METANEPHRINES

METANEPHRINES ANALYSIS IN PLASMA LC-MS/MS KIT

Normetanephrine and metanephrine are the O-methylated derivatives of the catecholamine hormones norepinephrine and epinephrine, respectively. Metanephrines are created by means of catecholamine metabolism but also generated excessively in pheochromocytoma tumors of the chromaffin cells, most of which are localized to the adrenal gland. The majority of patients with a pheochromocytoma tumor have plasma normetanephrine and metanephrine concentrations at least 2–3 times the upper reference limits. The presence of elevated concentrations of the analytes represents as a confirmation tool for the diagnosis of the tumor. In this context, the measurement of free (unconjugated) plasma metanephrines has been proved to be a highly sensitive (With a diagnostic sensitivity of 98% and specificity of 92%) test for the detection of pheochromocytoma.

  • Highlights

    • Analysis of metanephrine, normetanephrine and 3-metoxytyramine without derivatization
      rapid and easy sample preparation-just protein precipitation without SPE

    • Including stable labelled isotopes of each analyte as internal standards

    • 5.0 min. analysis time

  • PARAMETERS

    • Metanephrine
    • Normetanephrine
    • 3-Methoxytyramine
  • MATRIX

    • Plasma

Sample Preparation


  • Step 1

    Pipette 150 μl plasma sample into a centrifuge tube

  • Step 2

    Add 20 μl of Internal std and 250 μl Reagent-1 then vortex for 15 sec.

  • Step 3

    Centrifuge at 4000 rpm for 5 min

  • Step 4

    Decant the clear supernatant into HPLC vial

  • Extracted ion chromatogram of free metanephrines

  • Calibration Curves

    Normetanephrine

    Metanephrine

    Methoxytyramine

  • Linearity & Limit of Quantitation

          

     

    PLASMA

    Analyte

    R2

    LOQ

    (µg/mL)

    Normetanephrine

    0.999

    5.52

    Metanephrine

    0.999

    14.82

    3-Methoxytyramine

    0.998

    3.81

  • Recovery

    Analyte

    Concentration µg/L (plasma)

    Recovery %

    Normetanephrine

    381

    98.2

    Metanephrine

    238

    95.2

    3-Methoxytyramine

    186

    104.1

  • Repeatability

     

    Analyte

    Concentration µg/L (plasma)

    CV %

    Normetanephrine

    38.5

    1.59

    Metanephrine

    29.1

    2.81

    3-Methoxytyramine

    19.3

    1.46

ADAPTING DIAGNOSTICS IN CHROMATOGRAPHY COUPLED MASS SPECTROMETRY..

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