Development of a chromatographic method for the isolation and detection of hygromycin B in biological fluids.
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.
Campbell
N F
NF
Eastern Regional Research Center, Agricultural Research Service, US Department of Agriculture, Wyndmoor, PA 19038, USA.
Hubbard
L E
LE
Mazenko
R S
RS
Medina
M B
MB
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
D000900:Anti-Bacterial Agents; D006921:Hygromycin B