Improved colorimetric assay for glycosylated hemoglobin

Area of Science:

• Clinical Chemistry
• Biochemical Assays

Background:

• Glycosylated hemoglobin (GHb) is a key biomarker for long-term glycemic control in diabetes.
• Accurate and efficient measurement of GHb is crucial for diabetes management.

Purpose of the Study:

• To develop and validate a novel colorimetric assay for quantifying glycosylated hemoglobin.
• To assess the assay's performance characteristics, including speed, precision, and correlation with a standard method.

Main Methods:

• A colorimetric method involving the conversion of glycosylated hemoglobin to 5-hydroxymethylfurfural using oxalic acid at 124°C.
• Quantification of the 5-hydroxymethylfurfural adduct with 2-thiobarbituric acid via photometry.
• Assay validation using clinical specimens and comparison with liquid chromatography for hemoglobin A1c.

Main Results:

• The assay provides results within 120 minutes using standard clinical laboratory equipment.
• Excellent precision was observed with within-assay and between-assay coefficients of variation <2% and <3%, respectively.
• High correlation (r = 0.98) was achieved when comparing results with liquid chromatography-based hemoglobin A1c analysis.

Conclusions:

• The developed colorimetric assay is a rapid, reliable, and accurate method for measuring glycosylated hemoglobin.
• This assay is suitable for routine clinical laboratory use, aiding in effective diabetes monitoring.