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Hb HPLC • Thalassemia • Hemoglobinopathies

HPLC Blood Test (Hb HPLC): Precise Detection of Hemoglobin Variants

The HPLC blood test is essential for identifying and diagnosing disorders related to abnormal hemoglobin production—such as sickle cell disease, thalassemia, and other hemoglobinopathies. By separating and measuring different hemoglobin types, it detects variants like HbS (sickle cell) or HbC, and quantifies normal fractions such as HbA and HbF with high analytical precision.

HPLC Blood Test sample chromatogram and report preview

Quick facts & benefits

Hemoglobin variants & fractions

Separates and quantifies HbA, HbF, HbA2, and detects variants like HbS, HbC, and HbE with clear, interpretable peaks.

The HPLC blood test is essential for identifying and diagnosing disorders related to abnormal hemoglobin production, such as sickle cell disease, thalassemia and other hemoglobinopathies. By separating and measuring different types of hemoglobin in the blood, this test can detect variations like hemoglobin S (sickle cell) or hemoglobin C, as well as quantify normal types like hemoglobin A and F.

HPLC is an excellent, powerful diagnostic tool for the direct identification of hemoglobin variants with a high degree of precision in the quantification of major and minor, normal and abnormal, hemoglobin fractions. HPLC is suitable for the routine investigation of hemoglobin variants, hemoglobinopathies, and thalassemia.

What is the HPLC Blood Test?

The HPLC Blood Test (High Performance Liquid Chromatography) is a gold-standard diagnostic tool for analyzing and quantifying hemoglobin variants.

This test is crucial in the identification and monitoring of thalassemia syndromes, sickle cell disease, and other hemoglobinopathies, offering precise, automated results using state-of-the-art technology.

By separating and measuring different types of hemoglobin present in the blood, the HPLC Blood Test provides clear insights into both normal (e.g., Hemoglobin A, F) and abnormal (e.g., Hemoglobin S, C, E) hemoglobin fractions, making it an essential investigation in hematological diagnostics.

The HPLC Blood Test uses the principle of high-performance liquid chromatography to analyze the composition of hemoglobin in red blood cells. This method is highly accurate and preferred globally for the detection and quantification of hemoglobin variants, offering both clinical sensitivity and laboratory precision.

This test is conducted using fully automated HPLC systems, specifically Bio-Rad Variant II, ensuring standardized, consistent results with minimal manual error.

Sample Report:

HPLC Blood Test - Hb HPLC - Thalassemia test report

Clinical Relevance of the HPLC Blood Test

The test is specifically recommended for:

  • Suspected cases of thalassemia (alpha or beta)
  • Sickle cell trait or disease screening
  • Evaluation of unexplained anemia
  • Family history of hemoglobinopathies
  • Prenatal screening and carrier detection

The HPLC Blood Test helps detect variations such as:

  • Hemoglobin S (HbS): Indicates sickle cell trait or disease
  • Hemoglobin C (HbC): Associated with mild hemolytic anemia
  • Hemoglobin E (HbE): Common in certain ethnic populations, may cause mild anemia
  • Hemoglobin A2 and F: Key markers in diagnosing thalassemia and other blood disorders
Interpretation highlights (typical adult ranges; correlate clinically)
Fraction / VariantTypical findingClinical utility
HbA2~2.2–3.2% (adults)≥3.5% suggests β-thalassemia trait (consider iron status, recent transfusion)
HbF<1% (adults)Elevated in some hemoglobinopathies and HPFH; also in pregnancy/hematologic stress
HbS / HbC / HbEDistinct peaks/retention windows on HPLCHelps classify trait vs disease; may need confirmatory/clinical correlation

Why Choose HPLC for Hemoglobin Analysis?

  • High diagnostic accuracy in identifying normal and abnormal hemoglobins
  • Quantification of minor hemoglobin fractions such as HbA2 and HbF
  • Superior resolution compared to traditional electrophoresis techniques
  • Automated analysis minimizes human error and ensures reproducibility
  • Fast turnaround time and clinically actionable results

Performance Specifications

The precision of the HPLC Blood Test makes it a reliable diagnostic tool:

Intra-assay precision:

  • HbF: As low as 0.6%
  • HbA2: As low as 1.0%

Inter-assay precision:

  • HbF: 1.4%
  • HbA2: 1.9%

Thalassemia and Hemoglobinopathies – A Growing Concern

Thalassemia syndromes represent a group of inherited conditions caused by defective synthesis of one or more globin chains, components of hemoglobin.

Clinical Impact:

  • Inadequate hemoglobin production leads to microcytic hypochromic anemia.
  • Imbalanced globin chain accumulation causes hemolysis and ineffective erythropoiesis.
  • Severity ranges from asymptomatic microcytosis to life-threatening anemia.
  • In severe cases, it can result in fetal loss in utero.

Screening Guidelines:

  • All ethnic groups except Northern European Caucasians should be screened for variant hemoglobin.
  • All ethnic groups should be tested for beta-thalassemia trait.
  • Selected ethnic groups should be screened for alpha-thalassemia trait.

Definitive diagnosis may sometimes require DNA testing or amino acid sequencing, but the HPLC Blood Test is the most effective first-line screening tool.

How the HPLC Blood Test Works

Methodology: Fully Automated HPLC using Bio-Rad Variant II

  • Red blood cells are lysed to release hemoglobin.
  • The sample is injected into the HPLC system.
  • Different hemoglobin types are separated based on charge and structure.
  • A detector quantifies each variant based on the time it elutes.
  • Results are analyzed against reference patterns for precise diagnosis.

This process is fully automated, ensuring high throughput and reduced risk of error.

Please Correlate with Clinical Findings

While the HPLC Blood Test provides detailed laboratory insight, results must be interpreted in clinical context.

Your healthcare provider will consider symptoms, medical history, family history, and other blood parameters (like complete blood count or iron studies) before concluding any diagnosis.

Who Should Take the HPLC Blood Test?

  • Children with unexplained anemia or poor growth
  • Couples planning for children in high-prevalence zones
  • Individuals with family history of blood disorders
  • Those with borderline CBC reports showing microcytosis or low MCV
  • Pregnant women (especially in high-risk ethnic groups)
  • Individuals undergoing anemia profile tests

Now Available Across 50+ Indian Cities

You can now book the HPLC Blood Test in over 50 major Indian cities at leading NABL-accredited diagnostic centers. Cities include:

Delhi, Mumbai, Bengaluru, Hyderabad, Chennai, Kolkata, Pune, Ahmedabad, Jaipur, Lucknow, Surat, Bhopal, Patna, Indore, Kanpur, Vadodara, Thane, Visakhapatnam, Nashik, Ludhiana, Faridabad, Agra, Amritsar, Varanasi, Allahabad, Coimbatore, Madurai, Raipur, Ranchi, Chandigarh, Guwahati, Vijayawada, Gwalior, Jodhpur, Nagpur, Bhubaneswar, Aurangabad, Meerut, Noida, Ghaziabad, Dhanbad, Kota, Solapur, Bareilly, Jabalpur, Hubli, Mangalore, Mysuru, Dehradun, Navi Mumbai, Jamshedpur

HPLC Blood Test: Know More

The HPLC Blood Test is a highly sensitive, fully automated diagnostic test used for the quantitative and qualitative analysis of hemoglobin variants.

Whether you are screening for thalassemia, managing anemia, or diagnosing a suspected hemoglobinopathy, this test delivers trustworthy results with unmatched accuracy.

Available in India’s leading cities, this test is an essential part of modern diagnostic care—especially for families with a genetic predisposition to blood disorders.

You may also need a Viral Marker Test panel with the HPLC test to understand whether there is a genetic health issue or it’s due to certain viral infections.

If your doctor has recommended it, book your HPLC Blood Test today and gain clarity on your hemoglobin health.

Preparation, Limitations & After-Care

Test preparation
  • No fasting typically required for Hb HPLC.
  • Inform the lab about recent blood transfusions (can alter chromatogram for weeks).
  • Share any ongoing iron therapy; iron deficiency can affect HbA2 levels.
Analytical considerations & limitations
  • Co-elution of rare variants may require confirmatory tests (electrophoresis or molecular assays).
  • Borderline HbA2 should be interpreted with iron status and clinical context.
  • Pregnancy and certain hematologic conditions may elevate HbF.
Report discussion & follow-up

Discuss your results with a clinician. For couples, consider genetic counseling when a variant or carrier state is identified.

Book Your HPLC Blood Test

NABL-accredited partner labs • Home sample collection in most cities • Transparent reporting

Frequently Asked Questions

What is the HPLC Blood Test and why is it important?

The HPLC Blood Test (High Performance Liquid Chromatography) is a highly accurate diagnostic test used to detect and quantify various hemoglobin variants in the blood. It is essential for diagnosing blood disorders such as thalassemia, sickle cell disease, and other hemoglobinopathies by identifying abnormal hemoglobin types like HbS, HbC, HbE, and elevated HbA2 or HbF levels.

Which conditions can be diagnosed with the HPLC Blood Test?

The HPLC Blood Test helps identify disorders like beta thalassemia, alpha thalassemia, sickle cell trait or disease, and other inherited hemoglobin abnormalities. It is especially useful in evaluating anemia, detecting carrier states, and performing prenatal screening in high-risk populations.

How accurate is the HPLC Blood Test for detecting hemoglobin variants?

The HPLC Blood Test is extremely precise, with intra-assay precision as low as 0.6% for HbF and 1.0% for HbA2, and inter-assay precision as low as 1.4% and 1.9% respectively. Using fully automated Bio-Rad Variant II systems, it offers reliable, reproducible results for routine screening and diagnosis.

Who should get an HPLC Blood Test done?

This test is recommended for individuals with unexplained anemia, a family history of thalassemia or sickle cell disease, couples planning pregnancy, and those with abnormal red blood cell indices. It’s also suggested for prenatal screening and for people from ethnic groups where hemoglobinopathies are more prevalent.

What is the method used for the HPLC Blood Test?

The HPLC Blood Test is performed using a fully automated Bio-Rad Variant II system, which separates and quantifies hemoglobin fractions with high precision. This method ensures accurate diagnosis of both normal and abnormal hemoglobin types by analyzing their unique retention times and peak patterns.

References

These resources support clinical understanding; always correlate with local laboratory SOPs and clinician judgment.