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A major challenge for LC-MS analysis is the ability to compare data between laboratories and across instrument platforms. Currently, mycotoxin determination relies on dereplication strategies based on physicochemical properties such as the m/z of the precursor and product ions. Unlike these intrinsic properties, retention time (tR) is an extrinsic property impacted by LC conditions, including mobile phases and column chemistry, making exchange of data between groups difficult. To address this, we are introducing the concept of incorporating an electrospray compatible, retention index (RI) system based on a series of N-alkylpyridinium-3-sulfonates (NAPS) into routine mycotoxin determination. These standards of differing alkyl chain length span RI units from 100 to 2000, are UV active and have fixed positive and negative charges for electrospray ionization in either mode. Using high resolution LC-MS/MS, the RIs of 96 mycotoxins and fungal secondary metabolites were normalized as a proof of concept with the NAPS RI system under multiple pH, column and gradient chromatographic conditions. This method was then applied to the analysis of a crude extract of Penicillium roqueforti, where we were able to decrease the number of false positives by implementing an RI filter as well as a secondary correction factor. Additionally, we developed software that allows users to convert published RIs to a predicted tR values. Integration of the NAPS RI system into routine LC-MS analysis will improve compound identifications and help facilitate ease of data sharing.
Justin B. Renaud; Shawn Hoogstra; Michael A. Quilliam; Mark W. Sumarah. Normalization of LC-MS mycotoxin determination using the N-alkylpyridinium-3-sulfonates (NAPS) retention index system. Journal of Chromatography A 2021, 1639, 461901 .
AMA StyleJustin B. Renaud, Shawn Hoogstra, Michael A. Quilliam, Mark W. Sumarah. Normalization of LC-MS mycotoxin determination using the N-alkylpyridinium-3-sulfonates (NAPS) retention index system. Journal of Chromatography A. 2021; 1639 ():461901.
Chicago/Turabian StyleJustin B. Renaud; Shawn Hoogstra; Michael A. Quilliam; Mark W. Sumarah. 2021. "Normalization of LC-MS mycotoxin determination using the N-alkylpyridinium-3-sulfonates (NAPS) retention index system." Journal of Chromatography A 1639, no. : 461901.
Paralytic shellfish toxins (PSTs) in shellfish are regulated by most countries and are a focus for research. Major inconsistencies exist in how PST concentrations are calculated and expressed. Resulting errors can result in adverse human health and trade implications. The reporting format in the Codex standard should be rigorously and uniformly followed.
Alison R. Turnbull; D. Tim Harwood; Michael J. Boundy; Patrick T. Holland; Gustaaf Hallegraeff; Navreet Malhi; Michael A. Quilliam; Michael Quilliam. Paralytic shellfish toxins – Call for uniform reporting units. Toxicon 2020, 178, 59 -60.
AMA StyleAlison R. Turnbull, D. Tim Harwood, Michael J. Boundy, Patrick T. Holland, Gustaaf Hallegraeff, Navreet Malhi, Michael A. Quilliam, Michael Quilliam. Paralytic shellfish toxins – Call for uniform reporting units. Toxicon. 2020; 178 ():59-60.
Chicago/Turabian StyleAlison R. Turnbull; D. Tim Harwood; Michael J. Boundy; Patrick T. Holland; Gustaaf Hallegraeff; Navreet Malhi; Michael A. Quilliam; Michael Quilliam. 2020. "Paralytic shellfish toxins – Call for uniform reporting units." Toxicon 178, no. : 59-60.
[D-Leu1]MC-LY (1) ([M + H]+ m/z 1044.5673, Δ 2.0 ppm), a new microcystin, was isolated from Microcystis aeruginosa strain CPCC-464. The compound was characterized by 1H and 13C NMR spectroscopy, liquid chromatography–high resolution tandem mass spectrometry (LC–HRMS/MS) and UV spectroscopy. A calibration reference material was produced after quantitation by 1H NMR spectroscopy and LC with chemiluminescence nitrogen detection. The potency of 1 in a protein phosphatase 2A inhibition assay was essentially the same as for MC-LR (2). Related microcystins, [D-Leu1]MC-LR (3) ([M + H]+ m/z 1037.6041, Δ 1.0 ppm), [D-Leu1]MC-M(O)R (6) ([M + H]+ m/z 1071.5565, Δ 2.0 ppm) and [D-Leu1]MC-MR (7) ([M + H]+ m/z 1055.5617, Δ 2.2 ppm), were also identified in culture extracts, along with traces of [D-Leu1]MC-M(O2)R (8) ([M + H]+ m/z 1087.5510, Δ 1.6 ppm), by a combination of chemical derivatization and LC–HRMS/MS experiments. The relative abundances of 1, 3, 6, 7 and 8 in a freshly extracted culture in the positive ionization mode LC–HRMS were ca. 84, 100, 3.0, 11 and 0.05, respectively. These and other results indicate that [D-Leu1]-containing MCs may be more common in cyanobacterial blooms than is generally appreciated but are easily overlooked with standard targeted LC–MS/MS screening methods.
Patricia Leblanc; Nadine Merkley; Krista Thomas; Nancy I. Lewis; Khalida Békri; Susan LeBlanc Renaud; Frances R. Pick; Pearse McCarron; Christopher O. Miles; Michael A. Quilliam. Isolation and Characterization of [D-Leu1]microcystin-LY from Microcystis aeruginosa CPCC-464. Toxins 2020, 12, 77 .
AMA StylePatricia Leblanc, Nadine Merkley, Krista Thomas, Nancy I. Lewis, Khalida Békri, Susan LeBlanc Renaud, Frances R. Pick, Pearse McCarron, Christopher O. Miles, Michael A. Quilliam. Isolation and Characterization of [D-Leu1]microcystin-LY from Microcystis aeruginosa CPCC-464. Toxins. 2020; 12 (2):77.
Chicago/Turabian StylePatricia Leblanc; Nadine Merkley; Krista Thomas; Nancy I. Lewis; Khalida Békri; Susan LeBlanc Renaud; Frances R. Pick; Pearse McCarron; Christopher O. Miles; Michael A. Quilliam. 2020. "Isolation and Characterization of [D-Leu1]microcystin-LY from Microcystis aeruginosa CPCC-464." Toxins 12, no. 2: 77.
The commercial demand for domoic acid (DA), the phycotoxin responsible for Amnesic Shellfish Poisoning, is currently met by extraction from a diminishing supply of stockpiled contaminated mussels (Mytilus edulis). As this supply becomes scarce, a more reliable source is needed. Purification of the toxin from an algal source would be easier and more economical than from shellfish tissue if algal growth and yield of toxin were maximized. This project was initiated to determine if DA could be produced using large-scale semi-continuous algal cultures, which should reduce labour and shorten the time required for biomass production. Pseudo-nitzschia multiseries was grown in 300-L fibreglass photobioreactors called a Brite-Box™. The effect of temperature and nutrient depletion on the yield of DA by P. multiseries was examined. A decline in maximum cell number without a substantial increase in cellular DA was associated with increased temperature. Maximum total cellular DA (8.8 pg cell−1) was achieved at 20 °C. Semi-continuous culture of P. multiseries is accompanied by increasing amounts of DA lost to the medium. The process was deemed to be feasible for growing P. multiseries but methods to recover this extracellular DA are necessary for this process to be economical.
Nancy I. Lewis; Stephen S. Bates; Michael A. Quilliam. Production of domoic acid from large-scale cultures of Pseudo-nitzschia multiseries: A feasibility study. Harmful Algae 2018, 79, 58 -63.
AMA StyleNancy I. Lewis, Stephen S. Bates, Michael A. Quilliam. Production of domoic acid from large-scale cultures of Pseudo-nitzschia multiseries: A feasibility study. Harmful Algae. 2018; 79 ():58-63.
Chicago/Turabian StyleNancy I. Lewis; Stephen S. Bates; Michael A. Quilliam. 2018. "Production of domoic acid from large-scale cultures of Pseudo-nitzschia multiseries: A feasibility study." Harmful Algae 79, no. : 58-63.
Polar marine toxins are more challenging to analyze by mass spectrometry-based methods than lipophilic marine toxins, which are now routinely measured in shellfish by multiclass reversed-phase liquid chromatography–tandem mass spectrometry (MS/MS) methods. Capillary electrophoresis (CE)–MS/MS is a technique that is well suited for the analysis of polar marine toxins, and has the potential of providing very high resolution separation. Here, we present a CE–MS/MS method developed, with use of a custom-built interface, for the sensitive multiclass analysis of paralytic shellfish toxins, tetrodotoxins, and domoic acid in seafood. A novel, highly acidic background electrolyte (5 M formic acid) was designed to maximize protonation of analytes and to allow a high degree of sample stacking to improve the limits of detection. The method was applied to a wide range of regulated and less common toxin analogues, and exhibited a high degree of selectivity between toxin isomers and matrix interference. The limits of detection in mussel tissue were 0.0052 mg/kg for tetrodotoxins, 0.160 mg/kg for domoic acid, and between 0.0018 and 0.120 mg/kg for paralytic shellfish toxins, all of which showed good linearity. Minimal ionization suppression was observed when the response from neat and mussel-matrix-matched standards was corrected with multiple internal standards. Analysis of shellfish matrix reference materials and spiked samples demonstrated good accuracy and precision. Finally, the method was transferred to a commercial CE–MS/MS system to demonstrate its widespread applicability for use in both R & D and routine regulatory settings. The approach of using a highly acidic background electrolyte is of broad interest, and can be considered generally applicable to simultaneous analysis of other classes of small, polar molecules with differing pKa values.
Daniel G. Beach; Elliott S. Kerrin; Krista Thomas; Michael Quilliam; Pearse McCarron. Capillary electrophoresis–tandem mass spectrometry for multiclass analysis of polar marine toxins. Analytical and Bioanalytical Chemistry 2018, 410, 5405 -5420.
AMA StyleDaniel G. Beach, Elliott S. Kerrin, Krista Thomas, Michael Quilliam, Pearse McCarron. Capillary electrophoresis–tandem mass spectrometry for multiclass analysis of polar marine toxins. Analytical and Bioanalytical Chemistry. 2018; 410 (22):5405-5420.
Chicago/Turabian StyleDaniel G. Beach; Elliott S. Kerrin; Krista Thomas; Michael Quilliam; Pearse McCarron. 2018. "Capillary electrophoresis–tandem mass spectrometry for multiclass analysis of polar marine toxins." Analytical and Bioanalytical Chemistry 410, no. 22: 5405-5420.
Contamination of economic bivalves with paralytic shellfish toxins (PST) occurs frequently in many parts of the world, which potentially threatens consumer health and the marine aquaculture economy. It is the objective of this study to develop a suitable technology for accelerating detoxification of PST from shellfish using activated carbon (AC). The adsorption efficiency of PST by eight different AC materials and by different particle sizes of wood-based AC (WAC) were tested and compared. Then WAC particles (37-48µm) were fed to mussels Mytilus galloprovincialis and scallops Chlamys farreri previously contaminated with PST through feeding with dinoflagellate Alexandrium tamarense ATHK. Results showed that the maximum adsorption ratio (65%) of PST was obtained by WAC. No significant differences in adsorption ratios were found between different particle sizes of WAC. The toxicity of mussels decreased by 41% and 68% after detoxification with WAC for 1 d and 3 d, respectively. Meanwhile, the detoxification ratio of mussels was approximately 3 times higher than that of scallops. This study suggests that the WAC could be used to accelerate the detoxification of PST by shellfish.
Jiangbing Qiu; Hua Fan; Ting Liu; Xia Liang; Fanping Meng; Michael Quilliam; Aifeng Li. Application of activated carbon to accelerate detoxification of paralytic shellfish toxins from mussels Mytilus galloprovincialis and scallops Chlamys farreri. Ecotoxicology and Environmental Safety 2018, 148, 402 -409.
AMA StyleJiangbing Qiu, Hua Fan, Ting Liu, Xia Liang, Fanping Meng, Michael Quilliam, Aifeng Li. Application of activated carbon to accelerate detoxification of paralytic shellfish toxins from mussels Mytilus galloprovincialis and scallops Chlamys farreri. Ecotoxicology and Environmental Safety. 2018; 148 ():402-409.
Chicago/Turabian StyleJiangbing Qiu; Hua Fan; Ting Liu; Xia Liang; Fanping Meng; Michael Quilliam; Aifeng Li. 2018. "Application of activated carbon to accelerate detoxification of paralytic shellfish toxins from mussels Mytilus galloprovincialis and scallops Chlamys farreri." Ecotoxicology and Environmental Safety 148, no. : 402-409.
The non-protein amino acid β-methylamino-L-alanine (BMAA) has been linked to neurodegenerative disease and reported throughout the environment. Proposed mechanisms of bioaccumulation, trophic transfer and chronic toxicity of BMAA rely on the hypothesis of protein misincorporation. Poorly selective methods for BMAA analysis have led to controversy. Here, a recently reported highly selective method for BMAA quantitation using hydrophilic interaction liquid chromatography-differential mobility spectrometry-tandem mass spectrometry (HILIC-DMS-MS/MS) is expanded to include proteinogenic amino acids from hydrolyzed biological samples. For BMAA quantitation, we present a double spiking isotope dilution approach using D3-BMAA and 13C15N2-BMAA. These methods were applied to study release of BMAA during acid hydrolysis under a variety of conditions, revealing that the majority of BMAA can be extracted along with only a small proportion of protein. A time course hydrolysis of BMAA from mussel tissue was carried out to assess the recovery of BMAA during sample preparation. The majority of BMAA measured by typical methods was released before a significant proportion of protein was hydrolyzed. Little change was observed in protein hydrolysis beyond typical hydrolysis times but the concentration of BMAA increased linearly. These findings demonstrate protein misincorporation is not the predominant form of BMAA in cycad and shellfish.
Daniel G. Beach; Elliott S. Kerrin; Sabrina D. Giddings; Michael Quilliam; Pearse McCarron. Differential Mobility-Mass Spectrometry Double Spike Isotope Dilution Study of Release of β-Methylaminoalanine and Proteinogenic Amino Acids during Biological Sample Hydrolysis. Scientific Reports 2018, 8, 1 -11.
AMA StyleDaniel G. Beach, Elliott S. Kerrin, Sabrina D. Giddings, Michael Quilliam, Pearse McCarron. Differential Mobility-Mass Spectrometry Double Spike Isotope Dilution Study of Release of β-Methylaminoalanine and Proteinogenic Amino Acids during Biological Sample Hydrolysis. Scientific Reports. 2018; 8 (1):1-11.
Chicago/Turabian StyleDaniel G. Beach; Elliott S. Kerrin; Sabrina D. Giddings; Michael Quilliam; Pearse McCarron. 2018. "Differential Mobility-Mass Spectrometry Double Spike Isotope Dilution Study of Release of β-Methylaminoalanine and Proteinogenic Amino Acids during Biological Sample Hydrolysis." Scientific Reports 8, no. 1: 1-11.
Paralytic shellfish toxins (PSTs) are potent neurotoxins produced by marine dinoflagellates that are responsible for paralytic shellfish poisoning (PSP) in humans. This work highlights our ongoing efforts to develop quantitative methods for PSTs using hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS). Compared with the commonly used method of liquid chromatography with post-column oxidation and fluorescence detection (LC-ox-FLD), HILIC-MS/MS has the potential of being more robust, sensitive and straightforward to operate, and provides unequivocal confirmation of toxin identity. The main driving force for the present work was the need for a complementary method to LC-ox-FLD to assign values to shellfish tissue matrix reference materials for PSTs. Method parameters that were optimized included LC mobile and stationary phases, electrospray ionization (ESI) conditions, and MS/MS detection parameters. The developed method has been used in the detection and identification of a wide range of PSTs including less common analogues and metabolites in a range of shellfish and algal samples. We have assessed the matrix effects of shellfish samples and have evaluated dilution, standard addition and matrix matched calibration as means of mitigating them. Validation on one LC-MS/MS system for nine common PST analogues (GTX1-4, dcGTX2&3, STX, NEO, and dcSTX) was completed using standard addition. The method was then transferred to a more sensitive LC-MS/MS system, expanded to include five more PSTs (C1&2, dcNEO and GTX5&6) and validated using matrix matched calibration. Limits of detection of the validated method ranged between 6 and 280 nmol/kg tissue using standard addition in extracts of blue mussels, with recoveries between 92 and 108%. Finally, this method was used in combination with the AOAC Official Method based on LC-ox-FLD to measure PSTs in a new mussel tissue matrix reference material.
Krista M. Thomas; Daniel G. Beach; Kelley L. Reeves; Ryan S. Gibbs; Elliott S. Kerrin; Pearse McCarron; Michael A. Quilliam. Hydrophilic interaction liquid chromatography-tandem mass spectrometry for quantitation of paralytic shellfish toxins: validation and application to reference materials. Analytical and Bioanalytical Chemistry 2017, 409, 5675 -5687.
AMA StyleKrista M. Thomas, Daniel G. Beach, Kelley L. Reeves, Ryan S. Gibbs, Elliott S. Kerrin, Pearse McCarron, Michael A. Quilliam. Hydrophilic interaction liquid chromatography-tandem mass spectrometry for quantitation of paralytic shellfish toxins: validation and application to reference materials. Analytical and Bioanalytical Chemistry. 2017; 409 (24):5675-5687.
Chicago/Turabian StyleKrista M. Thomas; Daniel G. Beach; Kelley L. Reeves; Ryan S. Gibbs; Elliott S. Kerrin; Pearse McCarron; Michael A. Quilliam. 2017. "Hydrophilic interaction liquid chromatography-tandem mass spectrometry for quantitation of paralytic shellfish toxins: validation and application to reference materials." Analytical and Bioanalytical Chemistry 409, no. 24: 5675-5687.
Following heavy precipitation, we observed an intense algal bloom in the St. Lawrence Estuary (SLE) that coincided with an unusually high mortality of several species of marine fish, birds and mammals, including species designated at risk. The algal species was identified as Alexandrium tamarense and was determined to contain a potent mixture of paralytic shellfish toxins (PST). Significant levels of PST were found in the liver and/or gastrointestinal contents of several carcasses tested as well as in live planktivorous fish, molluscs and plankton samples collected during the bloom. This provided strong evidence for the trophic transfer of PST resulting in mortalities of multiple wildlife species. This conclusion was strengthened by the sequence of mortalities, which followed the drift of the bloom along the coast of the St. Lawrence Estuary. No other cause of mortality was identified in the majority of animals examined at necropsy. Reports of marine fauna presenting signs of neurological dysfunction were also supportive of exposure to these neurotoxins. The event reported here represents the first well-documented case of multispecies mass mortality of marine fish, birds and mammals linked to a PST-producing algal bloom.
Michel Starr; Stéphane Lair; Sonia Michaud; Michael Scarratt; Michael Quilliam; Denis Lefaivre; Michel Robert; Andrew Wotherspoon; Robert Michaud; Nadia Ménard; Gilbert Sauvé; Sylvie Lessard; Pierre Béland; Lena Measures. Multispecies mass mortality of marine fauna linked to a toxic dinoflagellate bloom. PLoS ONE 2017, 12, e0176299 .
AMA StyleMichel Starr, Stéphane Lair, Sonia Michaud, Michael Scarratt, Michael Quilliam, Denis Lefaivre, Michel Robert, Andrew Wotherspoon, Robert Michaud, Nadia Ménard, Gilbert Sauvé, Sylvie Lessard, Pierre Béland, Lena Measures. Multispecies mass mortality of marine fauna linked to a toxic dinoflagellate bloom. PLoS ONE. 2017; 12 (5):e0176299.
Chicago/Turabian StyleMichel Starr; Stéphane Lair; Sonia Michaud; Michael Scarratt; Michael Quilliam; Denis Lefaivre; Michel Robert; Andrew Wotherspoon; Robert Michaud; Nadia Ménard; Gilbert Sauvé; Sylvie Lessard; Pierre Béland; Lena Measures. 2017. "Multispecies mass mortality of marine fauna linked to a toxic dinoflagellate bloom." PLoS ONE 12, no. 5: e0176299.
Recent reports of the widespread occurrence of the neurotoxin β-N-methylamino-L-alanine (BMAA) in cyanobacteria and particularly seafood have raised concerns for public health. LC-MS/MS is currently the analytical method of choice for BMAA determinations but incomplete separation of isomeric and isobaric compounds, matrix suppression and conjugated forms are plausible limitations. In this study, capillary electrophoresis (CE) coupled with MS/MS has been developed as an alternative method for the quantitative determination of free BMAA. Using a bare fused silica capillary, a phosphate buffer (250 mM, pH 3.0) and UV detection, it was possible to separate BMAA from four isomers, but the limit of detection (LOD) of 0.25 μg mL proved insufficient for analysis of typical samples. Coupling the CE to a triple quadrupole MS was accomplished using a custom sheath-flow interface. The best separation was achieved with a 5 M formic acid in water/acetonitrile (9:1) background electrolyte. Strong acid hydrolysis of lyophilized samples was used to release BMAA from conjugated forms. Field-amplified stacking after injection was achieved by lowering sample ionic strength with a cation-exchange cleanup procedure. Quantitation was accomplished using isotope dilution with deuterium-labelled BMAA as internal standard. An LOD for BMAA in solution of 0.8 ng mL was attained, which was equivalent to 16 ng g dry mass in samples using the specified extraction procedure. This was comparable with LC-MS/MS methods. The method displayed excellent resolution of amino acid isomers and had no interference from matrix components. The presence of BMAA in cycad, mussel and lobster samples was confirmed by CE-MS/MS, but not in an in-house cyanobacterial reference material, with quantitative results agreeing with those from LC-MS/MS. Graphical Abstract CE-MS separation and detection of BMAA, its isomers and the internal standard BMAA-d3.
Elliott S. Kerrin; Robert L. White; Michael A. Quilliam. Quantitative determination of the neurotoxin β-N-methylamino-l-alanine (BMAA) by capillary electrophoresis–tandem mass spectrometry. Analytical and Bioanalytical Chemistry 2016, 409, 1481 -1491.
AMA StyleElliott S. Kerrin, Robert L. White, Michael A. Quilliam. Quantitative determination of the neurotoxin β-N-methylamino-l-alanine (BMAA) by capillary electrophoresis–tandem mass spectrometry. Analytical and Bioanalytical Chemistry. 2016; 409 (6):1481-1491.
Chicago/Turabian StyleElliott S. Kerrin; Robert L. White; Michael A. Quilliam. 2016. "Quantitative determination of the neurotoxin β-N-methylamino-l-alanine (BMAA) by capillary electrophoresis–tandem mass spectrometry." Analytical and Bioanalytical Chemistry 409, no. 6: 1481-1491.
A freeze-dried mussel tissue (Mytilus edulis) reference material (CRM-FDMT1) was produced containing multiple groups of shellfish toxins. Homogeneity and stability testing showed the material to be fit for purpose. The next phase of work was to assign certified values and uncertainties to 10 analytes from six different toxin groups. Efforts involved optimizing extraction procedures for the various toxin groups and performing measurements using liquid chromatography-based analytical methods. A key aspect of the work was compensating for matrix effects associated with liquid chromatography-mass spectrometry through standard addition, dilution, or matrix-matched calibration. Certified mass fraction values are reported as mg/kg of CRM-FDMT1 powder as bottled for azaspiracid-1, -2, and -3 (4.10 ± 0.40; 1.13± 0.10; 0.96 ± 0.10, respectively), okadaic acid, dinophysistoxin-1 and -2 (1.59 ± 0.18; 0.68 ± 0.07; 3.57± 0.33, respectively), yessotoxin (2.49 ± 0.28), pectenotoxin-2 (0.66 ± 0.06), 13-desmethylspirolide-C (2.70 ± 0.26), and domoic acid (126 ± 10). Combined uncertainties for the certified values include contributions from homogeneity, stability, and characterization experiments. The commutability of CRM-FDMT1 was assessed by examining the extractability and matrix effects for the freeze-dried material in comparison with its equivalent wet tissue homogenate. CRM-FDMT1 is the first shellfish matrix CRM with certified values for yessotoxins, pectenotoxins or spirolides, and is the first CRM certified for multiple toxin groups. CRM-FDMT1 is a valuable tool for quality assurance of phycotoxin monitoring programs and for analytical method development and validation.
Pearse McCarron; Elliott Wright; Hakan Emteborg; Michael Quilliam. A mussel tissue certified reference material for multiple phycotoxins. Part 4: certification. Analytical and Bioanalytical Chemistry 2016, 409, 95 -106.
AMA StylePearse McCarron, Elliott Wright, Hakan Emteborg, Michael Quilliam. A mussel tissue certified reference material for multiple phycotoxins. Part 4: certification. Analytical and Bioanalytical Chemistry. 2016; 409 (1):95-106.
Chicago/Turabian StylePearse McCarron; Elliott Wright; Hakan Emteborg; Michael Quilliam. 2016. "A mussel tissue certified reference material for multiple phycotoxins. Part 4: certification." Analytical and Bioanalytical Chemistry 409, no. 1: 95-106.
Okadaic acid (OA) and its analogs, dinophysistoxins-1 (DTX1) and -2 (DTX2) are lipophilic biotoxins produced by marine algae that can accumulate in shellfish and cause the human illness known as diarrhetic shellfish poisoning (DSP). Regulatory testing of shellfish is required to protect consumers and the seafood industry. Certified reference materials (CRMs) are essential for the development, validation, and quality control of analytical methods, and thus play an important role in toxin monitoring. This paper summarizes work on research and development of shellfish tissue reference materials for OA and DTXs. Preliminary work established the appropriate conditions for production of shellfish tissue CRMs for OA and DTXs. Source materials, including naturally incurred shellfish tissue and cultured algae, were screened for their DSP toxins. This preliminary work informed planning and production of a wet mussel (Mytilus edulis) tissue homogenate matrix CRM. The homogeneity and stability of the CRM were evaluated and found to be fit-for-purpose. Extraction and LC-tandem MS methods were developed to accurately certify the concentrations of OA, DTX1, and DTX2 using a combination of standard addition and matrix-matched calibration to compensate for matrix effects in electrospray ionization. The concentration of domoic acid was also certified. Uncertainties were assigned following standards and guidelines from the International Organization for Standardization. The presence of other toxins in the CRM was also assessed and information values are reported for OA and DTX acyl esters.
Pearse McCarron; Kelley L Reeves; Sabrina D Giddings; Daniel G Beach; Michael Quilliam. Development of Certified Reference Materials for Diarrhetic Shellfish Poisoning Toxins, Part 2: Shellfish Matrix Materials. Journal of AOAC INTERNATIONAL 2016, 99, 1163 -1172.
AMA StylePearse McCarron, Kelley L Reeves, Sabrina D Giddings, Daniel G Beach, Michael Quilliam. Development of Certified Reference Materials for Diarrhetic Shellfish Poisoning Toxins, Part 2: Shellfish Matrix Materials. Journal of AOAC INTERNATIONAL. 2016; 99 (5):1163-1172.
Chicago/Turabian StylePearse McCarron; Kelley L Reeves; Sabrina D Giddings; Daniel G Beach; Michael Quilliam. 2016. "Development of Certified Reference Materials for Diarrhetic Shellfish Poisoning Toxins, Part 2: Shellfish Matrix Materials." Journal of AOAC INTERNATIONAL 99, no. 5: 1163-1172.
The implementation of instrumental analytical methods such as LC-MS for routine monitoring of toxins requires the availability of accurate calibration standards. This is a challenge because many toxins are rare, expensive, dangerous to handle, and/or unstable, and simple gravimetric procedures are not reliable for establishing accurate concentrations in solution. NMR has served as one method of qualitative and quantitative characterization of toxin calibration solution Certified Reference Materials (CRMs). LC with chemiluminescence N detection (LC-CLND) was selected as a complementary method for comprehensive characterization of CRMs because it provides a molar response to N. Here we report on our investigation of LC-CLND as a method suitable for quantitative analysis of nitrogenous toxins. It was demonstrated that a wide range of toxins could be analyzed quantitatively by LC-CLND. Furthermore, equimolar responses among diverse structures were established and it was shown that a single high-purity standard such as caffeine could be used for instrument calibration. The limit of detection was approximately 0.6 ng N. Measurement of several of Canada's National Research Council toxin CRMs with caffeine as the calibrant showed precision averaging 2% RSD and accuracy ranging from 97 to 102%. Application of LC-CLND to the production of calibration solution CRMs and the establishment of traceability of measurement results are presented.
Krista Thomas; Dominik Wechsler; Yi-Min Chen; Sheila Crain; Michael Quilliam. Analysis of Natural Toxins by Liquid Chromatography-Chemiluminescence Nitrogen Detection and Application to the Preparation of Certified Reference Materials. Journal of AOAC INTERNATIONAL 2016, 99, 1173 -1184.
AMA StyleKrista Thomas, Dominik Wechsler, Yi-Min Chen, Sheila Crain, Michael Quilliam. Analysis of Natural Toxins by Liquid Chromatography-Chemiluminescence Nitrogen Detection and Application to the Preparation of Certified Reference Materials. Journal of AOAC INTERNATIONAL. 2016; 99 (5):1173-1184.
Chicago/Turabian StyleKrista Thomas; Dominik Wechsler; Yi-Min Chen; Sheila Crain; Michael Quilliam. 2016. "Analysis of Natural Toxins by Liquid Chromatography-Chemiluminescence Nitrogen Detection and Application to the Preparation of Certified Reference Materials." Journal of AOAC INTERNATIONAL 99, no. 5: 1173-1184.
Okadaic acid (OA) and its analogs dinophysistoxins-1 (DTX1) and -2 (DTX2) are lipophilic polyethers produced by marine dinoflagellates. These toxins accumulate in shellfish and cause diarrhetic shellfish poisoning (DSP) in humans. Regulatory testing of shellfish is essential to safeguard public health and for international trade. Certified reference materials (CRMs) play a key role in analytical monitoring programs. This paper presents an overview of the interdisciplinary work that went into the planning, production, and certification of calibration-solution CRMs for OA, DTX1, and DTX2. OA and DTX1 were isolated from large-scale algal cultures and DTX2 from naturally contaminated mussels. Toxins were isolated by a combination of extraction and chromatographic steps with processes adapted to suit the source and concentration of each toxin. New 19-epi-DSP toxin analogs were identified as minor impurities. Once OA, DTX1, and DTX2 were established to be of suitable purity, solutions were prepared and dispensed into flame-sealed glass ampoules. Certification measurements were carried out using quantitative NMR spectroscopy and LC-tandem MS. Traceability of measurements was established through certified external standards of established purity. Uncertainties were assigned following standards and guidelines from the International Organization for Standardization, with components from the measurement, stability, and homogeneity studies being propagated into final combined uncertainties.
Daniel G Beach; Sheila Crain; Nancy Lewis; Patricia LeBlanc; William R Hardstaff; Ruth A Perez; Sabrina D Giddings; Camilo F Martinez-Farina; Roumiana Stefanova; Ian W Burton; Jane Kilcoyne; Jeremy Melanson; Michael Quilliam; Pearse McCarron. Development of Certified Reference Materials for Diarrhetic Shellfish Poisoning Toxins, Part 1: Calibration Solutions. Journal of AOAC INTERNATIONAL 2016, 99, 1151 -1162.
AMA StyleDaniel G Beach, Sheila Crain, Nancy Lewis, Patricia LeBlanc, William R Hardstaff, Ruth A Perez, Sabrina D Giddings, Camilo F Martinez-Farina, Roumiana Stefanova, Ian W Burton, Jane Kilcoyne, Jeremy Melanson, Michael Quilliam, Pearse McCarron. Development of Certified Reference Materials for Diarrhetic Shellfish Poisoning Toxins, Part 1: Calibration Solutions. Journal of AOAC INTERNATIONAL. 2016; 99 (5):1151-1162.
Chicago/Turabian StyleDaniel G Beach; Sheila Crain; Nancy Lewis; Patricia LeBlanc; William R Hardstaff; Ruth A Perez; Sabrina D Giddings; Camilo F Martinez-Farina; Roumiana Stefanova; Ian W Burton; Jane Kilcoyne; Jeremy Melanson; Michael Quilliam; Pearse McCarron. 2016. "Development of Certified Reference Materials for Diarrhetic Shellfish Poisoning Toxins, Part 1: Calibration Solutions." Journal of AOAC INTERNATIONAL 99, no. 5: 1151-1162.
The concentration of the saxitoxin analogue LWTX-1 was quantified in samples of the benthic filamentous cyanobacterium Lyngbya wollei (Farlow ex Gomont) Speziale and Dyck collected in two fluvial lakes of the St. Lawrence River (Canada) over the 2006\u20132013 period. The study was aimed at documenting the spatial (between fluvial lakes, between sites within each lake) and temporal (interannual, monthly) variations of toxin concentration in relation with hydrological (water level), physical (water temperature, conductivity, transparency), chemical (nutrients in overlying water) and biological (L. wollei biomass and mat condition) characteristics. Toxin concentration was hypothesized to vary seasonally with biomass accumulation and environmental conditions. Toxin concentrations measured in Lake Saint-Louis (51 +/- 40 mg LWTX-1 g-1 DM, N = 29 days in 2007, 2009\u20132011) were double those in Lake Saint-Pierre (25 +/- 31 mg LWTX-1 g-1 DM, N = 26 days in 2006\u20132008, 2012\u20132013); however, August 2007 measurements taken from both lakes did not differ significantly. Ten of the twelve highest values (>100 mg LWTX-1 g-1 DM) were obtained from Lake Saint-Louis, between April and October in 2007, 2010 or 2011. Under ice samples showed intermediate concentrations of LWTX-1 (42 +/- 9 mg LWTX-1 g-1 DM, N = 2). Concentrations of LWTX-1 were positively correlated with Secchi depth (r = 0.59, p < 0.001), L. wollei biomass (Spearman r = 0.31, p < 0.01) and %N in filaments (r = 0.48, p < 0.001), suggesting toxin production was linked to mat growth and metabolism rather than water quality. Although LWTX-1 has been reported to have a low toxicity, monitoring of L. wollei abundance is required to assess the environmental and human health risks posed by this taxon in the St. Lawrence - Great Lakes system.Peer reviewed: YesNRC publication: Ye
C. Hudon; P. Gagnon; S. Poirier Larabie; C. Gagnon; A. Lajeunesse; M. Lachapelle; Michael Quilliam. Spatial and temporal variations of a saxitoxin analogue (LWTX-1) in Lyngbya wollei (Cyanobacteria) mats in the St. Lawrence River (Québec, Canada). Harmful Algae 2016, 57, 69 -77.
AMA StyleC. Hudon, P. Gagnon, S. Poirier Larabie, C. Gagnon, A. Lajeunesse, M. Lachapelle, Michael Quilliam. Spatial and temporal variations of a saxitoxin analogue (LWTX-1) in Lyngbya wollei (Cyanobacteria) mats in the St. Lawrence River (Québec, Canada). Harmful Algae. 2016; 57 ():69-77.
Chicago/Turabian StyleC. Hudon; P. Gagnon; S. Poirier Larabie; C. Gagnon; A. Lajeunesse; M. Lachapelle; Michael Quilliam. 2016. "Spatial and temporal variations of a saxitoxin analogue (LWTX-1) in Lyngbya wollei (Cyanobacteria) mats in the St. Lawrence River (Québec, Canada)." Harmful Algae 57, no. : 69-77.
Derivatisation with a labelled reagent allows for isotope dilution calibration of algal toxins for which no labelled standard is available. Two methods were developed for the analysis of algal biotoxins in complex biological and environmental samples to demonstrate the concept of isotope-labelling derivatisation for quantitation. These methods are based on dansyl chloride derivatisation of samples and dansyl-d 6 chloride derivatisation of toxin standards. Derivatised sample and standard are then mixed to achieve isotope dilution calibration in liquid chromatography–tandem mass spectrometry analyses. Quantitation of the marine toxin domoic acid (DA) in mussel tissues and the freshwater toxins anatoxin-a (ATX) and homoanatoxin-a (hATX) in cyanobacteria is demonstrated. For DA, isotope-labelling was incorporated into existing dansylation methodology using inexpensive and commercially available reagents. For ATXs, a novel sample preparation procedure is presented that involves solid phase extraction on a mixed reverse phase/weak anion exchange column that facilitates simultaneous clean-up of the derivatised toxins and removal of excess dansylation reagent through covalent bonding. The challenge of achieving co-elution in LC between deuterated and non-deuterated dansylated toxins was addressed by modifying separation conditions from the usual reverse phase (RP) separation to hydrophilic interaction liquid chromatography in the case of DA and a shortened RP separation with high organic modifier content in the case of the ATXs. The new methods gave limits of detection between 10 and 60 μg kg −1 and allowed for precise, accurate and fast determination of toxins in spiked control samples and matrix reference materials. This work demonstrates that isotope-labelling derivatisation is broadly applicable to the field of algal toxin analysis where derivatisation is well established but isotopically-labelled standards are not available.
Daniel G. Beach; Christie Hollingdale; Michael A. Quilliam. Isotope-labelling derivatisation: a broadly applicable approach to quantitation of algal toxins by isotope dilution LC-MS/MS. Analytical Methods 2016, 8, 2872 -2879.
AMA StyleDaniel G. Beach, Christie Hollingdale, Michael A. Quilliam. Isotope-labelling derivatisation: a broadly applicable approach to quantitation of algal toxins by isotope dilution LC-MS/MS. Analytical Methods. 2016; 8 (14):2872-2879.
Chicago/Turabian StyleDaniel G. Beach; Christie Hollingdale; Michael A. Quilliam. 2016. "Isotope-labelling derivatisation: a broadly applicable approach to quantitation of algal toxins by isotope dilution LC-MS/MS." Analytical Methods 8, no. 14: 2872-2879.
The neurotoxin β-N-methylamino-l-alanine (BMAA) has been reported in cyanobacteria and shellfish, raising concerns about widespread human exposure. However, inconsistent results for BMAA analysis have led to controversy. Liquid chromatography–tandem mass spectrometry (LC–MS/MS) is the most appropriate method for analysis of BMAA, but the risk of interference from isomers, other sample components, and the electrospray background is still present. We have investigated differential mobility spectrometry (DMS) as an ion filter to improve selectivity in the hydrophilic interaction liquid chromatographic (HILIC)–MS/MS determination of BMAA. We obtained standards for two BMAA isomers not previously analyzed by HILIC–MS, β-amino-N-methylalanine and 3,4-diaminobutanoic acid, and the typically used 2,4-diaminobutanoic acid and N-(2-aminoethyl)glycine. DMS separation of BMAA from these isomers was achieved and optimized conditions were used to develop a sensitive and highly selective multidimensional HILIC–DMS–MS/MS method. This work revealed current technical limitations of DMS for trace quantitation, and practical solutions were implemented. Accurate control of low levels of DMS carrier gas modifier was essential, but required external metering. The linearity of our optimized method was excellent from 0.01 to 6 μmol L−1. The instrumental LOD was 0.4 pg BMAA injected on-column and the estimated method LOD was 20 ng g−1 dry weight for BMAA in sample matrix. The method was used to analyze cycad plant tissue, a cyanobacterial reference material, and mussel tissues, by use of isotope-dilution quantitation with deuterated BMAA. This confirmed the presence of BMAA and several of its isomers in cycad and mussel tissues, including commercially available mussel tissue reference materials certified for other biotoxins.
Daniel G. Beach; Elliott S. Kerrin; Michael Quilliam. Selective quantitation of the neurotoxin BMAA by use of hydrophilic-interaction liquid chromatography–differential mobility spectrometry–tandem mass spectrometry (HILIC–DMS–MS/MS). Analytical and Bioanalytical Chemistry 2015, 407, 8397 -8409.
AMA StyleDaniel G. Beach, Elliott S. Kerrin, Michael Quilliam. Selective quantitation of the neurotoxin BMAA by use of hydrophilic-interaction liquid chromatography–differential mobility spectrometry–tandem mass spectrometry (HILIC–DMS–MS/MS). Analytical and Bioanalytical Chemistry. 2015; 407 (28):8397-8409.
Chicago/Turabian StyleDaniel G. Beach; Elliott S. Kerrin; Michael Quilliam. 2015. "Selective quantitation of the neurotoxin BMAA by use of hydrophilic-interaction liquid chromatography–differential mobility spectrometry–tandem mass spectrometry (HILIC–DMS–MS/MS)." Analytical and Bioanalytical Chemistry 407, no. 28: 8397-8409.
A new species of Pseudo‐nitzschia (Bacillariophyceae) is described from plankton samples collected from Port Dickson (Malacca Strait, Malaysia) and Manzanillo Bay (Colima, Mexico). The species possesses a distinctive falcate cell valve, from which they form sickle‐like colonies in both environmental samples and cultured strains. Detailed observation of frustules under TEM revealed ultrastructure that closely resembles P. decipiens, yet the new species differs by the valve shape and greater ranges of striae and poroid densities. The species is readily distinguished from the curve‐shaped P. subcurvata by the presence of a central interspace. The morphological distinction is further supported by phylogenetic discrimination. We sequenced and analyzed the nuclear ribosomal RNA genes in the LSU and the second internal transcribed spacer, including its secondary structure, to infer the phylogenetic relationship of the new species with its closest relatives. The results revealed a distinct lineage of the new species, forming a sister cluster with its related species, P. decipiens and P. galaxiae, but not with P. subcurvata. We examined the domoic acid (DA) production of five cultured strains from Malaysia by Liquid chromatography‐mass spectrometry (LC‐MS), but they showed no detectable DA. Here, we present the taxonomic description of the vegetative cells, document the sexual reproduction, and detail the molecular phylogenetics of Pseudo‐nitzschia sabit sp. nov.
Sing Tung Teng; Po Teen Lim; Hong Chang Lim; Maria Rivera; Sonia Quijano; Yoshinobu Takata; Michael Quilliam; Matthias Wolf; Stephen S. Bates; Chui Pin Leaw. A non-toxigenic but morphologically and phylogenetically distinct new species of Pseudo-nitzschia , P. sabit sp. nov. (Bacillariophyceae). Journal of Phycology 2015, 51, 706 -725.
AMA StyleSing Tung Teng, Po Teen Lim, Hong Chang Lim, Maria Rivera, Sonia Quijano, Yoshinobu Takata, Michael Quilliam, Matthias Wolf, Stephen S. Bates, Chui Pin Leaw. A non-toxigenic but morphologically and phylogenetically distinct new species of Pseudo-nitzschia , P. sabit sp. nov. (Bacillariophyceae). Journal of Phycology. 2015; 51 (4):706-725.
Chicago/Turabian StyleSing Tung Teng; Po Teen Lim; Hong Chang Lim; Maria Rivera; Sonia Quijano; Yoshinobu Takata; Michael Quilliam; Matthias Wolf; Stephen S. Bates; Chui Pin Leaw. 2015. "A non-toxigenic but morphologically and phylogenetically distinct new species of Pseudo-nitzschia , P. sabit sp. nov. (Bacillariophyceae)." Journal of Phycology 51, no. 4: 706-725.
The worldwide increase in cyanobacterial contamination of freshwater lakes and rivers is of great concern as many cyanobacteria produce potent hepatotoxins and neurotoxins (cyanotoxins). Such toxins pose a threat to aquatic ecosystems, livestock, and drinking water supplies. In addition, dietary supplements prepared from cyanobacteria can pose a risk to consumers if they contain toxins. Analytical monitoring for toxins in the environment and in consumer products is essential for the protection of public health. Reference materials (RMs) are an essential tool for the development and validation of analytical methods and are necessary for ongoing quality control of monitoring operations. Since the availability of appropriate RMs for cyanotoxins has been very limited, the present study was undertaken to examine the feasibility of producing a cyanobacterial matrix RM containing various cyanotoxins. The first step was large-scale culturing of various cyanobacterial cultures that produce anatoxins, microcystins, and cylindrospermopsins. After harvesting, the biomass was lyophilized, blended, homogenized, milled, and bottled. The moisture content and physical characteristics were assessed in order to evaluate the effectiveness of the production process. Toxin levels were measured by liquid chromatography with tandem mass spectrometry and ultraviolet detection. The reference material was found to be homogeneous for toxin content. Stability studies showed no significant degradation of target toxins over a period of 310 days at temperatures up to +40 °C except for the anatoxin-a, which showed some degradation at +40 °C. These results show that a fit-for-purpose matrix RM for cyanotoxins can be prepared using the processes and techniques applied in this work.
Christie Hollingdale; Krista Thomas; Nancy Lewis; Khalida Békri; Pearse McCarron; Michael A. Quilliam. Feasibility study on production of a matrix reference material for cyanobacterial toxins. Analytical and Bioanalytical Chemistry 2015, 407, 5353 -5363.
AMA StyleChristie Hollingdale, Krista Thomas, Nancy Lewis, Khalida Békri, Pearse McCarron, Michael A. Quilliam. Feasibility study on production of a matrix reference material for cyanobacterial toxins. Analytical and Bioanalytical Chemistry. 2015; 407 (18):5353-5363.
Chicago/Turabian StyleChristie Hollingdale; Krista Thomas; Nancy Lewis; Khalida Békri; Pearse McCarron; Michael A. Quilliam. 2015. "Feasibility study on production of a matrix reference material for cyanobacterial toxins." Analytical and Bioanalytical Chemistry 407, no. 18: 5353-5363.
The marine dinoflagellate Gymnodinium catenatum has been associated with paralytic shellfish poisoning (PSP) outbreaks in Portuguese waters for many years. PSP syndrome is caused by consumption of seafood contaminated with paralytic shellfish toxins (PSTs), a suite of potent neurotoxins. Gymnodinium catenatum was frequently reported along the Portuguese coast throughout the late 1980s and early 1990s, but was absent between 1995 and 2005. Since this time, G. catenatum blooms have been recurrent, causing contamination of fishery resources along the Atlantic coast of Portugal. The aim of this study was to evaluate the toxin profile of G. catenatum isolated from the Portuguese coast before and after the 10-year hiatus to determine changes and potential impacts for the region. Hydrophilic interaction liquid chromatography tandem mass spectrometry (HILIC-MS/MS) was utilized to determine the presence of any known and emerging PSTs in sample extracts. Several PST derivatives were identified, including the N-sulfocarbamoyl analogues (C1–4), gonyautoxin 5 (GTX5), gonyautoxin 6 (GTX6), and decarbamoyl derivatives, decarbamoyl saxitoxin (dcSTX), decarbamoyl neosaxitoxin (dcNeo) and decarbamoyl gonyautoxin 3 (dcGTX3). In addition, three known hydroxy benzoate derivatives, G. catenatum toxin 1 (GC1), GC2 and GC3, were confirmed in cultured and wild strains of G. catenatum. Moreover, two presumed N-hydroxylated analogues of GC2 and GC3, designated GC5 and GC6, are reported. This work contributes to our understanding of the toxigenicity of G. catenatum in the coastal waters of Portugal and provides valuable information on emerging PST classes that may be relevant for routine monitoring programs tasked with the prevention and control of marine toxins in fish and shellfish.
Pedro R. Costa; Alison Robertson; Michael A. Quilliam. Toxin Profile of Gymnodinium catenatum (Dinophyceae) from the Portuguese Coast, as Determined by Liquid Chromatography Tandem Mass Spectrometry. Marine Drugs 2015, 13, 2046 -2062.
AMA StylePedro R. Costa, Alison Robertson, Michael A. Quilliam. Toxin Profile of Gymnodinium catenatum (Dinophyceae) from the Portuguese Coast, as Determined by Liquid Chromatography Tandem Mass Spectrometry. Marine Drugs. 2015; 13 (4):2046-2062.
Chicago/Turabian StylePedro R. Costa; Alison Robertson; Michael A. Quilliam. 2015. "Toxin Profile of Gymnodinium catenatum (Dinophyceae) from the Portuguese Coast, as Determined by Liquid Chromatography Tandem Mass Spectrometry." Marine Drugs 13, no. 4: 2046-2062.