HPLC‐FL/ED in the Analysis of Biogenic Amines and their Metabolites in Urine

Abstract The analysis of biogenic amine neurotransmitters (adrenaline, noradrenaline, dopamine, serotonin) and their metabolites (metanephrine, normetanephrine, 3,4‐dihydroxyphenylacetic acid, vanillylmandelic acid, homovanillic acid, 5‐hydroxyindoleacetic acid, 3,4‐dihydroxyphenylglycol, 3‐methoxy‐4‐hydroxyphenylglycol, 3‐methoxytyramine) in body fluids may provide valuable information important in the diagnosis of the sympathoadrenomedullary system disturbances. An HPLC‐FL/ED system using two Chromolith RP‐18e columns and a mixture of acetate buffer (pH=4.66) and methanol (97∶3%, v/v) as the mobile phase was developed for the separation and determination of these compounds. Adrenaline, noradrenaline, dopamine and their synthetic analogs: dobutamine and isoproterenol, used in the treatment of clinical shock, were analyzed in urine of post‐operative cardiosurgical patients and in urine of healthy controls. The extraction and pre‐concentration step was performed on aluminium oxide SPE cartridges. Both detection techniques agreed well in final concentrations. Within‐day coefficients of variation were below 2.3% and between‐day coefficients of variation did not exceed 5%. The developed HPLC method is easy and reliable and could have a positive effect on the safety of severely ill patients treated with catecholamines and their synthetic analogues. Keywords: Biogenic amine neurotransmittersHPLC‐FL/EDSPEurine Acknowledgements This work was supported by the State Committee for Scientific Research, Project No. 4 T09A 08922.