title: High performance liquid chromatography of biological polyamines using immobilized enzyme as post-column reactor followed by electrochemical detection.
abstract: A novel analytical method for biological polyamines (putrescine, spermidine and spermine) was developed. Polyamines were separated by ion-pair reversed phase chromatography using a polymer-based octadecyl bonded column. A polyamine oxidase immobilized column worked effectively as a post-column reactor to convert polyamines to hydrogen peroxide which was eventually detected by electrochemical oxidation on platinum electrode. This method required neither tedious derivatization nor gradient elution, permitting us to perform simple and rapid analysis of polyamines. The detection limits were 0.3, 0.6, and 4 pmol injected for putrescine, spermidine, and spermine, respectively with a linear range of two to three orders of magnitude. Chromatograms obtained with samples from human urine and rat brain homogenates demonstrated the high sensitivity and selectivity of the method.
authors: Department of Industrial Chemistry, Faculty of Engineering, University of Tokyo, Japan.
authors: N
authors: N
authors: Watanabe
authors: M
authors: M
authors: Asano
authors: K
authors: K
authors: Yamamoto
authors: T
authors: T
authors: Nagatsu
authors: T
authors: T
authors: Matsumoto
authors: K
authors: K
authors: Fujita
mesh_terms: D000818:Animals; D001923:Brain Chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004563:Electrochemistry; D004800:Enzymes, Immobilized; D006801:Humans; D008297:Male; D000587:Oxidoreductases Acting on CH-NH Group Donors; D011073:Polyamines / Q000032:analysis*; D051381:Rats; D011919:Rats, Inbred Strains; D014556:Urine / Q000032:analysis; D000097667:Polyamine Oxidase
chemical_list: D004800:Enzymes, Immobilized; D000587:Oxidoreductases Acting on CH-NH Group Donors; D011073:Polyamines; D000097667:Polyamine Oxidase