Single-Laboratory Validation of a Multitoxin Ultra-Performance LC-Hydrophilic Interaction LC-MS/MS Method for Quantitation of Paralytic Shellfish Toxins in Bivalve Shellfish

Abstract

A single-laboratory validation study was conducted for the hydrophilic interaction-LC-MS/MS analysis of paralytic shellfish toxins (PSTs) in bivalve shellfish. The method was developed as an alternative to the precolumn oxidation AOAC 2005.06 and postcolumn oxidation AOAC 2011.02 LC with fluorescence detection methods, receptor binding assay AOAC 2011.27, as well as the mouse bioassay AOAC 959.08. PSTs assessed were saxitoxin, neosaxitoxin, deoxydecarbamoylsaxitoxin, decarbamoylsaxitoxin, decarbamoylneosaxitoxin, gonyautoxins 1-6, decarbamoylgonyautoxins 2-3, and N-sulfocarbamoyl gonyautoxins 2&3. The method also included the determination of decarbamoylgonyautoxins 1&4, N-sulfocarbamoyl gonyautoxins 1&4, and M toxins. Twelve commercially produced bivalve species from both New Zealand and the United Kingdom were assessed, including mussels, oysters, scallops, and clams. Validation studies demonstrated acceptable method performance characteristics for specificity, linearity, recovery, repeatability, and within-laboratory reproducibility. LOD and LOQ were significantly improved in comparison to current fluorescence-based detection methods, and the method was shown to be rugged. The method performed well in comparison to AOAC 2005.06, with evidence obtained from both comparative analysis of 1141 PST-contaminated samples and successful participation in proficiency testing schemes. The method is suitable for use in regulatory testing and will be submitted for an AOAC collaborative study.

Recent Trends in Marine Phycotoxins from Australian Coastal Waters

“…These have been the subject of several recent reviews [108,109]. In addition, new methods for the detection of these toxin groups have been developed to utilise LC-MS/MS equipment that is currently in use in shellfish monitoring programs [37,110]. …”

Biological toxins of potential bioterrorism risk: Current status of detection and identification technology

“…The method was subjected to an extensive intra-laboratory validation based on harmonized IUPAC validation guidelines with various matrices and PSP analogues. The results were in accordance with official reference methods and the sensitivity was better than that obtained by the official PSP toxin methods [78]. Another HILIC LC-MS/MS method for the analysis of PSP toxins was recently tested for several different food matrices from a forensic perspective [79].…”

“…Oxidation of PSP toxins into fluorescent iminopurine derivatives followed by LC separation and FLD detection [66] Post-column oxidation HPLC-FLD Official PSP toxin method for shellfish: LC separation and post-column oxidation of PSP toxins into fluorescent iminopurine derivatives followed by FLD detection [67] Spectrometric assay HILIC LC-MS/MS: Positive ESI with SRM detection of PSP toxins in shellfish [70] Positive or negative ESI with MRM transitions of PSP toxins in shellfish [77,78] Positive ESI and product ion scan of STX in algal and mussel matrices [72] * To the best of our knowledge, the thoroughly characterized [30]. The materials of highest relevance for humans (BoNT/A, B and E) were spiked in different concentrations (0.5-1000 ng/mL) into buffer, serum, milk, and meat extract.…”

Differential Mobility Spectrometry for Improved Selectivity in Hydrophilic Interaction Liquid Chromatography-Tandem Mass Spectrometry Analysis of Paralytic Shellfish Toxins

“…Method 1 uses a 1.8 μm particle size BEH amide column, a rapid 10 min gradient, and has been the validated with an activated carbon SPE cleanup for routine food safety testing applications [8,36]. Method 2 uses a 5 μm particle size amide 80 column and has been widely used for a range of research purposes since 2005 [6].…”