HbA1c Levels On High Performance Liquid Chromatography (HPLC) And Fluorescence Immunoassay (FIA) Methods

Abstract

Hemoglobin A1c (HbA1c) is an ideal parameter for the diagnosis and control of diabetes mellitus, reflecting the average blood glucose level through the non-enzymatic reaction between glucose and hemoglobin. Although HPLC is the gold standard of HbA1c measurement, this method is expensive and complex. This study aims to evaluate HbA1c measurement using Fluorescence Immunoassay (FIA), which is faster, simpler, and cost-effective. This study is a comparative study with a cross-sectional design to compare HbA1c levels using HPLC and FIA methods. The sample consisted of 30 patients of Klinik Utama Pramita Surabaya, with venous blood taken using EDTA tubes. Data analysis was performed using paired t-test or Wilcoxon test based on data distribution. Method validation included correlative analysis, sensitivity, specificity, and accuracy using IBM SPSS Statistics 25. The results showed that HPLC and FIA methods produced similar HbA1c distributions. Wilcoxon test showed no significant difference between the two methods (p-value =0.094). The sensitivity of the FIA method was 94.4%, specificity 100%, PPV 100%, NPV 92.9%, and accuracy 96.7. This study shows that the HPLC and FIA methods provide similar results in the measurement of HbA1c levels, with high levels of sensitivity, specificity, PPV, NPV and accuracy. The overall results support the use of the FIA method in the measurement of HbA1c for diabetes diagnosis and management, given its good analytical performance and compatibility with the HPLC method.