Objectives: An HPLC method was developed to quantify serum concentrations of phenylalanine and tyrosine simultaneously using fluorescence detection without derivatization.
Methods: Serum protein is precipitated with trichloroacetic acid, 0.015mM dihydrogen-phosphate solution is used for separation on reversed-phase C18 material, and acetonitrile is avoided. Both amino acids are monitored utilizing their natural fluorescence at 210nm excitation and 302nm emission wavelengths.
Results: One analytical run is completed within 7min. Lower detection limit for Phe and Tyr is 0.3μM. Comparison of the new method with a classical HPLC method for total amino acids and using UV-absorption detection reveals a highly significant relationship for Phe and Tyr.
Conclusion: The new HPLC method allows rapid and very sensitive measurement of phenylalanine and tyrosine concentrations.
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