Table 4. Validation parameters of the analytical method for to quantify ifosfamide in DBS at 45% HTC.
| LLQ (100 ng/mL) | |||
|---|---|---|---|
| Intra-day variability | Day 1 | Day 2 | Day 3 |
| Mean± SD (n = 5) | 105.8±6.0 | 103.7± 3.1 | 96.43± 3.9 |
| % CV | 5.7 | 3.0 | 4.1 |
| % Deviation | -5.8 | -3.7 | 3.5 |
| Inter-day variability | |||
| Mean± SD (n = 15) | 101.2 ± 5.7 | N/R | N/R |
| % CV | 5.6 | N/R | N/R |
| % Deviation | -1.2 | N/R | N/R |
| Inter-day variability | QC1 (ng/mL) | QC2 (ng/mL) | QC3 (ng/mL) |
| Mean± SD (n = 15) | 320.9 ± 20.7 | 4006.5 ± 279.4 | 7343.2 ± 463.5 |
| % CV | 6.4 | 6.9 | 6.3 |
| % Deviation | -6.9 | -0.1 | 8.2 |
| Intra-day variability | |||
| Mean± SD (n = 5) a | 322.1±17.5 | 3782.5± 84.7 | 6981.62± 114.3 |
| % CV | 5.4 | 2.2 | 1.6 |
| % Deviation | -6.0 | 5.4 | 12.7 |
| Mean± SD (n = 5) b | 307.5±12.6 | 4061.8± 161.8 | 7534.5± 545.6 |
| % CV | 4.1 | 3.9 | 7.2 |
| % Deviation | -2.51 | -1.55 | 5.8 |
| Mean± SD (n = 5) c | 337.1±21.6 | 4217.4± 381.9 | 7513.6± 442.3 |
| % CV | 6.4 | 9.0 | 5.8 |
| % Deviation | -12.3 | -5.4 | 6.0 |
| Room temperature stability (25 o C)/ 3 days | |||
| Mean± SD (n = 5) | 313.1± 25.4 | N/R | 7797.5± 179.7 |
| % CV | 8.1 | N/R | 2.3 |
| % Deviation | -4.4 | N/R | 2.5 |
| Long term stability (52 d at -80 ° C) | |||
| Mean± SD (n = 5) | 311.6± 15.3 | N/R | 7839.7±182.9 |
| % CV | 4.9 | N/R | 2.3 |
| % Deviation | -3.8 | N/R | 2.0 |
The intra-day variability with quality controls was performed in three consecutive days, data for days 1 to 3 are depicted as a, b and c, respectively. SD: Standard Deviation. CV: Coefficient Variation. N/R: Not required according to the guidelines. Amount claimed for QC1, QC2 and QC3 were 300, 4000 and 8000 ng/mL, respectively.