Original article / research
Year : 2026 Month : April Volume : 15 Issue : 2 Page : BO01 - BO04 Full Version

Analytical Performance of HPLC vs Immunoturbidimetric HbA1c Methods and their Clinical Utility in Type 2 Diabetes Mellitus: A Case-control Study
Haresingh Makwane, Pawanawanawan Kumar Kare, Tripti Saxena, Anil Sejwar, Aastha Sanadhya
1. Associate Professor, Department of Biochemistry, Gandhi Medical College, Bhopal, Madhya Pradesh, India. 2. Demonstrator, Department of Biochemistry, Gandhi Medical College, Bhopal, Madhya Pradesh, India. 3. Professor and Head, Department of Biochemistry, Gandhi Medical College, Bhopal, Madhya Pradesh, India. 4. Associate Professor, Department of Medicine, Gandhi Medical College, Bhopal, Madhya Pradesh, India. 5. MBBS Student, Department of Biochemistry, Gandhi Medical College, Bhopal, Madhya Pradesh, India.
 
Correspondence Address :
Dr. Pawan Kumar Kare,
Department of Biochemistry, Gandhi Medical College, Royal Market, Bhopal,
Madhya Pradesh, India.
E-mail: pawankare4@gmail.com
 
ABSTRACT
: Introduction: Glycated Haemoglobin (HbA1c) is the primary marker for long-term glycaemic control and is indispensable for the management of diabetes. While High-Performance Liquid Chromatography (HPLC) is considered a gold standard, Particle-Enhanced Immunoturbidimetric Assays (PEIA) offer a more rapid and cost-effective alternative. Despite the gold-standard status of HPLC and the practical benefits of PEIA, there is a continuous need to assess the analytical agreement and performance characteristics of these two methods, especially within the specific context of Type 2 Diabetes Mellitus (T2DM) patients. This comparison is critical to validate the clinical utility of the more rapid immunoturbidimetric method as a reliable alternative for routine monitoring and diagnosis, ensuring accurate patient management.

Aim: To compare the analytical performance and clinical utility of HPLC versus immunoturbidimetric HbA1c methods.

Materials and Methods: The present case-control study was conducted in the Department of Biochemistry in collaboration with the Department of Medicine, Gandhi Medical College, Bhopal, Madhya Pradesh, India, from January to February 2021. The present study included 50 patients with T2DM and 50 healthy controls. Demographic and clinical parameters, including age, gender, serum glucose, and haemoglobin levels, were recorded. HbA1c was measured in all participants using both the HPLC and immunoturbidimetric methods. Additionally, a Quality Control (QC) sample was subjected to repeated measurements by both methods to assess analytical precision and bias. Statistical analysis was performed using student’s t-test for group comparisons and a Chi-square test for categorical data.

Results: As expected, the T2DM group demonstrated significantly higher mean serum glucose and HbA1c values compared to the healthy controls (p<0.001). A significant difference in mean haemoglobin levels was also noted between the T2DM and healthy control groups (11.6±1.4 gm% vs. 13.3±1.2 gm%; p<0.001). A method comparison on patient samples showed that while both assays correlated strongly, the immunoturbidimetric method yielded slightly lower mean HbA1c values compared to the HPLC method in the healthy control group. The analysis of the QC sample confirmed these findings, revealing that both methods were highly precise (CV<3.0%), with HPLC showing slightly superior precision. A small but statistically significant systematic bias was identified between the two methods (p<0.001) and indicating a consistent, non-random difference between the HbA1c values reported by the HPLC method and the PEIA method.

Conclusion: Both HPLC and immunoturbidimetric methods are reliable for the routine measurement of HbA1c in a clinical setting. While they provide clinically similar results for diabetic patients, a small but significant systematic bias exists between them. Consequently, it is crucial for laboratories to use a single method for serial monitoring of individual patients to ensure consistency and prevent misinterpretation of treatment efficacy.
Keywords : Anaemia, Glycated haemoglobin, High-performance liquid chromatography, Serum glucose
DOI and Others : DOI: 10.7860/NJLM/2026/84447.2958 Date of Submission: Sep 18, 2025 Date of Peer Review: Dec 01, 2025 Date of Acceptance: Dec 31, 2025 Date of Publishing: Apr 01, 2026 AUTHOR DECLARATION: • Financial or Other Competing Interests: None • Was Ethics Committee Approval obtained for this study? Yes • Was informed consent obtained from the subjects involved in the study? Yes • For any images presented appropriate consent has been obtained from the subjects. NA PLAGIARISM CHECKING METHODS: • Plagiarism X-checker: Sep 19, 2025 • Manual Googling: Dec 24, 2025 • iThenticate Software: Dec 29, 2025 (7%) ETYMOLOGY: Author Origin EMENDATIONS: 5
 
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