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At least 19 recordsLinked to original sources
PubMed2005

Contamination of shellfish from Shanghai seafood markets with paralytic shellfish poisoning and diarrhetic shellfish poisoning toxins determined by mouse bioassay and HPLC.

This paper reports the results of investigations of shellfish toxin contamination of products obtained from Shanghai seafood markets. From May to October 2003, 66 samples were collected from several major seafood markets. Paralytic shellfish poisoning (PSP) and diarrhetic shellfish poisoning (DSP) toxins in shellfish samples were monitored primarily by a mouse bioassay, then analysed by HPLC for the chemical contents of the toxins. According to the mouse bioassay, eight samples were detected to be contaminated by PSP toxins and seven samples were contaminated by DSP toxins. Subsequent HPLC analysis indicated that the concentrations of the PSP toxins ranged from 0.2 to 1.9 microg/100 g tissues and the main components were gonyautoxins 2/3 (GTX2/3). As for DSP, okadaic acid was detected in three samples, and its concentration ranged from 3.2 to 17.5 microg/100 g tissues. Beside okadaic acid, its analogues, dinophysistoxins (DTX1), were found in one sample. According to the results, gastropod (Neverita didyma) and scallop (Argopecten irradians) were more likely contaminated with PSP and DSP toxins, and most of the contaminated samples were collected from Tongchuan and Fuxi markets. In addition, the contaminated samples were always found in May, June and July. Therefore, consumers should be cautious about eating the potential toxic shellfish during this specific period.

Animals↗
PubMedDate not supplied

Microtitre plate assay for paralytic shellfish toxins using saxiphilin: gauging the effects of shellfish extract matrices, salts and pH upon assay performance.

Saxiphilin is a hydrophilic protein with a high affinity and specificity for paralytic shellfish toxins (PSTs) found in the circulatory fluid of many invertebrates and ectothermic vertebrates. Saxiphilin has been found to be closely related to the iron binding transferrins, a group of proteins that range in molecular weight between 70 and 90 kDa. One saxiphilin isoform, that from the centipede Ethmostigmus rubripes, has been used to develop a microtitre plate assay for PSTs which relies upon detection of bound tritiated saxitoxin (STX). In this study, this assay was challenged with differing conditions of salt concentration and identity, pH and addition of non-toxic extracts of commercial shellfish prepared following the Association of Official Analytical Chemists endorsed procedure, elements the assay would encounter if used for PST monitoring and may compromise assay performance. The assay tolerated up to 15% of the total reaction, volume being shellfish extract before assay signal started to diminish. The presence of these extract matrices had little effect upon assay accuracy and precision when measuring STX, as a typical PST. Also, the detection of STX in the presence of shellfish matrices could be confidently reproduced on different days by different experimenters. The elements present in the shellfish extracts were measured by inductively coupled plasma spectroscopy, with the most common cationic elements being Na followed by K, Mg and Ca. Only trace amounts of other cationic elements were also present. From these results, the effects upon the assay by the four most common salts of these elements, NaCl, KCl, CaCl(2) and MgCl(2) were measured. Both NaCl and KCl did not impair assay performance at concentrations as high as 550 mM. It should be noted, however, that greater than 80 mM of either of these salts must be present in the assay for it to achieve the maximal signal. Adding CaCl(2) and MgCl(2) to the assay had dramatic effects upon performance. In the case of CaCl(2), the NaCl that was present in standard assay conditions enhanced its negative impact upon the assay. With MgCl(2), NaCl counteracted its inhibitory effect to some extent. After taking into account sample dilutions of shellfish extracts however, the potential for an interfering effect by Ca or Mg is minimal. A pH of 5.4 or less is necessary for there to be any significant impact upon the assay, with the assay signal being stable up to an alkaline pH as high as 9. Using the conditions herein, this assay can be used to reliably detect 1.3 microg STXeq/100 g shellfish tissue if it were to be used for monitoring for PST contaminated shellfish. These results demonstrate that this assay is a highly robust diagnostic tool for the measurement of PSTs in shellfish extracts.

Amphibian Proteins↗
PubMed2003

[Paralytic shellfish toxins in shellfish from the coast of high frequent harmful algae blooms occurrence areas in East China Sea and South China Sea].

The paralytic shellfish poisoning (PSP) toxicity levels of shellfish collected from shellfish growing zones in Zhoushan areas of East China Sea and Shenzhen areas of South China Sea during January-November 2002 were determined with AOAC bioassay method. The results showed that the toxicity levels of shellfish from Zhoushan were very low, and few shellfish samples could be detected the PSP toxicities. The same levels appeared in the shellfish from Shenzhen, but the percentage of shellfish samples contaminated PSP was up to 30%, and the scallop Chalmys nobilis was dominated in the contaminated shellfish species. Analyses on the toxic profiles of digest gland and edible tissue extraction of scallop from Shenzhen showed that a similar suite of toxins presented in the gland and edible tissues. The high potency neoSTX and low potency GTX5 had a high proportions, up to 60.8% and 11.4%, respectively in the tissue, and the high potency GTX1 + 2 and GTX5 were the dominant toxins, up to 38.9% and 20%, respectively in the gland. Most of the toxin were accumulated in ingested gland of scallop, and the toxic content in gland was about 8 folds more than that in the edible tissues.

Animals↗
PubMed2006

Hybridisation of F+ RNA coliphages detected in shellfish samples with oligonucleotide probes to assess the origin of microbiological pollution of shellfish.

Current measures for controlling the public health risks associated with bivalve molluscan shellfish consumption rely on the use of Escherichia coli to indicate the sanitary quality of shellfish harvesting areas. However, it has been demonstrated that E. coli is an inadequate indicator of the viral risk associated with shellfish. An alternative indicator, male-specific B+ coliphages, have been investigated as viral indicators of faecal contamination that may provide source-specific information for impacted environmental waters. This study compared the distribution of E. coli and F+ RNA bacteriophages in shellfish grown in harvesting areas of Greece and also examined the presence and proportions of the different subgroups of F+ RNA coliphages in shellfish. F+ RNA bacteriophages were present in shellfish at higher concentrations than E. coli. Elevated numbers of F+ RNA bacteriophages observed in the winter concur with the known increased viral risk associated with shellfish harvested at that time of year in Greece. The majority of F+ RNA coliphages detected in shellfish samples belonged to group IV which indicated the possible presence of animal faecal material in sample harvesting areas. Phages of groups II and III (human waste and human faecal material, respectively) were present at low levels. Finally, 8% of the phages hybridised were found to belong to group I. The presence of group IV showed seasonal distribution (more in winter, less in summer) whereas the other groups did not show any difference. Monitoring of F+ coliphage subgroups may indicate the presence and major sources of microbial inputs to surface waters; however, environmental effects on the relative occurrence of different groups need to be considered.

Animals↗
PubMed1998

Distribution of indicator bacteria and bacteriophages in shellfish and shellfish-growing waters.

Shellfish (mussels and clams) and shellfish-growing waters were examined for indicator bacteria according to the EC regulations, Salmonella spp., coliphages and anti-Salmonella phages. Samples were collected both from natural-growing areas along the coast and from authorized shellfish-harvesting beds. The coastal area was affected by organic pollution and extensive faecal contamination and, according to the legal requirements, was unsuitable for shellfish farming. The shellfish collected along the coast also showed faecal contamination at levels which did not conform to legal standards. No significant differences were observed between the frequency of isolation of somatic coliphages and indicator bacteria from sea water. In contrast, both the authorized and wild coastal shellfish were contaminated by coliphages at a significantly higher level than the corresponding bacterial indicators for faecal contamination (chi 2 test, P < 0.01). Coliphage concentrations were significantly correlated with faecal indicators in marine waters (P < 0.001) and sediments (P < 0.05), but no correlation was found in shellfish, thus showing their low specificity as indicators of faecal pollution of human origin in shellfish of economic importance.

Analysis of Variance↗
PubMedDate not supplied

Comparison evaluation of liquid chromatographic and bioassay methods of analysis for determination of paralytic shellfish poisons in shellfish tissues.

A liquid chromatographic (LC) method was compared with the AOAC mouse bioassay method (18.086-18.092) for determination of paralytic shellfish toxins in shellfish tissues. Shellfish samples were collected from Massachusetts coastal waters as part of a state surveillance program, and extracts of shellfish meat were analyzed for toxins by using both analytical methods. Overall correlation of the LC and bioassay methods is good (r = 0.943), but for samples with toxicities less than 100 micrograms saxitoxin/100 g shellfish meat, the correlation is significantly less (r = 0.531). Limits of detection are 10 micrograms saxitoxin/100 g shellfish meat and 40 micrograms saxitoxin/100 g shellfish meat for the LC and bioassay methods, respectively. Analytical capacity of the LC method is limited to 12 samples/person-day compared with 30 samples/person-day for the bioassay. Sampling capacity of the LC method could be increased by using a fluorescence detector with a wider response range, which would eliminate the need for dilution of concentrated samples.

Animals↗
PubMed2006

[Analysis of paralytic shellfish poisoning and diarrhetic shellfish poisoning of bivalves in seafood market of Guangzhou].

OBJECTIVE: Paralytic shellfish poisoning (PSP) and diarrhetic shellfish poisoning (DSP) in 7 species of economical shellfishes were analyzed for 1 year, which collected from Huangsha seafood market of Guangzhou from Apr, 2004 to Mar, 2005. METHODS: The levels of PSP and DSP in bivalves were determined with mouse bioassay of AOAC. The risk assessment of PSP and DSP in bivalves was conducted according to FAO and Chinese Administration Organization of Fish Culture and Seaport. RESULTS: PSP was detected in 2 species of the shellfishes assayed and DSP was found out in 6 species. The content of PSP was lower than 4MU/g tissue, whereas the level of PSP in glands was higher than in muscles. DSP toxin was detected in 36 samples of 6 species, what is more, DSP level in 10 samples exceeded the safety threshold. The levels in PSP and DSP of bivalves were all higher in spring and winter with some characteristic of season. CONCLUSION: The results suggested that PSP in economical shellfish in Guangzhou market was lower, and shellfish was safe to eat in term of the PSP level if glands were discarded, but DSP contamination in bivalves was severe. It is essential to detect and assess the risk of DSP and PSP in bivalves from seafood market in the future.

Animals↗
PubMed2002

Detection of paralytic shellfish poisoning (PSP) toxins in shellfish tissue using MIST Alert, a new rapid test, in parallel with the regulatory AOAC mouse bioassay.

In parallel trials with the mouse bioassay, MIST Alert for Paralytic Shellfish Poisoning (PSP), a rapid diagnostic test for PSP, detected 100% of the toxic extracts in over 2100 regulatory samples. Toxic extracts contained at least 80 microg saxitoxin equivalents (STX equiv.) in 100 g of shellfish tissue, or more, as measured by the regulatory AOAC mouse bioassay. Only one potentially toxic sample, which contained 78 and 86 microg STX equiv./100 g shellfish tissue in two different mouse bioassays, was recorded as negative in one replicate of MIST Alert. All other toxic extracts among more than 2100 regulatory shellfish tissue samples were detected by MIST Alert for PSP. The MIST Alert for PSP also detected the majority of extracts containing PSP toxin greater than 32 microg STX equiv./100 g, which is the mouse bioassay detection limit. The MIST Alert for PSP gave a false positive result compared to the mouse bioassay at an average rate of about 14% over all sites, although some differences were seen between sites. Further analysis by high performance liquid chromatography (HPLC) of the (false positive) extracts showed that many contained PSP toxicity in the range of 20-40 microg STX equiv./100 g, below the level detectable by the mouse bioassay. The MIST Alert for PSP gave false positive results from extracts containing less than 20 microg STX equiv./100 g shellfish tissue only about 6% of the time. The PSP family of toxin analogues can occur in any combination in naturally contaminated shellfish tissue and the antibody mixture in the MIST Alert tests detect each of the different PSP toxin analogues with different efficacy. It is therefore impossible to provide an exact detection limit for the MIST Alert that would be applicable for all possible toxin profiles. Through the experience of comparison testing with the regulatory mouse bioassay in many parts of the world, with over 2100 different samples, the MIST Alert for PSP has proven its ability to detect all types of profiles of the PSP toxin analogues. The detection limit for MIST Alert for PSP was about 40 microg STX equiv./100 g for the 'average' profile of PSP toxin analogues. Since the detection limit depends on the toxin profile in the individual extract, it will also vary depending on the profile of analogues most commonly found at each geographic location. This was observed in our study. Over all sites in the trials, approximately 5% of samples below 40 microg STX equiv./100 g were positive, and 5% of samples between 40-80 microg STX equiv./100 g were negative. This is a reflection of the different analogue profiles found in naturally contaminated extracts, even after acid hydrolysis using the AOAC extraction method.

Animals↗
PubMedDate not supplied

Comparison of mouse bioassay and sodium channel cytotoxicity assay for detecting paralytic shellfish poisoning toxins in shellfish extracts.

A neuroblastoma cell culture assay was used to analyze shellfish extracts for presence of paralytic shellfish poisoning toxins (saxitoxins). Results were compared with mouse bioassays performed as part of a screening program for shellfish toxins in New Zealand. Twenty-nine samples gave negative results in both assays. Fifty-seven samples gave positive results in at least one assay. The correlation between the assays for saxitoxin equivalent levels in shellfish was 0.867. In spiking studies on shellfish extracts, the neuroblastoma assay showed a good response to added saxitoxin. Although these results support use of the neuroblastoma assay as a screening procedure for shellfish toxicity, results close to regulatory limits should be confirmed by mouse bioassay.

Animals↗
PubMed2006

Assessment of fitness for purpose of an insect bioassay using the desert locust (Schistocerca gregaria L.) for the detection of paralytic shellfish toxins in shellfish flesh.

We have developed a bioassay using 5th instar desert locusts (Schistocerca gregaria L.) for the detection of saxitoxin-the paralytic shellfish poison in shellfish flesh. The bioassay procedure is to inject 10 locusts with a shellfish extract, and assess their conditions at time points up to 2h post injection, looking for an endpoint of paralysis. From the proportion responding, the equivalent dose of pure saxitoxin could be estimated. Performance characteristics of the bioassay were assessed using shellfish samples spiked with saxitoxin, and we found the bioassay could detect and quantify toxin levels in the range of regulatory relevance. Relative toxicities of selected saxitoxin analogues differed from those reported in mammalian systems. Variation for repeatability conditions was acceptable but variation was higher under reproducibility conditions. This was related to (a) batches of insects from different suppliers, (b) different operators, and (c) different observers assessing the endpoint. We also noted adverse reactions with some shellfish species. These problems may be resolved by further refinement of the method and operator training, before formal validation. Nevertheless, we suggest the method potentially offers a simple, ethically acceptable, broad-specificity functional bioassay, which is desirable in any toxin-monitoring programme.

Animals↗
PubMed2001

Determination of paralytic shellfish toxins in Portuguese shellfish by automated pre-column oxidation.

Automated pre-column oxidation (the method of Lawrence) was implemented on a routine basis since the end of 1996 to study paralytic shellfish poisoning (PSP) toxins in Portuguese shellfish. Liquid chromatography confirmed the presence of PSP toxins when the known toxic algae were present: Gymnodinium catenatum and/or Alexandrium cf. lusitanicum. On the other side, it has eliminated PSP toxins as a possible recurrent contaminant in oysters from Sado estuary. These oysters were already known to contain high levels of some metals (mainly zinc, copper and cadmium) due to their location in a contaminated area and their particular physiology prone to accumulate metals. The presence of PSP toxins in Scrobicularia plana from Mondego estuary and Tellina crassa from the northern coast, during the absence of the above toxic microalgae in the water column, was confirmed. Unlike other shellfish, these two genera have the feeding habit of aspirating more sediment than organisms in suspension, and probably ingest from the sediment resting cysts of PSP producing microalgae. This is another route of contamination that may help to explain why after a bloom certain shellfish species maintain toxicity for long periods. The method revealed to have a fast implementation on a daily basis, short analysis time (around 20 min between samples), high sensitivity and robustness, and therefore, it is one of the best HPLC methods for screening a large number of shellfish samples for monitoring purposes.

Animals↗
PubMed2006

Cell bioassay for paralytic shellfish poisoning (PSP): comparison with postcolumn derivatization liquid chromatographic analysis and application to the monitoring of PSP in shellfish.

We performed a neuroblastoma cell (Neuro2a) culture assay modified slightly from a method reported previously to provide a simple and sensitive evaluation of paralytic shellfish poisoning (PSP) toxicity in shellfish. The cell bioassay was just as sensitive for C-toxins as for gonyautoxins. The sensitivity of our cell bioassay was 4 times that of the current standard mouse bioassay. Using the cell bioassay, we evaluated PSP toxicity in 361 shellfish samples collected from Mikawa Bay and Ise Bay, Aichi Prefecture, Japan, from April 1999-March 2002. The results were compared with those obtained in a postcolumn derivatization liquid chromatographic analysis. PSP toxins were detected in 236/361 samples by both assays, and there was a fairly good correlation (r = 0.9001, n = 236, p < 0.001) between the results from the two assays. We applied this cell bioassay when short-necked clams in the bay turned poisonous in 2001. The chronological changes in PSP toxicity in the short-necked clams were analyzed and compared with those of the cell density of poisonous plankton (Alexandrium tamarense) occurring in the bay. The PSP toxicity in shellfish peaked 2 weeks after the cell density reached a maximum. We recommend using the cell bioassay for routine monitoring of PSP toxicity in shellfish living in natural marine environments.

Animals↗
PubMedDate not supplied

Determination and confirmation of the amnesic shellfish poisoning toxin, domoic acid, in shellfish from Scotland by liquid chromatography and mass spectrometry.

During 1998 and early 1999, shellfish samples from sites in Scotland were found to contain the amnesic shellfish poisoning toxin, domoic acid (DA). Two different techniques, liquid chromatography (LC) with UV diode-array detection and LC with mass spectrometric (MS) detection, were used to detect and confirm DA in shellfish extracts. The LC/UV method was validated for routine monitoring by recovery experiments on spiked mussel and scallop tissues with a certified mussel tissue used as reference material. Crude extracts of selected samples as well as extracts cleaned with strong anion exchange (SAX) were analyzed by both LC/UV and LC/MS. Good correlation (linear regression r2 = 0.996, slope = 0.93) between the 2 methods was found for cleaned extracts. Analyses of crude extracts by LC/UV produced false-positive results in 2 crab samples, whereas LC/MS analyses gave accurate results. It was concluded that LC/UV is a valid approach for routine monitoring of DA in shellfish when cleanup is performed with a SAX cartridge to prevent false positives. A variety of shellfish species were surveyed for DA content, including Pecten maximus (king scallops), Chlamys opercularis (queen scallop), Mytilus edulis (blue mussels), Cancer pugaris (crab), and Ensis ensis (razor fish). The highest concentration of DA was 105 microg/g in Pecten maximus.

Amnesia↗
PubMed1999

Monitoring brevetoxins during a Gymnodinium breve red tide: comparison of sodium channel specific cytotoxicity assay and mouse bioassay for determination of neurotoxic shellfish toxins in shellfish extracts.

In October of 1996, a Gymnodinium breve bloom occurred in shellfish harvesting waters of Alabama, Mississippi and Louisiana, Gulf of Mexico, USA. Bloom densities reached 5.6x10(5) cells liter(-1) and bloom residence at shellfish sampling stations ranged from 3 to 28 days. Brevetoxin-2 dominated G. breve toxin profiles in bloom seawater extracts. Shellfish toxicity, assessed by mouse bioassay, exceeded the guidance level for up to 75 days after the bloom had dissipated. Cytotoxicity assays and mouse bioassays showed similar temporal patterns of shellfish toxicity, but the two methods differed in estimations of brevetoxin-3 equivalent toxicity by a factor of 93 to 1. LC-ESI-MS showed the temporal patterns in shellfish toxicity reflected metabolism of G. breve toxins. The molecular ions m/z 1004, 1017 and 1033 dominated LC-ESI-MS spectra of toxic chromatographic fractions from the extracts and were identified as brevetoxin metabolites on the basis of LC-APCI-MS-MS. The discrepancy between cytotoxicity and mouse bioassay estimates of brevetoxin-3 equivalent toxicity resulted from the difference in extraction efficiency of solvents used in the respective methods and the relative sensitivity of the assays to toxin metabolite mixtures present in the extracts. The normalized cytotoxicity assay showed 75% agreement with mouse bioassay positive test samples and 64% agreement with mouse bioassay negative test samples. Published in 1999 by John Wiley & Sons, Ltd.

Animals↗
PubMed2001

Study of paralytic shellfish poisoning toxin profile in shellfish from the Mediterranean shore of Morocco.

Since 1992, a monitoring program for bivalve molluscs contaminated by algal toxins was established at different stations along the Mediterranean Moroccan shores. The monitored stations were tested every 2 weeks. The presence of toxicity was determined using the mouse bioassay method. Toxin profile was carried out by HPLC/FD in selected contaminated tissues. According to the outcomes of this surveillance from 1994 to 1999, reliable information on toxicity of shellfish was obtained. They indicate that PSP is a recurrent toxicity in molluscs along the Mediterranean shore of Morocco. It has been noted a difference of PSP accumulation among individual shellfish. The cockle (Achanthocardia tuberculatum) presents toxicity throughout the year, while other specimens from the same area such as clam (Callista chione), warty venus (Venus gallina) and marine beans (Donax trunculus) accumulate it seasonally from January to April, after which they depurate the toxin. Moreover, the study of toxin profiles among individual shellfish was undertaken. It was found that shellfish presented a complex profile pointing to contamination by Gymnodinium catenatum.

Animals↗
PubMed1997

Comparison of a protein phosphatase inhibition assay, HPLC assay and enzyme-linked immunosorbent assay with the mouse bioassay for the detection of diarrhetic shellfish poisoning toxins in European shellfish.

Consumption of shellfish contaminated with algal toxins produced by marine dinoflagellates can lead to diarrhetic shellfish poisoning (DSP). UK legislation necessitates toxin detection by mouse bioassay but this method is non-specific and lacks sensitivity. As an alternative method, an HPLC technique has been optimized, with detection limits of 0.26 micrograms of toxin/g of shellfish hepatopancreas for both Okadaic Acid (OA) and Dinophysistoxin-1 (DTX-1). A colorimetric protein phosphatase inhibition (PPI) assay has also been optimized. This assay detects inhibition of protein phosphatase 1 (PPI gamma) by OA and DTX-1 with detection limits of 1.5 ng of total toxin/g of hepatopancreas. Contaminated shellfish from several European sources, the UK monitoring programmes and mussels associated with an outbreak of DSP poisoning in the UK, have been analyzed and assessed using the two alternative methods and a commercially available enzyme-linked immunosorbent assay (ELISA) kit. The results indicate that both the HPLC and PPI assays correlate well with each other and with the UK standard mouse bioassay. In contrast, and not withstanding its advantages of rapidity and ease, the ELISA kit did not accurately and consistently detect low toxin concentrations, although it may be useful as a screening tool.

Animals↗
PubMed2004

Proficiency studies on the determination of paralytic shellfish poisoning toxins in shellfish.

Paralytic shellfish poisoning toxins are produced by dinoflagellates. Shellfish filtering these unicellular algae will accumulate the toxins and pose a health risk when consumed by man. In the European Union, paralytic shellfish poisoning toxins in bivalve molluscs are regulated at a maximum content of 80 microg/100 g (91/492/EEC). The current reference method in the European Union is the mouse bioassay, but alternative methods including the liquid chromatography methodology are preferred for ethical reasons. Analyses of suspected shellfish batches revealed, however, unacceptable differences in results reported by a small group of Dutch laboratories all using liquid chromatography methods with precolumn derivatization, followed by fluorescence detection. Therefore, a series of proficiency studies were undertaken among these laboratories. In the first three studies, participants were more or less allowed their own choice of method execution details. This approach yielded unsatisfactory results. A fourth study was then initiated in which a standardized method was mandatory. Two types of test material were used in the fourth study: lyophilized Cardium tuberculatum material containing saxitoxin (STX) and decarbamoyl-saxitoxin (dc-STX), and lyophilized mussel material containing dc-STX. The latter material was investigated in an interlaboratory study involving 15 participants and was considered as the reference material. Among the four laboratories, coefficients of variation (ANOVA) for C. tuberculatum material were 10% (n = 11) and 9% (n = 12) for STX and dc-STX, respectively, and for the reference material was 8% (n = 12) for dc-STX. The joint efforts showed that variability in analysis results between laboratories that all apply more or less the same method can be drastically improved if the methodology is rigorously standardized.

Chromatography, Liquid↗
PubMed1989

Liquid chromatographic determination of domoic acid in shellfish products using the paralytic shellfish poison extraction procedure of the association of official analytical chemists.

Domoic acid, the recently discovered toxic substance found in contaminated mussels from an area in eastern Prince Edward Island (Canada) was extracted from mussel tissue using the procedure of the Association of Official Analytical Chemists for paralytic shellfish poisons. This involved a 5-min boiling of the sample with 0.1 M hydrochloric acid then cooling and centrifuging. An aliquot of the supernatant was diluted ten to one-hundred times with water, filtered and analysed by reversed-phase liquid chromatography with a mobile phase consisting of acetonitrile-water (12:88) at pH 2.5 and an absorption wavelength of 242 nm. The detection limit was about 0.5 mg/kg domoic acid in seafood samples. The technique was successfully applied to a variety of commercially purchased shellfish and shellfish products.

Animals↗