MassTox® Immunosuppressants ONEMinute Test - LC-MS/MS

MassTox® Immunosuppressants ONEMinute Test - LC-MS/MS

   
  • Cyclosporin A
  • Everolimus
  • Sirolimus
  • Tacrolimus

Order no.: 93900/400, 93900/1200
400 / 1200 Tests

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

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Description

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® kit 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, light protected, 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, light protected, 12 x 100 pcs.       
Analytical column, 1 pc.           
Trap column, 2 pcs.                   
6PLUS1® Multilevel Calibrator Set Immunosuppressants (XL), 7 x 2 ml      

Chromatogram

Technical Data

Method of Analysis

LC-MS/MS

Parameter

Cyclosporin A, Everolimus, Sirolimus, Tacrolimus

Analysis Time

1-2 min (mass spectrometer dependent)

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

Information

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

Specimen

Whole blood

Pre-analytic Treatment

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

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

Injection Volume

5–50 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.

Kit Components

Components available separately

Mobile Phase A


Order no.: 93911

Mobile Phase B


Order no.: 93922

Precipitation Reagent


Order no.: 93003

Rinsing Solution


Order no.: 93909

Internal Standard Set


Order no.: 93946

Extraction Buffer


Order no.: 93005

Calibrators

Calibrators available separately

Controls

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