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1. WO1999003970 - PROCEDE CHROMATOGRAPHIQUE DE COURTE DUREE, A HAUTE TEMPERATURE POUR LE DOSAGE DES PROTEINES SERIQUES GLYCOSYLEES

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[ EN ]

I claim:
1. In a method of analyzing a sample containing glycated and non-glycated hemoglobin fractions, including the steps of passing the sample through an HPLC column, successively separating non-glycated and glycated fractions, and analyzing at least the glycated fraction to determine the amount of glycated hemoglobin present in the sample, the improvement which comprises the step of maintaining said HPLC column at a temperature of above 50°C and up to about 60°C.

2. The method of claim 1, including the steps of passing mobile phases through said column for separating said non-glycated and glycated fractions, said mobile phases being passed through the column at a flow rate of from about 1-4 ml/min.

3. The method of claim 2, said flow rate being from about 1-3 ml/min.

4. The method of claim 1 , said temperature being from above 50 °C up to about 55 °C.

5. The method of claim 1 , said sample containing from about 3-38 micrograms of blood-derived analyte containing said glycated and non-glycated hemoglobin fractions.

6. In a method of continuously and sequentially analyzing a plurality of individual samples each containing glycated and non-glycated hemoglobin fractions, including the steps of (a) passing a respective sample through an HPLC column, (b) successively separating non-glycated and glycated fractions of the sample, (c) analyzing at least the glycated fraction of the sample to determine the amount of glycated hemoglobin present therein, and (d) repeating steps (a)-(c) for each individual sample, the improvement which comprises the steps of maintaining said HPLC column at a temperature of above 50 °C and up to about 60 °C, and carrying out the analysis of said samples with the total time between successive initial contacts of respective samples with the HPLC column being less than 2 minutes.

7. The method of claim 6, said time being from about 1.25-1.9 minutes.

8. The method of claim 6, said time being from about 1.4-1.75 minutes.

9. The method of claim 6, including the steps of passing mobile phases through said column for separating said non-glycated and glycated fractions, said mobile phases being passed through the column at a flow rate of from about 1-4 ml/min.

10. The method of claim 9, said flow rate being from about 1-3 ml/min.

11. The method of claim 6, said temperature being from above 50°C up to about 55 °C.

12. The method of claim 6, each of said samples containing from about 3-38 micrograms of blood-derived analyte containing said glycated and non-glycated hemoglobin fractions.