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HPLC Application Sheet
Catecholamines: Analysis by LC-MS
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 on Cogent Bidentate C18
column is shown.

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Method Conditions
| Column |
Cogent Bidentate C18, 4µm, 100A |
| Catalog No. |
40018-75P |
| Dimensions |
4.6 x 75mm |
| Mobile Phase |
Acetonitrile/25 mM ammonium formate 5:95, pH = 4
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| Flow rate |
0.5 mL/minute |
| Injection Volume |
1 µL |
| Samples |
L-alpha-methyl-dopaNorepinephrineEpinephrineDopamine hydrochloride - 10 µg were dissolved in 1 mL of the mobile phas, except the concentration of norephinephrine was 3 µg/mL in mobile phase |
| Detection |
Mass Spec: Atmospheric Pressure Chemical Ionization in the Positive mode: APCI+ |
Discussion: Major Peaks
Table below: Major peaks in mass spectra of slutes used in the study.
| Solute |
m/z |
Peak Identity |
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| L-alpha-methyl-dopa |
212 194 |
MH+ MH+ -H20 |
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| Norepinephrine |
152 135 |
MH+ -H20 MH+ -2H20 |
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| Epinephrine |
184 166 148 |
MH+ MH+ -H20 MH+ -2H20 |
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| Dopamine hydrochloride |
154 136 |
MH+ MH+ -H20 |
This method is easy and robust and very effective. |
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