A simple flow injection-reduced volume column system for hemoglobin typing

Area of Science:

• Biochemistry
• Analytical Chemistry
• Hematology

Background:

• Thalassemia screening is crucial for early diagnosis and management.
• Conventional hemoglobin (Hb) typing methods can be time-consuming and reagent-intensive.
• There is a need for rapid, cost-effective screening methods for common thalassemia types.

Purpose of the Study:

• To develop and validate a flow injection (FI)-reduced volume column system for hemoglobin typing.
• To assess the system's efficacy as an initial screening method for thalassemia.
• To evaluate the system's advantages over conventional column techniques.

Main Methods:

• Development of a reduced volume column packed with DEAE-Sephadex A-50 ion exchange beads.
• Hb separation using a Tris-HCl buffer with a pH gradient (8.5-6.5).
• Spectrophotometric monitoring at 415 nm for HbA(2) and HbE quantification in 40 blood samples.

Main Results:

• The FI system successfully identified samples with beta, E-trait, and EE homozygous thalassemia.
• Results correlated with conventional methods used in hospital laboratories.
• The system required only 80 microl of diluted packed cells per analysis.
• Each analysis was completed within 35 minutes.

Conclusions:

• The developed FI-reduced volume column system is an economic and efficient alternative for initial Hb typing.
• This method is suitable for screening common thalassemia types like beta-trait, E-trait, and EE-homozygous, particularly in regions like Thailand.
• The system's advantages include reduced reagent and sample volume, shorter analysis time, and lower cost.