Evaluation of hemoglobin A_1c measurement from filter paper using high-performance liquid chromatography and immunoturbidimetric assay

Abstract

Glycated hemoglobin (HbA_1c) measurement from whole blood (WB) samples is inconvenient for epidemic surveillance and self-monitoring of glycemic level. We evaluated HbA_1c measurement from WB blotted on filter paper (FP), which can be easily transported to central laboratories, with high-performance liquid chromatography (HPLC) and immunoturbidimetric assay (ITA). WB was applied to Whatman filter paper. By using HPLC and WB samples as reference methods, these FP samples were evaluated on HPLC and ITA. Inter- and intra-assay variation, WB vs. FP agreement and sample stability at 20–25 °C and −70 °C were assessed by statistical analysis. Results showed that the coefficient of variation (CV, %) of FP samples for HPLC and ITA were 0.44–1.02% and 1.47–2.72%, respectively (intra-assay); 2.13–3.56% and 3.21–4.82%, respectively (inter-assay). The correlation of WB HPLC with FP analyzed using HPLC and ITA are both significant (p < 0.001). Sample stability showed that FP method up to 5 days at 20–25 °C and 5 weeks at −70 °C is accurate and reproducible. In conclusion, FP samples analyzed by HPLC and ITA can both provide an alternative to WB for HbA_1c measurement, supporting the use of FP method in epidemic surveillance and healthcare units.

Keywords:

• Diabetes
• filter paper
• HbA_1c
• high-performance liquid chromatography
• immunoturbidometric assay

Acknowledgements

We wish to thank our phlebotomy staff for their assistance with the specimen collection.

Disclosure statement

The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper. There was no external funding.