MassTox® Immunosuppressants in Whole Blood – ONEMinute Test - LC-MS/MS

Order No.: 93900/400 & 93900/1200
Parameters:
Cyclosporin A, Everolimus, Sirolimus, Tacrolimus

Reduced matrix effects through online sample preparation with trap column technology and isotope-labelled internal standards
High sensitivity and selectivity

Grouped product items
Product Name Qty
MassTox® Immunosuppressants in Whole Blood – ONEMinute Test - LC-MS/MS
Order No.: 93900/400
MassTox® Immunosuppressants in Whole Blood – ONEMinute Test - LC-MS/MS
Order No.: 93900/1200

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 sensitivity and specificity
  • Very short analysis time of 1-2 minutes
  • Optional: Automated sample preparation

The ONEMINUTE MassTox® assay for the analysis of the four immunosuppressants leads to robust, precise and reproducible results with a run time of only 1-2 minutes per sample. The ONEMINUTE method saves time, especially for laboratories with high sample throughput. Manual sample preparation is reduced to a simple and effective protein precipitation step. The concentration of the analytes and the elimination of other interfering substances is performed ONLINE using a trap column, which massively minimises the matrix effects. The isotopically labelled internal standards compensate entirely for remaining ion suppression. The trap column is connected by a simple switching system to the HPLC column and the MS/MS system. The additional use of 6PLUS1® multilevel calibrators and the MassCheck® controls ensures highly precise results.

This test is also available as an automated method which is comprehensively validated for all established tandem mass spectrometers.

Additionally, Chromsystems also offers a standard method for the determination of immunosuppressants in whole blood (order no. 93000).

 

Kit content 93900/400
Mobile Phase A, 900 ml
Mobile Phase B, 450 ml
Precipitation Reagent, 100 ml
Internal Standard Set, 1 box
Extraction Buffer, 40 ml
Rinsing Solution, 1000 ml
Reaction Vials, transparent, 4 x 100 pcs.

 

Kit content 93900/1200
Mobile Phase A, 5 x 900 ml
Mobile Phase B, 3 x 450 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, transparent, 12 x 100 pcs.
Analytical Column, 1 pc.
Trap Column, 2 pcs.
6PLUS1® Multilevel Calibrator Set Immunosuppressants (XL), 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 1-2 min (mass spectrometer dependent)
Sample Preparation
  • Place 50 µl sample/calibrator/MassCheck® into a 1.5 ml reaction vial
  • Add 25 µl reconstituted Internal Standard Mix
  • Add 100 µl Extraction Buffer
  • Add 250 µl Precipitation Reagent, mis 1 min  (vortex) and incubate for 2 min at room temperature
  • Centrifuge for 5 min at 15000 x g
  • Transfer the supernatant to an autosampler vial and inject 5 to 50 µl of the supernatant into the LC-MS/MS system

 

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 5–50 µl
Ionisation ESI positive
MS/MS-Mode MRM
Information

This Chromsystems product is comprehensively validated. User re-validation is not required. For validation data (instrument specific) please see instruction manual.

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. 93911 Mobile Phase A OneMinuteTest Immunosuppressants
    Mobile Phase A
    Order no.: 93911

    MassTox® Immunosuppressants ONEMinute Test - LC-MS/MS

    Details
  2. 93122 Trap Column Immunosuppressants
    Mobile Phase B
    Order no.: 93922

    MassTox® Immunosuppressants ONEMinute Test - LC-MS/MS

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

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

    Details
  4. 93909 LCMS MassTox Immunosuppressants Whole Blood OneMinute Rinsing Solution
    Rinsing Solution
    Order no.: 93909

    MassTox® Immunosuppressants ONEMinute Test - LC-MS/MS

    Details
  5. 93946 Internal Standard Mix Immunosuppressants
    Internal Standard Set
    Order no.: 93946

    MassTox® Immunosuppressants ONEMinute Test - LC-MS/MS

    Details
  6. 93005 Extraction Buffer
    Extraction Buffer
    Order no.: 93005

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

    Details
  7. Reaction Vials, transparent, 1.5 ml
    Reaction Vials, transparent, 1.5 ml
    Order no.: 33006
    Transparent
    Details
  1. 28039/XL Immunosuppressants Multilevel Calibrator
    6PLUS1® Multilevel Whole Blood Calibrator Set Immunosuppressants
    Order no.: 28039/XL

    MassTox® Immunosuppressants in Whole Blood ONEMinute Test - 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. 93122 Trap Column Immunosuppressants
    Trap column
    Order no.: 93122

    MassTox® Immunosuppressants ONEMinute Test - LC-MS/MS

    Details
  4. 93915 LCMS MassTox Immunosuppressants Whole Blood OneMinute Tuning Mix
    Tuning Mix Immunosuppressants ONEMinute Test
    Order no.: 93915

    MassTox® Immunosuppressants ONEMinute Test - LC-MS/MS

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