Liquid chromatography method for simultaneous analysis of amino acids and biogenic amines in biological fluids with simultaneous gradient of pH and acetonitrile

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 860; no. 1; pp. 92 - 97
• Main Authors: Lozanov, Valentin, Benkova, Bistra, Mateva, Lyudmila, Petrov, Stefan, Popov, Elenko, Slavov, Chavdar, Mitev, Vanio
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01-12-2007; Elsevier Science
• Subjects: Acetonitriles; Amino acids; Amino Acids - blood; Amino Acids - urine; Analysis; Analytical, structural and metabolic biochemistry; Biogenic Amines - blood; Biogenic Amines - urine; Biological and medical sciences; Catecholeamines; Chromatography, High Pressure Liquid - methods; Fluorenes - chemistry; Fmoc; Fundamental and applied biological sciences. Psychology; General pharmacology; Humans; Hydrogen-Ion Concentration; Liquid chromatography; Medical sciences; Metanephrines; Pharmacology. Drug treatments; Polyamines; Amino acids; Catecholeamines; Liquid chromatography; Fmoc; Polyamines; Metanephrines; Biological fluid; Polyamine; Chemical analysis; pH gradient; Metadrenaline; Analysis method; Aminoacid; Biogenic amine; Simultaneous measurement
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2007.10.020

A liquid chromatography method for simultaneous analysis of amino acids, polyamines, catecholeamines and metanephrines in human body fluids after derivatization with 9-fluorenylmethyloxycarbonyl chloride was developed. The chromatographic behavior of analytes at different pH of mobile phase was studied. Successful baseline resolution of all analyzed compounds was achieved using simultaneous gradient of pH and organic modifier in reverse phase mode of HPLC within 36
min. The repeatability of the proposed procedure in respect of retention time and peak area, expressed as RSD, ranges from 0.06 to 1.64% and 0.4 to 7.6%, respectively. The method linearity in the range of 1–200
μM for amino acids and in the range of 0.1–20
μM for polyamines, catecholeamines and metanephrines was found to be with correlation coefficients higher than 0.994. The limit of quantification (LOQ) was assessed to be in the range of 2.6–10
pmol for amino acids and 2–4
pmol for polyamines, catecholeamines and metanephrines.