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, 4.6x150 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 Analytes: