Paralytic shellfish toxins (PSTs) and tetrodotoxin (TTX) of Korean pufferfish

이가정,하광수,정연중,목종수,손광태,Hee-Chung Lee,김지회 한국수산과학회 2021 Fisheries and Aquatic Sciences Vol.24 No.11

Paralytic shellfish toxins (PSTs) and tetrodotoxin (TTX) are neurotoxins that display pharmacological activity that is similar to that of specific sodium channel blockers; they are the principle toxins involved in shellfish and puffer fish poisoning. In Korea, puffer fish is a very popular seafood, and several cases of accidental poisoning by TTX have been reported. Therefore, it is necessary to determine whether puffer fish poisoning incidents are caused by PSTs or by TTX. In this study, we used mouse bioassay (MBA) and liquid chromatograph-tandem mass spectrometry (LC-MS/MS) to determine the presence of PSTs and TTX in puffer fish from an area near Mireuk-do, Tong-Yeong on the southern coast of Korea from January through March, 2014. The toxicity of PSTs and TTX extracts prepared from three organs of each specimen was analyzed by MBA. Most of the extracts killed mice with typical signs of TTX and PSTs. The LC-MS/MS analysis of seven specimens of Takifugu pardalis and Takifugu niphobles, each divided into muscles, intestines, and liver, were examined for TTX. In T. pardalis, the TTX levels were within the range of 1.3–1.6 μg/g in the muscles, 18.8–49.8 μg/g in the intestines, and 23.3–96.8 μg/g in the liver. In T. niphobles, the TTX levels were within the range of 2.0–4.5 μg/g in the muscles, 23.9–71.5 μg/g in the intestines, and 28.1–114.8 μg/g in the liver. Additionally, the toxicity profile of the detected PSTs revealed that dcGTX3 was the major component in T. pardalis and T. niphobles. When PSTs were calculated as saxitoxin equivalents the levels were all less than 0.5 μg/g, which is below the permitted maximum standard of 0.8 μg/g. These findings indicate that the toxicity of T. pardalis and T. niphobles from the southern coast of Korea is due mainly to TTX and that PSTs do not exert an effect.

41 마비성패류독소 검출을 위한 분석법 비교

이가정,정연중,손광태,하광수,목종수,이태식,김지회 한국수산과학회 양식분과 2017 한국수산과학회 양식분과 학술대회 Vol.2017 No.5

마비성패류독소 검출을 위한 분석법 비교

이가정,권순재,정연중,손광태,하광수,목종수,김지회 한국수산과학회 2017 한국수산과학회지 Vol.50 No.6

Paralytic shellfish toxins (PSTs) are produced by marine dinoflagellate phytoplankton Alexandrium spp. and Gymnodiniumspp. These toxins accumulate in filter feeding organisms such as bivalves and the ingestion of contaminatedshellfish can cause illness in humans. The mouse bioassay (MBA) has been the preferred PST testing methodworldwide for more than 50 years. However, this assay has several disadvantages, such as detection limits, non-toxicprofiles,and the ethical issues of using animals. The aim of this study was to establish an alternative to the MBAmethod for testing for PSTs. We optimized the analysis conditions of a post-column oxidation-high performanceliquid chromatography (PCOX-HPLC) method and the Scotia Rapid Test Kit, and then compared the accuracy ofthese methods to the MBA method. The results demonstrated a strong correlation between the PCOX-HPLC methodand the MBA, although the PCOX-HPLC method required expensive equipment and standard material, and wastime consuming. The Scotia Rapid Test Kit promises to be a useful tool, as it provided rapid and qualitative results,although the method sometimes gave a false positive result that could not be explained by toxin profiles.

Concentration of heavy metals in shellfishes and health risk assessment from Korean coastal areas

이가정,강은혜,윤민철,조미라,유홍식,손광태 한국수산과학회 2022 Fisheries and Aquatic Sciences Vol.25 No.12

Shellfish are exoskeleton-bearing aquatic invertebrates that consume various organic and inorganic substances floating in seawater through filter feeding. Heavy metals are known as absorbed and accumulated in seawater. Some of the toxic heavy metals are highly accumulated in seawater, and exposure to them can cause a variety of risks to the human body. Since Koreans like to eat seafood, they are more likely to be exposed to contaminated seafood with heavy metals. In this study, nine types of heavy metals were analyzed on ten different shellfish species in the coastal area of South Korea. The risk assessment was also done on shellfish in which heavy metals were detected. Zinc (Zn) and copper (Cu) were identified at an average of 56.7 mg/kg (6.70 to 466 mg/kg) and 13.2 mg/kg (0.064 to 143 mg/kg), respectively. Lead (Pb) average of 0.208 mg/kg (0.000750 to 1.02 mg/kg), cadmium (Cd) average of 0.454 mg/kg (0.0388 to 1.56 mg/ kg) and mercury (Hg) average of 0.0266 mg/kg (0.00548 to 0.174 mg/ kg) were identified. Additionally, arsenic (As), chromium (Cr), nickel (Ni), and silver (Ag) were also identified as average concentrations of 4.02 (0.460 to 15.0 mg/kg), 0.167 (< limit of quantification [LOQ] to 0.820 mg/kg), 0.281 (< LOQ to 1.46 mg/kg), and 0.158 mg/kg (< LOQ to 1.15 mg/kg). The result indicates that the monitoring results of heavy metals in most shellfish satisfied the Korean standard. However, Pb and Cd have exceeded some foreign standards, such as the United States and the EU. The permissible human exposure calculated using the heavy metal intake and detection amount was lower than the Joint FAO/WHO Expert Committee on Food Additives human safety standard, and the risk of heavy metals from shellfish consumption was at an acceptable level.

새로운 생산방식의 한국도입 실태와 노동조합의 역할

이가정 淑明女子大學校 1993 숙명 - 淑明女子大學校 Vol.33 No.-

LC-MS/MS를 이용한 설사성패류독소의 분석조건 확립

이가정(Ka Jeong Lee),스즈키 도시유키(Toshiyuki Suzuki),김풍호(Poong Ho Kim),오은경(Eun Gyoung Oh),송기철(Ki Cheol Song),김지회(Ji Hoe Kim) 한국식품과학회 2009 한국식품과학회지 Vol.41 No.4

설사성패류독의 신속정밀 분석조건 확립을 위하여 LC-MS/MS를 사용하여 이동상, 분석용 column 및 collision energy 등을 변화시키면서 시험하였다. 50 mM formic acid와 2 mM ammonium formate가 함유된 acetonitrile 수용액을 이동상으로 사용하였을 때 OA와 DTX1이 검출되었다. Collision energy는 독소 성분에 따라 달리하는 것이 다성분 동시분석에 적합하였으며 OA와 DTX1 고유의 fragment ion들은 48 V 정도에서 최적의 intensity로 확인되었다. Column의 종류에 따라서는 C8 column의 경우 OA, DTX1, DTX3, PTX2 및 YTX 모두 검출 가능하였으나 실제 검출 대상이 OA와 DTX1인 경우에는 일반적으로 사용되는 Csub{18} column도 적합한 것으로 확인되었다. 본 연구에서 확립한 LC-MS/MS 분석 조건의 검출한계는 OA와 DTX1 모두 1 ng/g, 정량한계는 각각 3ng/g이었고, 표준독 성분을 첨가한 시료에서 process efficiency는 굴의 경우 91-118%, 진주담치에서는 96-117%이었고, matrix의 영향은 거의 없었다. 마우스 시험에서 양성을 나타낸 시료를 LCMS/MS법으로 분석한 결과, 일부 시료에서만 OA 및 DTX1이 검출되어 두 시험법의 독성은 일치하지 않았으며 LC-MS/MS법은 마우스 시험법보다 하루 이상 분석시간을 단축할 수 있었다. To establish and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the rapid and accurate quantitation of diarrhetic shellfish poisoning (DSP) toxins, we compared the results from different mobile phases and columns used for their analysis and collision energies for MS/MS experiments. Clear peaks of okadaic acid (OA) and dinophysistoxin-1 (DTX1) were obtained by using a mobile phase comprising aqueous acetonitrile containing 2 mM ammonium formate and 50 mM formic acid. The collision energies were optimized to facilitate the most sensitive detection for each toxin, namely, OA, DTX1, pectenotoxin-2 (PTX2), or yessotoxin (YTX). Further, the maximum ion response was obtained at a collision energy of 48 V for OA and DTX1. We compared the analytical performance of Csub{8} and Csub{18} columns. A wide range of toxins namely, OA, DTX1, PTX2, and YTX, except DTX3, were detected by both the columns. Although DTX3 was only detected by the Csub{8} column, we found that the Csub{18} column was also suitable for the quantitation of OA and DTX1 the toxins responsible for inducing diarrhea. The limit of detection of OA and DTX1 by the established LC-MS/MS conditions was 1 ng/g, and the limit of quantitation of the toxins under the same conditions was 3 ng/g. The process efficiencies were 91-118% for oysters (Crassostrea gigas) and 96-117% for mussels (Mytilus galloprovincialis) further, we observed no significant effect of matrix during the ionization process in LC-MS/MS. The comparison between mouse bioassay (MBA) and LC-MS/MS yielded varying results because low concentrations of OA and DTX1 were detected by LC-MS/MS in some shellfish samples, which provided positive results on MBA for DSP. The analysis time required by MBA for DSP analysis can be reduced by LC-MS/MS.

경남 진해만해역의 마비성패류독소 모니터링 빈도에 대한 위해성평가

하광수,이가정,손광태,김지회,김풍호,김연계,김영목,노맹석,이태식 한국수산과학회 양식분과 2017 한국수산과학회 양식분과 학술대회 Vol.2017 No.5

경남 한산·거제만해역에서 생산된 패류의 위생학적 안전성 평가

하광수,이가정,정연중,목종수,김풍호,김연계,이희정,김동욱,손광태,Ha, Kwang-Soo,Lee, Ka-Jeong,Jeong, Yeon-Jung,Mok, Jong-Soo,Kim, Poong-Ho,Kim, Yeon-Kye,Lee, Hee-Jung,Kim, Dong-Wook,Son, Kwang-Tae 한국식품위생안전성학회 2018 한국식품위생안전성학회지 Vol.33 No.5

본 연구는 2013년부터 2017년까지 경남 한산 거제만 해역에서 생산되는 패류에 대한 위생지표세균, 마비성 패류독소, 설사성 패류독소를 분석하여 세균학적 독물학적 안전성을 평가하였다. 굴 시료 404점에 대한 분변계대장균은 < 18~330 MPN/100 g의 범위를 나타내었으나, 대장균 230 E. coli MPN/100 g을 초과한 시료는 없었다. 굴 수확시기 동안 분석한 대장균의 기하학적 평균치는 24.3 MPN/100 g으로 매우 낮게 나타났다. 한산 거제만해역에서 생산되는 굴의 세균학적 안전성을 평가한 결과, 식품위생법의 생식용 굴에 대한 위생기준을 만족하였고, EU의 패류생산해역 A등급 기준에 부합하였다. 한산 거제만해역의 독물학적 평가를 위해 굴 532개 시료와 패류독소 지표종인 지중해담치 268개 시료에 대한 분석을 실시하였다. 마비성패류독소는 2013년 4월에 지중해담치 3개 시료에서 1.20~2.29 mg/kg 범위로 기준을 초과하여 검출되었으나, 굴시료에서는 전혀 검출되지 않았다. 총 120개 시료에서 설사성패류독소를 분석하였으나, 정량한계 이하로 매우 낮게 나타났다. 독물학적 안전성 평가결과, 굴 시료에서는 마비성 및 설사성패류독소가 전혀 검출되지 않았으나, 지표종인 지중해담치에서는 마비성패류독소 기준치를 초과한 것으로 나타나 지속적인 모니터링과 안전관리가 요구되었다. To evaluate bacteriological and toxicological safety hygienic indicator bacterium and paralytic and diarrhetic shellfish toxins in the shellfish produced in Hansan Geojeman 2013-2017 were investigated. Fecal coliforms were < 18~330 MPN/100 g in 404 oyster samples. But all samples tested, did not exceed 230 E. coli MPN/100 g. Geometric mean of E. coli for oyster samples collected during major shellfish production period was 24.3 MPN/100 g, considerde stable results. Bacteriological quality of oysters collected from Hansan Geojeman meets the standard value based on shellfish hygiene of the Food Sanitation Act of Korea and also meets Grade A, according to classification of shellfish harvesting areas of the European Union. For toxicological evaluation of Hansan Geojeman, 532 oyster samples and 268 mussel samples as an indicator, were analyzed. Paralytic shellfish toxins were detected in the range of 0.42~2.29 mg/kg in eight mussel samples, and exceeded criteria in three samples from early to late April 2013. Diarrhetic shellfish toxin was detected in three of 120 samples, but it was revealed to be under regulation value (0.16 mg Okadaic Acid equ./kg). As a result of toxicological evaluation, paralytic and diarrhetic shellfish toxins were not detected in oyster samples, but it was found that mussel as an indicator species, exceeded the threshold value of paralytic shellfish toxin. Accordingly, sanitary surveys were continuously requested for food safety management of shellfish.

경남 한산·거제만해역에서 생산된 패류의 위생학적 안전성 평가

하광수,이가정,정연중,목종수,김풍호,김연계,이희정,김동욱,손광태 한국식품위생안전성학회 2018 한국식품위생안전성학회지 Vol.33 No.5

본 연구는 2013년부터 2017년까지 경남 한산·거제만 해역에서 생산되는 패류에 대한 위생지표세균, 마비성 패 류독소, 설사성 패류독소를 분석하여 세균학적·독물학적 안전성을 평가하였다. 굴 시료 404점에 대한 분변계대장균은 < 18~330 MPN/100 g의 범위를 나타내었으나, 대장 균 230 E. coli MPN/100 g을 초과한 시료는 없었다. 굴 수확시기 동안 분석한 대장균의 기하학적 평균치는 24.3 MPN/100 g으로 매우 낮게 나타났다. 한산·거제만해역에서 생산되는 굴의 세균학적 안전성을 평가한 결과, 식품위 생법의 생식용 굴에 대한 위생기준을 만족하였고, EU의 패 류생산해역 A등급 기준에 부합하였다. 한산·거제만해역 의 독물학적 평가를 위해 굴 532개 시료와 패류독소 지표 종인 지중해담치 268개 시료에 대한 분석을 실시하였다. 마비성패류독소는 2013년 4월에 지중해담치 3개 시료에서 1.20~2.29 mg/kg 범위로 기준을 초과하여 검출되었으나, 굴 시료에서는 전혀 검출되지 않았다. 총 120개 시료에서 설사성패류독소를 분석하였으나, 정량한계 이하로 매우 낮게 나타났다. 독물학적 안전성 평가결과, 굴 시료에서는 마비성 및 설사성패류독소가 전혀 검출되지 않았으나, 지 표종인 지중해담치에서는 마비성패류독소 기준치를 초과 한 것으로 나타나 지속적인 모니터링과 안전관리가 요구 되었다. To evaluate bacteriological and toxicological safety hygienic indicator bacterium and paralytic and diarrhetic shellfish toxins in the shellfish produced in Hansan·Geojeman 2013-2017 were investigated. Fecal coliforms were < 18~330 MPN/100 g in 404 oyster samples. But all samples tested, did not exceed 230 E. coli MPN/ 100 g. Geometric mean of E. coli for oyster samples collected during major shellfish production period was 24.3 MPN/100 g, considerde stable results. Bacteriological quality of oysters collected from Hansan·Geojeman meets the standard value based on shellfish hygiene of the Food Sanitation Act of Korea and also meets Grade A, according to classification of shellfish harvesting areas of the European Union. For toxicological evaluation of Hansan·Geojeman, 532 oyster samples and 268 mussel samples as an indicator, were analyzed. Paralytic shellfish toxins were detected in the range of 0.42~2.29 mg/kg in eight mussel samples, and exceeded criteria in three samples from early to late April 2013. Diarrhetic shellfish toxin was detected in three of 120 samples, but it was revealed to be under regulation value (0.16 mg Okadaic Acid equ./kg). As a result of toxicological evaluation, paralytic and diarrhetic shellfish toxins were not detected in oyster samples, but it was found that mussel as an indicator species, exceeded the threshold value of paralytic shellfish toxin. Accordingly, sanitary surveys were continuously requested for food safety management of shellfish.

가막만해역 패류의 세균학적·독물학적 안전성 평가

하광수,신순범,이가정,정상현,오은경,이희정,김동욱,김연계 한국식품위생안전성학회 2017 한국식품위생안전성학회지 Vol.32 No.6

To evaluate bacteriological and toxicological safety, the hygienic indicator bacterium and paralytic and diarrhetic shellfish toxins in the shellfish produced in the Kamakman Area from 2012 to 2016 were investigated. Fecal coliforms and E. coli of all 194 oyster samples tested did not exceed 230 MPN/100 g. The geometric mean of the fecal coliform analyzed with the oyster samples of harvesting period was 19.6 MPN/100 g, which was more stable than the non-harvesting period (26.5 MPN/100 g). For the toxicological evaluation of the Kamakman Area, 77 oyster samples and 350 mussel samples as an indicator were analyzed. Paralytic shellfish toxins were detected very low in the range of 40~46 μg/100 g in 13 mussel samples during late April and early June, but not in oyster samples. Diarrhetic shellfish toxin was detected in 2 of 180 samples, but it was found to be below the regulation value (0.16 mg OA equ./kg). Based on the bacteriological studies, it was confirmed that the shellfish produced in Kamakman area meets the standard of shellfish hygiene of the Food Sanitation Act and meets the Grade A of the shellfish production area of EU. As the results of the paralytic and diarrhetic shellfish toxin evaluation, it was confirmed that the Kamakman Area is also toxicologically safe for shellfish production.