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HPLC Application Sheet
Catecholamines: Analysis by LC-MS

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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
Flow rate 0.5 mL/minute
Injection Volume  1 µL
Samples
  • L-alpha-methyl-dopa
  • Norepinephrine
  • Epinephrine
  • Dopamine 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

    L-alpha-methyl-dopa 212
    194
    MH+
    MH+ -H20

    Norepinephrine 152
    135
    MH+ -H20
    MH+ -2H20

    Epinephrine 184
    166
    148
    MH+
    MH+ -H20
    MH+ -2H20

    Dopamine hydrochloride 154
    136
    MH+
    MH+ -H20


    This method is easy and robust and very effective.







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