Andrew D Turner1, Monika Dhanji-Rapkova1, Sum Y T Fong1, James Hungerford2, Paul S McNabb3, Michael J Boundy4, D Tim Harwood4. 1. Centre for Environment, Fisheries and Aquaculture Science, Barrack Rd, The Nothe, Weymouth, Dorset DT4 8UB, United Kingdom. 2. U.S. Food and Drug Administration, 22201 23rd Dr, S.E., Bothell, WA 98021. 3. Private Consultancy, 7 Elliott St, Nelson 7010, New Zealand. 4. Cawthron Institute, 98 Halifax St East, Nelson 7010, Private Bag 2, Nelson 7042, New Zealand.
Abstract
BACKGROUND: An ultrahigh-performance LC (UHPLC)-tandem MS (MS/MS) method for determination of paralytic shellfish poisoning toxins and tetrodotoxin (TTX) in bivalve molluscs was developed. To be used for regulatory testing, it needed to be validated through collaborative study. OBJECTIVE: The aim was to conduct a collaborative study with 21 laboratories, using results to assess method performance. METHODS: Study materials incorporated shellfish species mussels, oysters, cockles, scallops, and clams and were assessed to demonstrate stability and homogeneity. Mean concentrations determined by participants for blind duplicate samples were used to assess reproducibility, repeatability, and trueness. RESULTS: Method performance characteristics were excellent following statistical assessment of participant data, with method trueness showing excellent method accuracy against expected values. No significant difference was found in the trueness results determined by different chromatographic column types. Acceptability of the between-laboratory reproducibility for individual analytes was evidenced by >99% of valid Horwitz ratio values being less than the 2.0 limit of acceptability. With excellent linearity and sensitivity fit-for-purpose over a range of mass spectrometer instruments, the UHPLC-MS/MS method compared well against other detection methods. It includes additional paralytic shellfish toxin (PST) analogues as well as TTX, which, to date, have not been incorporated into any other hydrophilic marine toxin official method of analysis. CONCLUSIONS: The results from this study demonstrate that the method is suitable for the analysis of PST analogues and TTX in shellfish tissues and is recommended as an official alternative method of analysis for regulatory control. HIGHLIGHTS: A new mass spectrometric method for PST and TTX has been validated successfully through collaborative study.
BACKGROUND: An ultrahigh-performance LC (UHPLC)-tandem MS (MS/MS) method for determination of paralytic shellfish poisoning toxins and tetrodotoxin (TTX) in bivalve molluscs was developed. To be used for regulatory testing, it needed to be validated through collaborative study. OBJECTIVE: The aim was to conduct a collaborative study with 21 laboratories, using results to assess method performance. METHODS: Study materials incorporated shellfish species mussels, oysters, cockles, scallops, and clams and were assessed to demonstrate stability and homogeneity. Mean concentrations determined by participants for blind duplicate samples were used to assess reproducibility, repeatability, and trueness. RESULTS: Method performance characteristics were excellent following statistical assessment of participant data, with method trueness showing excellent method accuracy against expected values. No significant difference was found in the trueness results determined by different chromatographic column types. Acceptability of the between-laboratory reproducibility for individual analytes was evidenced by >99% of valid Horwitz ratio values being less than the 2.0 limit of acceptability. With excellent linearity and sensitivity fit-for-purpose over a range of mass spectrometer instruments, the UHPLC-MS/MS method compared well against other detection methods. It includes additional paralytic shellfish toxin (PST) analogues as well as TTX, which, to date, have not been incorporated into any other hydrophilic marine toxin official method of analysis. CONCLUSIONS: The results from this study demonstrate that the method is suitable for the analysis of PST analogues and TTX in shellfish tissues and is recommended as an official alternative method of analysis for regulatory control. HIGHLIGHTS: A new mass spectrometric method for PST and TTX has been validated successfully through collaborative study.
Authors: Monika Dhanji-Rapkova; Andrew D Turner; Craig Baker-Austin; Jim F Huggett; Jennifer M Ritchie Journal: Mar Drugs Date: 2021-02-02 Impact factor: 5.118
Authors: Karl J Dean; Robert G Hatfield; Vanessa Lee; Ryan P Alexander; Adam M Lewis; Benjamin H Maskrey; Mickael Teixeira Alves; Benjamin Hatton; Lewis N Coates; Elisa Capuzzo; Jim R Ellis; Andrew D Turner Journal: Mar Drugs Date: 2020-07-29 Impact factor: 5.118
Authors: Karl J Dean; Ryan P Alexander; Robert G Hatfield; Adam M Lewis; Lewis N Coates; Tom Collin; Mickael Teixeira Alves; Vanessa Lee; Caroline Daumich; Ruth Hicks; Peter White; Krista M Thomas; Jim R Ellis; Andrew D Turner Journal: Mar Drugs Date: 2021-12-07 Impact factor: 5.118