Detection and quantification of melatonin in a dinoflagellate, Gonyaulax polyedra: solutions to the problem of methoxyindole destruction in non-vertebrate material.
Preservative procedures are described for the extraction and quantification of melatonin in cell material from the dinoflagellate Gonyaulax polyedra, an organism in which this indoleamine is rapidly degraded due to interaction with free oxygen radicals and photooxidation. Cells were shock-frozen in liquid nitrogen and pulverized. Various extraction methods were applied to the powder, and rates of recovery were compared. For the determination of melatonin by high performance liquid chromatography (HPLC), extractions with acetone or perchloric acid were more suitable than the use of other solvents. For purposes of radioimmunoassay (RIA), extraction with acetone gave the best results. Several other inorganic solvents, which are often applied in melatonin research, such as chloroform, dichloromethane, and diethyl ether, led to considerable losses of the indoleamine. The procedures developed for HPLC with either electrochemical or fluorescence detection also allow the quantification of other indolic compounds, in particular, tryptophan and 5-methoxytryptamine. The methods described may be of value in the further search for melatonin and related indoleamines in non-vertebrate material, especially, from unicells, multicellular plants, and invertebrates.
Poeggeler
B
B
Department of Cellular and Structural Biology, University of Texas Health Science Center, San Antonio 78284-7762.
Hardeland
R
R
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D015925:Cryopreservation; D004141:Dinoflagellida / Q000737:chemistry*; D007211:Indoles / Q000737:chemistry*; D008550:Melatonin / Q000032:analysis* / Q000302:isolation & purification; D009584:Nitrogen; D011863:Radioimmunoassay; D017382:Reactive Oxygen Species; D014021:Tissue Preservation / Q000379:methods
D007211:Indoles; D017382:Reactive Oxygen Species; D008550:Melatonin; D009584:Nitrogen