CATECHOLAMINES

CATECHOLAMINES IN PLASMA LC-MS/MS ANALYSIS KIT

Plasma catecholamines including epinephrine (E), norepinephrine (NE), and dopamine (DA) are used to evaluate adrenosympathetic function in patients. Quantification of plasma catecholamine concentrations is clinically important for the diagnosis of pheochromocytoma and paraganglioma during dynamic clonidine tests. The low circulating concentrations of catecholamines and analytical interferences require tedious sample preparation and long chromatographic runs to ensure their accurate quantification by HPLC with electrochemical detection. The reliable and simultaneous analysis of a broad panel of catecholamines is a powerful tool for the investigation of the hormone status, which is relevant for a variety of clinical questions and diagnoses

  • Features

    •  Analysis of norepinephrine, epinephrine and dopamine without derivatization

    • No need of SPE!

    •  Including stable labelled isotopes of each analyte as internal standards

    • 10 minutes analysis time from injection to injection

  • Matrix

    • Plasma
  • PARAMETERS

    • Epinephrine
    • Norepinephrine
    • Dopamine

Sample Preparation


  • Step 1

    300 µl of plasma sample is pipetted into the glass tube.

  • Step 2

    20 µl internal standard is added to the sample and vortexed. Then 600 µl of Reagent-1 is added and swirled for 15 seconds.

  • Step 3

    1 ml of Reagent-2 is added to the mixture in the same tube. After stirring in the shaker for 3 minutes, it is centrifuged for 2 minutes.

  • Step 4

    700 µl of the upper phase is pipetted and discarded. Solid Reagent-3 is added over the remaining part in the tube and vortexed for 30 seconds.

  • Step 5

    200 µl of Reagent-4 and 2 ml of Reagent-5 are added respectively and allow to stir for 7 minutes, then centrifuge at 4000 rpm for 3 minutes.

  • Step 6

    1.6 ml of the supernatant is evaporated to dryness under nitrogen stream.

  • Step 7

    Following evaporation, residue is reconstituted using 100 µl of Reagent-1 and vortex 5 seconds prior to analysis.

  • Chromatogram of Catecholamines

  • Retention times of the analytes

                   Analytes

    Retention Times (min)

    Norepinephrine

              3.35

    Epinephrine

              3.90

    Dopamine

              4.3

  • Limit of quantification

    LOQ: Limit of quantification, 10 SD

          

     

    PLASMA

               Compound

    R2

    LOQ ng/L

    Epinephrine

    0.997

    29.1

    Norepinephrine

    0.998

    38.5

    Dopamine

    0.995

    19.3




  • Recovery

     

    Concentration ng/L (plasma)

    Recovery %

    Concentration ng/L (plasma)

    Recovery %

    Compound

    Level-I

    Level-I 

    Level-II

    Level-II

    Epinephrine

    311.8

    97.63

    551.9

    93.45

    Norepinephrine

    1345

    98.52

    2098

    95.78

    Dopamine

    335

    99.25

    512.7

    101.2

     

           
  • Repeatability

     

    Analyte

    Concentration µg/L (plasma)

    CV %

    Epinephrine

    311.8

    5.3

    Norepinephrine

    1345

    6.2

    Dopamine

    335

    4.4

     

ADAPTING DIAGNOSTICS IN CHROMATOGRAPHY COUPLED MASS SPECTROMETRY..

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