Simultaneous determination of phenylalanine and tyrosine in peripheral capillary blood by HPLC with ultraviolet detection

Phenylalanine (Phe) and tyrosine (Tyr) are the most reliable indicators for the diagnosis of phenylketonuria (PKU). The purpose of this study is to establish a simple and rapid method for the determination of Phe and Tyr in peripheral capillary blood from newborns and children by high performance li...

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Bibliographic Details
Published in:Clinical biochemistry 2013-08, Vol.46 (12), p.1074-1078
Main Authors: Mo, Xi-ming, Li, Ying, Tang, Ai-guo, Ren, Ya-ping
Format: Article
Language:English
Subjects:
Anticoagulants - pharmacology
Calibration
Child
Child, Preschool
Chromatography, High Pressure Liquid - methods
Female
Health
High performance liquid chromatography
Humans
Infant
Infant, Newborn
Male
Phenylalanine
Phenylalanine - blood
Phenylketonuria
Reference Standards
Temperature
Time Factors
Tyrosine
Tyrosine - blood
Ultraviolet detection
Ultraviolet Rays
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Summary:Phenylalanine (Phe) and tyrosine (Tyr) are the most reliable indicators for the diagnosis of phenylketonuria (PKU). The purpose of this study is to establish a simple and rapid method for the determination of Phe and Tyr in peripheral capillary blood from newborns and children by high performance liquid chromatography with ultraviolet detection (HPLC-UV). Peripheral blood specimens were taken from newborns or children by heel stick or skin puncture on the fingers. Blood samples were deproteinized by equal volume of 5% perchloric acid. Amino acid separation was carried out on a Hypersil C8 column. The mobile phase containing 5% acetonitrile (v/v) was run at a flow rate of 1.5mL/min. Phe and Tyr were determined in the ultraviolet detector at 210nm. The determination was within 10.0min. The linear range was from 6.0 to 1512.0μmol/L for Phe and 5.5 to 1250.0μmol/L for Tyr. Although Phe and Tyr levels from peripheral blood samples were relatively lower than those from serum or plasma, the data showed a good agreement between them. The method we developed is simple, rapid and convenient. It is useful for screening and diagnosis of PKU in newborns and children. •We establish a method for determination of Phe and Tyr in peripheral blood by HPLC-UV.•This method is suitable for diagnosing PKU and monitoring PKU therapeutic efficacy.•There were good correlation between Phe and Tyr levels in plasma and capillary blood.
ISSN:0009-9120
1873-2933
DOI:10.1016/j.clinbiochem.2013.05.047