Premium
Amperometric determination of amino group on a solid support using glutaraldehyde
Author(s) -
Ukeda Hiroyuki,
Nakzono Yukika,
Matumoto Kiyoshi,
Osajima Yutaka
Publication year - 1991
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260380816
Subject(s) - glutaraldehyde , amperometry , chemistry , chromatography , group (periodic table) , electrode , organic chemistry , electrochemistry
A simple amperometric method has been developed for the determination of amino groups on a solid support. The method is based on oxygen consumption during the reaction between the bifunctional reagent gluaraldehyde and the amino groups on the solid support. Oxygen consumption was monitored with a Clark oxygen electrode. AHSepharose 4B, Affi‐Gel 102, and Amino‐Celluofine were used as amino‐terminal solid supports. The concentration of amino groups on each support was calculated from a calibration curve of the standard compound. When hexamethylenediamine was used as the standard, the concentration of amino groups on AH‐Sepharose 4B and Affi‐Gel 102 by the present method showed good agreement with that by the conventional method. In the case of Amino‐Cellulofine, ethanolamine had to be selected as the standard. The relative standard deviation for six successive determinations of AH‐Sepharose 4B was 3.2%; the polymerization of glutaraldehyde showed no effect on the result. The spectral property of AH‐Sepharose 4B treated with glutataraldehyde showed no effect on the result. The spectral property of AH‐Sepharose 4B treated with lutaraldehyde suggested that the reaction mechanism of the present method was based on the oxygen consumption during the formation of the pyridinium salt on the support. A comparative study of the present method with a colorimetric method with Traut's regent is also included.