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Chromatogram

Description

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.

Method Parameters

Mobile PhaseMeCN/H2O – 80/20%
BufferTFA – 0.05%
Flow Rate1.0 ml/min
DetectionUV, 250 nm
Class of CompoundsDrug, Acid, Hydrophilic, Ionizable, Hormone
Analyzing Compoundsdl-DOPA, Creatinine, Hydroxytryptophan (5-HTP), Serotonin

HPLC Column Used

Primesep 200, 4.6×50 mm, 5 µm, 100A