Analytical Sciences
Online ISSN : 1348-2246
Print ISSN : 0910-6340
ISSN-L : 0910-6340
Determination of 2, 3-Diphosphoglycerate in Red Blood Cells by High Performance Liquid Chromatography
Yukihiro TAKEBAYASHIRumiko MITSUMAToshio IMANARI
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1987 Volume 3 Issue 6 Pages 569-572

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Abstract

A method for the determination of 2, 3-diphosphoglycerate(2, 3-DPG) was established by high performance liquid chromatography with spectrophotometric detection using the ferrous ion as a postcolumn labeling reagent. Chromatographic conditions were as follows: Column, Asahipak BEST 502N (5mm i.d.×100mm); eluent, 0.3M acetate buffer (pH 5.0) containig 15mM Mg(NO3)2 at a flow rate of 1ml/min; complex forming reagent, 20 mM FeSO4(NH4)2SO4 containing 1.5mM dithiothreitol and 5% sodium dodecylsulfate (0.5ml/min); detection wavelength, 260nm. The calibration curve for 2, 3-DPG was linear in the range of 1.0 to 20.0nmol, and the relative standard deviations were 2.05 and 0.79% (n=5) at 2.7 and 16.0nmol, respectively. The correlation of the present method to the conventional enzymatic method was satisfactory (r=0.999).

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