HPLC Separation of Biogenic Amines on Primesep 100 Column

HPLC Method for Tryptophan, Serotonin, N,N-Dimethyltryptamine, Tryptamine, N,N-Dimethyl-5-methoxytryptamine, N,N-dimethyltryptophan on Primesep 100 by SIELC Technologies

High Performance Liquid Chromatography (HPLC) Method for Analysis of Tryptophan, Serotonin, N,N-Dimethyltryptamine, Tryptamine, N,N-Dimethyl-5-methoxytryptamine, N,N-dimethyltryptophan.

Tryptophan is an essential amino acid used in the synthesis of proteins. It cannot be produced by the body and therefore must be consumed through a diet of meats, eggs, dairy, seeds, and nuts. The proteins that it helps synthesize are important for muscle growth and repair. Tryptophan is also converted into serotonin, which is responsible for mood, appetite, and pain reception. It is further down converted into melatonin. Tryptophan is also used in treatments for severe PMS symptoms, depression, and insomnia. It is naturally found in red meat, poultry eggs, and dairy. It’s chemical formula is C11H12N2O2. You can find detailed UV spectra of Tryptophan and information about its various lambda maxima by visiting the following link.

Tryptamine is an indolamine derived from tryptophan with the chemical formula  C10H12N2.  It is naturally found in plants, fungi, and animals. It helps with regulating the activity of dopaminergic, serotonergic, and glutamatergic systems. You can find detailed UV spectra of Tryptamine and information about its various lambda maxima by visiting the following link.

Serotonin is a neurotransmitter derived from tryptophan with the chemical formula  C₁₀H₁₂N₂O. Serotonin is a chemical messenger that is crucial for bodily functions. It helps regulate mood and reduce anxiety, digestion, blood clotting, and contributes to the sleep-wake cycle. Low serotonin levels are often associated with a variety of mental illnesses including but not limited to Obsessive Compulsive Disorder (OCD), Post-Traumatic Stress Disorder (PTSD), Depression, and Anxiety.

Dimethyltryptamine is a psychedelic substance derived from tryptamine with the chemical formula C12H16N2. It works as an agonist for Serotonin. It is not approved for medical use. Due to it’s high risk of abuse, it is a Schedule I drug in the United States.

5-MeO-DMT, also known as 5-methoxy-N,N-dimethyltryptamine), O-methylbufotenin, and mebufotenin, is a psychedelic substances derived from tryptamine found in plants and toad secretions. Between 1970s to 1990s, a certain religious sect believed the use of mebufotenin to be sacred to their religious practice. It is currently considered a Schedule I controlled substance in the United States, however.

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.

Condition

Column Primesep 100, 4.6 x 150 mm, 5 µm, 100 A, dual ended
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

 

Description

Class of Compounds
Drug, Hydrophilic, Supplements, Monoamine,  Neurotransmitter
Analyzing Compounds Tryptophan, Serotonin, N,N-Dimethyltryptamine, Tryptamine, N,N-Dimethyl-5-methoxytryptamine, N,N-dimethyltryptophan

 

Application Column

Primesep 100

Column Diameter: 4.6 mm
Column Length: 150 mm
Particle Size: 5 µm
Pore Size: 100 A
Column options: dual ended

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Application Analytes:
N,N-Dimethyl-5-methoxytryptamine
N,N-Dimethyltryptamine
N,N-dimethyltryptophan
Serotonin
Tryptamine
Tryptophan

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 research@sielc.com so we can ensure you get optimal results for your compound/s of interest.