MassTox® Immunosuppressants in Whole Blood – ONEMinute Test - LC-MS/MS
Reduced matrix effects through online sample preparation with trap column technology and isotope-labelled internal standards
High sensitivity and selectivity
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.
- 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).
Content for 400 tests [93900/400)
Mobile Phase A, 900 ml
Mobile Phase B, 450 ml
Precipitation Reagent, 100 ml
Internal Standard Set, 1 Set
Extraction Buffer, 40 ml
Rinsing Solution, 1000 ml
Reaction Vials, transparent, 4 x 100 pcs.
Content for 1200 tests (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 Set
Extraction Buffer, 3 x 40 ml
Rinsing Solution, 2 x 1000 ml
Reaction Vials, transparent, 12 x 100 pcs.
6PLUS1® Multilevel Calibrator Set Immunosuppressants (XL), 7 x 2 ml
Analytical Column, 1 pc.
Trap Column, 2 pcs.
|Method of Analysis||LC-MS/MS|
|Limit of quantification||
Cyclosporin A: < 5.0 µg/l
Cyclosporin A: up to 2000 µg/l
|Analysis Time||1-2 min (mass spectrometer dependent)|
Stored at +2 to +8 °C patient samples are stable up to 7 days.
|Sample Stability||up to 7 days at +2°C to 8°C|
|Injection Volume||5–50 µl|
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.|
|Parameter||Cyclosporin A, Everolimus, Sirolimus, Tacrolimus|
6PLUS1® Multilevel Whole Blood Calibrator Set ImmunosuppressantsOrder no.: 28039/XL
MassCheck® Immunosuppressants Whole Blood ControlsOrder no.: 0081/0082/0083/0084/0085/0089