Improved Method for Analysis of Glycated Haemoglobin by Ion Exchange Chromatography

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Abstract
The Pharmacia Mono S column for glycated haemoglobin (HbA1c) was compared with Mono S HR 5/5 and three other cation exchange columns to optimize analysis of HbA1c. This column allows a significant decrease in separation time, resulting in a sample throughput of 12 samples per hour. The choice of parameters for integration of peak areas in the chromatograms is very important for reproducible results. Since pure calibrators of HbA1c are not available, the relative contribution of HbA1c to the total amount of haemoglobin is calculated. We have compared valley-to-valley integration with baseline integration to assess reproducibility. Valley-to-valley integration significantly enhances the reproducibility of the method and is therefore preferable for routine analyses of HbA1c.