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Hemoglobin variant detection from dried blood specimens by high performance liquid chromatography
P D Roa1, E A Turner, M del P Aguinaga
1Comprehensive Sickle Cell Center, Meharry Medical College, Nashville, TN 37208.
Annals of Clinical and Laboratory Science
|November 1, 1993
Summary
Dried blood filter paper is stable for hemoglobin identification using high-performance liquid chromatography (HPLC) for up to six weeks at room temperature. This method accurately screens for hemoglobinopathies, even with stored samples.
Area of Science:
- Biochemistry
- Genetics
- Clinical Diagnostics
Background:
- Dried blood spot (DBS) specimens offer convenience for genetic screening.
- Hemoglobinopathies require reliable diagnostic methods for early detection.
Purpose of the Study:
- To evaluate the stability of dried blood filter paper specimens for hemoglobin identification and quantitation using cation exchange high-performance liquid chromatography (HPLC).
- To determine the feasibility of using DBS-HPLC for hemoglobinopathy screening under ambient storage conditions.
Main Methods:
- Cation exchange high-performance liquid chromatography (HPLC) was employed.
- Dried blood filter paper specimens were stored at room temperature.
- Hemoglobin types (e.g., Hb AA, Hb AS, Hb SS) were identified and quantified.
Main Results:
- Hemoglobin identification was successful for various hemoglobin types, including Hb AA, Hb AF, Hb AS, Hb FAS, Hb AJ, Hb FJ, Hb EF, and Hb SS, after six weeks of room temperature storage.
- Accurate hemoglobin quantitation was achievable from the same samples within three weeks of room temperature storage.
- The DBS-HPLC method demonstrated robustness for hemoglobin analysis over extended storage periods.
Conclusions:
- Dried blood filter paper specimens are stable for hemoglobin identification by HPLC for at least six weeks at room temperature.
- Accurate hemoglobin quantitation is possible from DBS within three weeks of room temperature storage.
- The combination of DBS and HPLC provides a reliable and accurate system for screening hemoglobinopathies, even with samples stored at ambient conditions.
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