Hemoglobin Quantitation by HPLC


Specimen Collection Criteria

Collect: One Lavender-top EDTA tube. (Minimum Whole Blood: 1.0 mL)
Also acceptable (Pediatric): One 500 mcL Lavender-top EDTA Microtainer®. (Minimum Whole Blood: 250 mcL) 

Physician Office/Drawsite Specimen Preparation

Do not freeze specimen. Store whole blood refrigerated (2-8°C or 36-46°F) prior to transport. (Minimum Whole Blood: 1.0 mL)

Preparation for Courier Transport

Transport: Whole blood specimen, refrigerated (2-8°C or 36-46°F). (Minimum Whole Blood: 1.0 mL)

Rejection Criteria

  • Frozen specimens.
  • Severely hemolyzed specimens.


Routine results available within 3–5 days. Upon request, results may be available on the same day Monday – Friday, or on Saturday – Sunday if the sample is received before 1:00 pm.

Reference Range

Hemoglobin F
0-30 Days: 60-90%.
31 Days - 23 Months: Less than or equal to 60%.
Greater than or Equal to 2 Years: 0-2%.

Hemoglobin A2
Less than 1 Year: 0.0 - 3.3%.
1 Year - Adult: 2.0 - 3.3%.

Hemoglobin S, C, D, E: 0%.

Test Methodology

High Performance Liquid Chromatography (HPLC).


Presumptive diagnosis and quantitation of Hemoglobin F, A2, S, and other variants.
Any abnormal hemoglobin variants identified must be confirmed by another method.

Clinical Utility

Adult blood contains primarily hemoglobin A (HbA), a small percentage of hemoglobin A2 (HbA2) and trace amounts of fetal hemoglobin (HbF). β-thalassemia trait is usually associated with an elevated HbA2 (greater than 3.3%) and either a normal or slightly elevated HbF. Together with hematological with hematological information, determination of hemoglobin's A2 and F has become the most practical means to diagnose β-thalassemia trait (minor).

The most commonly occurring hemoglobin variants include hemoglobin's D, S, C, and E. Presumptive identification and quantitation of these variants is made using retention time windows.

CPT Code


Test Codes


Last Updated