β - THALASSEMİA FOR HPLC
KR109 reagent kit for the hplc determination of β-thalassemia in whole blood.
CONTENTS FOR 1000 ANALYSES:
Beta-thalassemias (β-thalassemias)are a group of inherited blood disorders caused by reduced or absent synthesis of the beta chains of hemoglebin resulting in variablephenotypes ranging from severe anemia to clinically asymptomatic individuals. The total annual incidence of symptomatic individuals is estimated at 1 in 100,000 throughout the world. Three main forms have been described: thalassemia major,thalassemia intemedia and thalassemia mimor. Individuals with beta thalassemia major usually present within the fırst two years of life with severe anemia, poor growth and skeletal abnrmalities during infancy. Affected children will require regular lifelong blood transfusions. Beta thalassemia intermedia is less severe than beta thalassemia major and may require episodic blood transfusion-dependent patients will develop iron overload and requirechelation therapy to remove the excess iron. Bone marrow transplants can be curative for some shildren with beta thalassemia major.transmission is autosomal recessive; however, dominant mutations have also been reported.genetic counseling is recommended and prenatal diagnosis may be offered.
EQUIPMENT AND INSTRUMENTAL PARAMETERS
the analysis of hemoglobin variants requires a hplc ıncluding gradient system with hplc pump, autosampler, auyoinjector and uv detector. The use of a solvent degasser prevents air bubbles and ensure a stable baseline.
Sampler : injection volume 10µl,analytical run time 6.5 min.
hplc pump : flow rate 1.1-1.4 ml/min is recommended
column temperature : romm temperature(25°c)
uv detector : 415 nm
The following table shows the retention times of the analytes at a flow rate of 1.1-1.4 ml/min:
RECONSTITUTION OF THE CONTROLS
the controls level I and level II are prepare with the other sample preparation. the lyophilized controls are dıssolved 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 Reconstitued Controls:
the reconstituted conrol is stable for 30 days when stored at 2-8 0c.the reconstituted control should not be frozen.
COLLECTION AND STORAGE OF PATIENT SPECIMENS
the sample material is whole blood. Storage life at room temoperature is about 3 days,refrigerated (+2 to +8 0c) up to 1 week.
SAMPLE PREPARATION PROCEDURE
Dilute 5µl of whole blood with 1 ml hemolysis reagent.
Inject 10 µl of 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 36 h,frozen below -18 °c up to 3 months.
Linearity : R2 = 0.99
Specifity : results of (Rt) hbA2 are given below: