15330_Hemoglobin Variants - HPLC

Hemoglobin Variants - HPLC

Order no.: 15330
1000 Tests
  • HbA
  • HbA1c
  • HbA2
  • HbC
  • HbD
  • HbE
  • HbF
  • HbS

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

Description

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.

Chromatogram

Technical Data

Method of Analysis

HPLC

Number of Tests

1000

Parameter

HbA, HbA1c, HbA2, HbC, HbD, HbE, HbF, HbS

Analysis Time

9 min

Limit of quantification

HbA2: 2 %
HbF: 1 %

Linearity

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

Recovery

HbA2: 100 %; HbF: 99.2 %

Intraassay

CV < 8 %

Interassay

CV < 4 %

Column temperature

ambient temperature ( ~ 25 °C)

Flow rate

1.5 ml/min

Specimen

Whole blood

Sample Preparation

  • Dilute 5 to 8 µl whole blood with 1 ml Hemolysis Reagent, mix well.
  • Inject 10 to 20 µl of the hemolysis mixture into the HPLC system.

Samples prepared as above can be stored at ambient temperature for about 24 hours, at +2 to +8 °C up to 36 h, frozen below -18 °C up to 3 months.

Sample Stability

At ambient temperature approx. 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 refrozen.

Injection Volume

10-20 µl

Additional Info

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

Wavelength

415 nm

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.

Kit Components

Components available separately

Buffer A - 2.1 l


Order no.: 15331

Buffer A - 5 l


Order no.: 15331/C

Buffer B - 3 l


Order no.: 15332

Buffer B - 5 l


Order no.: 15332/C

Hemolysis Reagent


Order no.: 15334

Wash Buffer


Order no.: 15335

HPLC column


Order no.: 15390

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