Table 3.
Validation: stability data for thymoquinone in different biomatrixes
| Exposure condition | LQC(290.0 ng/mL or ng g−1) | MQC(1000.0 ng/mL or ng g−1) | HQC (2000.0 ng/mL or ng g−1) | ||||||
|---|---|---|---|---|---|---|---|---|---|
| Brain homogenate | Lungs homogenate | Plasma | Brain homogenate | Lungs homogenate | Plasma | Brain homogenate | Lungs homogenate | Plasma | |
| Long term stability; recovery (ng) after storage (− 80 °C) | |||||||||
| Previous day | 288.9 ± 0.93 | 288.6 ± 0.50 | 288.6 ± 0.54 | 994.6 ± 13.36 | 992.6 ± 14.6 | 993.4 ± 17.4 | 1993.6 ± 26.4 | 1992.6 ± 27.8 | 1989.1 ± 27.5 |
| 30th day | 281.2 ± 0.91 (97.33%) | 287.1 ± 0.45 (99.48%) | 284.1 ± 0.91 (98.44%) | 982.4 ± 15.6 (98.77%) | 988.6 ± 15.4 (99.60%) | 984.6 ± 17.1 (99.11%) | 1981.4 ± 24.1 (99.39%) | 1988.2 ± 26.1 (99.78%) | 1970.4 ± 24.6 (99.06%) |
| Freeze-thaw stress; recovery (ng) after freeze-thaw cycles (− 80 °C to 25 °C) | |||||||||
| Pre-cycle | 287.9 ± 0.67 | 289.3 ± 0.81 | 289.1 ± 0.64 | 993.7 ± 15.6 | 993.4 ± 14.6 | 991.5 ± 16.4 | 1990.4 ± 27.0 | 1991.8 ± 27.3 | 1990.6 ± 26.4 |
| First cycle | 284.1 ± 0.90 (98.68%) | 286.6 ± 0.86 (99.07%) | 288.2 ± 0.74 (99.69%) | 985.4 ± 13.9 (99.16%) | 988.4 ± 2.46 (99.50%) | 987.9 ± 17.5 (99.64%) | 1981.7 ± 26.9 (99.56%) | 1988.4 ± 26.4 (99.83%) | 1981.4 ± 27.4 (99.54%) |
| Second cycle | 282.3 ± 0.92 (98.05%) | 283.4 ± 0.94 (97.96%) | 286.47 ± 0.66 (99.09%) | 979.4 ± 14.6 (98.56) | 985.7 ± 14.2 (99.22%) | 986.4 ± 16.3 (99.49%) | 1975.4 ± 23.8 (99.25%) | 1981.5 ± 25.8 (99.48%) | 1975.4 ± 26.1 (99.24%) |
| Third cycle | 280.3 ± 0.93 (97.36%) | 280.1 ± 0.93 (96.82%) | 285.94 ± 0.91 (98.91%) | 975.6 ± 15.4 (98.18%) | 982.4 ± 14.5 (98.89%) | 979.9 ± 17.1 (98.83%) | 1971.1 ± 24.7 (99.03%) | 1974.2 ± 26.7 (99.12%) | 1970.9 ± 25.4 (99.01%) |
| Bench top stability; recovery (ng) at room temperature (25 °C) | |||||||||
| 0 h | 288.4 ± 0.91 | 287.8 ± 0.91 | 288.47 ± 0.71 | 992.8 ± 14.6 | 991.4 ± 14.7 | 989.9 ± 16.8 | 1992.4 ± 25.4 | 1989.1 ± 28.1 | 1991.5 ± 26.7 |
| 24 h | 286.3 ± 0.78 (99.27%) | 282.3 ± 0.81 (98.09%) | 286.44 ± 0.84 (99.30%) | 986.4 ± 15.0 (99.36%) | 984.6 ± 13.5 (99.31%) | 981.4 ± 15.8 (99.14%) | 1981.5 ± 27.1 (99.45%) | 1977.6 ± 27.5 (99.42%) | 1979.1 ± 27.4 (99.38%) |
| Post processing stability; recovery (ng) after storage in auto sampler (4 °C) | |||||||||
| 0 h | 289.3 ± 0.34 | 288.6 ± 0.81 | 289.21 ± 0.64 | 993.4 ± 14.9 | 993.4 ± 14.5 | 988.7 ± 17.6 | 1991.4 ± 26.3 | 1988.9 ± 28.3 | 1988.3 ± 23.4 |
| 4 h | 287.5 ± 0.62 (99.38%) | 281.94 ± 0.49 (97.69%) | 287.3 ± 0.84 (99.34%) | 989.6 ± 13.6 (99.62%) | 986.6 ± 13.9 (99.32%) | 981.2 ± 18.0 (99.24%) | 1983.1 ± 25.7 (99.58%) | 1971.5 ± 26.4 (99.13%) | 1970.1 ± 22.9 (99.08%) |
Values (Mean ± SD) are derived from six replicates. Figures in parenthesis represent analyte concentration (%) relative to time zero. Theoretical contents; LQC: 290.0 ng mL−1; MQC: 1000.0 ng mL−1; and HQC: 2000.0 ng mL−1