Method Description Describes how the test is performed and provides a method-specific reference
Hemoglobin A2 and F:
Hemolysate of whole blood is injected into an analysis stream passing through a cartridge containing diethylaminoethyl-resin using HPLC. A preprogrammed gradient controls the elution buffer mixture that also passes through the analytical cartridge. The ionic strength of the elution buffer is raised by increasing the percentage of a second buffer. As the ionic strength of the buffer increases the more strongly retained hemoglobins elute from the cartridge. Absorbance changes are detected by a dual-wavelength filter photometer. Changes in absorbances are displayed as a chromatogram of absorbances versus time.(Huismann TH, Scroeder WA, Brodie AN, et al: Microchromotography of hemoglobins. III. A simplified procedure for the determination of hemoglobin A2. J Lab Clin Med 1975;86:700-702; Ou CN, Buffone GJ, Reimer GL, Alpert AJ: High-performance liquid chromatography of human hemoglobins on a new cation exchanger. J Chromatogr 1983;266:197-205)
The CAPILLARYS System is an automated system that uses capillary electrophoresis to separate charged molecules by their electrophoretic mobility in an alkaline buffer. Separation occurs according to the electrolyte pH and electro-osmotic flow. A sample dilution with hemolysing solution is injected by aspiration. A high-voltage protein separation occurs and direct detection of the hemoglobin protein fractions is at 415 nm, which is specific to hemoglobins. The resulting electrophoregrams peaks are evaluated for pattern abnormalities and are quantified as a percentage of the total hemoglobin present. Examples of position of commonly found hemoglobin fractions are, from cathode to anode: Hb A2', C, A2/O-Arab, E, S, D, G-Philadelphia, F, A, Hope, Bart, J, N-Baltimore, and H.
Oxygen Dissociation, P50:
The operating principle of the Hemox-Analyzer is based on dual wave-length spectrophotometry for the measurement of the oxygen saturation of hemoglobin (in percent) and a Clark electrode for measuring the oxygen partial pressure in millimeters of mercury. The resulting output signals from both measuring systems are fed into a computer, which plots the curve of the resulting function on paper.(Guarnone R, Centenara E, Barosi G: Performance characteristics of Hemox-Analyzer for assessment of the hemoglobin dissociation curve. Haematologica 1995;80:426-430)
Isoelectric focusing: Hemolyzed blood is placed on a polyacrylamide gel containing ampholytes pH 6 to 8. An electrical current is applied to the gel. When hemoglobin (Hb) is in this pH gradient, it moves to its isoelectric point, the pH where its net charge is zero. Once this happens, diffusion is counteracted by the electric field and Hb variants are thus separated as bands at their different isoelectric point.(Hoyer JD, Hoffman DR: The thalassemia and hemoglobinopathy syndromes. In Clinical Laboratory Medicine. Second edition. Edited by KD McMlatchey. Philadelphia, Lippincott Williams and Wilkins, 2002, pp 866-892)
Day(s) and Time(s) Test Performed Outlines the days and times the test is performed. This field reflects the day and time the sample must be in the testing laboratory to begin the testing process and includes any specimen preparation and processing time required before the test is performed. Some tests are listed as continuously performed, which means assays are performed several times during the day.
Monday through Friday; Varies