Radio-detection high-performance liquid chromatographic enzyme assay for inhibitors of fungal sterol delta 14-reductase.
An enzyme assay for inhibitors of fungal sterol delta 14-reductase employing isocratic reversed-phase high-performance liquid chromatography is described. A Hypersil 5-microns octadecylsilyl (ODS) column (250 mm x 4.6 mm I.D.) was used and a mobile phase consisting of methanol-water-ethanol (86:4:10, v/v) was pumped at a flow-rate of 1.5 ml/min. Typical analysis times were 15 min. Using [4-14C]ignosterol as a substrate and an enzyme preparation from Saccharomyces cerevisiae, this method was used to compare the inhibition of sterol delta 14-reductase by the fungicides fenpropidin and fenpropimorph with three N-substituted 8-azadecaline compounds.
Department of Agrobiological Research, Dielsdorf, Switzerland.
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D002250:Carbon Radioisotopes; D002851:Chromatography, High Pressure Liquid; D007700:Kinetics; D009025:Morpholines / Q000494:pharmacology*; D010088:Oxidoreductases / Q000037:antagonists & inhibitors*; D010880:Piperidines / Q000494:pharmacology*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*
D002250:Carbon Radioisotopes; D009025:Morpholines; D010880:Piperidines; C050435:fenpropimorph; C063198:fenpropidine; D010088:Oxidoreductases; C022672:delta(14)-sterol reductase