• 한국수산과학회지
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• 제43권4호
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• pp.293-297
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• 2010

To prevent amnesic shellfish poisoning (ASP) resulting from the consumption of shellfish contaminated with domoic acid, the quantitative analysis of domoic acid is very important. We validated a high performance liquid chromatography (HPLC) method for accurate and precise quantification of domoic acid. A clear peak and the isolation of domoic acid resulted on injecting a 50% methanol extract of CRM-ASP-Mus-c mussel reference material using HPLC. The limit of detection of domoic acid under the established HPLC conditions was $0.10\;{\mu}g/g$, and the limit of quantification of the toxin under the same conditions was $0.25\;{\mu}g/g$. The intra-accuracy and precision for domoic acid in CRM-ASP-Mus-c were 90.7-95.7% and 0.28-22.25%, respectively. The inter-accuracy and precision for domoic acid were 89.1-97.1% and 1.7-4.1%, respectively. The mean recovery of domoic acid in methanol extracts from ten species of marine invertebrates was 88.6-1105.1%.

• Fisheries and Aquatic Sciences
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• 제8권2호
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• pp.76-82
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• 2005

We analyzed the paralytic shellfish poisoning (PSP) toxins of the toxic marine dinoflagellate Alexandrium tamarense collected from Dadaepo and Gaduck-do in Busan and from Sujeong-ri in Jinhae Bay, Korea, in April 2003. We also analyzed the PSP toxin of mussels (Mytilus galloprovincialis) and oysters (Crassostrea gigas) collected around Busan and Jinhae Bay. PSP toxin analyses were conducted by high performance liquid chromatography (HPLC). Fifteen cultured A. tamarense isolates contained 2.78 to 57.47 fmol/cell, with nearly identical toxin profiles: major components C2, GTX4; minor components C1, GTX1, NEO; and trace components GTX2, GTX3, STX. PSP toxin contents were 0 to $492\;\mu{g}$ STXeq/100 g in mussels and 0 to $48\;\mu{g}$ STXeq/100 g in oysters. Mussels at Gijang and Sujeong-ri contained the most PSP toxin contents ($492\;\mu{g}\;STXeq/100\;g\;and\;252\;\mu{g}\;STXeq/100\;g,\;respectively$), exceeding the quarantine level ($80\;\mu{g}$ STXeq/100 g). Their dominant toxin components were C2, C1, GTX2, and GTX3; the minor components GTX1, GTX4, GTX5, and NEO were sporadically detected. Phytoplankton contained 0.774 fmol/L seawater and 1.228 fmol/L seawater at Gijang and Sujeong-ri in April. At that time, Alexandrium cells were present in the water column at Gijang at 2,577 cells/mL and at Sujeong-ri at 6,750 cells/mL. Overall, we found the high and similar PSP toxin contents in AZexandrium isolates and mussels, and a correlation between occurrence of toxic Alexandrium cells in the water column and mussel intoxification. High densities of toxic Alexandrium cells in the water column immediately preceded shellfish intoxification at Gijang and Sujeong-ri in April.

• 한국수산과학회지
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• 제46권2호
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• pp.154-159
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• 2013

The AOAC Mouse Bioassay method (MBA) has been widely used for routine monitoring of paralytic shellfish poisoning (PSP) for more than 50 years. However, this method has low sensitivity and experiences interference from other components in the extract. Also, ethical issues have been raised against the continued use of this live-mouse assay. To establish an alternative method for PSP analysis, we attempted to develop PSP analysis conditions using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The LC-MS/MS analysis of reference material showed very reasonable accuracy, and the analysis time was just 15 min. However, the recovery rate of toxin spike samples using the LC-MS/MS analysis was 59.4-91.0%. We also attempted to remove the matrix effect using shellfish extracts, but recoveries of C1 and C2 did not improve. A comparison between the results of MBA and LC-MS/MS analysis revealed good correlations, with values of 0.8878 and 0.9211 for oyster and mussel matrices, respectively.

• 한국수산과학회지
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• 제42권4호
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• pp.366-372
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• 2009

This study looked at toxicity of Mediterranean mussels, Mytilus galloprovincialis, which had accumulated paralytic shellfish toxins (PST) from early March to late May 2005 at Jinhae Bay, Korea. Alexandrium sp. was observed in low densities (< 1,000 cells/L) at the beginning of the study in March, increased rapidly in April, declined rapidly and disappeared in May. Although low densities of Alexandrium sp. were observed in March, mussel toxicity exceeded regulation level ($80{\mu}g$ STXeq. /100 g). Peak PSP (Paralytic Shellfish Poisoning) toxicity in the mussels occurred during high Alexandrium sp. cell densities in April. Mussels toxicity decreased with decline of Alexandrium sp. cell density. Major toxin components identified were $GTX_1$, $GTX_4$, followed by $C_1$, $C_2$, $GTX_2$, $GTX_3$ and neoSTX. Trace or sporadic toxin components were STX, $GTX_5$, $dcGTX_2$, $dcGTX_3$ and dcSTX. Toxin component analysis from the middle to end of the study showed that $11{\beta}$-epimers ($GTX_{3,4}$, $C_2$) were converted into $11{\alpha}$-epimers ($GTX_{1,2}$, $C_1$) and started to determine STX.

• Fisheries and Aquatic Sciences
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• 제16권3호
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• pp.159-164
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• 2013

The mouse bioassay and high performance liquid chromatography (HPLC) post-column oxidation method are different methods of quantifying paralytic shellfish poisoning toxins. In this study, we compared their ability to accurately quantify the toxicity levels in two types of field sample (oysters and mussels) with different toxin profiles for routine regulatory monitoring. A total of 72 samples were analyzed by both methods, 44 of which gave negative results, with readings under the limit of detection of the mouse bioassay ($40{\mu}g/100g$ saxitoxin [STX] eq). In 14 oysters, the major toxin components were gonyautoxin (GTX) 1, -2, -3, -4, -5, decarbamoylgonyautoxin-2 (dcGTX2), and decarbamoylsaxitoxin (dcSTX), while 14 mussels tested positive for dcSTX, GTX2, -3, -4, -5, dcGTX2, neosaxitoxin (NEO), STX, and dcSTX. When the results obtained by both methods were compared in two matrices, a better correlation ($r^2=0.9478$) was obtained for mussels than for oysters ($r^2=0.8244$). Additional studies are therefore needed in oysters to investigate the differences in the results obtained by both methods. Importantly, some samples with toxin levels around the legal limit gave inconsistent results using HPLC-based techniques, which could have a strong economic impact due to enforced harvest area closure. It should therefore be determined if all paralytic shellfish poisoning toxins can be quantified accurately by HPLC, and if the uncertainties of the method lead to doubts regarding regulatory limits.

• Fisheries and Aquatic Sciences
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• 제16권3호
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• pp.137-142
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• 2013

To understand critical aspects of paralytic shellfish poisoning (PSP) in a chief area of bivalve production in Korea, seasonal variation in PSP toxins in bivalves collected from Jinhae Bay, Korea in 2009 was surveyed by the pre-column high-performance liquid chromatography oxidation method. We also confirmed the profiles of major bivalves such as oysters Crassostrea gigas and mussels Mytilus galloprovincialis in Jinhae Bay. PSP toxins in the bivalves showed remarkable seasonal variation. PSP toxin levels were detected from April to May in 2009, and the highest total toxin levels at all stations were recorded in May. The major toxins in bivalves were gonyautoxin [GTX] 1&4 and C 1&2; in oysters GTX 2&3 were also detected as major components. GTX 1&4, which showed the highest PSP toxin levels at each station, accounted for the highest proportions of toxin components in mussels and oysters (64.5-71.3% and 41.4-42.4%, respectively). It was also confirmed that the highest toxicity (in ${\mu}g$ saxitoxin [STX] eq/g) was derived from GTX 1&4. The highest total toxicity (in ${\mu}g$ STX eq/g) was approximately 2-8-fold higher in mussels than in oysters collected from the same station. PSP toxin levels in bivalves differed significantly according to the sample collection station. However, the profiles of toxins in the bivalves did not show significant differences during the survey period according to sample collection station. This study shows that PSP toxin levels in some samples from Jinhae Bay were above the regulatory limit in Korea during a specific period in spring.

• 한국수산과학회지
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• 제39권1호
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• pp.49-54
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• 2006

Cultured cells of the toxic Alexandrium tamarense were fed to four mussel species, Mytilus coruscus, M. edulis, M. galloprovincialis and Septifer vulgatus, to examine the interspecies and interlocality differences in the ability to accumulate paralytic shellfish poisoning (PSP) toxins. Toxin content of A. tamarense cells varied during culture period. In contrast, toxin composition in the cell (C1,2, GTX1-4 and neoSTX) was constantly stable. In feeding experiment, the four mussel species collected from Geoje intoxicated after uptake of A. tamarense. Toxin content ($average{\pm}SD\;{\mu}g$ STXeq/100 g) of M. coruscus, M. edulis, M. galloprovincialis and Septifer vulgatus were $1,660{\pm}79,\;3,914{\pm}2,242,\;5,626{\pm}1,620\;and\;958{\pm}163$, respectively. Toxin profiles included C1,2, GTX1,4 and neoSTX as the major components, and dcGTX2,3, GTX2,3, neoSTX and STX as the minor ones. Toxin accumulation of three mussel species collected from Pohang, Geoje and Anmyon-do showed interspecies and interlocality differences. Toxin content ($average{\pm}SD\;{\mu}g$ STXeq/100 g) were $91{\pm}4,\;151{\pm}14,\;39{\pm}3$ in M coruscus, $189{\pm}1,\;231{\pm}11,\;206{\pm}15$ in M edu/is and $214{\pm}28,\;326{\pm}30,\;291{\pm}26$ in M. galloprovincialis in order of Anmyon-do, Geoje and Pohang.

• 한국식품과학회지
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• 제44권4호
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• pp.390-392
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• 2012

LC/MS/MS로 패류 114건에서 설사성패독인 OA, DTX1, PTX2, YTX을 정량한 결과 110건(96.5%)에서는 설사성패독이 불검출이었으며, 굴 2건(0.004, 0.001 ${\mu}g/g$), 진주담치 1건(0.001 ${\mu}g/g$), 바지락 1건(0.001 ${\mu}g/g$)에서 OA(Okadaic acid)가 검출되어 3.5%의 검 출률을 나타내었으며 동일 시료를 마우스시험법으로 마비성 패독을 실험한 결과 설사성 패독과의 유의성은 보이지 않았다.

• 해양환경안전학회지
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• 제27권1호
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• pp.119-126
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• 2021

본 연구에서는 실내에서 다양한 수온과 염분 조건에서 유독와편모조류 Alexandrium catenella(Group I)의 생장과 함께 마비성패독의 함량을 조사하였다. A. catenella의 수온과 염분 최적생장은 각각 20~30℃과 20~30 psu였으며, 광온성과 광염성종으로 나타났다. A. catenella는 낮은 수온 구간(10℃와 15℃)에서 독함량과 독성이 높게 나타났으며, 염분에 따라서는 차이가 크지 않았다. 따라서 15℃이하의 수온에서 본 종주와 같은 생리적인 특성을 가진 유독와편모조류가 우점한다면, 이매패류는 빠르게 독화될 가능성이 있다. 향후 A. catenella의 출현에 따른 마비성패독 예찰·예보를 위해 다양한 환경변화에 따른 A. catenella의 다른 종주와 상업적인 이매패류 독화에 대한 연구가 더 필요한 것으로 판단된다.

• 한국수산과학회지
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• 제42권1호
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• pp.8-14
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• 2009

To prevent the food poisoning originated by consumption of shellfish contaminated with domoic acid, the quantitative analysis of domoic acid is to be very important. The stability of domoic acid at different temperature, pH and light was investigated using high performance liquid chromatography (HPLC). The mean recoveries of domoic acid in the methanol extracts from oyster (Crassostrea gigas), blue mussel (Mytilus edulis), short neck clam (Ruditapes philippinarum) and ark shell (Scapharca broughtonii) were 85.4-104.5%, 94.8-101.2%, 91.0-104.6%, and 95.7-109.6%, respectively. The working solutions of domoic acid standard were very stable for one month at $-18^{\circ}C$, $4^{\circ}C$, and room temperature. And domoic acid in the methanol extract from oyster was stable for a day at $4^{\circ}C$ and room temperature, and for a week at $-18^{\circ}C$. Therefore, this implies that quantitative analysis for domoic acid must consider the storage conditions of the standard solutions and the methanol extracts from shellfish. The standard solutions adjusted to pH 3-9 were also stable after heating at $121^{\circ}C$ for 30 min. The effect of light exposure on domoic acid was tested by exposing the methanol extracts to light. Domoic acid degraded slowly when the samples were kept in the dark (brown vial). However, following the light exposure the photodegradation became more rapid; no detectable domoic acid remained in $1.0{\mu}g/mL$ of methanol extract after 5 hours.