title: Development of a chromatographic method for the isolation and detection of hygromycin B in biological fluids.
abstract: An affinity chromatography method was developed for the purification of hygromycin B from biological fluids. Lysozyme and alpha-lactalbumin were immobilized on an N-hydroxysuccinimide activated agarose support. Hygromycin B solubilized in water was bound by the proteins and subsequently eluted using 10 mM sodium citrate buffer, pH 4.0. Hygromycin B was purified from swine plasma, bovine serum and bovine milk samples using a combination of ion-exchange chromatography for initial clean-up of spiked biological samples followed by affinity chromatography. Thin layer chromatographic analysis of the isolated hygromycin B revealed one band with the same R(F) value as the hygromycin B standard.
authors.affiliation: Eastern Regional Research Center, Agricultural Research Service, US Department of Agriculture, Wyndmoor, PA 19038, USA.
authors.forename: N F
authors.initials: NF
authors.lastname: Campbell
authors.forename: L E
authors.initials: LE
authors.lastname: Hubbard
authors.forename: R S
authors.initials: RS
authors.lastname: Mazenko
authors.forename: M B
authors.initials: MB
authors.lastname: Medina
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000097:blood / Q000302:isolation & purification*; D001826:Body Fluids / Q000737:chemistry*; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D002855:Chromatography, Thin Layer; D006921:Hygromycin B / Q000097:blood / Q000302:isolation & purification*; D008892:Milk / Q000737:chemistry*; D013552:Swine
chemical_list: D000900:Anti-Bacterial Agents; D006921:Hygromycin B