Haemoglobin is a helix of globin chains composed of amino acids. A haemoglobin variant refers to an alteration in the amino acid composition of the globin chain that can occur through genetic mutation and may result in changes not only to the net charge on the molecule but also changes in polarity due to alterations in the helical structure. These changes in physical characteristics may, in some cases, produce clinically significant effects and have been used to fractionate haemoglobin species.
A decreased capacity for the synthesis of a globin chain results in clinical conditions known as thalassemias. Beta thalassemia is characterised by an increase in haemoglobin A2. Alpha thalassemia is characterized by a decrease in hemoglobin A2 and delta-beta thalassemia is characterised by an increase in hemoglobin F. Accurate quantitation of haemoglobins A2 and F is required for the evaluation of these conditions.
High performance liquid chromatographic (HPLC) methods with high sensitivity and specificity have been developed for hemoglobin fractionation. Trinity Biotech’s ultra2 Resolution system for the separation and quantitation of normal and variant hemoglobin species in blood is designed to provide the maximum amount of information in the shortest possible time. This method combines the sensitivity and specificity of HPLC with sample preparation automation and assistance in evaluating the result.
The user friendly software for the ultra2 is a Windows XP™ based program especially designed for haemoglobin variant analysis.
As the sample is chromatographed, it is displayed on the monitor in real time.
The computer produces printed reports with the sample identification information, date and time, followed by the chromatogram with retention times indicated at the apex of each peak.
A peak summary report is printed after the chromatogram with the retention time, relative retention time, % area and comments for each peak in the chromatogram.
Correlated to column chromatography for A2 quantitation
Correlated to alkali denaturation for F quantitation
Provides a screening and confirmatory testing
Extensive variant library: detection of over 200 variants
Variety of sample formats concurrently:
- Whole blood
- Dried blood spots for neonatal / newborn testing
- Large sample capacity:
- Microtiter plates
Barcode reading – positive sample ID from specimen to end result
Automated – walk-away, cost-effective operation
Selectable Test Methods:
- Quick Scan (screening) followed by High Resolution(confirmatory)
- Quick Scan only
- High Resolution only
Relative retention times: 0.001 minutes
Hb measurement starts at time zero:
- No calibration for A2 and F quantitation necessary
- Sensitivity of 0.1%, Principle Ion-exchange high performance liquid chromatography