Table 1.
Validation parameters for determination of treosulfan and S,S-EBDM in rat plasma and RBC
| Parameter | Treosulfan | S,S-EBDM | ||
|---|---|---|---|---|
| Plasma | RBC | Plasma | RBC | |
| Linearity (μM)a | 1.1–110 (curve I) | 1.9–93 | ||
| 110–2209 (curve II) | ||||
| Precision—CV (%)b | ||||
| Within-run (n = 5) | 2.8–7.6 (2.8) | 3.1–9.0 (9.0) | 3.7–19.5 (19.5) | 3.1–5.1 (4.1) |
| Between-run (n = 3) | 1.0–6.1 (6.1) | 2.9–11.1 (4.1) | 3.0–6.0 (6.0) | 2.4–4.0 (4.0) |
| Accuracy (%)b | ||||
| Within-run (n = 5) | 94.7–119.5 (119.5) | 94.3–109.5 (100) | 101.8–109.2 (109.2) | 91.4–109.1 (91.4) |
| Between-run (n = 3) | 97.8–104.6 (104.6) | 85.7–102.9 (85.7) | 102.4–113.2 (113.2) | 95.2–103.9 (95.2) |
| Stability (%) (n = 3)c | 90.7–108.8 | 85.8–104.8 | 88.3–89.4 | 86.7–89.8 |
CV coefficient of variation, LLOQ lowest limit of quantitation, RBC red blood cells, S,S-EBDM (2S,3S)-1,2-epoxy-3,4-butanediol 4-methanesulfonate (treosulfan monoepoxide)
aThe calibration curves were established using the calibration standards at 7–8 concentration levels. For the analysis of treosulfan, two calibration curves were prepared that covered low and high concentrations. The LLOQ was the lowest concentration of the calibration standard
bThe ranges shown in the table include the mean results obtained for the quality control samples containing treosulfan at 1.1, 22, 55, 110, 1104, and 2209 μM, and S,S-EBDM at 1.9, 37, and 94 μM. The values in the parentheses show the results obtained for the LLOQ level
cThe stability values were calculated as a ratio of the final to the initial concentration of the analyte in the samples used in the freeze–thaw stability test. The ranges shown in the table include the mean results obtained for the quality control samples containing treosulfan at 5.5, 55, and 800 μM, and S,S-EBDM at 9.4 and 94 μM