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

Abstract

Glycated hemoglobin (HbA) measurement from whole blood (WB) samples is inconvenient for epidemic surveillance and self-monitoring of glycemic level. We evaluated HbA 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 measurement, supporting the use of FP method in epidemic surveillance and healthcare units.

Determination of haemoglobin A1c levels using high-performance liquid chromatography of bloodstains

“…21 Wu et al allowed bloodstains prepared on filter paper to sit at a room temperature of between 20 C and 25 C, and found that HbA1c values remained stable. 17 The present study differs from other studies in its preparation of the bloodstains on glass surfaces and cotton fabrics, as these are surfaces that are more likely to be encountered at a crime scene.…”

“…14 Clinical studies testing HbA1c levels in dried blood samples in the follow-up of patients with diabetes showed a perfect correlation with the test results obtained from an analysis of whole blood and the results obtained from dried blood samples. [15][16][17] The present study aimed to investigate the feasibility of HbA1c testing in bloodstains recovered from a crime scene in forensic cases, for which bloodstains were prepared from the whole blood samples of patients undergoing routine HbA1c testing. The samples were left in uncontrolled room conditions to simulate a real crime scene.…”

A Comparison Between the Therapeutic Effect of Metformin Alone versus a Combination Therapy with Insulin in Uncontrolled, Non-Adherence Patients with Type 2 Diabetes: Six Months Follow-Up

“… 23 Glycated hemoglobin (HbA1c) was measured by high-performance liquid chromatography and by an immunoturbidimetric method. 24 TC, LDL-C, HDL-C, and TG were measured with commercially available assay kits (Dimension ® , Siemens, Germany). High-density lipoprotein cholesterol- precipitated using dextran sulfate, was measured enzymatically, low-density lipoprotein cholesterol- was calculated using the Friedewald equation (LDL-Ch= TCh- (TG/5)- HDL-Ch.…”