Beta Thalassemia on G7 HPLC Analyzer

(April 28, 2009) … HPLC Testing Video focusses on switching to beta thalassemia testing mode on G7 HPLC Analyzer.  The Tosoh G7 Automated HPLC Analyzer has both hemoglobin A1c and beta thalassemia programs to provide full patient profile. The beta-thalassemia program quantifies hemoglobins F and A2, which are used in screening for beta-thalassemia. In addition, the beta-thalassemia program is used for presumptive identification of hemoglobins S, C and D and for screening for other hemoglobinopathies.  

The G7 provides time and labor-savings with primary tube sampling, automated cap piercing and positive sample identification. Most importantly, with less than 2% CVs, the NGSP-certified G7 offers a high level of both precision and accuracy.  

Tosoh G7 HPLC Analyzer

HPLC Testing by Tosoh Bioscience

(March 3, 2009) - High performance liquid chromatography (HPLC) is the gold standard methodology for the HbA1c test.  By manipulating the flow rate of the liquid (i.e. mobile phase) and the temperature of the HPLC column (i.e. stationary phase), it is possible to change the chromatographic display.

The Tosoh G8 HPLC Analyzer provides direct determination of stable HbA1c. The system is used for in vitro diagnostic measurement of HbA1c in blood specimens. The G8 uses a non-porous column and microcomputer technology to quickly and accurately measure the stable portion of HbA1c as a percentage of the total amount of hemoglobin present in the sample. The G8 provides accurate and precise separation of HbA1c from other hemoglobin fractions. Off-line pretreatment is not required, and there is no interference from the labile portion of HbA1c.

The analyzer dilutes the whole blood specimen with Hemolysis & Wash Solution and then injects a small volume of the treated specimen onto the TSKgel Glyco HSi Variant Column. Separation is achieved by utilizing differences in ionic interactions between the cation exchange group on the column resin surface and the hemoglobin components in a step gradient elution. The hemoglobin fractions (designated as A1a, A1b, F, LA1c , SA1c, A0, and H-V0, H-V1, H-V2) are subsequently removed from the column material by performing a step-wise elution using Elution Buffers HSi Variant 1, 2, and 3 that have specific salt and pH concentrations.

The separated hemoglobin components pass through the LED photometer flow cell where changes in absorbance are measured at 415nm. The G8 software integrates and reduces the raw data, and then calculates the relative percentages of each hemoglobin fraction. The print-out consists of the numerical results and the chromatogram. This represents the changes in absorbance versus retention time for each peak fraction. An analysis requires only 1.6 minutes.

HPLC Solutions: Tosoh Bioscience