Separation type: Liquid Chromatography Mixed-mode

High Performance Liquid Chromatography (HPLC) Method for Analysis of Dopamine and Serotonin

Dopamine is a key neurotransmitter and medical stimulant used to treat low blood pressure, low heart rate, and heart attacks. Serotonin is another key neurotransmitter that has pharmaceutical applications – namely as the key ingredient in antidepressant and anti-anxiety medications.These two neurotransmitters can be retained, separated, and analyzed on a Primesep 200 mixed-mode column using an isocratic analytical method with a simple mobile phase of water, Acetonitrile (MeCN), and an Ammonium Formate (AmFm) buffer. This analysis method can be UV detected at 280 nm with high resolution and peak symmetry.

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.

HPLC Method for Serotonin on Primesep 200 by SIELC Technologies

High Performance Liquid Chromatography (HPLC) Method for Analysis of Serotonin.

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.

Serotonin can be retained and analyzed using the Primesep 200 stationary phase column. The analysis utilizes an isocratic method with a simple mobile phase consisting of water and acetonitrile (MeCN) with an ammonium formate buffer. Detection is performed using UV.

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.

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.).

Dopamine, norphenylephrine and serotonin are separated on Primesep P HPLC column. Base line separation of these neurotransmitters can be achieved using mix-mode approach. Primesep P provides retention by reverse phase, pi-pi and cation-exchange mechanisms. Method can be used for fast quantitation of these compounds in various sample matrices. Detection technique ranges from UV to ELSD, LC/MS, RI and CAD. Primesep P column can be used with tetrahydrofuran or with 100% aqueous mobile phase to enhance pi-pi interaction.