Separation of Serotonin, Dopamine, and Related Compounds

Catecholamines are chemical compounds derived from the amino acid tyrosine containing catechol and amine groups. Some of them are biogenic amines. Retention of compounds of the catecholamine pathway is achieved on Obelisc N column. All polar compounds are well retained by combination of HILIC and ion-exchange mechanisms. Obelisc N columns produce very good peak shapes for all analytes. The method is very sensitive to amount of ACN, buffer and buffer pH. The retention time changes with variation of the main parameters. This method can be used for quantitation of biogenic amines and related compounds (homovanillic acid, dihydroxyphenyl acetic acid, serotonin, dopamine, epinephrine, hydroxytryptophan, epinephrine and DOPA) in urine, blood and other biological fluids. Further optimization of this HPLC method can be used during screening and validation. Amines and acids can be analyzed in the same run and retained by a combination of polar organic mode, cation-exchange and anion-exchange modes. Various buffers within specified pH can be employed (ammonium formate, ammonium acetate, sodium phosphate, etc.).


Column Obelisc N, 4.6x150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Buffer AmAc pH 4.0- 30 mM
Flow Rate 1.0 ml/min
Detection UV, 270 nm


Class of Compounds Drug, Acid, Monocarboxylic acid,  Hydrophilic, Ionizable, Hormone
Analyzing Compounds Homovanillic acid, Dihydroxyphenyl acetic acid, Serotonin, Dopamine, Epinephrine, Hydroxytryptophan, DOPA
Application Analytes:
DOPA (3,4-dihydroxy-L-phenylalanine)
DOPAC (Dihydroxyphenylacetic Acid)
Homovanillic Acid