Table 1. Analytical Validation Parameters of nanoLC-HRMS Method for OXA and OXB Measurementa.
| |
intraday (N = 5) |
interday (N = 15) |
|||||
|---|---|---|---|---|---|---|---|
| analyte | matrix | theoretical concentration (pg/mL) | RE (%) | CV (%) | RE (%) | CV (%) | |
| OXA | surrogate | LLOQ | 2.5 | –3.3 | 7.8 | 1.9 | 12.8 |
| LQC | 7.5 | –4.9 | 4.8 | –1.5 | 6.6 | ||
| MQC | 120 | 0.4 | 2.6 | 0.3 | 2.1 | ||
| HQC | 200 | 0.6 | 1.8 | –0.5 | 1.9 | ||
| real | LQC | 57.7 | –9.3 | 7.7 | –8.8 | 14.0 | |
| HQC | 157.7 | –0.8 | 5.0 | 0.3 | 6.4 | ||
| OXB | surrogate | LLOQ | 2.5 | 0.0 | 10.3 | 2.0 | 9.6 |
| LQC | 7.5 | 0.1 | 4.6 | –4.3 | 8.2 | ||
| MQC | 120 | –7.6 | 2.4 | –3.3 | 2.7 | ||
| HQC | 200 | –2.1 | 3.3 | –3.9 | 2.9 | ||
| real | LQC | 28.2 | –3.5 | 8.6 | –5.6 | 9.7 | |
| HQC | 128.2 | 1.6 | 2.6 | –4.0 | 5.4 | ||
a
Intraday and interday assay precision (% CV) and accuracy (% RE) were evaluated in the surrogate matrix at 2.5 pg/mL (LLOQ), 7.5 pg/mL (lower quality control [LQC]), 120 pg/mL (middle quality control [MQC]), and 200 pg/mL (higher quality control [HQC]), and in real matrix spiked with orexins at 25 pg/mL (LQC) and 125 pg/mL (HQC).