title: Improved high-performance liquid chromatographic method for the determination of domoic acid and analogues in shellfish: effect of pH.
abstract: Domoic acid (DA) is a naturally-occurring amino acid that causes a form of human intoxication called amnesic shellfish poisoning (ASP) following the consumption of shellfish. A rapid and sensitive HPLC-UV method has been developed for analysis of DA and analogues in shellfish without the need for SPE clean-up. Isocratic chromatographic separation of DA and its isomers from shellfish matrix interferences and from the prevalent amino acid, tryptophan, was achieved by careful control of the mobile phase pH. The optimised pH was found to be 2.5 when using a Luna(2) C18 column. Sample extraction was verified with control extracts from shellfish spiked at 5.0 and 10.0 microg/g of DA and with certified reference material. The average extraction efficiency was 98.5%. The calibration, based on mussel tissue spiked with DA standard, was linear in the range 0.05-5.0 microg/ml (r = 0.9999) and the detection limit (signal:noise 3:1) was better than 25 ng/ml. The DA assay achieved good precision; %RSD = 1.63 (intra-day, n = 6) and %RSD = 3.7 (inter-day, n = 8). This method was successfully applied to a variety of shellfish species, allowing the rapid screening of a large number of samples per day (20-30), without the need for SPE clean-up. Quantitative data were obtained for shellfish samples containing domoic acid in the concentration range 0.25-330 microg/g. Using the same chromatographic conditions, LC-MS3 was used to determine DA and its isomers, isodomoic acid D and epi-domoic acid, in scallop tissues.
authors.affiliation: Laboratory of Marine Toxins, Faculty of Medicine, Institute of Biomedical Sciences, University of Chile, Independencia 1027, Santiago, Chile. amlopez@med.uchile.cl
authors.forename: A
authors.initials: A
authors.lastname: López-Rivera
authors.forename: B A
authors.initials: BA
authors.lastname: Suárez-Isla
authors.forename: P P
authors.initials: PP
authors.lastname: Eilers
authors.forename: C G
authors.initials: CG
authors.lastname: Beaudry
authors.forename: S
authors.initials: S
authors.lastname: Hall
authors.forename: M
authors.initials: M
authors.lastname: Fernández Amandi
authors.forename: A
authors.initials: A
authors.lastname: Furey
authors.forename: K J
authors.initials: KJ
authors.lastname: James
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D007608:Kainic Acid / Q000031:analogs & derivatives* / Q000032:analysis*; D008387:Marine Toxins / Q000032:analysis*; D015394:Molecular Structure; D012758:Shellfish / Q000032:analysis*; D057096:Shellfish Poisoning; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods
chemical_list: D008387:Marine Toxins; C012301:domoic acid; D007608:Kainic Acid