Effect of hemoglobin variants on the derermination of glycohemoglobin by two high performance liquid chromatography-based methods

Objective To compare the results of HbAlc in blood samples with various configuration of hemoglobin(Hb) by affinity chromatogragh-high performance liquid chromatography(AC-HPLC)system and ion exchange-high performance liquid chromatography(IE-HPLC)system,and evaluate the effect of glycohemoglobin variants on AC-HPLC and IE-HPLC.MethodsThe levels of HbA1c in 30 blood samples with the normal Hb configuration and the HbA1c levels were 5%-11%,the blood samples with different HbF levels and the blood samples of patients with α-Mediterranean anemia(α-MA)and β-Mediterranean anemia(β-MA) were determined by AC-HPLC and IE-HPLC,respectively.ResultsThere was not difference between the AC-HPLC and IE-HPLC for the blood sanples with normal Hb configuration and the HbA1c levels were 5%-11%(P0.05).There was not interference with HbF for HbA1c determation by AC-HPLC.However,the bias of HbA1c determination by IE-HPLC was occurred when HbF level 8.8%,and the HbA1c was undetectable when HbF level was 70%.For blood samples with patients with α-MA and mitis β-MA,the results of HbA1c deteremind by IE-HPLC were significantly higher than AC-HPLC(P0.01 and P0.05,respectively).The level of HbA1c in patient with gravis type β-MA was undetected by IE-HPLC.ConclusionsThe results of HbA1c in blood samples containing Hb variants or HbF may be interfered when analyzed by IE-HPL.However,there was not interferences with AC-HPLC.