Evaluation of a Colorimetric Method in Glycated Hemoglobin measurment
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R . Mohammadi Ph.D , M.Sheikhzadeh B.S , V.Norouzi B.S |
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Abstract: (17878 Views) |
Objective:Measuring of hemoglobin A1c( HbA1c )as the main glycated hemoglobin in circulation, is the most reliable method for evaluation of hyperglycemia control in diabetic patients.
Method:Colorimetric measuring method for HbA1c is based on releasing glucose in 100°C as5-hydroxymethylfurfural(5HMF)and then measuring it in a colorimetric reaction with thiobarbituric acid (TBA).This method is affected by different variables.
Results: This study shows that TBA solution is stable at 4°c at least for one month. But by reducing the time of boiling from 5 hours to 1 hour, color intensity would decrease as much as 42% . On the other hand, reducing the time and temperature of colorimetric reaction from 40 to 15 minutes and 50 ° C to 25° C would result in 29% and 63% reduction in color intensity, respectively. In addition, developed color is unstable at room temprature and would decerase as much as 53% within 40 minutes.
Conclusion: With the colorimetric measurment of HbA1c , it is recommended to use 5 hours boiling , colorimetric reaction in 50 ° C for 40 minutes,and rapid reading of optical density. |
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Keywords: Diabetes, Glycated hemoglobin / HbA1c, Glycated hemoglobin Determination |
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Subject:
Cilinical Biochemistry Received: 2006/09/6 | Accepted: 2013/12/17 | Published: 2013/12/17
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