Table 2.
Validation parameters of the HPLC method.
| Calibration Data for the Seven Bromophenol Standards | |||||||
|---|---|---|---|---|---|---|---|
| Substance | Regression Equation | Coefficient of Determination | Range (μg/mL) | LOD 1 (μg/mL) | LOQ 2 (μg/mL) | ||
| 1 | y = 83407x + 5.5502 | R² = 1.0000 | 102–0.05 | 0.017 | 0.053 | ||
| 2 | y = 98794x + 18.324 | R² = 0.9999 | 181–0.06 | 0.038 | 0.116 | ||
| 3 | y = 156713x + 34.799 | R² = 0.9991 | 47–0.02 | 0.008 | 0.024 | ||
| 4 | y = 126302x + 44.688 | R² = 0.9990 | 49–0.02 | 0.011 | 0.034 | ||
| 5 | y = 164091x + 25.645 | R² = 0.9998 | 56–0.03 | 0.014 | 0.043 | ||
| 6 | y = 126669x - 28.684 | R² = 0.9995 | 49–0.05 | 0.032 | 0.097 | ||
| 7 | y = 114941x + 42.574 | R² = 0.9996 | 95–0.02 | 0.015 | 0.047 | ||
| Accuracy and Precision of the Assay | |||||||
| Accuracy, High Spike (n = 3) | Accuracy, Medium Spike (n = 3) | ||||||
| Substance | Measured Values 3 | Theoretical Values 3 | Displace-ment 4 | Substance | Measured Values 3 | Theoretical Values 3 | Displace-ment 4 |
| 1 | 89.5 | 97.3 | 2.67 | 1 | 69.0 | 71.3 | 3.29 |
| 2 | 135.1 | 142.1 | 4.93 | 2 | 49.5 | 51.3 | 3.51 |
| 3 | 41.1 | 41.6 | 1.23 | 3 | 28.1 | 26.8 | 4.68 |
| 4 | 48.3 | 49.5 | 2.45 | 4 | 27.9 | 27.3 | 1.96 |
| 5 | 49.8 | 51.7 | 3.60 | 5 | 29.8 | 28.5 | 4.80 |
| 6 | 43.9 | 45.2 | 2.85 | 6 | 25.3 | 25.1 | 1.12 |
| 7 | 87.9 | 87.7 | 0.24 | 7 | 46.0 | 45.0 | 2.15 |
| Accuracy, Low Spike (n = 3) 3 | Precision (n = 5) 5 | ||||||
| Substance | Measured Values 3 | Theoretical Values 3 | Displace-ment 4 | Day 1 | Day 2 | Day 3 | Inter-day |
| 1 | 50.7 | 51.4 | 1.33 | 1.83 | 0.71 | 1.21 | 2.33 |
| 2 | 29.1 | 30.4 | 4.07 | 1.26 | 0.76 | 1.16 | 2.35 |
| 3 | 12.5 | 13.0 | 3.57 | 2.15 | 0.62 | 1.06 | 2.93 |
| 4 | 9.7 | 10.1 | 4.30 | 2.59 | 1.71 | 3.86 | 2.93 |
| 5 | 6.9 | 7.0 | 0.54 | 3.83 | 6.28 | 2.56 | 5.21 |
| 6 | 5.8 | 5.6 | 3.56 | 1.23 | 0.84 | 2.87 | 1.74 |
| 7 | 23.6 | 23.5 | 0.51 | 1.23 | 0.43 | 2.28 | 2.11 |
1 LOD: limit of detection determined with the purified standards. 2 LOQ: limit of quantification determined with the purified standards. 3 Values in µg/mL. 4 Displacement in percent (sample Vertebrata lanosa, Brittany, 2018). 5 Maximum relative standard deviation based on the peak area given in percent.