Neutral amino acids monitoring in phenylketonuric plasma microdialysates using micellar electrokinetic chromatography and laser-induced fluorescence detection.
Neutral and non-polar amino acids such as phenylalanine (Phe), valine (Val), tyrosine (Tyr), threonine (Thre) and GABA are hard to resolve by capillary zone electrophoresis (CZE). Their separation is possible by adding a surfactant to the mobile phase. This method is called micellar electrokinetic chromatography (MEKC). We used MEKC with laser-induced fluorescence detection (LIFD) to separate and quantitate these amino acids in plasma microdialysates of patients with phenylketonuria (PKU). This disease is an inborn enzymatic defect with decreased conversion of Phe to Tyr that causes severe neurological damage and mental deterioration, which is diagnosed by measuring plasma Phe and Phe/Tyr ratio. The amino acids tested had linear concentration-signal relation. PKU patients had significantly higher Phe, lower Tyr, 21 times higher Phe/Tyr ratio and decreased values of Val and Thre than controls. These results show that microdialysis of biological fluids coupled with MEKC-LIFD is a convenient technique to measure neutral amino acids in clinical disorders such as PKU.
Páez
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Laboratory of Behavioral Physiology, School of Medicine, Universidad de los Andes, Mérida, Venezuela. pacap@ing.ula.ve
Rada
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Hernández
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D000596:Amino Acids / Q000097:blood*; D020374:Chromatography, Micellar Electrokinetic Capillary / Q000379:methods*; D006801:Humans; D007834:Lasers; D017551:Microdialysis; D010661:Phenylketonurias / Q000097:blood*; D013050:Spectrometry, Fluorescence / Q000379:methods*
D000596:Amino Acids