Question

in your own idea, summarize the principle of the glycohemoglobin assay

Answer 1

Ans- Summary of the glycohemoglobin assay is as follow-

Hemoglobin subfractions formed by the glycation of the alpha or beta chains of hemoglobin A1 (HbA) are collectively known as glycosylated or glycated hemoglobins.
Hemoglobin A1c, the best-defined of these, is formed by the reversible condensation of the carbonyl group of glucose and the amino group at the N-terminus of the beta chain of hemoglobin A, resulting in a labile aldimine or Schiff base. As the red cell circulates, some of the aldimine undergoes a slow, irreversible conversion (Amadori rearrangement) to a stable ketoamine form (HbA1c). As blood glucose levels rise, the increase in
glycated hemoglobin is proportional to both the level of glucose and the lifespan of the red cell. Hemoglobin A1c measurements are used in the clinical management of
diabetes to assess the long-term efficacy of diabetic control. The glycated hemoglobin result is a reflection of the mean daily blood glucose concentration and the degree of carbohydrate imbalance over the preceding two to three months.
In the past, accurate measurement of stable HbA1c was possible only after removing liabile HbA1c by pretreatment. In this assay, the stable (SA1c) and labile (LA1c) 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 Hemolysis & Wash 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 1, 2 and 3, each with a differing 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.

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