TITLE

Improved high-performance liquid chromatographic method for the determination of domoic acid and analogues in shellfish: effect of pH

AUTHOR(S)
López-Rivera, A.; Suárez-Isla, B. A.; Eilers, P. P.; Beaudry, C. G.; Hall, S.; Fernández Amandi, M.; Furey, A.; James, K. J.
PUB. DATE
April 2005
SOURCE
Analytical & Bioanalytical Chemistry;Apr2005, Vol. 381 Issue 8, p1540
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
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 µg/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 µg/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 µg/g. Using the same chromatographic conditions, LC-MS3 was used to determine DA and its isomers, isodomoic acid D andepi-domoic acid, in scallop tissues.
ACCESSION #
16865356

 

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