University of North Florida
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Stuart Chalk, Ph.D.
Department of Chemistry
University of North Florida
Phone: 1-904-620-1938
Fax: 1-904-620-3535
Website: @unf

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Infant Blood

Classification: Biological fluid -> blood -> baby

Citations 1

"HPLC Assay Of Phenylalanine And Tyrosine In Blood Spots On Filter-paper"
Clin. Chim. Acta 1986 Volume 156, Issue 1 Pages 91-96
R. August Roesel, Paul R. Blankenship and Frits A. Hommes

Abstract: Capillary blood was applied to filter-paper (S. and S. No. 903) and dried at room temperature A disc (6.4 mm diameter) was punched from the paper and placed in 0.5 mL of 60% ethanol containing 3 µL mL-1 of 30% Brij 35 and 20 mM in 4-amino-3-hydroxybutyric acid (internal standard). The mixture was shaken for 2 h at 25°C and the extracted amino-acids were analyzed by HPLC on a column (15 cm x 4 mm) of Dionex ion-exchange resin DC-4A operated at 60°C with 0.554 M Li citrate buffer (pH 3.97) as mobile phase (0.3 mL min-1). The eluate was subjected to post-column derivatization with phthalaldehyde and subsequent fluorimetric detection at 450 nm (excitation at 350 nm). The calibration graph for phenylalanine was rectilinear up to 2 nmol with a limit of detection of 20 pmol and a recovery of 98 to 104%; the analysis time was 20 min. The coefficient of variation were 3.4 and 3.3% for tyrosine and phenylalanine, respectively; other amino-acids did not interfere with the determination. The method has been applied to the screening of newborn babies for phenylketonuria.
Phenylalanine Tyrosine Clinical analysis HPLC Fluorescence Heated reaction Interferences Post-column derivatization