Separation of Norepinephrine, Epinephrine, and Dopamine hydrochloride

This Method shows separation of similar Catecholamines analyzed with Atmospheric Pressure Chemical Ionization (APCI) in Positive mode MS due to the lack of suitable chromophores on these molecules.  Proper separation of these compounds is important in assessment of biological samples.

Peaks:
1. L-alpha– methyl – dopa 
2. Norepinephrine
3. Epinephrine
4. Dopamine hydrochloride

Method Conditions
Column: Cogent Bidentate C18™, 4µm, 100Å
Catalog No.: 40018-75P
Dimensions: 4.6 x 75mm
Mobile Phase:  Acetonitrile / 25 mM Ammonium Formate 5:95, pH=4
Injection vol.: 1µL
Flow rate: 0.5mL / minute
Detection: Atmospheric Pressure Chemical Ionization in the Positive mode: APCI
Sample Preparation: 10μg were dissolved in 1 mL of the Mobile Phase, except the concentration of Norepinephrine was 3μg / mL in Mobile Phase.

Note: The concentration of Catecholamines and their metabolites in plasma or brain are useful for diagnosis and evaluation of therapeutic and pharmacodynamic effects for  psychiatric, neurological and cardiovascular disorders. The use of Mass Spectroscopy for detection in the analysis of Catecholamines is an effective approach due to the lack of a suitable chromophore on these molecules making UV detection difficult. In addition Mass Spectrometry provides positive identification of each component. The analysis (Total Ion Chromatogram) of a four- component Catecholamine mixture. 

Attachment

Catecholamines Analyzed with LCMS – AppNote 0.3 Mb Download File