Hemoglobin Variants - HPLC

Order No.: 15330, for 1000 tests
Parameters:
HbA, HbA1c, HbA2, HbC, HbD, HbE, HbF, HbS

All important hemoglobin variants within 9 min.
Improved sample throughput and efficiency

HbA

HbA1c

HbA2

HbC

HbD

HbE

HbF

HbS

Clinical relevance

The haemoglobin molecule is responsible for transporting oxygen inside of the human organism, and consists of four globin polypeptide chains and one haem molecule. In humans, the globin polypeptide chains are further subdivided in α, β, γ and δ chains. Human haemoglobin usually consists of two α chains and two additional globin chains (β, γ or δ). In adults, the major component of haemoglobin is HbA (ααββ). Its glycated forms are referred to as HbA1a, HbA1b and HbA1c. HbA2 consists of two α chains and two δ chains. Foetal haemoglobin (HbF) consists of two α chains and two β chains.

Genetic disorders of haemoglobin production are called thalassaemias – they most commonly affect the β chain. This leads to β-thalassaemia. It is especially common in populations in the Mediterranean region, the Middle East and East Asia, but it also occurs in Middle Europe and North America as a result of demographic change.

 

Product advantages

  • Measurement of all key haemoglobin variants in just 9 minutes
  • Clear identifiable peaks and reliable values

 

By making use of the high separating capacities of the analytical column, the Chromsystems kit for the analysis of haemoglobin variants can determine the most common variants HbA1c, HbA2, HbC, HbD, HbE, HbF and HbS within 9 minutes. A column material specially designed for this specific separation process also allows a stable separation of HbA2 from HbD and HbE even with aged columns. This helps to eliminate false quantifications and qualifications.

More Information
Number of Tests 1000
Limit of quantification

HbA2: 2 %
HbF: 1 %

Linearity

HbA2: up to at least 75 %
HbF: up to at least 70 %

Recovery HbA2: 100 %; HbF: 99 %
Intraassay

CV < 8 %

Interassay

CV < 4 %

Analysis Time 9 min
Sample Preparation
  • Dilute 5 to 8 µl whole blood with 1 ml Hemolysis Reagent, mix well (vortex).
  • Inject 10 to 20 µl of the hemolysis mixture into the HPLC system.

Sample Stability Prepared samples can be stored at ambient temperature for about 24 h, at +2 to +8 °C up to 36 h, below -18 °C up to 3 months.
Specimen Whole blood
Pre-analytic Treatment The Patient samples are stable at room temperature for 3 days, at +2 to +8 °C up to 1 week and below -18 °C up to 6 months. Once thawed, samples should not be re-frozen.
Injection Volume 10-20 µl
Flow rate 1.5 ml/min
Column temperature ambient temperature (~ 25 °C)
Wavelength 415 nm
Gradient binary
Please note The freely available information on this website, in particular on the sample preparation, are not sufficient to work with our products. Please read instructions and warning notices on products and/or instruction manuals.
Method of Analysis HPLC
Parameter HbA, HbA1c, HbA2, HbC, HbD, HbE, HbF, HbS
Complete Specification
Linearity: up to at least 75 % (HbA2)
up to at least 70 % (HbF)
Limit of quantification: 2 % (HbA2)
1 % (HbF)
Intra-assay: < 8 %
Inter-assay: < 4 %
Recovery: 100 % (HbA2)
99.2 % (HbF)
Run time: approx. 9 min

HPLC parameters

For the analysis of hemoglobin variants any HPLC gradient system with UV/VIS detector is suitable.

Injection volume: 10-20 µl
Flow rate: 1.5 ml/min
Wavelength: 415 nm
Column temperature: ambient (approx. 25 °C)
Additional Info Any HPLC gradient system with UV/VIS detector is suitable.
Pressure Maximum pressure: 140 bar
  1. Buffer A - 2.1 l
    Buffer A - 2.1 l
    Order no.: 15331
    Hemoglobin Variants - HPLC
    Details
  2. Buffer A - 5 l
    Buffer A - 5 l
    Order no.: 15331/C
    Hemoglobin Variants - HPLC
    Details
  3. Buffer B - 3 l
    Buffer B - 3 l
    Order no.: 15332
    Hemoglobin Variants - HPLC
    Details
  4. Buffer B - 5 l
    Buffer B - 5 l
    Order no.: 15332/C
    Hemoglobin Variants - HPLC
    Details
  5. Hemolysis Reagent
    Hemolysis Reagent
    Order no.: 15334
    Hemoglobin Variants - HPLC
    Details
  6. Wash Buffer
    Wash Buffer
    Order no.: 15335
    Hemoglobin Variants - HPLC
    Details
  7. HPLC column
    HPLC column
    Order no.: 15390
    Hemoglobin Variants
    Details
  1. PEEK Prefilter Housing
    PEEK Prefilter Housing
    Order no.: 15010
    Details
  2. Reaction Vials
    Reaction Vials
    Order no.: 15007/B
    Details
  3. PEEK-encased Prefilter, 5 µm
    Details
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