HPLC Separation of Biogenic Amines on Primesep 100 Column

Tryptophan is an essential amino acid used in the synthesis of proteins. Tryptamine is structurally similar to tryptophan. Serotonin is a neurotransmitter derived from tryptophan. Dimethyltryptamine and 5-MeO-DMT are psychedelic substances derived from tryptamine. All the compounds have similar structures and can present difficulties to separation in reverse-phase HPLC. They can be separated using the Primesep 100 mixed-mode column in gradient analysis with acetonitrile (ACN) and water mobile phase with ammonium formate (AmFm) buffer, making the method MS-compatible. The amines can also be UV detected at 280nm.


Column Primesep 100, 4.6x150 mm, 5 µm, 100A
Mobile Phase Gradient MeCN - 40-50%, 15 min
Buffer Gradient AmFm pH 3.0- 20-80 mM, 15 min
Flow Rate 1.0 ml/min
Detection UV 280 nm,  MS-compatible mobile phase


Class of Compounds Drug, Hydrophilic, Supplements, Monoamine,  Neurotransmitter
Analyzing Compounds Serotonin, Tryptophan, N,N-dimethyltryptophan, N,N-Dimethyl-5-methoxytryptamine, Tryptamine, N,N-dimethyltryptamine
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Application Analytes: