Analysis element and method for analyzing glycated hemoglobin content ratio

Claim 1

An analysis method for simultaneously analyzing both amounts of glycated hemoglobin and total hemoglobin in a single aqueous liquid sample so as to calculate glycated hemoglobin content ratio in the sample, the method comprising the steps of: (a) mixing the entire single aqueous liquid sample containing hemoglobin including glycated hemoglobin with an enzyme-labelled antibody against the glycated hemoglobin to allow the antigen-antibody reaction between the glycated hemoglobin and the enzyme-labelled antibody; (b) supplying the reaction mixture after the completion of said antigen-antibody reaction to a substrate layer containing a non-diffusible substrate which forms a diffusible material in the presence of the enzyme of said enzyme-labelled antibody, the activity of said enzyme being effected relative to the steric hindrance due to said antigen-antibody reaction; (c) allowing said diffusible material formed in said substrate layer to migrate to a reagent layer which contains a reagent composition for reacting with the diffusible material to form a dye, the formed dye being detectable colorimetrically in a wavelength range which is not affected by an absorption spectrum of the hemoglobin, the diffusion resistance of said reagent layer being larger than the diffusion resistance of said substrate layer; (d) measuring colorimetrically the amount of the formed dye in the reagent layer to analyze quantitatively the amount of glycated hemoglobin contained in the sample; (e) measuring the amount of the total hemoglobin retained in the substrate layer by the colorimetrical determination at a wavelength range in which the hemoglobin has its inherent absorption, to analyze quantitatively the amount of total hemoglobin in the sample; and (f) calculating the ratio of the glycated hemoglobin amount to the total hemoglobin amount so as to determine the glycated hemoglobin content ratio in the sample.