Hemoglobin Variants

HEMOGLOBIN VARIANTS FOR HPLC

INTENDED USE

kr118 reagent kit for the hplc determination of hemoglobin variants in whole blood.

KIT SPECIFICATION

Contents for 1000 analyzes:

INTRODUCTION

Hemoglobinopathy is a kind of genetic defect that results in abnormal structure of one of the globin chains of the hemoglobin molecule.

Hemoglobin is a protein in erythrocytes that oxygen from the lungs to the rest of the body and gives blood its red color. hb consists of four subunits.each of them contain a polypeptide chain and a prosthetic group called”hem”. Six different polypeptide chains namely alpha(α),beta (β),gamma(µ),delta(δ),epsilon( ε)and zeta (ζ)have been identified in the globin portion of human hb. The genes coding for and a-polypeptide chains are located on chromosome 16,£,y,s and b-polypeptide chains are coded on chromosome 11. These genes are inherited according to mendel laws and are co-dominant.normal hbs in the red blood cells are hb a, hb a2 and hb f. Most of the structural changes in hb are resulted from the mutations on the globin genes. Up to now, about 890 different variants of hb have been described. While some of these variants are not cause health problems, some of them cause severe hemolytic anemia an life-threatening complications. hb is one of the systems use in population screening and neonatal screening programs due to clinical importance and high frequency of hb variants.

EQUIPMENT AND INSTRUMENTAL PARAMETERS

The analysis of hemoglobin variants requires a hplc including gradient 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 10pl, analytical run time 8.5min.

Hplc pump: Flow rate 1.5 ml/min is recommended

Column temperature : ambient( ~25 `.c)

UV detector: 415 nm

RECONSTITUTION OF THE CONTROLS

the controls level I and level II are prepared with the other sample preparation. The lyophilized controls are dissolved 0.2 ml distilled water to each control bottle and replace caps. The controls should be allowed to stand at room temperature for 15 minutes.

Storage Life Of The Reconstituted Controls:
the reconstituted control is stable for 30 days when stored at 2-8.c. the reconstituted control should not be frozen.it is recommended to use kreon medical contols and calibrator.

COLLECTION AND STORAGE OF PATIENT SPECIMENS

the sample material is whole blood. storage life at room temperature is about 3 days,refrigerated (+2 to +8 .c)up to 1 week.

SAMPLE PREPARATION PROCEDURE

dilute 5 pl of whole blood with 1 ml hemolysis reagent.

inject 10 pl of the hemolysis mixture into the hplc system.

storage life of the preared as above can be stored at ambient temperature for about 24 hours,at+2to+8 .c up to 36 h,frozen below -18.c up to 3months.

GRADIENT PROFILE