15440_β-Thalassemia Testing - HPLC

β-Thalassemia Testing - HPLC

Order no.: 15440
1000 Tests
  • HbA1c
  • HbA2
  • HbC
  • HbF
  • HbS

Simultaneous quantifications of HbS and HbC within 5.5 minutes
HbA2 will be quantified in presence of HbE and HbD

 

Description

Clinical relevance

Thalassaemias are caused by impaired synthesis of one or more globin chains of the haemoglobin (consisting of 4 polypeptide chains), resulting in much less oxygen being bound to the haemoglobin molecules and transported through the body. Depending on which polypeptide chains are affected, the thalassaemias are named α, β, γ or δ thalassaemia. It is the β chain that is most frequently affected, so that this disorder is called β-thalassaemia. β-thalassaemia major is a homozygous form in which both alleles are severely mutated so that β chain synthesis is stopped completely, whereas β-thalassaemia minor is a heterozygous form resulting in an approximately 20 % reduction of polypeptide synthesis. To compensate this reduction, more HbA2 and HbF are produced: in β-thalassaemia major it is more HbF, and in β-thalassaemia minor primarily HbA2.

Depending on severity, patients may suffer from enlargement of the liver and spleen, growth disorders, severe inner organ damages and bone deformities. Further mutations are known that modify the peptide chains in other ways, leading to abnormal haemoglobin variants. The most common and clinically relevant variants are HbS, HbC and HbE. HbD also occurs relatively often. Combinations of various thalassaemias with different genetically related haemoglobin variants are also possible.

 

Product advantages

  • Simultaneous determination of HbS and HbC in less than 6 minutes
  • Quantification of HbA2 without interferences from HbD and HbE variants

 

The Chromsystems kit for β-thalassaemia testing allows the reliable quantification of HbA2 and HbF. The column’s high separation capability allows the determination of the most common variants HbA1c, HbA2, HbC, HbF and HbS in less than 6 minutes. A column material specially designed for this specific separation process allows a stabilised separation of HbA2 from the other Hb-molecules even with aged columns. This helps to avoid misinterpretations and eliminate false quantifications and qualifications.

Chromatogram

Technical Data

Method of Analysis

HPLC

Number of Tests

1000

Parameter

HbA1c, HbA2, HbC, HbF, HbS

Analysis Time

< 5.5 min

Limit of quantification

HbA2: 2 %
HbF: 1 %

Linearity

HbA2: up to 75 %
HbF: up to 82 %

Recovery

101 %

Intraassay

CV < 5 %

Interassay

CV < 10 %

Column temperature

ambient ( ~ 25 °C)

Flow rate

2.3 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.

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 ß-thalassemia screening 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.: 15441

Buffer A - 5 l


Order no.: 15441/C

Buffer B - 2.8 l


Order no.: 15442

Buffer B - 5 l


Order no.: 15442/C

Hemolysis Reagent


Order no.: 15444

Wash Buffer


Order no.: 15445

HPLC column


Order no.: 15490

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