HPLC as well as immunoturbidimetric HbA1c assays may be affected by hemoglobinopathies which are frequent in certain patient groups. The extraordinary specificity of DiaSys' HbA1c net FS ensures reliable results for patients, which makes it the new standard in HbA1c determination.
The HbA1c net FS test is based on a colorimetric, enzymatic method. The concentrations of HbA1c and total hemoglobin are determined separately. Calculation of the HbA1c ratio from total hemoglobin is done by the instrument automatically.
Pretreatment and hemoglobin measurement
Whole blood samples are lysed with hemolyzing solution. Hemoglobin is released from erythrocytes. The absorbance of hemoglobin is measured at 570 nm after addition of reagent 1 (R1) and is proportional to the total hemoglobin concentration in the sample.
After addition of reagent 2 (R2), fructosylated dipeptides from the N-terminal hemoglobin β-chain are released by a protease. Hydrogen peroxide (H2O2) is produced after oxidative cleavage of fructosylated dipeptides by FPOX (fructosyl peptide oxidase). H2O2 is determined colorimetrically by reaction with a chromogen in presence of peroxidase at 660 nm. The absorbance increase is proportional to the HbA1c concentration.