Purification of fusaproliferin from cultures of Fusarium subglutinans by preparative high-performance liquid chromatography.
Thirty-six Fusarium strains were grown on cracked yellow corn and evaluated for optimum fusaproliferin production, with Fusarium subglutinans E-1583 producing the highest levels (1600 microg/g). Three solvent systems were tested for extracting fusaproliferin from the cultures of F. subglutinans E-1583. Methanol gave the highest fusaproliferin recovery, followed by methanol/1% aqueous NaCl (55:45, v/v) and acetonitrile/methanol/H(2)O (16:3:1, v/v/v). Hexane partitioning was effective in removing many impurities from the crude fusaproliferin extracts prior to the liquid chromatography step. Fusaproliferin samples were further purified by high-performance liquid chromatography (HPLC) with a C18 preparatory column using a mobile phase of acetonitrile/H(2)O (80:20, v/v). The purity of the fusaproliferin was verified by analytical HPLC, GC/MS, (1)H NMR spectroscopy, and electrospray ionization (ESI) MS. The isolated fusaproliferin was shown to be free of impurities and can be used as a standard for routine analysis. Fusaproliferin was shown to be temperature-sensitive when samples were stored at room temperature (20-24 degrees C) for more than several days. After 30 days at 4 degrees C, approximately 8% of the fusaproliferin had been transformed to deacetyl-fusaproliferin; however, samples stored at -20 degrees C for 1 year contained only trace amounts of the deacetylated form.
Wu
Xiaorong
X
Food Science Institute and Department of Plant Pathology, Kansas State University, Manhattan, KS 66506, USA.
Leslie
John F
JF
Thakur
Rohan A
RA
Smith
J Scott
JS
D000097:Acetonitriles; D002851:Chromatography, High Pressure Liquid; D004355:Drug Stability; D005670:Fusarium / Q000254:growth & development / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry; D009682:Magnetic Resonance Spectroscopy; D000432:Methanol; D012965:Sodium Chloride; D012997:Solvents; D021241:Spectrometry, Mass, Electrospray Ionization; D013696:Temperature; D013729:Terpenes / Q000302:isolation & purification*; D014867:Water
D000097:Acetonitriles; D012997:Solvents; D013729:Terpenes; C093577:fusaproliferin; D014867:Water; D012965:Sodium Chloride; D000432:Methanol; C032159:acetonitrile