Quantification of immunosuppressants in human blood by liquid chromatography-tandem mass spectrometry

by | Bioclinical, Biopharma, Pharmaceutical

This technical note describes the implementation of an analytical method for the quantification of four immunosuppressants (tacrolimus, sirolimus, everolimus, and cyclosporine A) in human blood on a Thermo Scientific™ TSQ Quantis™ triple quadrupole mass spectrometer.

Fast and robust LC-MS methods for analysis and quantitation of immunosuppressants are critical for ensuring quality data within a short period of time. In this report, an analytical method covering four commonly monitored immunosuppressants in human blood for clinical research is reported. The analytes include tacrolimus, sirolimus, everolimus, and cyclosporine A. Blood samples were extracted by offline internal standard addition and protein precipitation. Extracted samples were injected onto a Thermo Scientific™ Transcend™ LX-2 system. Detection by mass spectrometry was performed using a TSQ Quantis triple quadrupole mass spectrometer with heated electrospray ionization by selected-reaction monitoring (SRM) using one isotopically labeled internal standard for each analyte.

Full experimental conditions can be obtained by downloading the technical note.

Results and Discussion
A linear interpolation with 1/x weighting was used for all the analytes. The percentage bias between nominal and back-calculated concentration was always within ±10% for all the calibrators in all the runs. The correlation factor (R2) was always above 0.999 for all the analytes. 

A robust, reproducible, and sensitive LC-MS method for clinical research for quantification of
immunosuppressants in human blood was developed and implemented. The method was analytically validated on a Transcend LX-2 system connected to a TSQ Quantis triple quadrupole mass spectrometer.
Calibrators and controls from Chromsystems were used. The data obtained with the described method
successfully met sensitivity, reliability, accuracy, and precision expectations typically found in clinical research laboratories.

Download now and achieve the following application benefits:

  • Simple offline sample preparation by protein precipitation
  • Robust and easy to use quantitative method for four immunosuppressants in the same run
  • Best-in-class sensitivity with mid-range triple quadrupole MS.

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Published  Aug 6, 2019

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