Hemoglobin A1c

KR117 Hemoglobin A1c For Short Program

Intended Use

Reagent kit for the hplc determination of hemoglobin a1c in whole blood.

KIT SPECIFICATION 

contents for 1000 analyses:

INTRODUCTION

Hemoglobin (hb)is one of the many proteins subjected to non-enzymatıc glycosylation.97% of adult hemoglobin is hba,25%hba2,0.5%hbf.hba is composed of 2a and 2β chains.hba0 is non-glycolise hemoglobin fraction.hba1c  is a major fraction that accur 80% of hba1.glycosylated hemoglobins are the result of simple chemical reactions between hemoglobin and sugars.the reaction proceeds in twostages;enzymes are not involved.first stage,glucose combines with the amino group of the valine residue at the n-terminus of 18 globin chains to form an aldimine compound(schiff base).this reaction is reversible ,and dissociation to native hemoglobin and glucose occurs readily.second stage,internal rearrangement of the aldimine intermediate by the amadori reaction yields a stable ketoamine derivative.

Life span of red blood cells are about 2-3 months,hba1c shows mean glucose level of exposure for 2-3 months.glycosylated hemoglobin concentrations have been show to be related to:physicians’ratings of diabetic control,fasting blood glucose concentrations,postrandial blood glucose  concentrations,and home monitoring of blood glucose concentrations.the unresolved labile a1c and inadequate storage conditions may have a significant effect on ghb results.labile a1c may accoumt for a2-3% increase in total a1c in normal subjects and up to 10% increase in diabetic individuals.

EQUIPMENT AND INSTRUMENTAL PARAMETERS 

the analysis of hemoglobin variants requires a hplc including system with hplc pump,autosampler,autoinjector and uv detector.the use of a solvent degasser prevents air bubbles and ensure a stable baseline.

Sampler : injection volume 10 µl,analytical run tıme 2 min.

Hplc pump : flow rate 2 ml/min is recommended

Column temperature  : 28°c

Uv detector :   415 nm

Collection And Storage Of Patient Specimens 

The sample material is whole blood. All analytes will be stable for 7 days when stored tightlycapped at 2-8°c.

Preparation Of Calibrator

The calibrator is injected directly,without dilution or pre-incubation.

Reconstitution Of The Controls

The controls level I and level II are prepared with the other sample preparation. The lyophilised controls are dissolved 0.5 ml distilled water to each control bottle and replace capsthe controls should be allowed to stand at room temperature for 5-10 minutes.before sampling, gently swirl the vial sveral times to ensure homogeneity.

Storage life of the reconstitute controls:
This product will be stabled until the expiation date when stored unopened at 2-8°c. Once the control is reconstitued, all analytes will be stable for 7 dyas when stored tightly capped at 2-8°c.it is recommended to use Kreon Medical control and calibrators.

• Dilute 5 µl of whole blood with 1 ml hemolysis reagent,mix well.
• Incubate 20min at 37 °c,cool down immediately.
• Inject 10 µlof the hemolysis mixture into the hplc system.

Storage life of the prepared samples:
Samples prepared as above can be stored at ambient temperature for about 24 hours, at+2 to +8 °c up to 7 days, frozen below -18°c up to 3 months.

Gradient Profile