Validation parameters of the developed method for detection and quantification of four antibiotics in animal feed - repeatability, within-laboratory reproducibility, recovery and uncertainty
| Target compound | Fortification level (mg kg−1) | Repeatability, CV (%) | Reproducibility, CV (%) | Recovery (%) | Uncertainty (%) |
|---|---|---|---|---|---|
| Lincomycin | 0.25 | 18.33 | 11.62 | 91.23 | 23.4 |
| 1.0 | 9.44 | 8.76 | 89.60 | ||
| 2.0 | 6.92 | 11.69 | 90.31 | ||
| Spiramycin | 0.25 | 4.73 | 11.20 | 96.29 | 22.4 |
| 1.0 | 5.66 | 7.84 | 101.05 | ||
| 2.0 | 3.80 | 7.18 | 97.51 | ||
| Tylosin | 0.25 | 6.47 | 6.98 | 86.60 | 16.9 |
| 1.0 | 4.30 | 7.04 | 104.44 | ||
| 2.0 | 8.26 | 8.45 | 96.63 | ||
| Tiamulin | 0.25 | 7.62 | 7.29 | 102.81 | 14.6 |
| 1.0 | 2.57 | 6.25 | 105.07 | ||
| 2.0 | 2.93 | 6.22 | 100.27 |
The gradient programme followed for the elution of the examined analytes
| t (min) | A: 0.05M CH2O2 in water (%) | B: 0.05M CH2O2 in acetonitrile (%) |
|---|---|---|
| 0 | 95 | 5 |
| 3 | 73 | 27 |
| 11 | 73 | 27 |
| 13 | 50 | 50 |
| 20 | 50 | 50 |
| 22 | 95 | 5 |
| 35 | 95 | 5 |
Validation parameters of the developed method for detection and quantification of four antibiotics in animal feed - linearity, decision limit (CCα), detection capability (CCβ), limit of detection (LOD) and limit of quantification (LOQ)
| Target compound | Linearity R2 | CCα mg kg−1 | CCβ mg kg−1 | LOD mg kg−1 | LOQ mg kg−1 |
|---|---|---|---|---|---|
| Lincomycin | 0.9977 | 0.318 | 0.535 | 0.151 | 0.223 |
| Spiramycin | 0.9986 | 0.242 | 0.286 | 0.103 | 0.107 |
| Tylosin | 0.9989 | 0.294 | 0.317 | 0.029 | 0.069 |
| Tiamulin | 0.9970 | 0.213 | 0.259 | 0.065 | 0.147 |
Monitored ion (m/z) and retention time of analysed antibiotics
| Antibiotic | Monitored ion (m/z) | Retention time (min) |
|---|---|---|
| Lincomycin | 407 | 10–11 |
| Spiramycin | 422 | 11–12 |
| Tylosin | 916.5 | 20–21 |
| Tiamulin | 494 | 21–22 |