Grade Number 3 Chromatography Paper

VIII. Methods and materials : hemoglobin types determined by cellulose acetate and citrate agar electrophoresis methods based on different electrical charges when place the sample in electrophoresis, when electrical current is passed through the hemoglobin blood sample, causes separation of hemoglobin type at different rates and form bands .(14)(15)

materials

1. Tris-EDTA Boric Acid buffer (TEB), pH 8.4.

Tris hydroxymethyl amino methane (TRIS): 10.2 g

Ethylene diamine tetracetic acid (EDTA): 0.6 g

Boric Acid: 3.2 g

1 litre with distilled water

2. Whatman No. 3 chromatography paper.

3. Cellulose acetate membranes

4. HbA2 control .(13)

Method

Cellulose acetate electrophoresis :

1. Prepare Haemolysate from whole blood (in K2EDTA) .

2. Prepare the electrophoresis tank by filling the tank with 900ml of TEB buffer wicks are cut from Grade Number 3 chromatography paper and were placed along the 22 cm long bridges in the tank.

3. The cellulose acetate membranes are cut in 40xl00 mm each and soaked in TEB buffer for 5 minutes. 5 strips are plotted then placed on the electrophoresis tanks.

4. Supply the voltage current at 250 V for 5 minutes to the membranes to equilibrate the membranes with the buffer.

5. The current is turned off and apply 8-10 μl haemolysate (10 g/μl on

Methods: A 10-year retrospective study , between 2001, and 2010 was conducted at King Khalid University Hospital in Riyadh , Which included 392 pregnant women with sickle cell disease and 784 controls group . Results: The major complications in the pregnant group were anemia (86.2%); sickle cell crisis (64.8%); bacterial infection (8.8%); preterm delivery (15.3%). Conclusion: Pregnant Saudi women associated with sickle cell disease are at increased risk for complications . the need for exists of multidisciplinary approach to the management of complications in pregnancy is