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Quantitative analysis of amino acids by HPLC in dried blood and urine in the neonatal period: Establishment of reference values
Author(s) -
Nolasco Daniela M.,
Fortes Isabel C. P.,
Valadares Eugênia R.
Publication year - 2020
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.4931
Subject(s) - chromatography , urine , repeatability , chemistry , high performance liquid chromatography , filter paper , amino acid , quantitative analysis (chemistry) , extraction (chemistry) , detection limit , dried blood , sample preparation , amino acid analysis , biochemistry
Quantitative analysis of amino acids in blood and urine is primarily indicated for the diagnosis of amino acid disorders. The high‐performance liquid chromatography (HPLC) technique is frequently used for this detection. The frequency of sample collection on filter paper has been increasing exponentially, and there are many advantages attributed to processing biological samples in this way. The aim of this study was to validate a quantitative analysis of amino acids by HPLC in blood and urine collected on filter paper and to establish reference values in the neonatal period. Dried blood and dried urine samples of respectively 58 and 45 healthy newborns (2–9 days) were collected. Pre‐treatment and extraction of samples were done according to the literature. Separation and analysis of amino acids were carried out by HPLC with fluorescence detection. The developed method demonstrated excellent separation, linearity, limits of detection and quantification, repeatability and recovery. The reference values for 17 amino acids were defined in dried blood and urine samples of newborns. This work presents a simple, fast and effective method for the simultaneous analysis of 17 amino acids in blood and urine collected on filter paper in a single run. The reference values were established and validated.