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High performance liquid chromatography of biological polyamines using immobilized enzyme as post‐column reactor followed by electrochemical detection
Author(s) -
Watanabe Noriyuki,
Asano Masao,
Yamamoto Katsunobu,
Nagatsu Toshiharu,
Matsumoto Takatoshi,
Fujita Keisuke
Publication year - 1989
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1130030502
Subject(s) - chemistry , chromatography , putrescine , spermidine , spermine , derivatization , polyamine , polyamine oxidase , hydrogen peroxide , amperometry , detection limit , elution , high performance liquid chromatography , cadaverine , electrochemistry , enzyme , electrode , biochemistry
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.