Analysis of Urinary Amino Acids by High-Performance Liquid Chromatography with Fluorescence Detection Using 2,3-Naphthalenedicarboxaldehyde as Fluorescence Derivatization Reagent

Amino acids are involved in various chemical reactions vivo, and changes several amino urine related to certain disease states. Therefore, developing an efficient method analyze the is useful timely diagnosis of diseases. In this study, we developed a high-performance liquid chromatography (HPLC) fluorescence for quantitative analysis urinary using derivatization reagent 2,3-naphthalenedicarboxaldehyde (NDA). NDA was selected because it does not require heating reaction can react within short time, rendering its use clinical settings feasible. The temperature, other conditions were optimized, found be completed 5 min at 25 °C. separation NDA–amino investigated on octadecylsilyl (ODS) column under gradient conditions. mobile phase mixture water–acetonitrile–trifluoroacetic acid. Eighteen (histidine (His), arginine (Arg), asparagine (Asn), glutamine (Gln), citrulline (Cit), serine (Ser), aspartic acid (Asp), threonine (Thr), glutamic (Glu), glycine (Gly), tyrosine (Tyr), alanine (Ala), tryptophan (Trp), valine (Val), phenylalanine (Phe), isoleucine (Ile), leucine (Leu), 5-aminovaleric (internal standard)) separated 100 optimal calibration curves showed good linearity range 0.25–25 pmol per injection with correlation coefficients >0.998. limits quantification 16.7–74.7 fmol. analytical applied human sample 16 (His, Arg, Asn, Gln, Cit, Ser, Thr, Glu, Gly, Tyr, Ala, Trp, Val, Phe, Ile, Leu) quantified. concentrations 5–960 μM. Urinary expected clinically applicable as novel biomarker diseases affecting bladder, tract, kidneys.