MassTox® Immunosuppressants in Whole Blood - LC-MS/MS

Order No.: 93000 & 93000/1200
Parameters:
Cyclosporin A, Everolimus, Sirolimus, Tacrolimus

Matrix effects are compensated by online sample preparation using the trap column technique and deuterated internal standards

Grouped product items
Product Name Qty
MassTox® Immunosuppressants in Whole Blood - LC-MS/MS
Order No.: 93000
MassTox® Immunosuppressants in Whole Blood - LC-MS/MS
Order No.: 93000/1200

Cyclosporin A

Everolimus

Sirolimus

Tacrolimus

Clinical relevance

Immunosuppressants suppress the activity of the immune system in order to prevent organ transplant rejection. They are also used as therapeutic drugs for the treatment of autoimmune diseases. Therapeutic drug monitoring of immunosuppressants is indispensable for therapy following after organ transplantation. Besides mycophenolic acid, the immunosuppressants most commonly used following an organ transplantion include cyclosporin A (INN: Ciclosporin), tacrolimus (FK506), sirolimus and everolimus. Cyclosporin A and tacrolimus block the cellular signal transduction, which is required for the immune response, by inhibiting the enzyme calcineurin. For instance, it controls the synthesis of interleukin in the T cells during an immune response. In contrast, sirolimus, and its derivative everolimus, are calcineurin-independent and inhibit mTOR (mammalian target of rapamycin), an important enzyme of the intracellular signal transduction during the immune response.

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. The compensation of matrix effects also plays a key role in the diagnostic analysis of immunosuppressants from a complex matrix such as whole blood. It is therefore essential that interferences are reduced significantly to achieve precise and reliable quantification.

 

Product advantages

  • Reliable elimination of matrix effects
  • High selectivity and sensitivity
  • 6PLUS1® Multilevel Calibrator Set and MassCheck® Controls available

The sample preparation is reduced to a simple and effective protein precipitation step and ensures a major minimisation of matrix effects. After this, the analytes are concentrated online using a trap column and interfering matrix substances are separated. The trap column is connected by a simple switching system to the HPLC column and the MS/MS system. The isotopically labelled internal standards compensate very effectively for the remaining ion suppression. Therefore, the analysis of the four immunosuppressants using this kit is robust, precise and reproducible. The use of 6PLUS1® multilevel calibrators from human whole blood and our MassCheck® controls ensures highly accurate results.

For accelerated analysis and very high sample throughput, we also offer the ONEMINUTE Method for LC-MS/MS (order no. 93900).

 

Kit content 93000
Mobile Phase A, 1000 ml
Mobile Phase B, 1000 ml
Precipitation Reagent, 100 ml
Internal Standard Set, 1 box
Extraction Buffer, 40 ml
Rinsing Solution, 1000 ml
Reaction Vials, light protected, 4 x 100 pcs.

 

Kit content 93000/1200
Mobile Phase A, 3 x 1000 ml
Mobile Phase B, 3 x 1000 ml;
Precipitation Reagent, 3 x 100 ml
Internal Standard Set, 3 x 1 box
Extraction Buffer, 3 x 40 ml
Rinsing Solution, 2 x 1000 ml
Reaction Vials, light protected, 12 x 100 pcs.
Analytical Column, 1 pc.
Trap Column, 2 pcs.
6PLUS1® Multilevel Calibrator Set Immunosuppressants, 7 x 2 ml

More Information
Limit of quantification

Cyclosporin A: < 5.0 µg/l
Everolimus: < 0.5 µg/l
Sirolimus: < 0.5 µg/l
Tacrolimus: < 0.5 µg/l

Linearity

Cyclosporin A: up to 2000 µg/l
Everolimus: up to 100 µg/l
Sirolimus: up to 100 µg/l
Tacrolimus: up to 100 µg/l

Analysis Time 4.5 min
Sample Preparation
  • Place 50 µl sample/calibrator/control into a 1.5 ml reaction vial.
  • Add 25 µl reconstituted Internal Standard Mix.
  • Add 100 µl Extraction Buffer mix 10 s (vortex) and incubate for 2 min at ambient temperature.
  • Add 250 µl Precipitation Reagent,  mix 1 min (vortex) and incubate for 2 min at ambient temperature.
  • Centrifuge 5 min at 15000 x g.
  • Transfer the supernatant for analysis into the LC-MS/MS systems.
Sample Stability up to 7 days at +2°C to 8°C
Specimen Whole blood
Pre-analytic Treatment

Stored at +2 to +8 °C patient samples are stable up to 7 days.

Injection Volume 5–25 µl
Ionisation ESI positive
MS/MS-Mode MRM
Please note The freely available information on this website, in particular on the sample preparation, are not sufficient to work with our products. Please read instructions and warning notices on products and/or instruction manuals.
Method of Analysis LC-MS/MS
Parameter Cyclosporin A, Everolimus, Sirolimus, Tacrolimus
  1. 93001 Immunosuppressants Mobile Phase A
    Mobile Phase A
    Order no.: 93001

    MassTox® Immunosuppressants in Whole Blood - LC-MS/MS

    Details
  2. 93002 LCMS MassTox Immunosuppressants Whole Blood Mobile Phase B
    Mobile Phase B
    Order no.: 93002

    MassTox® Immunosuppressants in Whole Blood - LC-MS/MS

    Details
  3. 93003 Precipitation Reagent
    Precipitation Reagent
    Order no.: 93003

    MassTox® Immunosuppressants in Whole Blood - LC-MS/MS

    Details
  4. 93005 Extraction Buffer
    Extraction Buffer
    Order no.: 93005

    MassTox® Immunosuppressants in Whole Blood - LC-MS/MS

    Details
  5. 93009  LCMS MassTox Immunosuppressants Whole Blood Rinsing Solution
    Rinsing Solution
    Order no.: 93009

    MassTox® Immunosuppressants in Whole Blood - LC-MS/MS

    Details
  6. 93046 Internal Standard
    Internal Standard Set
    Order no.: 93046

    MassTox® Immunosuppressants in Whole Blood - LC-MS/MS

    Details
  7. Reaction Vials, 1.5 ml
    Reaction Vials, 1.5 ml
    Order no.: 33005
    Amber colour, light protected
    Details
  1. 28039 Immunosuppressants Multilevel Calibrator
    6PLUS1® Multilevel Whole Blood Calibrator Set Immunosuppressants
    Order no.: 28039

    MassTox® Immunosuppressants in Whole Blood - LC-MS/MS

    Details
  1. MassCheck® Immunosuppressants Whole Blood Controls
    MassCheck® Immunosuppressants Whole Blood Controls
    Order no.: 0081/0082/0083/0084/0085/0089

    MassTox® Immunosuppressants in Whole Blood - LC-MS/MS

    Details
  1. PEEK-encased Prefilter, 2 µm
    Details
  2. 93100 Analytical Column Immunosuppressants
    Analytical column
    Order no.: 93100

    MassTox® Immunosuppressants in Whole Blood - LC-MS/MS

    Details
  3. 93110 Trap Column Immunosuppressants
    Trap column
    Order no.: 93110

    MassTox® Immunosuppressants in Whole Blood - LC-MS/MS

    Details
  4. 93015 LCMS MassTox Immunosuppressants Whole Blood Tuning Mix
    Tuning Mix Immunosuppressants
    Order no.: 93015

    MassTox® Immunosuppressants in Whole Blood - LC-MS/MS

    Details
  5. PEEK Prefilter Housing
    PEEK Prefilter Housing
    Order no.: 15010
    Details
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