This is an Application Brief and does not contain a detailed Experimental section.
For in vitro diagnostic use. Not available in all countries.
This article demonstrates the ACQUITY UPLC I-Class/Xevo TQ Absolute IVD System’s analytical performance for the analysis of everolimus, sirolimus, tacrolimus, and cyclosporine in whole blood specimens collected with the Capitainer® B Device.
Experimental
The system was controlled using MassLynx™ IVD (v4.2) while all data was processed via the TargetLynx™ Application Manager. Whole-blood calibrators, EQA samples, and Quality Controls were processed according to the conditions outlined here.
Sample preparation conditions
A 30 µL sample was placed on a Capitainer B device. This resulted in the generation of a 10 µL dried blood spot, in line with the manufacturer’s stated performance. The sample was then processed with water/methanol, with 0.05 M hydrochloric acid/tert-butyl methyl ether added prior to analysis.
LC conditions
Source: Waters Advanced Diagnostics
| . |
. |
| Column |
ACQUITY UPLC HSS C18 SB 1.8 µm, 2.1 mm x 30 mm |
| Mobile phase A |
0.05 mM Ammonium fluoride in water |
| Mobile phase B |
0.05 mM Ammonium fluoride in methanol |
| Flow rate |
0.8 mL/min |
| Gradient |
50 % B for 0.2 minutes, 50–100 % B over 0.4 minutes, hold 100 % B for 0.6 minutes, equilibrate with 50 % B for 0.3 minutes |
MS conditions
Source: Waters Advanced Diagnostics
| . |
. |
| Resolution |
MS1 (0.75 FWHM), MS2 (0.75 FWHM) |
| Acquisition mode |
MRM |
| Polarity |
ESI+ |
Results and discussion
Table 1 features the performance characteristics of everolimus, sirolimus, tacrolimus, and cyclosporine on the ACQUITY UPLC I-Class/Xevo TQ Absolute IVD System. Figure 1 highlights the chromatographic separation's analytical sensitivity.
Table 1. Performance characteristics of cyclosporine, everolimus, sirolimus, and tacrolimus. Range defined by linear fit where r2>0.995. LLOQ is defined by S/N (PtP)>10. Total precision and repeatability of QCs performed over five occasions in whole blood (n=25), with the exception of cyclosporine, for which there were four occasions (n=20). EQA mean bias determined by comparison of obtained values to the LC-MS all laboratories trimmed mean (LC-MS ALTM) values (n=20 for cyclosporine, n=19 for everolimus, n=25 for sirolimus, and n=25 for tacrolimus). Source: Waters Advanced Diagnostics
Figure 1. Chromatogram showing the analysis of 25 ng/mL cyclosporine and 1 ng/mL everolimus, sirolimus, and tacrolimus in whole blood collected using a Capitainer B Device using the ACQUITY UPLC I-Class/ Xevo TQ Absolute IVD System.
Conclusion
The ACQUITY UPLC I-Class/Xevo TQ Absolute IVD System was shown to deliver the necessary analytical precision and sensitivity for the analysis of sirolimus, cyclosporine, everolimus, and tacrolimus in whole blood collected via a Capitainer B device.
For In vitro diagnostic use. Not available in all countries.
Disclaimer
The analytical performance data presented here is for illustrative purposes only. Waters does not recommend or suggest analysis of the analytes described herein. These data are intended solely to demonstrate the performance capabilities of the system for analytes representative of those commonly analyzed using liquid chromatography and tandem mass spectrometry. Performance in an individual laboratory may differ due to a number of factors, including laboratory methods, materials used, intra-operator technique, and system conditions. This document does not constitute a warranty of merchantability or fitness for any particular purpose, express or implied, including for the testing of the analytes in this analysis.
Acknowledgments
Produced from materials originally authored by Stephen Balloch, Lisa J. Calton, and Gareth Hammond from Waters Corporation.
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