KING, C.; ANGELES, C.; NDUKA, E.; CATAPANE, E.J.; Kingsborough Community College; Kingsborough Community College; Medgar Evers College; Medgar Evers College: Identification and Distribution of Biogenic Amines in Tissues of Crassostrea virginica

Biogenic amines are neurotransmitters and hormones in animals. They are well studied in the bivalve mollusc Mytilus edulis, but not as well in oysters, particularly Crassostrea virginica. We studied their presence in C. virginica using an isocratic, ion-pairing HPLC analysis with fluorescence detection to resolved norepinephrine, epinephrine, dopamine, 3,4-dihydroxyphenylacetic acid, 3,4-dihydroxyphenylalanine, tryptophan, tyrosine, serotonin, 5-hydroxytryptophan, N-acetyl serotonin, tyramine and octopamine in a twenty minute run. It lacked sufficient sensitivity to identify amines in small tissue samples. We explored pre- and post-column derivitization methods with benzylamine and terbidium chloride to increase detection limits and purchased a more modern spectrofluorometer. The newer instrument was more sensitive allowing detection of lower levels of amines using native fluorescence. Tissues were dissected, weighed, homogenized, centrifuged, filtered and injected into the HPLC system fitted with a BDS Hypersil C18 column and a Hitachi F-1050 Spectrofluorometer with a 12 µL flow cell. The mobile phase was 50 mM acetate buffer (pH 4.7) with 1 mM EDTA and 1.1 mM SOS, and methanol (85%/15%, v/v). We identified and quantified norepinephrine, epinephrine, dopamine and serotonin in mantle, gill, heart, palps, posterior adductor muscle and visceral ganglia in low amounts (100s of ng/g) which correlates well with published reports of related bivalves. We believe the study will be an important step in elucidating neurobiological and neuroendocrine functions in C. virginica and of these biogenic amines in general. This work was supported by grants 1R25GM62003 of NIGMS and the Groundworks Program of CUNY.We thank Frank M. Flower and Sons, Inc., Oyster Bay, NY for supplying oysters.