Reference no: EM132328631
Description of Laboratory Methodology
Glycohemoglobin (GHB_I)
In this assay, the stable (SA1c) and labile (LA1c) A1c forms can be individually resolved on the chromatogram without manual pretreatment, allowing accurate measurement of the stable form of HbA1c. The analyzer dilutes the whole blood specimen with a hemolysis solution, and then injects a small volume of the treated specimen onto the HPLC analytical column. Separation is achieved by utilizing differences in ionic interactions between the cation exchange group on the column resin surface and the hemoglobin components. The hemoglobin fractions (A1c, A1b, F, LA1c, SA1c, A0 and H-Var) are subsequently removed from the column material by step-wise elution using elution buffers each with a different salt concentration. The separated hemoglobin components pass through the photometer flow cell where the analyzer measures changes in absorbance at 415 nm. The analyzer integrates and reduces the raw data, and then calculates the relative percentages of each hemoglobin fraction. Analysis requires three minutes.
Description of Laboratory Methodology
Plasma Fasting Glucose (GLU_I)
Glucose
In this enzymatic method glucose is converted to glucose-6-phosphate (G-6-P) by hexokinase in the presence of ATP, a phosphate donor.
Glucose-6-phosphate dehydrogenase then converts the G-6-P to gluconate-6-P in the presence of NADP+. As the NADP+ is reduced to NADPH during this reaction, the resulting increase in absorbance at 340 nm (secondary wavelength = 700 nm) is measured. This is an endpoint reaction that is specific for glucose.
Refer to the Laboratory Method Files section for a detailed description of the laboratory methods used.
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Description of laboratory methodology
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