Abstract
A flow-injection assay for inhibition of diamine oxidase (DAO) is described. Histamine is decomposed fish is detected and used as the enzyme substrate. Sample extracts are doubly injected; one of the two resulting sample zones is passed through a reactor containing DAO immobilized to Sepharose. Histamine is detected sequentially in both the pristine and enzymatically treated zones. The detection chemistry (kinetics-optimized condensation with phthalic dicarboxaldehyde) allows the selective detection of histamine. Subsequent calibration with histamine standard and comparison of the two peak heights allows generic detection of compounds inhibiting the DAO-mediated conversion of histamine. Preliminary data show that the system can detect inhibitors in extracts of decomposed Mahi-Mahi (associated with a Scombroid poisoning outbreak) and that far lower levels of inhibition are observed in fresh fish. Several known inhibitors and competitive substrates of DAO are also tested.
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