30/08/2025
HbA1c Test
1. Objective
The objective of the HbA1c test was to measure the average blood glucose concentration over the previous two to three months by detecting the proportion of glycated hemoglobin in a blood sample. This evaluation provided long-term glycemic control status for diabetic patients.
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2. Principle
The test was based on the principle that glucose in the bloodstream bound irreversibly to hemoglobin within red blood cells, forming glycosylated hemoglobin (HbA1c). Since red blood cells had an average lifespan of 120 days, the amount of HbA1c reflected the average blood glucose concentration over that period. Various analytical methods such as ion-exchange chromatography, immunoassay, or high-performance liquid chromatography (HPLC) were used to separate and quantify HbA1c.
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3. Materials
The materials that were used included:
• Sterile syringes and vacutainer tubes (with EDTA anticoagulant) for blood collection
• Microscopic slides and cover slips
• Stains (as required for microscopic preparation, e.g., Wright’s stain for morphology verification)
• Pipettes and micropipette tips
• Centrifuge machine
• HbA1c testing kit (HPLC or immunoassay reagents)
• Microscope for examining red blood cells and confirming sample integrity
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4. Procedure (Microscopic)
1. A venous blood sample was collected in an EDTA tube.
2. The sample was centrifuged, and red blood cells were separated for analysis.
3. A smear was prepared on a microscopic slide to assess red cell morphology, ensuring no hemolysis or abnormality interfered with the test.
4. The prepared slide was air-dried and stained, then examined under a microscope to confirm red cell integrity.
5. The HbA1c concentration was then determined using the chosen method (HPLC or immunoassay). The microscopic step ensured sample quality before analytical processing.
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5. Result
The results were expressed as a percentage of HbA1c relative to total hemoglobin.
• Normal range: 4% – 5.6%
• Prediabetes: 5.7% – 6.4%
• Diabetes: ≥ 6.5%
The test report provided both the HbA1c percentage and an estimated average glucose (eAG) level.
6. Uses
The HbA1c test was used for:
• Diagnosis of diabetes mellitus
• Monitoring long-term glycemic control in diabetic patients
• Assessing the effectiveness of treatment regimens
• Predicting risks of diabetic complications (neuropathy, retinopathy, nephropathy)
7. Consultation
After obtaining the results, patients were advised to consult a physician or endocrinologist. The healthcare provider interpreted the values, adjusted medication if necessary, and gave dietary, lifestyle, or insulin therapy recommendations. The consultation also included counseling on preventing long-term diabetic complications through strict glycemic control.
