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, 4.6×150 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 Column

Obelisc N

SIELC has developed the Obelisc™ columns, which are mixed-mode and utilize Liquid Separation Cell technology (LiSC™). These cost-effective columns are the first of their kind to be commercially available and can replace multiple HPLC columns, including reversed-phase (RP), AQ-type reversed-phase, polar-embedded group RP columns, normal-phase, cation-exchange, anion-exchange, ion-exclusion, and HILIC (Hydrophilic Interaction Liquid Chromatography) columns. By controlling just three orthogonal method parameters - buffer concentration, buffer pH, and organic modifier concentration - users can adjust the column properties with pinpoint precision to separate complex mixtures.

Select options
Application Analytes:
DOPA (3,4-dihydroxy-L-phenylalanine)
DOPAC (Dihydroxyphenylacetic Acid)
Homovanillic Acid

Application Detection:
UV Detection
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to so we can ensure you get optimal results for your compound/s of interest.