This is an Application Brief and does not contain a detailed Experimental section.
For in vitro diagnostic use. Not available in all countries.
The ACQUITY™ UPLC™ I-Class/Xevo™ TQ Absolute IVD System from Waters is designed to facilitate the accurate quantification of organic compounds in human biological liquid matrices.
This document describes a test of the analytical performance of the ACQUITY UPLC I-Class/Xevo TQ Absolute IVD System for the analysis of cyclosporine, everolimus, sirolimus, and tacrolimus in whole blood.
Experimental
This work saw the ACQUITY UPLC I-Class/Xevo TQ Absolute IVD System controlled by MassLynx™ IVD (v4.2); data processing was acquired using the TargetLynx™ Application Manager. Whole blood Quality Controls and calibrators were processed according to the conditions outlined here.
Sample preparation conditions
A 50 µL sample was processed with acetonitrile and zinc sulfate before being centrifuged.
LS conditions
Source: Waters Advanced Diagnostics
| . |
. |
| Column |
ACQUITY UPLC HSS C18 SB 1.8 µm, 2.1 mm x 30 mm, |
| Mobile phase A |
2 mM Ammonium acetate + 0.1 % formic acid in water |
| Mobile phase B |
2 mM Ammonium acetate + 0.1 % formic acid in methanol |
| Flow rate |
0.4 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 0.6 minutes at 0.6 mL/min |
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, cyclosporine, and tacrolimus on the ACQUITY UPLC I-Class/Xevo TQ Absolute IVD System, while Figure 1 illustrates 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 defined by S/N (PtP) >10. Total precision and repeatability of QCs performed over five occasions in whole blood (n=25). EQA mean bias determined by comparison of obtained values to the LC-MS all laboratories trimmed mean (LC-MS ALTM) values (n=27 for cyclosporine, n=25 for everolimus, n=28 for sirolimus, and n=27 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 using the ACQUITY UPLC I-Class/Xevo TQ Absolute IVD System. Image Credit: Waters Advanced Diagnostics
Conclusion
The Waters ACQUITY UPLC I-Class/Xevo TQD IVD System has demonstrated the capability to deliver analytical sensitivity, accuracy, and precision for the analysis of cyclosporine, everolimus, sirolimus, and tacrolimus in whole blood.
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, Gareth Hammond, and Elizabeth Embrey from Waters Corporation.
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