefficient between the mouse bioassay and LC/MS/MS method was higher than 0.95.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.1
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• pp.31-34
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• 1995

Ten specimens (5 males and 5 females) of the pufferfish, fugu stictonotus ('gachilbog'), were collected at a fish market of Pusan, Korea in July 1993, and examined for anatomical distribution of toxicity by mouse assay method. The frequency of toxic specimens was $40\%\;for\;liver,\;60\%$ for ovary, $40\%\;for\;skin\;and\;60\%$ for bile in female puffers. The highest toxicities were 107, 107, 29 and 93MU/g for liver, ovary, skin and bile, respectively; and average toxicity $\pm S.E.\;values\;were\;14\pm11,\;48\pm22.4\pm3\;and\;12\pm9MU/g,$ respectively. The range of total toxicity was shown to be from 0 to 35,316MU. The characteristic pattern of toxin distribution observed on these specimens was exhibited; both muscle and testis were non-toxic, but others were weakly toxic. Also, there was significant difference for toxicity between male and female specimens.