New fluorimetric method of liquid chromatography for the determination of the neurotoxin domoic acid in seafood and marine phytoplankton.
Domoic acid (DA) is a neurotoxic amino acid that is responsible for the human toxic syndrome, amnesic shellfish poisoning (ASP). A new rapid, sensitive liquid chromatographic (LC) method has been developed for the determination of DA in various marine samples. DA in marine biological materials was derivatised with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) and analysed using isocratic reversed-phase LC with fluorimetric detection. The calibration, based on standard DA solutions, was linear in the range 0.04-2 microg/ml (r2=0.998) and the detection limit (3:1, signal/noise) was better than 1 ng/ml. Using the certified reference material (MUS-1B), recoveries of DA from shellfish tissue were >95% (n=5). When a strong anion exchange SPE cartridge was used for sample clean-up the detection limit was 6 ng DA/g mussel tissue. Good reproducibility was achieved with RSD values ranging from 3% for 8 microg DA/g (n=5), to 5% for 0.04 microg DA/g (n=5). This new method was successfully applied to the determination of DA in naturally contaminated shellfish and in marine phytoplankton cultures of Pseudonitzschia sp.
James
K J
KJ
Chemistry Department, Cork Institute of Technology, Ireland. kjames@cit.ie
Gillman
M
M
Lehane
M
M
Gago-Martinez
A
A
D002853:Chromatography, Liquid / Q000379:methods*; D005470:Fluorometry / Q000379:methods*; D007608:Kainic Acid / Q000031:analogs & derivatives* / Q000032:analysis; D009498:Neurotoxins / Q000032:analysis*; D010839:Phytoplankton / Q000737:chemistry*; D017747:Seafood / Q000032:analysis*; D012680:Sensitivity and Specificity
D009498:Neurotoxins; C012301:domoic acid; D007608:Kainic Acid