HPLC determination of salinomycin and related compounds in fermentation media of Streptomyces albus and premixes.
A rapid, sensitive and selective HPLC method with post-column derivatization was proposed for the determination of salinomycin and related products in fermentation broths and premixes. The solvent extracts of samples were analysed on a reversed-phase monolithic type column. The mobile phase consisted of methanol/zinc acetate (0.05 M) adjusted to pH 4.0 with acetic acid (85/15, v/v). Post-column derivatization with vanillin at 85 degrees C was used for simultaneous, selective detection of salinomycin at 520 nm and related products at 460 nm. Optimal ratio of mobile phase/reagent flow rate was 2:1. Alternatively, pre-column derivatization of salinomycin and related products with three different reagents (2,4-dinitrophenylhydrazine, p-bromophenacyl bromide and p-nitrobenzoyl chloride) was examined. Suitable derivates for HPLC separation and UV detection were prepared using p-nitrobenzoyl chloride. Extraction ability of various solvents for extracting of salinomycin and co-products from premix samples was also tested. Acetone, ethanol and pyridine were found to be the best extraction solvents for these compounds.
QC Department, Biotika a.s., SK-976 13 Slovenská Lupca, Slovakia. milos.blazsek@zentiva.sk
Milos
M
Blazsek
Marián
M
Kubis
D019342:Acetic Acid / Q000737:chemistry; D000096:Acetone / Q000032:analysis; D001547:Benzaldehydes / Q000737:chemistry; D002626:Chemistry, Pharmaceutical / Q000379:methods; D002845:Chromatography; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004345:Drug Industry / Q000379:methods; D000431:Ethanol / Q000032:analysis; D005285:Fermentation; D006863:Hydrogen-Ion Concentration; D000432:Methanol / Q000737:chemistry; D008956:Models, Chemical; D008958:Models, Molecular; D011714:Pyrans / Q000032:analysis* / Q000737:chemistry; D011725:Pyridines / Q000032:analysis; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D013302:Streptomyces / Q000378:metabolism*; D013696:Temperature; D014466:Ultraviolet Rays; D019345:Zinc Acetate / Q000737:chemistry; D000095662:Polyether Polyketides
D019342:Acetic Acid; D000096:Acetone; D001547:Benzaldehydes; D000431:Ethanol; D000432:Methanol; D011714:Pyrans; D011725:Pyridines; D019345:Zinc Acetate; C010327:salinomycin; D000095662:Polyether Polyketides; C100058:vanillin; C023666:pyridine