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Immunosuppressive Drugs in Whole Blood

Immunosuppressive Drugs in Whole Blood LC-MS/MS Analysis 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, Everolimus, Sirolimus and Cyclosporine A.
  • Matrix

    Whole blood

Sample Preparation

  • Step 1
    Pipette 250 µL of whole blood patient’s sample (calibrator/control) into a glass centrifuge tube.
  • Step 2
    Add 20 µL of internal standard solution and vortex for 5 sec.
  • Step 3
    Add 480 µL of Reagent-1, vortex for 5 sec. Then, centrifuge at 4000 rpm for 5 min.
  • Step 4
    Decant the clear supernatant into HPLC vial prior to injection.
  • Chromatogram


    Analyte LOQ (µg/L)   6460 QQQ LOQ (µg/L)   Ultivo QQQ Accuracy % 6460 QQQ Repeatabilty % CV 6460 QQQ Linearity R2 6460 QQQ
    Tacrolimus 0,09 0,05 101,8 1,6 0,999
    Sirolimus 0,54 0,71 89,5 4,9 0,997
    Evrolimus 0,35 0,19 107,1 1,5 0,999
    Cyclosporine A 6,07 0,02 96,5 2 0,999
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