Assays for Monitoring Apixaban and Rivaroxaban in Emergency Settings, State-of-the-Art Routine Analysis, and Volumetric Absorptive Microsamples Deliver Discordant Results
Abstract
:1. Introduction
2. Materials and Methods
- -
- methanol,
- -
- methanol-water 1:1 (vol/vol%),
- -
- methanol-acetonitrile 3:1 (vol/vol%), 1:1 (vol/vol%), or 1:3 (vol/vol%).
3. Results
3.1. Relative Recovery of the Analytes from VAMS Samples
3.2. Technical Validation of the LC-MS/MS Assays
3.3. Hematocrit- and Concentration-Dependence of the Relative Analyte Recovery from VAMS Samples
3.4. Cross-Validation Experiments Using Leftover Patient Samples
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Appendix A
References
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Performance Characteristics | Within-Run Experiment | Between-Run Experiment, Day 1 | Between-Run Experiment, Day 2 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
QC 1 | QC 2 | QC 3 | QC 4 | QC 5 | QC 6 | QC 2 | QC 3 | QC 5 | QC 2 | QC 3 | QC 5 | |
Apixaban in plasma | ||||||||||||
Accuracy (%) | 99.9 | 105.3 | 95.6 | 103.5 | 114.0 | 111.3 | 102.1 | 98.1 | 104.4 | 97.2 | 94.4 | 104.1 |
RSD (%) | 4.9 | 4.3 | 12.8 | 6.8 | 4.1 | 3.9 | 2.8 | 5.2 | 5.4 | 5.1 | 6.2 | 5.6 |
Apixaban in VAMS, whole blood calibrators | ||||||||||||
Accuracy (%) | 94.1 | 94.7 | 93.8 | 91.7 | 95.8 | 90.6 | 99.3 | 95.9 | 96.9 | 97.0 | 94.0 | 98.3 |
RSD (%) | 8.0 | 6.5 | 4.8 | 7.8 | 3.4 | 7.1 | 4.8 | 5.3 | 2.6 | 5.1 | 6.3 | 4.7 |
Apixaban in VAMS, dried plasma calibrators | ||||||||||||
Accuracy (%) | 100.0 | 98.2 | 96.9 | 94.6 | 98.8 | 93.4 | 102.8 | 99.4 | 100.5 | 102.3 | 98.6 | 102.6 |
RSD (%) | 7.7 | 6.5 | 4.8 | 7.8 | 3.4 | 7.1 | 4.8 | 5.3 | 2.6 | 5.0 | 6.3 | 4.7 |
Apixaban in VAMS, liquid plasma calibrators | ||||||||||||
Accuracy (%) | 105.4 | 114.5 | 114.3 | 112.4 | 117.7 | 111.4 | 115.8 | 113.0 | 115.2 | 120.6 | 116.6 | 121.7 |
RSD (%) | 8.7 | 6.6 | 4.8 | 7.8 | 3.4 | 7.1 | 4.9 | 5.4 | 2.6 | 5.1 | 6.3 | 4.7 |
Rivaroxaban in plasma | ||||||||||||
Accuracy (%) | 98.1 | 103.8 | 107.8 | 105.7 | 111.7 | 107.0 | 101.2 | 101.2 | 104.4 | 100.5 | 100.1 | 100.8 |
RSD (%) | 3.8 | 4.3 | 2.2 | 1.9 | 2.4 | 1.7 | 6.9 | 3.3 | 5.7 | 4.5 | 3.3 | 1.7 |
Rivaroxaban in VAMS, whole blood calibrators | ||||||||||||
Accuracy (%) | 100.7 | 96.6 | 97.7 | 93.3 | 97.1 | 92.3 | 97.8 | 96.1 | 97.2 | 94.8 | 93.6 | 98.5 |
RSD (%) | 10.9 | 4.7 | 4.6 | 5.6 | 2.9 | 6.1 | 4.7 | 3.6 | 1.4 | 8.1 | 5.9 | 3.5 |
Rivaroxaban in VAMS, dried plasma calibrators | ||||||||||||
Accuracy (%) | 100.2 | 96.5 | 97.7 | 93.3 | 97.2 | 92.3 | 97.8 | 96.2 | 97.3 | 94.5 | 92.4 | 96.4 |
RSD (%) | 11.0 | 4.7 | 4.6 | 5.6 | 2.9 | 6.1 | 4.7 | 3.6 | 1.4 | 7.9 | 5.8 | 3.5 |
Rivaroxaban in VAMS, liquid plasma calibrators | ||||||||||||
Accuracy (%) | 116.5 | 121.3 | 124.0 | 119.1 | 124.3 | 118.2 | 117.5 | 117.4 | 120.4 | 126.2 | 122.1 | 126.2 |
RSD (%) | 12.1 | 4.7 | 4.6 | 5.7 | 2.9 | 6.1 | 4.9 | 3.6 | 1.4 | 7.8 | 5.7 | 3.5 |
Matrix | API | RIV | ||
---|---|---|---|---|
Low Spiking Level | High Spiking Level | Low Spiking Level | High Spiking Level | |
Plasma | 0.86 ± 0.05 (5.4%) | 1.01 ± 0.07 (7.2%) | 0.90 ± 0.13 (15.0%) | 1.05 ± 0.06 (6.1%) |
Dried whole blood | 1.00 ± 0.04 (3.6%) | 0.96 ± 0.05 (5.5%) | 1.03 ± 0.05 (4.9%) | 0.94 ± 0.06 (6.7%) |
Analyte | Spiking Level | Spiked Concentration (ng/mL) | Relative Recovery (%) | |
---|---|---|---|---|
Plasma | VAMS Samples | |||
API | Low | 53.2 | 94.3–118 | |
49.8 | 98.0–110 | |||
Middle | 107 | 93.4–113 | ||
99.6 | 91.3–105 | |||
High | 428 | 89.5–103 | ||
498 | 96.3–105 | |||
RIV | Low | 53.2 | 96.1–106 | |
25.2 | 94.0–106 | |||
Middle | 107 | 102–109 | ||
50.4 | 90.1–100 | |||
High | 428 | 100–107 | ||
252 | 95.1–99.2 |
Substance | Approaches Compared | Slope (95% CI) | Intercept (95% CI) | h Statistic |
---|---|---|---|---|
Apixaban | Plasma, LC-MS/MS and plasma, Anti-Xa | 1.11 (1.03–1.59) | 0.12 (−12.7–5.73) | 0.91 |
Plasma, LC-MS/MS and VAMS (calibrator: DP) | 0.98 (0.86–1.23) | −2.26 (−18.3–1.74) | 0.91 | |
Plasma, LC-MS/MS and VAMS (calibrator: WB0) | 0.95 (0.87–1.16) | −4.25 (−13.6–1.80) | 1.21 | |
VAMS, DP calibrator and WB0 calibrator | 1.06 (0.94–1.09) | −0.91 (−4.92–0.23) | 0.91 | |
Rivaroxaban | Plasma, LC-MS/MS and plasma, Anti-Xa | 1.43 (1.34–1.75) | −1.14 (−8.39–0.85) | 0.89 |
Plasma, LC-MS/MS and VAMS (calibrator: DP) | 1.12 (0.97–1.26) | −2.27 (−7.20–1.76) | 0.89 | |
Plasma, LC-MS/MS and VAMS (calibrator: WB0) | 1.15 (0.97–1.27) | −2.09 (−6.85–1.40) | 1.12 | |
VAMS, DP calibrator and WB0 calibrator | 1.02 (1.00–1.03) | −0.05 (−1.21–0.87) | 0.89 |
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Fehér, A.; Vincze, I.; Rudge, J.; Domján, G.; Vásárhelyi, B.; Karvaly, G.B. Assays for Monitoring Apixaban and Rivaroxaban in Emergency Settings, State-of-the-Art Routine Analysis, and Volumetric Absorptive Microsamples Deliver Discordant Results. Diagnostics 2024, 14, 1939. https://doi.org/10.3390/diagnostics14171939
Fehér A, Vincze I, Rudge J, Domján G, Vásárhelyi B, Karvaly GB. Assays for Monitoring Apixaban and Rivaroxaban in Emergency Settings, State-of-the-Art Routine Analysis, and Volumetric Absorptive Microsamples Deliver Discordant Results. Diagnostics. 2024; 14(17):1939. https://doi.org/10.3390/diagnostics14171939
Chicago/Turabian StyleFehér, Adrienne, István Vincze, James Rudge, Gyula Domján, Barna Vásárhelyi, and Gellért Balázs Karvaly. 2024. "Assays for Monitoring Apixaban and Rivaroxaban in Emergency Settings, State-of-the-Art Routine Analysis, and Volumetric Absorptive Microsamples Deliver Discordant Results" Diagnostics 14, no. 17: 1939. https://doi.org/10.3390/diagnostics14171939