HPLC Separation of Catecholamines on Newcrom AH Column

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.


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



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:
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.