HPLC Separation of Neurotransmitters

The neurotransmitters dl-DOPA, creatinine, hydroxytryptophan (5-HTP), and serotonin are separated in less than 4 minutes on a short 50 mm Primesep 200 column. The HPLC separation uses a mobile phase of water, acetonitrile (MeCN, ACN) and trifluoroacetic acid (TFA) and UV detection at 250 nm. Ion-pair reagents were not needed for retention of these polar, hydrophilic compounds, instead, a combination of ion-exchange and reversed-phase interactions were used.


Column Primesep 200, 4.6x50 mm, 5 µm, 100A
Mobile Phase MeCN/H2O - 80/20%
Buffer TFA - 0.05%
Flow Rate 1.0 ml/min
Detection UV, 250 nm


Class of Compounds Drug, Acid, Hydrophilic, Ionizable, Hormone
Analyzing Compounds dl-DOPA, Creatinine, Hydroxytryptophan (5-HTP), Serotonin
Application Analytes:
DOPA (3,4-dihydroxy-L-phenylalanine)