ACQUITY™ UPLC™ I-Class/Xevo™ TQ Absolute IVD System: Analytical Performance for Immunosuppressive Agents using a Microsampling Device

Importance of the Topic

The quantification of immunosuppressive agents such as cyclosporine, everolimus, sirolimus, and tacrolimus in whole blood is essential for therapeutic drug monitoring to maintain efficacy and minimize adverse effects. Microsampling technologies like the Capitainer B device enable precise collection of small blood volumes, supporting patient convenience and stable sample handling.

Goals and Overview of the Study

This application note evaluates the analytical performance of the Waters ACQUITY UPLC I-Class/Xevo TQ Absolute IVD System for measuring key immunosuppressants in whole blood samples collected via the Capitainer B microsampling device. The aim is to demonstrate sensitivity, precision, and accuracy under standardized conditions.

Methodology and Used Instrumentation

• Sample Preparation: 30 µL whole blood applied to Capitainer B device yielding a 10 µL dried blood spot; extraction performed with water/methanol followed by 0.05 M HCl/tert-butyl methyl ether.
• Liquid Chromatography: ACQUITY UPLC HSS C18 SB column (1.8 µm, 2.1×30 mm); mobile phases: 0.05 mM ammonium fluoride in water (A) and in methanol (B); flow rate 0.8 mL/min; rapid gradient from 50% to 100% B within 0.4 min.
• Mass Spectrometry: Xevo TQ in ESI positive mode; multiple reaction monitoring; MS1 and MS2 resolution at 0.75 FWHM; controlled via MassLynx IVD v4.2 and processed with TargetLynx.

Main Results and Discussion

• Calibration curves exhibited excellent linearity for all analytes (r2 > 0.995).
• Lower limit of quantification achieved signal-to-noise ratios above 10, confirming analytical sensitivity.
• Total precision and repeatability of quality control samples remained within acceptable limits across multiple runs (n=20–25).
• External quality assessment bias was minimal relative to LC-MS all-laboratories trimmed mean, demonstrating method accuracy.

Benefits and Practical Applications

• The method provides high sensitivity and precision required for reliable therapeutic drug monitoring.
• Microsampling reduces blood volume requirements, facilitating patient-friendly sampling and remote collection.
• Fast UPLC gradient and streamlined workflow support high throughput in clinical laboratories.

Future Trends and Possibilities

• Integration of microsampling with telemedicine platforms for at-home monitoring.
• Extension of the workflow to additional pharmacological and diagnostic biomarkers.
• Automation of sample preparation and data analysis to enhance laboratory efficiency.
• Application of advanced data analytics and machine learning for personalized dosing recommendations.

Conclusion

The Waters ACQUITY UPLC I-Class/Xevo TQ Absolute IVD System combined with the Capitainer B microsampling device delivers robust sensitivity, precision, and accuracy for quantifying major immunosuppressive drugs in dried blood spots, offering significant advantages for clinical therapeutic drug monitoring.

Reference

No literature references were provided in the original text.