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1- Department of Biochemistry, Mamata Academy of Medical Sciences, Bachupally, Hyderabad - 500090 India
2- Department of Biochemistry, Mamata Academy of Medical Sciences, Bachupally, Hyderabad - 500090 India ,brneeharika@gmail.com
2- Department of Biochemistry, Mamata Academy of Medical Sciences, Bachupally, Hyderabad - 500090 India ,
Abstract: (2892 Views)
Abstract
Background
Accurate estimation of glycated haemoglobin (HbA1c) is critical for diagnosing and managing diabetes mellitus. Ion-exchange high-performance liquid chromatography (HPLC) is considered the gold standard for HbA1c measurement, while nephelometry is widely used in clinical laboratories due to its rapid turnaround time. This study compares the accuracy, reliability, and clinical applicability of these two methods.
Methods
A cross-sectional study was conducted at a tertiary care hospital, where HbA1c levels of 50 patients with diabetes mellitus were measured using both ion-exchange HPLC and nephelometry. The results were analysed using descriptive statistics, Pearson correlation, intraclass correlation coefficient (ICC), Bland–Altman analysis, and receiver operating characteristic (ROC) curves. A paired t-test was used to assess statistical significance, with p<0.05 considered significant.
Results
The mean HbA1c levels were comparable between HPLC (6.2% ± 1.5) and nephelometry (6.3% ± 1.4), with a high correlation (r = 0.96, p < 0.01) and excellent agreement (ICC = 0.96). Bland–Altman analysis showed minimal bias (mean difference = 0.09%). ROC curve analysis indicated high diagnostic performance for both methods, with HPLC demonstrating superior sensitivity (90%) and specificity (92%) compared to nephelometry (88% sensitivity, 91% specificity). Nephelometry had a faster turnaround time but slightly lower specificity.
Conclusion
Both HPLC and nephelometry provide reliable HbA1c measurements, with HPLC offering superior specificity and diagnostic precision, while nephelometry provides a quicker alternative for routine clinical use. The selection of the method should be based on clinical requirements, balancing accuracy and operational efficiency.
Background
Accurate estimation of glycated haemoglobin (HbA1c) is critical for diagnosing and managing diabetes mellitus. Ion-exchange high-performance liquid chromatography (HPLC) is considered the gold standard for HbA1c measurement, while nephelometry is widely used in clinical laboratories due to its rapid turnaround time. This study compares the accuracy, reliability, and clinical applicability of these two methods.
Methods
A cross-sectional study was conducted at a tertiary care hospital, where HbA1c levels of 50 patients with diabetes mellitus were measured using both ion-exchange HPLC and nephelometry. The results were analysed using descriptive statistics, Pearson correlation, intraclass correlation coefficient (ICC), Bland–Altman analysis, and receiver operating characteristic (ROC) curves. A paired t-test was used to assess statistical significance, with p<0.05 considered significant.
Results
The mean HbA1c levels were comparable between HPLC (6.2% ± 1.5) and nephelometry (6.3% ± 1.4), with a high correlation (r = 0.96, p < 0.01) and excellent agreement (ICC = 0.96). Bland–Altman analysis showed minimal bias (mean difference = 0.09%). ROC curve analysis indicated high diagnostic performance for both methods, with HPLC demonstrating superior sensitivity (90%) and specificity (92%) compared to nephelometry (88% sensitivity, 91% specificity). Nephelometry had a faster turnaround time but slightly lower specificity.
Conclusion
Both HPLC and nephelometry provide reliable HbA1c measurements, with HPLC offering superior specificity and diagnostic precision, while nephelometry provides a quicker alternative for routine clinical use. The selection of the method should be based on clinical requirements, balancing accuracy and operational efficiency.
Keywords: HbA1c, Diabetes Mellitus, Ion-Exchange HPLC, Nephelometry, Diagnostic Accuracy, Glycated Haemoglobin
Research Article: Original Paper |
Subject:
Biochemistry
Received: 2025/02/28 | Accepted: 2025/11/22
Received: 2025/02/28 | Accepted: 2025/11/22
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