Liquid chromatography method for simultaneous analysis of amino acids and biogenic amines in biological fluids with simultaneous gradient of pH and acetonitrile.
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 microM for amino acids and in the range of 0.1-20 microM 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.
Lozanov
Valentin
V
Department of Chemistry and Biochemistry, Medical University of Sofia, 2 Zdrave Street, Sofia 1431, Bulgaria. lozanov@medfac.acad.bg
Benkova
Bistra
B
Mateva
Lyudmila
L
Petrov
Stefan
S
Popov
Elenko
E
Slavov
Chavdar
C
Mitev
Vanio
V
D000097:Acetonitriles; D000596:Amino Acids / Q000097:blood* / Q000652:urine*; D001679:Biogenic Amines / Q000097:blood* / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005449:Fluorenes / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration
D000097:Acetonitriles; D000596:Amino Acids; D001679:Biogenic Amines; D005449:Fluorenes; C032159:acetonitrile