Stephen Merrigan (Presenter)
ARUP Institute for Clinical and Experimental Path
Authorship: Stephen D. Merrigan (1), Matthew Slawson (2), Serge Auger (3), and Kamisha L. Johnson-Davis (1,4)
(1)ARUP Laboratories, 500 Chipeta Way, Salt Lake City, Utah, USA 84108 (2)Utah Public Health Laboratory, Salt Lake City, Utah, USA (3)Phytronix Technologies Inc., Quebec,Canada (4)University of Utah Health Sciences Center, Department of Pathology, Salt Lake City, Utah, USA
| Short Abstract Accuracy, turnaround time, and analytical cost are important factors to consider when developing a therapeutic drug monitoring assay for immunosuppressive drug therapy. Sirolimus, Cyclosporin A, Tacrolimus, and Everolimus therapies are monitored to balance therapeutic efficacy and prevent organ rejection, while minimizing the adverse effects associated with high concentrations in whole blood. Laser Diode Thermal Desorption Tandem Mass Spectrometry (LDTD-MS/MS) technology can provide rapid results and reduced analytical costs associated with mobile phases and liquid chromatography columns. An eight second LDTD-MS/MS method was developed for the quantification of four immunosuppressive drugs, in whole blood. Method validation data and cost analysis are presented. |
Long Abstract
Introduction: Accuracy, turnaround time, and analytical cost are important factors to consider when developing a therapeutic drug monitoring assay for immunosuppressive drug therapy. Sirolimus, Cyclosporin A, Tacrolimus, and Everolimus therapies are monitored to balance therapeutic efficacy and to prevent organ rejection, while minimizing the adverse effects associated with high concentrations in whole blood. The local hospital expressed the need for faster turnaround time to help support patient care. Laser Diode Thermal Desorption Tandem Mass Spectrometry (LDTD-MS/MS) technology can provide rapid results and reduced analytical costs associated with mobile phases and high performance liquid chromatography columns.
Methods: An eight second LDTD-MS/MS method was developed and used for the quantification of Sirolimus, Cyclosporin A, Tacrolimus, and Everolimus in whole blood. Samples were prepared using a protein crash containing internal standard followed by filtration and reverse phase 96-well solid phase extraction. The mass spectrometry method monitored two transitions for each analyte and two transitions for each isotopically labeled internal standard in positive mode. The method described utilizes a six point calibration curve with two concentrations of quality control and a blank. The LDTD laser profile ramped from 0% to 65% of full power over six seconds and is held at 65% for two seconds before returning to initial conditions.
Results: Run time was significantly decreased using the LDTD-MS/MS method. Sample to sample time with the LDTD-MS/MS method was 10 seconds (8 seconds of mass spec time and 2 seconds to move well position) compared to 3.25 minutes with the LC-MS/MS assay (2.5 minutes of mass spec and HPLC time and 45 second of autosampler time). A 96 sample LDTD-MS/MS run time was decreased 95% at 16 minutes compared to 312 minutes with the LC-MS/MS assay. Method validation data for the LDTD-MS/MS quantitative immunosuppressant drug assay are presented including method comparison to LC-MS/MS, imprecision, accuracy, carryover, stability and interference. Cost analysis comparison to similar LC-MS/MS methods will be presented.
Conclusions: Laser Diode Thermal Desorption Tandem Mass Spectrometry technology can provide rapid results and reduced analytical costs. We developed an eight second LDTD-MS/MS method used for the quantification of Sirolimus, Cyclosporin A, Tacrolimus, and Everolimus in whole blood. Method validation data and cost analysis are presented.
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