In this novel procedure for determining ethylene glycol in plasma by liquid chromatography, benzoyl esters of ethylene glycol and of benzyl alcohol (used as the internal standard) are prepared directly in plasma. The benzoyl esters, highly ultraviolet-absorbing chromogens, are ideal compounds for analysis by reversed-phase liquid chromatography with methanol/water as the mobile phase. The benzoyl derivative of ethylene glycol is well separated from the derivative of the internal standard and from plasma constituents. The standard curve is linear to 400 mg of ethylene glycol per liter. As little as 10 mg of ethylene glycol per liter of plasma can be measured. Other commonly ingested alcohols do not interfere.
Introduction: Hb Hope is a clinically asymptomatic β-chain variant [beta136 (H14) Gly→Asp (GGT→GAT)]. It is more prevalent in Mediterranean region of the world than in Asian countries and extremely rare in India. Aim:To evaluate the interference of clinically silent and extremely rare variant; haemoglobin Hope (Hb Hope) on the final reported value of glycated haemoglobin (HbA1c) by high performance liquid chromatography (HPLC). Materials and Methods:Eight EDTA blood samples showed very high values of HbA1c (>50 gm%) when processed by Bio-Rad Variant II turbo (HPLC). These were further processed for by capillary electrophoresis to look for the abnormal haemoglobin. Glycated Hb was also measured by immunoturbidimetry.Results: On HPLC, single abnormally high peak was observed for all the samples in the chromatogram leading to very high glycated haemoglobin values while in immunoturbidimetric assay reportable values were obtained. Moreover, capillary electrophoresis showed the presence of haemoglobin Hope in all the samples. Conclusion:Hb hope might lead to overestimation and possible misinterpretation of glycated haemoglobin results. Hb Hope is extremely rare in India but the possibility of interference by such clinically silent variant should always be kept in mind while interpreting the HbA1c values. INTRODUCTIONGlycated haemogobin (HbA1c) is a result of non-enzymatic glycation of hemoglobin with plasma glucose and is known to be clinically important in identifying the average plasma glucose concentration over periods of time. HbA1c is a measure of the beta-N-1-deoxy fructosyl component of haemoglobin. The extent of glycation and the relative involvement of β chains of haemoglobin remain unclear [1, 2]. HbA1c is expressed as the percentage of total haemoglobin which is 50% higher than HbA1c alone. Depending on the method used for estimating, the level of HbA1c is approximately 4-6% in healthy patients without diabetes. In clinical practice, the measurement of HbA1c serves as the marker the glycaemic control over the last three months and to determine whether the patient's blood glucose levels have remained within the target range [3]. Various methods and techniques such as high performance liquid chromatography (HPLC), immunoagglutination, boronate affinity assays, and electrophoresis have been developed based on different principles to measure HbA1C clinically but the designated DCCT comparison method is a cation exchange HPLC [4,5]. Despite advances in the measurement of glycated haemoglobin, an increasing number of hemoglobinopathies have been reported to cause falsely elevated or falsely low HbA1c results [3].Hb Hope is a clinically asymptomatic [beta136 (H14) Gly→Asp (GGT→GAT)] unstable variant of the β-globin chain that is frequently found in the Thai population [6]. It is extremely rare in India. We report the interference of Hb Hope with glycated haemoglobin values in South India. MATERIALS AND METHODSIt is a clinical study conducted in Department of Biochemistry, Kasturba Hospital, Manipal from A...
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