Identification of aromatic moieties and mycosamine in antifungal heptaenes with high-performance liquid chromatography, high-performance liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry.
A high-performance liquid chromatographic (HPLC) method for the determination of the aromaticity of heptaene polyene antibiotics has been developed. The released aromatic moiety of the heptaene polyenes aureofungin, candicidin, candimycin, hamycin and trichomycin was assayed after alkaline hydrolysis. The presence of p-aminoacetophenone (PAAP) and N-methyl-p-aminoacetophenone (N-methyl-PAAP) in the hydrolysates was determined by HPLC, HPLC-mass spectrometry (HPLC-MS) and gas chromatography-MS (GC-MS). Candicidin and hamycin contained only the PAAP residue; aureofungin contained both PAAP and N-methyl-PAAP. Trichomycin contained PAAP and also some unknown component of molecular weight 179. The aromatic nature of the individual components of the heptaene complex was demonstrated using radioactivity flow detection for the determination of the incorporation of [14C]-p-aminobenzoic acid to individual candicidin components. Ammonia chemical ionization MS was successfully used for the GC-MS identification of the acetylated mycosamine moiety of heptaenes.
Raatikainen
O
O
Department of Pharmaceutical Chemistry, University of Kuopio, Finland.
Auriola
S
S
Tuomisto
J
J
D010129:4-Aminobenzoic Acid / Q000378:metabolism; D000098:Acetophenones / Q000737:chemistry*; D000935:Antifungal Agents / Q000737:chemistry*; D002174:Candicidin / Q000096:biosynthesis; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D008401:Gas Chromatography-Mass Spectrometry; D006595:Hexosamines / Q000737:chemistry*; D013058:Mass Spectrometry; D011090:Polyenes / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet; D013305:Streptomyces griseus / Q000378:metabolism
D000098:Acetophenones; D000935:Antifungal Agents; D006595:Hexosamines; D011090:Polyenes; D002174:Candicidin; C014692:mycosamine; C032065:4-aminoacetophenone; D010129:4-Aminobenzoic Acid