Table 2.
Summary of validation results.
| Property | QC | ATV | 2-OH ATV | ATV L | 2-OH ATV L | BSP | CLP-CA | |
|---|---|---|---|---|---|---|---|---|
| Selectivity | Analyte (% LLOQ) | – | 11.0 | 14.3 | 4.3 | 7.3 | 2.1 | 3.2 |
| Internal Standard (% IS) | – | 0.71 | 0.90 | 0.28 | 1.35 | 0.09 | 0.08 | |
| Calibration curve performance | Correlation coefficient (r) 1 | – | 0.9995 | 0.9995 | 0.9982 | 0.9975 | 0.9983 | 0.9977 |
| Dilution integrity | Accuracy (%)2 | – | 98.7 | 93.7 | 102.9 | 106.7 | 109.0 | 96.4 |
| Precision, CV (%) | – | 11.7 | 6.2 | 3.4 | 4.9 | 14.0 | 3.3 | |
| Carryover | Analyte (% LLOQ) | – | 6.2 | 3.7 | 2.7 | 10.9 | 1.1 | 5.2 |
| Internal Standard (% IS) | – | 0.1 | 0.5 | 0.1 | 0.1 | 0.1 | 0.1 | |
| IS-normalised Matrix effect | Mean (%)2 | Low | 103.9 | 98.0 | 113.1 | 88.9 | 107.4 | 106.5 |
| High | 112.2 | 98.4 | 114.7 | 102.8 | 106.3 | 100.9 | ||
| Precision, CV (%) | Low | 12.3 | 12.2 | 11.6 | 8.8 | 7.0 | 8.8 | |
| High | 5.8 | 4.7 | 3.5 | 8.8 | 4.3 | 2.2 | ||
| Recovery | Mean (%)2 | Low | 102.4 | 96.5 | 102.5 | 108.5 | 98.5 | 93.8 |
| High | 99.1 | 98.8 | 98.2 | 100.6 | 97.4 | 92.7 | ||
| Precision, CV (%) | Low | 13.3 | 13.9 | 8.9 | 5.4 | 11.0 | 12.4 | |
| High | 5.7 | 6.8 | 5.1 | 6.6 | 7.0 | 3.9 | ||
| Within run accuracy and precision | Accuracy (%)2 | LLOQ | 95.0 | 97.0 | 90.3 | 107.6 | 92.8 | 101.5 |
| Low | 104.5 | 102.2 | 98.8 | 108.6 | 102.8 | 102.7 | ||
| Medium | 99.0 | 99.8 | 99.4 | 93.8 | 99.1 | 99.7 | ||
| High | 96.1 | 94.3 | 99.0 | 97.8 | 96.0 | 95.0 | ||
| Precision, CV (%) | LLOQ | 11.1 | 6.0 | 9.5 | 12.1 | 2.3 | 3.3 | |
| Low | 8.2 | 7.7 | 6.0 | 8.7 | 4.3 | 3.4 | ||
| Medium | 1.6 | 2.2 | 2.3 | 4.2 | 2.0 | 2.3 | ||
| High | 4.5 | 4.0 | 4.7 | 5.8 | 4.3 | 5.5 | ||
| Between run accuracy and precision | Accuracy (%)2 | LLOQ | 95.3 | 96.1 | 100.0 | 100.5 | 94.4 | 97.0 |
| Low | 103.7 | 101.5 | 98.2 | 101.2 | 102.8 | 101.1 | ||
| Medium | 100.5 | 99.2 | 95.1 | 95.0 | 99.8 | 99.0 | ||
| High | 99.0 | 97.3 | 98.0 | 99.8 | 99.0 | 98.0 | ||
| Precision, CV (%) | LLOQ | 8.9 | 8.2 | 10.9 | 14.6 | 5.2 | 4.7 | |
| Low | 6.3 | 8.5 | 5.8 | 12.1 | 4.4 | 3.7 | ||
| Medium | 2.6 | 4.1 | 4.6 | 5.7 | 2.6 | 3.5 | ||
| High | 3.9 | 4.7 | 4.0 | 5.7 | 5.3 | 4.7 | ||
| RT Benchtop stability (% of nominal concentration) | Low | 114.3 | 107.7 | 100.3 | 105.6 | 98.5 | 107.7 | |
| High | 107.1 | 112.9 | 94.7 | 107.8 | 96.1 | 103.2 | ||
| Freeze-thaw stability (% of nominal concentration) | Low | 110.2 | 114.5 | 89.7 | 96.6 | 102.5 | 106.6 | |
| High | 103.6 | 102.4 | 88.2 | 96.0 | 100.9 | 99.2 | ||
| Long term stability (% of nominal concentration) | Low | 107.5 | 111.4 | 109.7 | 112.5 | 103.3 | 103.9 | |
| High | 104.4 | 106.0 | 106.4 | 103.2 | 104.5 | 99.8 | ||
| Autosampler stability (% of nominal concentration) | Low | 91.9 | 108.7 | 101.7 | 103.8 | 101.0 | 104.5 | |
| High | 103.9 | 101.1 | 97.4 | 99.0 | 99.7 | 103.9 | ||
Abbreviations: LLOQ = lower limit of quantification; IS = internal standard; QC = quality control; CV = coefficient of variation; RT = room temperature.
1
Average of 3 independent runs.
2
Accuracy is represented as % of nominal concentration.