A stability-indicating assay for the antibiotic fumagillin in technical materials has been developed. Measurement is by reversed-phase HPLC with detection at 351 nm. The assay is rapid, specific, and precise within the fumagillin concentration range of 0.000-0.035 mg/mL. It was also found that protection of fumagillin from room fluorescent lights is required to prevent photodegradation.Fumagillin is an antibiotic produced by the fermentation of certain strains of Aspergillus fumigatus. Fumagillin, first isolated in 1951 (Eble and Hanson, 1951), has been demonstrated to be effective against Nosema apis, a universally occurring protozoan parasite found in the gut of the adult honey been (Girardeau, 1972). Additionally, it has been found to control diseases caused by microorganisms in fish (Takeda Chemical Industries, 1983).Fumagillin has been assayed by thin-layer chromatography (Issaq et al., 1977), spectrophotometry (Garrett and Eble, 1954), and a microbial assay (Girolami, 1963). Each of these techniques has disadvantages limiting their utility in the analysis of fumagillin. Thin-layer chromatography yields only a limited amount of quantitative information and is not ideally suited for the assay of fumagillin powders. The spectrophotometric assay is subject to error due to interferences from UV-absorbing impurities, especially in complex samples (Girolami, 1963). Finally, the microbial assay requires approximately 16-18 h to complete an analysis.Described in this paper is an HPLC assay for fumagillin that is rapid and precise and separates fumagillin from possible impurities and degradation products.
EXPERIMENTAL SECTIONGlassware. Nonactinic glassware was used whenever possible. Standard glassware was wrapped in aluminum foil when its use was necessary.Reagents. The acetonitrile used in this study was HPLC grade. All other reagents were analytical grade.Fumagillin Standard and Samples. The fumagillin standard used was an Abbott Laboratories house reference standard. The standard had been previously characterized and assigned a purity of 98.5% (w/w). All samples of fumagillin powder were actual production lots supplied by Abbott Laboratories.Fumagillin Standard Solution Preparation. The fumagillin stock standard solution was prepared by accurately weighing 50 mg of fumagillin standard into a 100-mL volumetric flask. The fumagillin standard was dissolved in and diluted to volume with acetonitrile. The working standard solution was prepared by diluting the stock solution 1:25 with acetonitrile. Fumagillin Sample Preparation. About 100 mg of the fumagillin sample was accurately weighed into a 100-mL volumetric flask and dissolved in and diluted to volume with acetonitrile. This solution was diluted 1:50 with acetonitrile for the assay preparation.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.