Dopamine


Dopamine structural formula

HPLC separation of Neurotransmitters with Primesep 200

CAS Number51-61-6
Molecular FormulaC8H11NO2
Molecular Weight153.181
InChI KeyVYFYYTLLBUKUHU-UHFFFAOYSA-N
LogP-0.980
Synonyms
  • Dopamine
  • 4-(2-Aminoethyl)benzene-1,2-diol
  • 1,2-Benzenediol, 4-(2-aminoethyl)-
  • 51-61-6
  • 1,2-Benzenediol, 4-(2-aminoethyl)-
  • 2-(3,4-Dihydroxyphenyl)ethylamine
  • 3,4-Dihydroxyphenethylamine
  • 3,4-Dihydroxyphenylethylamine
  • 3-Hydroxytyramine
  • 4-(2-Aminoethyl)-1,2-benzenediol
  • 4-(2-Aminoethyl)catechol
  • 4-(2-Aminoethyl)pyrocatechol
  • Dopamin
  • dopamina
  • Dophamine
  • Hydroxytyramin
  • NSC 173182
  • Oxytyramine
  • Pyrocatechol, 4-(2-aminoethyl)-
  • α-(3,4-Dihydroxyphenyl)-β-aminoethane
  • alpha-(3,4-Dihydroxyphenyl)-beta-aminoethane
  • EINECS 200-110-0
  • UNII-VTD58H1Z2X
  • 4-(2-Aminoethyl)-Pyrocatechol
  • 4-(2-azanylethyl)benzene-1,2-diol
  • Deoxyepinephrine
  • Dopaminum
  • Dopastat
  • Dynatra
  • Hydroxytyramine
  • Intropin
  • Revivan
  • a-(3,4-Dihydroxyphenyl)-b-aminoethane
  • alpha-(3,4-Dihydroxyphenyl)-beta-aminoethane

Applications:

HPLC Method for Separation of Amines in Non Aqueous MP on BIST B+ Column

March 1, 2023

HPLC Method for Separation of of Amines in Non Aqueous MP on BIST B+ by SIELC Technologies

Separation type: Bridge Ion Separation Technology, or BIST™ by SIELC Technologies

HPLC Method for Separation of of Amines in Non Aqueous MP on BIST B+ Column by SIELC Technologies
HPLC Method for Separation of of Amines in Non Aqueous MP on BIST B+ by SIELC Technologies

Dopamine is a key neurotransmitter and medical stimulant used to treat low blood pressure, low heart rate, and heart attacks. m-Xylylenediamine (MXDA) is a popular curing agent used on epoxy resins. 2,4,6-Tris(dimethylaminomethyl)phenol is another amine-based curing agent used on epoxy resins. Using SIELC’s newly introduced BIST method, these 3 amines can be retained on a positively-charged anion-exchange BIST B+ column. There are two keys to this retention method: 1) a multi-charged, negative buffer, such as Sulfuric acid (H2SO4), which acts as a bridge, linking the positively-charged analytes to the positively-charged column surface and 2) a mobile phase consisting mostly of less polar organic solvent (such as IPA) to minimize the formation of a solvation layer around the charged analytes. This method uses an entirely non-aqueous mobile phase to drive BIST retention. The gradient starts with a high concentration of the less polar IPA to generate the initial BIST retention and progresses to the more polar MeOH to elute the longer-retaining amines in a reasonable time. Using this new and unique analysis method, these 3 amines can be retained and UV detected at 210 nm.

High Performance Liquid Chromatography (HPLC) Method for Analysis of of Amines in Non Aqueous MP

Condition

ColumnBIST B+, 4.6 x 150 mm, 5 µm, 100 A
Mobile PhaseGradient IPA/MeOH- 80/20% to 100% MeOH, 5 min , 5 min hold
BufferH2SO4 – 0.2%
Flow Rate1.0 ml/min
DetectionUV 210 nm
Peak Retention Time3.52 min, 6.12 min, 8.57 min

Description

Class of CompoundsAmines
Analyzing CompoundsDopamine, m-Xylylenediamine, 2,4,6-Tris(dimethylaminomethyl)phenol

Application Column

BIST B+

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

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Application Analytes:
2,4,6-Tris(dimethylaminomethyl)phenol
Dopamine
m-Xylylenediamine

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.

HPLC Method For Analysis Of Dopamine and Serotonin on Primesep 200 Column

March 25, 2022

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.

 

Condition

Column Primesep 200, 4.6×150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O – 20/80%
Buffer Ammonium Formate pH 3.0 – 40 mM
Flow Rate 1.0 ml/min
Detection UV, 280 nm,

 

Description

Class of Compounds
Neurotransmitters
Analyzing Compounds  Dopamine, Serotonin

 

Application Column

Primesep 200

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

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Application Analytes:
Dopamine
Serotonin
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.

HPLC Separation of Catecholamines on Newcrom AH Column

May 20, 2021

Separation type: Liquid Chromatography Mixed-mode



 




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Catecholamines are neurotransmitters that generate the ‘fight-or-flight’ responsein the body. The 3 main catecholamines are epinephrine (also known as adrenaline), norepinephrine, and dopamine, and they are produced by the adrenal glands.
These 3 catecholamines can be detected in the low UV regime. Using a Newcrom AH mixed-mode column and a mobile phase consisting of almost entirely water with either a phosphoric (H3PO4) acid or ammonium formate (AmFm) buffer, the catecholamines can be retained, separated, and measured. This analysis method can be UV detected at 275 nm with high resolution. The latter mobile phase (utilizing AmFm) is compatible with Mass Spectrometry.



Condition

Column Newcrom AH, 4.6×150 mm, 5 µm, 100A
Mobile Phase H2O
Buffer H3PO4  or AmFm pH 3.0
Flow Rate 1.0 ml/min
Detection UV, 275 nm

 

Description

Class of Compounds
Drug, Acid, Hydrophilic, Ionizable, Hormone
Analyzing Compounds  Norepinephrine, Epinephrine, Dopamine

 

Application Column

Newcrom AH

The Newcrom columns are a family of reverse-phase-based columns. Newcrom A, AH, B, and BH are all mixed-mode columns with either positive or negative ion-pairing groups attached to either short (25 Å) or long (100 Å) ligand chains. Newcrom R1 is a special reverse-phase column with low silanol activity.

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Application Analytes:
Dopamine
Epinephrine
Norepinephrine
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.

HPLC Determination of Dopamine in Human Plasma Sample

May 10, 2019

 


FlipLC™ is an alternative method to avoid the interference of most of the contaminants by the use of an isolation column and a high pressure switching valve before the separation column. This method allows sample cleaning and analyte separation in one automated process. The isolation column and the separation column should have orthogonal retention characteristics to operate efficiently in this setup. Mixed-mode columns with reverse phase and ion-exchange characteristics were used in this analysis.

Application Column

Primesep 200

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

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Primesep SB

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

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Application Analytes:
Dopamine
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.

HPLC Determination of Dopamine in Serum

May 6, 2019


FlipLC™ is an alternative method to avoid the interference of most of the contaminants by the use of an isolation column and a high pressure switching valve before the separation column. This method allows sample cleaning and analyte separation in one automated process. The isolation column and the separation column should have orthogonal retention characteristics to operate efficiently in this setup. Mixed-mode columns with reverse phase and ion-exchange characteristics were used in this analysis.

Condition

Column Primesep 200, 4.6×150 mm, 5 µm, 100A

 

Primesep SB, 4.6*50 mm, 5 µm, 100A

Mobile Phase MeCN/H2O
Buffer AmFm pH 3.0
Flow Rate  1.0 ml/min
Detection UV, 280 nm

Description

Class of Compounds
Drug, Hydrophilic, Ionizable, Hormone
Analyzing Compounds Dopamine

Application Column

Primesep 200

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

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Primesep SB

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

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Application Analytes:
Dopamine
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.

HPLC Separation of DOPAC, Homovanillic Acid and Dopamine on the Primesep 100 Column

July 10, 2012

 

Application Notes: Neurotransmitters DOPAC, homovanillic acid and dopamine were separated by mixed-mode chromatography on Primesep 100 and Primesep 200 HPLC columns. The method can be used for quantification of neurotransmitters with LC/MS compatible conditions. The compounds are retained by combination of reversed-phase, ion-exchange or ion-exclusion mechanisms. The retention time and selectivity of separation can be adjusted by variation of amount of acetonitrile, buffer pH and buffer concentration.

 
Application Columns: Primesep 100

Application compounds: Dopac, Homovanillic Acid, Dopamine

Detection technique: UV, LC/MS, ELSD/CAD

Condition

Column Primesep 100, 4.6×50 mm, 5 µm, 100A
Mobile Phase MeCN/H2O – 5/95%
Buffer AmFm pH 3.0- 30 mM
Flow Rate 1.0 ml/min
Detection UV, 270 nm

 

Description

Class of Compounds
Drug, Acid, Monocarboxylic acid,  Hydrophilic, Ionizable, Hormone
Analyzing Compounds DOPAC, Homovanillic Acid, Dopamine

 

Application Column

Primesep 100

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

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Primesep 200

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

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Application Analytes:
DOPAC (Dihydroxyphenylacetic Acid)
Dopamine
Homovanillic Acid

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.

HPLC Separation of DOPAC, Homovanillic Acid in Dopamine on the Primesep 200 Column

July 10, 2012

 

Application Notes: Neurotransmitters DOPAC, homovanillic acid and dopamine were separated by mixed-mode chromatography on Primesep 100 and Primesep 200 HPLC columns. The method can be used for quantification of neurotransmitters with LC/MS compatible conditions. The compounds are retained by combination of reversed-phase, ion-exchange or ion-exclusion mechanisms. The retention time and selectivity of separation can be adjusted by variation of amount of acetonitrile, buffer pH and buffer concentration.
Application Columns: Primesep 100, Primesep 200
Application compounds: Dopac, Homovanillic Acid, Dopamine
Detection technique: UV, LC/MS, ELSD/CAD

Condition

Column Primesep 200, 4.6×50 mm, 5 µm, 100A
Mobile Phase MeCN/H2O – 5/95%
Buffer AmFm pH 3.0- 30 mM
Flow Rate 1.0 ml/min
Detection UV, 270 nm

 

Description

Class of Compounds
Drug, Acid, Monocarboxylic acid,  Hydrophilic, Ionizable, Hormone
Analyzing Compounds DOPAC, Homovanillic Acid, Dopamine

 

Application Column

Primesep 100

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

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Primesep 200

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

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Application Analytes:
DOPAC (Dihydroxyphenylacetic Acid)
Dopamine
Homovanillic Acid
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.

USP Analysis of Dopamine Using a Legacy L1 column

June 21, 2012

 

 

Application Notes: Dopamine is a naturally occurring neurotransmitter found in the brain. Dopamine is a well studied compound because dopamine is an important neurotransmitter known to regulate many functions from pain response to behavior disorders. According to USP methods, dopamine hydrochloride contains not less than 98% and not more than 102% dopamine hydrochloride calculated on the dried basis. The USP HPLC method for the separation of hydrocortisone was developed on Legacy L1 column according to the US Pharmacopeia methodology. L1 classification is assigned to reversed-phase HPLC column containing C18 ligand. Support for the material is spherical silica gel with particles size 3-10 um and pore size of 100-120A. Resolution between critical pairs corresponds to rules and specifications of USP.

Application Columns: Legacy L1 C18 HPLC column

Application compounds: Dopamine hydrochloride

Mobile phase: Water with 1% acetic and 5mM octanesulfonic acid/MeCN

Detection technique: UV

Reference: USP35: NF30

 

Condition

Column Legacy L1, 4.6×150 mm, 5 µm, 100A
Mobile Phase Water with 1% Acetic and 5 mM Octanesulfonic acid/ MeCN ( 87-13)
Flow Rate 1.0 ml/min
Detection UV, 270 nm

 

Description

Class of Compounds
Drug,  Hydrophilic, Ionizable, Hormone, Acid
Analyzing Compounds Dopamine

 

Application Column

SIELC Legacy L1 HPLC column

Legacy L1

SIELC's family of Legacy columns is based on the United States Pharmacopeia's (USP) published chromatographic methods and procedures. Numerous brands have columns used in USP reference standards and methods. USP has created various designations to group together columns with similar types of packing and properties in the solid phase. SIELC's Legacy columns adhere to these strict requirements and properties, allowing you to easily replace older columns that are no longer available without needing to significantly modify your method or SOPs.

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Application Analytes:
Benzoic Acid
Dopamine

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.

HPLC Separation of Homovanillic Acid, Dopamine, and DOPAC using Hydrogen Bonding

June 18, 2012

Condition

Column Sharc 1, 3.2×50 mm, 5 µm, 100A
Mobile Phase Gradient MeCN/MeOH – 100/0 – 30/70%
Buffer Formic Acid – 0.5% AmFm 0.05% 7 min
Flow Rate 1.0 ml/min
Detection UV, 270 nm

 

Description

Class of Compounds
Drug, Acid, Monocarboxylic acid,  Hydrophilic, Ionizable, Hormone
Analyzing Compounds DOPAC, Homovanillic Acid, Dopamine

 

Application Column

SHARC 1

The SHARC™ family of innovative columns represents the first commercially available columns primarily utilizing separation based on hydrogen bonding. SHARC stands for Specific Hydrogen-bond Adsorption Resolution Column. Hydrogen bonding involves an interaction or attraction between a bound hydrogen atom and molecules containing electronegative atoms, such as oxygen, nitrogen, and fluorine.

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Application Analytes:
DOPAC (Dihydroxyphenylacetic Acid)
Dopamine
Homovanillic Acid

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.

HPLC Separation of Dopamine and Epinephrine in HILIC and Cation-Exchange Modes

July 14, 2011

chr_305.gif

Dopamine and epinephrine were separated on a Primesep S2 HILIC mixed-mode column. Dopamine and epinephrine are retained by HILIC and cation-exchange mechanisms. Retention time is controlled by the amount of ACN, buffer and buffer pH. Method is compatible with LC/MS and can be used for analysis of polar metabolites in biofluids.

Condition

Column Primesep S2, 4.6×150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Buffer AmFm
Flow Rate 1.0 ml/min
Detection UV, 250 nm

 

Description

Class of Compounds
Drug, Acid, Monocarboxylic acid,  Hydrophilic, Ionizable, Hormone
Analyzing Compounds Dopamine, Epinephrine

 

Application Column

Primesep S2

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

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Application Analytes:
Dopamine
Epinephrine

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.

HPLC Separation of Catecholamines on Primesep 100 Column with Phosphate Buffers

July 8, 2011

chr_284.gif

The catecholamine neurotransmitters are amino-acid derivatives of tyrosine. DOPA, tyrosine, phenylalanine, norepinephrine, epinephrine, and dopamine and baseline are resolved on a Primesep 100 column with UV-transparent phosphate buffer. This method can be used for analysis of catecholamines and related impurities in various matrices. Peak order and retention time can be changed by changing the amount of ACN, buffer concentration and buffer pH. Various buffers can be used to accommodate desired detection technique. Primesep 100 is a reversed-phase cation-exchange mixed-mode column that can be used for analysis of polar neutral, polar ionizable, polar zwitter-ionic, hydrophobic neutral, and hydrophobic ionic compounds in the same run. Column can be operated in reverse-phase, cation-exchange, anion-exclusion, HILIC and mixed-modes depending on the mobile phase selection and nature of analytes. Column is compatible with LC/MS and does not require use of ion-pairing reagents.

Condition

Column Primesep 100, 4.6×150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Buffer Na2HPO4
Flow Rate 1.0 ml/min
Detection UV, 210 nm

 

Description

Class of Compounds
Drug, Acid, Hydrophilic, Ionizable, Hormone
Analyzing Compounds Tyrosine, DOPA, Phenylalanine, Norepinephrine, Epinephrine, Dopamine

 

Application Column

Primesep 100

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

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Application Analytes:
DOPA (3,4-dihydroxy-L-phenylalanine)
Dopamine
Epinephrine
Norepinephrine
Phenylalanine
Tyrosine

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.

HPLC Analysis of Polar Basic and Acetic Compounds on Primesep AB Column

May 6, 2010


Mixture of polar acidic and basic compounds is separated on a Primesep AB mixed-mode HPLC column. Dopamine and tyrosine are retained by combination of reversed-phase and cation-exchange mechanisms. Maleic acid is retained by anion-exchange mechanism, and benzoic acid is retained by reversed-phase mechanism. Primesep AB is a trimodal column with a C12 hydrophobic chain and cation-exchange and anion exchange groups on the surface. Method utilizes UV detection but can be used with other detection techniques (ELSD, LC/MS, Corona).

Condition

Column Primesep AB, 4.6×150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Buffer TFA – 0.1%
Flow Rate 1.0 ml/min
Detection UV, 210 nm

 

Description

Class of Compounds
Drug, Acid, Monocarboxylic acid,  Hydrophilic, Ionizable, Hormone
Analyzing Compounds Tyrosine, Dopamine, Maleic Acid, Benzoic Acid

 

Application Column

Primesep AB

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

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Application Analytes:
Benzoic Acid
Dopamine
Maleic Acid
Tyrosine

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.

HPLC Separation of Polar Compounds

January 13, 2010


The separation of amino acids, the building blocks of proteins, can be challenging to separate on a reverse-phase column due to their high polarity. Using a mixed-mode HPLC column, allows the separation of amino acids by cation-exchange and ion-exclusion mechanisms as well as hydrophobicity. Fine tuning of separation can be achieved with changes in organic concentration of the mobile phase as well as choice of buffer and pH.

Condition

Column Primesep 200, 4.6×150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Buffer AcOH, AmFm
Flow Rate 1.0 ml/min
Detection ELSD

 

Description

Class of Compounds
Drug, Acid, Hydrophilic, Ionizable, Hormone
Analyzing Compounds Aspartic acid, Glutaric acid, Glycine, Hydroxytriptophan, GABA, Norepinephrine, Dopamine

 

Application Column

Primesep 200

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

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Application Analytes:
5-Hydroxytryptophan
Aspartic Acid
Dopamine
Glutamic Acid
Norepinephrine
gamma-Aminobutyric Acid (GABA)

Application Detection:
ELSD 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.

Separation of Serotonin, Dopamine, and Related Compounds

August 22, 2008

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.).

Condition

Column Obelisc N, 4.6×150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Buffer AmAc pH 4.0- 30 mM
Flow Rate 1.0 ml/min
Detection UV, 270 nm

 

Description

Class of Compounds
Drug, Acid, Monocarboxylic acid,  Hydrophilic, Ionizable, Hormone
Analyzing Compounds Homovanillic acid, Dihydroxyphenyl acetic acid, Serotonin, Dopamine, Epinephrine, Hydroxytryptophan, DOPA

 

Application Column

Obelisc N

SIELC has developed the Obelisc™ columns, which are mixed-mode and utilize Liquid Separation Cell technology (LiSC™). These cost-effective columns are the first of their kind to be commercially available and can replace multiple HPLC columns, including reversed-phase (RP), AQ-type reversed-phase, polar-embedded group RP columns, normal-phase, cation-exchange, anion-exchange, ion-exclusion, and HILIC (Hydrophilic Interaction Liquid Chromatography) columns. By controlling just three orthogonal method parameters - buffer concentration, buffer pH, and organic modifier concentration - users can adjust the column properties with pinpoint precision to separate complex mixtures.

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Application Analytes:
DOPA (3,4-dihydroxy-L-phenylalanine)
DOPAC (Dihydroxyphenylacetic Acid)
Dopamine
Epinephrine
Homovanillic Acid
Hydroxytryptophan
Norepinephrine
Serotonin

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.

HPLC Separation of Polar and Hydrophobic Drugs on Obelisc R and N

March 3, 2007

Condition

Column Obelisc N, Obelisc R 4.6×150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Buffer H3PO4
Flow Rate 1.0 ml/min
Detection UV, 220 nm

 

Description

Class of Compounds
Drug, Acid, Preservative,  Hydrophilic, Ionizable, Hormone
Analyzing Compounds Propyl paraben,  Dopamine, DOPA

 

Application Column

Obelisc R

SIELC has developed the Obelisc™ columns, which are mixed-mode and utilize Liquid Separation Cell technology (LiSC™). These cost-effective columns are the first of their kind to be commercially available and can replace multiple HPLC columns, including reversed-phase (RP), AQ-type reversed-phase, polar-embedded group RP columns, normal-phase, cation-exchange, anion-exchange, ion-exclusion, and HILIC (Hydrophilic Interaction Liquid Chromatography) columns. By controlling just three orthogonal method parameters - buffer concentration, buffer pH, and organic modifier concentration - users can adjust the column properties with pinpoint precision to separate complex mixtures.

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Obelisc N

SIELC has developed the Obelisc™ columns, which are mixed-mode and utilize Liquid Separation Cell technology (LiSC™). These cost-effective columns are the first of their kind to be commercially available and can replace multiple HPLC columns, including reversed-phase (RP), AQ-type reversed-phase, polar-embedded group RP columns, normal-phase, cation-exchange, anion-exchange, ion-exclusion, and HILIC (Hydrophilic Interaction Liquid Chromatography) columns. By controlling just three orthogonal method parameters - buffer concentration, buffer pH, and organic modifier concentration - users can adjust the column properties with pinpoint precision to separate complex mixtures.

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Application Analytes:
DOPA (3,4-dihydroxy-L-phenylalanine)
Dopamine
Propylparaben

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.

HPLC Separation of Compounds of Catecholamine Pathway

March 3, 2006

The catecholamine neurotransmitters are amino-acid derivatives of tyrosine. DOPA, tyrosine, phenylalanine, norepinephrine, epinephrine, and dopamine and baseline are resolved on a
Primesep 200 column with UV-transparent phosphate buffer. This method can be used for the analysis of catecholamines and related impurities in various matrices. The peak order and retention time can be changed by changing the amount of ACN, buffer concentration and buffer pH. Various buffers can be used to accommodate the desired detection technique. Primesep 200 is a reversed-phase cation-exchange mixed-mode column that can be used for analysis of polar neutral, polar ionizable, polar zwitterionic, hydrophobic neutral, and hydrophobic ionic compounds in the same run. Column can be operated in reverse-phase, cation-exchange, anion-exclusion, HILIC and mixed-modes depending on the mobile phase selection and the nature of the analytes. The column is compatible with LC/MS and does not require the use of ion-pairing reagents.

Condition

Column Primesep 200, 4.6×150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Buffer TFA, AmFm
Flow Rate 1.0 ml/min
Detection UV, 210 nm

 

Description

Class of Compounds
Drug, Acid, Hydrophilic, Ionizable, Hormone
Analyzing Compounds Tyrosine, DOPA, Phenylalanine, Norepinephrine, Epinephrine, Dopamine

 

Application Column

Primesep 200

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

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Application Analytes:
DOPA (3,4-dihydroxy-L-phenylalanine)
Dopamine
Epinephrine
Phenylalanine
Tyrosine

Application Detection:
ELSD 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.

HPLC Analysis of the Catecholamine Pathway

January 17, 2004

Primesep 200 separates catecholamines in the catecholamine pathway in 10 minutes. Phenylalanine, tyrosine, DOPA, dopamine, norepinephrine, and epinephrine are baseline resolved by a combination of reversed-phase, ion-exchange, and ion-exclusion mechanisms. Excellent peak shape results with a mass spec compatible mobile phase of water, acetonitrile (MeCN, ACN) and trifluoracetic acid (TFA) with UV detection at 210 nm.

Condition

Column Primesep 200, 3.2×150 mm, 5 µm, 100A
Mobile Phase MeCN/H2O – 10/90%
Buffer TFA – 0.1%
Flow Rate 0.5 ml/min
Detection UV, 210 nm

 

Description

Class of Compounds
Drug, Acid, Hydrophilic, Ionizable, Hormone
Analyzing Compounds Tyrosine, DOPA, Phenylalanine, Norepinephrine, Epinephrine, Dopamine

 

Application Column

Primesep 200

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

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Application Analytes:
DOPA (3,4-dihydroxy-L-phenylalanine)
Dopamine
Epinephrine
Phenylalanine
Tyrosine

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.

HPLC Separation of Catecholamines

January 8, 2004

Primesep 200 separates catecholamines in less than 8 minutes. Tyrosine, dl-DOPA, dopamine, epinephrine are baseline resolved by a combination of reversed-phase, ion-exchange, and ion-exclusion mechanisms. Excellent peak shape results with a mass spec compatible mobile phase of water, acetonitrile (MeCN, ACN) and trifluoracetic acid (TFA) with UV detection at 210 nm.

Condition

Column Primesep 200,4.6×50 mm, 5 µm, 100A
Mobile Phase MeCN/H2O
Buffer TFA
Flow Rate 1.0 ml/min
Detection UV, 210 nm

 

Description

Class of Compounds
Drug, Acid, Hydrophilic, Ionizable, Hormone
Analyzing Compounds Tyrosine, DOPA, Epinephrine, Dopamine

 

Application Column

Primesep 200

The Primesep family of mixed-mode columns offers a wide variety of stationary phases, boasting unprecedented selectivity in the separation of a broad array of chemical compounds across multiple applications. Corresponding Primesep guard columns, available with all stationary phases, do not require holders. SIELC provides a method development service available to all customers. Inquire about our specially-tailored custom LC-phases for specific separations.

Select options
Application Analytes:
Dopamine
Epinephrine

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.