Akumulasi dan Depurasi Toksin PSP (Paralytic Shellfish Poisoning)  oleh Kerang Hijau (Accumulation and Depuration of PSP Toxin (Paralytic Shellfish Poisoning) by Green Mussels)

Ledakan mikroalga sering dilaporkan terjadi di Teluk Jakarta, dimana di lokasi tersebut juga terdapat kegiatan budidaya kerang hijau (Perna viridis). Terkait dengan hal tersebut maka dilakukan studi akumulasi dan depurasi toksin PSP (Paralytic Shellfish Poisoning) pada kerang hijau. Studi akumulasi dilakukan di bagan kerang hijau perairan Cilincing Jakarta Utara, dengan memisahkan kerang hijau yang berukuran sama dan ditempatkan kembali ke bagan. Sampling dilakukan setiap minggu selama 2 bulan dan diukur juga kelimpahan fitoplankton, pH, suhu dan salinitas perairan. Depurasi dilakukan di Unit Depurasi Kekerangan KKP Panimbang Banten, yang dilakukan selama 24 jam. Pencuplikan sampel dilakukan setiap jam pada 4 jam pertama dan setiap 2 dan 3 jam pada waktu berikutnya. Penentuan konsentrasi toksin PSP dilakukan dengan menggunakan HPLC detektor fluoresensi. Prosedur preparasi, ekstraksi dan pengukuran konsentrasi toksin mengikuti Manual AOAC Official Method 2005.06 untuk toksin PSP dalam kekerangan. Akumulasi toksin PSP oleh kerang hijau di perairan Cilincing pada bulan Januari–Pebruari 2011 berkisar antara 4,11–11,96 µg STX eq. per 100 g dan tidak mempunyai korelasi dengan kelimpahan Dinoflagelata di perairan. Hal ini disebabkan uji akumulasi tidak dilakukan pada saat blooming mikroalga. Uji depurasi selama 24 jam mengeliminasi toksin PSP sebesar 60%, sehingga bisa diajukan sebagai sistem pemutus rantai toksin dari mikroalga ke manusia. Kata kunci: akumulasi, depurasi, PSP toksin, kerang hijau, Cilincing Microalgae blooms have been frequently reported in the Jakarta Bay, which is also the location of green mussel (Perna viridis) aquaculture. Accumulation and depuration of Paralytic Shellfish Poisoning (PSP) toxin in the green mussels were investigated in the field, where the toxin accumulation studies conducted in the mussel farming at Cilincing, North Jakarta. Accumulation test carried out by placing back the selected green mussel (equal size) into the mussel farming. Every week for 2 months, the green mussel were collected from mussel farming and transported to the laboratory. The fitoplankton abundance also was checked including pH, Suhue and salinitiy paramaters. Toxin depuration was conducted at Clams Sanitation Unit at Panimbang Banten. The depuration studies were conducted for 24 hours with sampling every hour in the first 4 hours and every 3 and 2 hours until the 24th hour. Preparation, extraction and toxin concentration measurements performed by following the Manual AOAC Official Method 2005.06 for PSP toxin in oyster. This research concluded that the accumulation of PSP toxin by green mussel, Perna viridis in the mussel farming at Cilincing, North Jakarta in ranged between 4,11–11,96 µg STX eq. per 100 g during January–February 2011. No correlation between PSP toxin concentration in the green mussel, Perna viridis with abundance of the PSP toxin sources phytoplankton, because the study wasnt done when microalgae blooming. The depuration processes was eliminate 60% the PSP toxins for 24 hours depuration processing. It can be proposes as a banded system the PSP toxin from algae to human being. Keywords: accumulation, depuration, PSP toxin, green mussel, Cilincing

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Extension of the Validation of AOAC Official MethodSM 2005.06 for dc-GTX2,3: Interlaboratory Study

Journal of AOAC International ◽

10.5740/jaoacint.10-446 ◽

2012 ◽

Vol 95 (1) ◽

pp. 111-121 ◽

Cited By ~ 22

Author(s):

Begoña Ben-Gigirey ◽

María L Rodríguez-Velasco ◽

Ana Gago-Martínez

Keyword(s):

Collaborative Study ◽

Paralytic Shellfish Poisoning ◽

Official Method ◽

Reference Laboratory ◽

The European Union ◽

Shellfish Poisoning ◽

Precision Data ◽

European Union Reference Laboratory ◽

Aoac Official ◽

Aoac Official Method

Abstract AOAC Official MethodSM 2005.06 for the determination of saxitoxin (STX)-group toxins in shellfish by LC with fluorescence detection with precolumn oxidation was previously validated and adopted First Action following a collaborative study. However, the method was not validated for all key STX-group toxins, and procedures to quantify some of them were not provided. With more STX-group toxin standards commercially available and modifications to procedures, it was possible to overcome some of these difficulties. The European Union Reference Laboratory for Marine Biotoxins conducted an interlaboratory exercise to extend AOAC Official Method 2005.06 validation for dc-GTX2,3 and to compile precision data for several STX-group toxins. This paper reports the study design and the results obtained. The performance characteristics for dc-GTX2,3 (intralaboratory and interlaboratory precision, recovery, and theoretical quantification limit) were evaluated. The mean recoveries obtained for dc-GTX2,3 were, in general, low (53.1–58.6%). The RSD for reproducibility (RSDr%) for dc-GTX2,3 in all samples ranged from 28.2 to 45.7%, and HorRat values ranged from 1.5 to 2.8. The article also describes a hydrolysis protocol to convert GTX6 to NEO, which has been proven to be useful for the quantification of GTX6 while the GTX6 standard is not available. The performance of the participant laboratories in the application of this method was compared with that obtained from the original collaborative study of the method. Intralaboratory and interlaboratory precision data for several STX-group toxins, including dc-NEO and GTX6, are reported here. This study can be useful for those laboratories determining STX-group toxins to fully implement AOAC Official Method 2005.06 for official paralytic shellfish poisoning control. However the overall quantitative performance obtained with the method was poor for certain toxins.

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Interlaboratory Comparison of Two AOAC Liquid Chromatographic Fluorescence Detection Methods for Paralytic Shellfish Toxin Analysis through Characterization of an Oyster Reference Material

Journal of AOAC International ◽

10.5740/jaoacint.sgeturner ◽

2014 ◽

Vol 97 (2) ◽

pp. 380-390 ◽

Cited By ~ 10

Author(s):

Andrew D Turner ◽

Adam M Lewis ◽

Wade A Rourke ◽

Wendy A Higman ◽

Z Amzil ◽

...

Keyword(s):

Reference Material ◽

Fluorescence Detection ◽

Official Method ◽

Shellfish Poisoning ◽

Paralytic Shellfish Toxin ◽

Paralytic Shellfish ◽

Shellfish Toxin ◽

Aoac Official ◽

Interlaboratory Reproducibility ◽

Aoac Official Method

Abstract An interlaboratory ring trial was designed and conducted by the Centre for Environment, Fisheries, and Aquaculture Science to investigate a range of issues affecting the analysis of a candidate Pacific oyster paralytic shellfish toxin reference material. A total of 21 laboratories participated in the study and supplied results using one or more of three instrumental methods, specifically precolumn oxidation (Pre-COX) LC with fluorescence detection (FLD; AOAC Official Method 2005.06), postcolumn oxidation (PCOX) LC-FLD (AOAC Official Method 2011.02), and hydrophilic interaction LC/MS/MS. Each participant analyzed nine replicate samples of the oyster tissue in three separate batches of three samples over a period of time longer than 1 week. Results were reported in a standardized format, reporting both individual toxin concentrations andtotal sample toxicity. Data were assessed to determine the equivalency of the two AOAC LC methods and the LC/MS/MS method as well as an assessment of intrabatch and interbatch repeatability and interlaboratory reproducibility of each method. Differences among the results reported using the three methods were shown to be statistically significant, although visualcomparisons showed an overlap between results generated by the majority of tests, the exception being the Pre-COX quantitation of N-hydroxylated toxins in post ion-exchange fractions. Intralaboratory repeatability and interlaboratory reproducibility were acceptable for most of the results, withthe exception of results generated from fractions. The results provided good evidence for the acceptableperformance of the PCOX method for the quantitation of C toxins. Overall the study showed the usefulnessof interlaboratory analysis for the characterizationof paralytic shellfish poisoning matrix reference materials, highlighting some issues that may need to be addressed with further method assessment at individual participant laboratories.

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Assessment of a Semiquantitative Liquid Chromatography- Fluorescence Detection Method for the Determination of Paralytic Shellfish Poisoning Toxin Levels in Bivalve Molluscs from Great Britain

Journal of AOAC International ◽

10.5740/jaoacint.13-381 ◽

2014 ◽

Vol 97 (2) ◽

pp. 492-497 ◽

Cited By ~ 3

Author(s):

Andrew D Turner ◽

Monika Dhanji-Rapkova ◽

Clothilde Baker ◽

Myriam Algoet

Keyword(s):

Great Britain ◽

Fluorescence Detection ◽

Detection Method ◽

Reference Method ◽

Paralytic Shellfish Poisoning ◽

Official Method ◽

Shellfish Poisoning ◽

Bivalve Molluscs ◽

Paralytic Shellfish

Abstract AOAC Official Method 2005.06 precolumn oxidation LC-fluorescence detection method has been used for many years for the detection and quantitation of paralytic shellfish poisoning (PSP) toxins in bivalve molluscs. After extensive single- and multiple-laboratory validation, the method has been slowly gaining acceptance worldwide as a useful and practical tool for official control testing. In Great Britain, the method has become routine since 2008, with no requirement since then for reverting back to the bioassay reference method. Although the method has been refined to be semiautomated, faster, and more reproducible, the quantitation step can be complex and time-consuming. An alternative approach was developed to utilize the qualitative screening results for generatinga semiquantitative results assessment. Data obtained over 5 years enabled the comparison of semiquantitative and fully quantitative PSP results in over 15 000 shellfish samples comprising eight different species showed that the semiquantitative approach resulted in over-estimated paralytic shellfish toxin levels by an average factor close to two in comparison with the fully quantified levels. No temporal trends were observed in the data or relating to species type, with the exception of surf clams. The comparison suggested a semiquantitative threshold of 800 μg saxitoxin (STX) eq/kg should provide a safe limitfor the determination of samples to be forwarded to full quantitation. However, the decision was taken to halve this limit to include an additional safety factor of 2, resulting in the use of a semiquantitative threshold of 400 μg STX eq/kg. Implementation of the semiquantitative method into routine testing would result in a significant reduction in the numbers of samples requiring quantitation and have a positive impact on the overall turnaround of reported PSP results. The refined method would be appropriate for any monitoring laboratory faced with high throughput requirements.

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Paralytic shellfish toxins in clinical matrices: Extension of AOAC official method 2005.06 to human urine and serum and application to a 2007 case study in Maine

Deep Sea Research Part II Topical Studies in Oceanography ◽

10.1016/j.dsr2.2012.08.001 ◽

2014 ◽

Vol 103 ◽

pp. 368-375 ◽

Cited By ~ 16

Author(s):

Stacey DeGrasse ◽

Victor Rivera ◽

John Roach ◽

Kevin White ◽

John Callahan ◽

...

Keyword(s):

Human Urine ◽

Official Method ◽

Shellfish Toxins ◽

Paralytic Shellfish Toxins ◽

Paralytic Shellfish ◽

Human Urine And Serum ◽

Aoac Official ◽

Urine And Serum ◽

Aoac Official Method

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Comparison of AOAC 2005.06 LC official method with other methodologies for the quantitation of paralytic shellfish poisoning toxins in UK shellfish species

Analytical and Bioanalytical Chemistry ◽

10.1007/s00216-010-4428-7 ◽

2010 ◽

Vol 399 (3) ◽

pp. 1257-1270 ◽

Cited By ~ 42

Author(s):

Andrew D. Turner ◽

Robert G. Hatfield ◽

Monika Rapkova ◽

Wendy Higman ◽

Myriam Algoet ◽

...

Keyword(s):

Paralytic Shellfish Poisoning ◽

Official Method ◽

Shellfish Poisoning ◽

Paralytic Shellfish

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Refinement of AOAC Official MethodSM 2005.06 Liquid Chromatography-Fluorescence Detection Method to Improve Performance Characteristics for the Determination of Paralytic Shellfish Toxins in King and Queen Scallops

Journal of AOAC International ◽

10.5740/jaoacint.11-184 ◽

2012 ◽

Vol 95 (1) ◽

pp. 129-142 ◽

Cited By ~ 28

Author(s):

Andrew D Turner ◽

Robert G Hatfield

Keyword(s):

Fluorescence Detection ◽

Periodate Oxidation ◽

Performance Characteristics ◽

Official Method ◽

Refined Method ◽

Paralytic Shellfish ◽

Aoac Official ◽

Psp Toxins ◽

Aoac Official Method

Abstract AOAC Official MethodSM 2005.06 LC-fluorescence detection (FLD) method is an official alternative to the mouse bioassay for the determination of paralytic shellfish poisoning (PSP) toxins in bivalve shellfish. To validate the method for species of relevance to the UK official control monitoring program, the method performance characteristics were tested for whole king and queen scallops. Validation showed that, while the performance was generally acceptable for the quantitation of non-N-hydroxylated toxins, poor toxin recovery and sensitivity was evident for the analysis of N-hydroxylated toxins following periodate oxidation. These effects occurred in a range of scallop samples with variable temporal and spatial sources. The effects were also noted in other laboratories following a small interlaboratory study. As a result, the method was refined to improve the recovery and sensitivity of analysis following the periodate oxidation step in the PSP method for scallops. Performance improved through alterations to the preparation of the periodate oxidant, use of higher volumes for C18 cleanup, and injection volumes in combination with the use of a king scallop matrix modifier for oxidation of N-hydroxylated toxin calibration standards. A single-laboratory validation of the refined method showed that the selectivity, linearity, sensitivity, recovery, and precision were acceptable and similar to values reported previously for AOAC Official Method 2005.06 in other bivalve species. Results showed the method to be rugged for all parameters investigated, including small changes to the composition of the new periodate reagent utilized in the refined method. The refined scallops LC method was subsequently compared with the European reference method. PSP-positive scallops showed an excellent agreement between the methods for queen and Atlantic scallops, with a small level of positive bias in the LC results for whole king scallops. These differences were related solely to the use of the highest toxicity equivalence factors for toxin epimeric pairs, with gonyautoxin (GTX)1,4 and GTX2,3 in particular present at high concentrations in the king scallops. Overall, the refined LC-FLD method improved the performance characteristics of AOAC Official Method 2005.06 for the determination of PSP toxins in whole king and queen scallops, and showed a good overall agreement between the official methodologies. It is, therefore, recommended as a more appropriate option for the routine monitoring of PSP toxins in these species.

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Paralytic Shellfish Poisoning (PSP) in Mussels from the Eastern Cantabrian Sea: Toxicity, Toxin Profile, and Co-Occurrence with Cyclic Imines

Toxins ◽

10.3390/toxins13110761 ◽

2021 ◽

Vol 13 (11) ◽

pp. 761

Author(s):

Tamara Rodríguez-Cabo ◽

Ángeles Moroño ◽

Fabiola Arévalo ◽

Jorge Correa ◽

Juan Pablo Lamas ◽

...

Keyword(s):

Basque Country ◽

Geographical Area ◽

Paralytic Shellfish Poisoning ◽

Monitoring Systems ◽

Mouse Bioassay ◽

Official Method ◽

Shellfish Poisoning ◽

Paralytic Shellfish ◽

First Time ◽

Psp Toxins

In the late autumn of 2018 and 2019, some samples taken by the official monitoring systems of Cantabria and the Basque Country were found to be paralytic shellfish poisoning (PSP)-positive using a mouse bioassay. To confirm the presence of PSP toxins and to obtain their profile, these samples were analyzed using an optimized version of the Official Method AOAC 2005.06 and using LC–MS/MS (HILIC). The presence of some PSP toxins (PSTs) in that geographical area (~600 km of coast) was confirmed for the first time. The estimated toxicities ranged from 170 to 983 µg STXdiHCl eq.·kg−1 for the AOAC 2005.06 method and from 150 to 1094 µg STXdiHCl eq.·kg−1 for the LC–MS/MS method, with a good correlation between both methods (r2 = 0.94). Most samples contained STX, GTX2,3, and GTX1,4, and some also had NEO and dcGTX2. All of the PSP-positive samples also contained gymnodimine A, with the concentrations of the two groups of toxins being significantly correlated. The PSP toxin profiles suggest that a species of the genus Alexandrium was likely the causative agent. The presence of gymnodimine A suggests that A. ostenfeldii could be involved, but the contribution of a mixture of Alexandrium species cannot be ruled out.

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