Immunosuppresıve Drugs

IMMUNOSUPPRESSIVE DRUGS IN WHOLE BLOOD LC-MS/MS KIT

Tacrolimus, sirolimus, everolimus and Cyclosporine A are immunosuppressive drugs used in organ transplantation in order to prevent the rejection of transplanted organs and tissues. In addition, they are used in the therapy of autoimmune diseases. Due to high pharmacokinetic variability, narrow therapeutic indexes and many potential drug-drug interactions, close monitoring of blood concentrations is required to prevent rejection and minimize toxicity. The quantitative determination of these immunosuppressants should be performed only with LC-MS/MS, because the immunological tests vary considerably in terms of selectivity and specificity, and sometimes also detect inactive metabolites.

  • Highlights

    • Simultaneous measurement the concentrations of 4 immunosuppressive drugs
    • Total run time is 7.5 min.
    • Simple sample preparation-protein precipitate and shoot approach, no need for online
      sample preparation using the trap column technique
    • Whole blood based multi-level calibrator and control
    • Long life span of HPLC column
  • Parameters

    • Tacrolimus
    • Sirolimus
    • Everolimus
    • Cyclosporine A
  • Matrıx

    • Whole blood

Sample Preparation for Whole Blood


  • Step 1

    Pipet 500 μl EDTA-treated whole blood sample into a centrifuge tube

  • Step 2

    Add 25 μL of internal standard mix onto sample and vortex for 5 seconds

  • Step 3

    Add 975 μL of Reagent-1 onto sample and additionally vortex for 15 seconds

  • Step 4

    Centrifuge at 4000 rpm for 5 minutes

  • Step 5

    Transfer the supernatant into HPLC vial and inject into the system

  • Valıdatıon Results

    Compound

    LOQ (µg/L)

    Recoveries (%)

    Repeatability (%)

    Tacrolimus

    1.45

    102

    1.12

    Sirolimus

    2.02

    90

    3.71

    Everolimus

    1.77

    108

    1.89

    Cyclosporine A

    31.11

    100

    7.02

  • Extracted LC-MS/MS Chromatogram of Immunosuppressıve Drugs

ADAPTING DIAGNOSTICS IN CHROMATOGRAPHY COUPLED MASS SPECTROMETRY..

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