Table 1.
Validation data for the analytical method used for the determination of pregabalin in human plasma by UPLC-MS/MS using gabapentin as the internal standard
| Parameters | At concentration of
|
||
|---|---|---|---|
| Low (30.00 ng/mL) | Medium (2,500.11 ng/mL) | High (4,500.19 ng/mL) | |
| Precisiona | |||
| Intra-assay CV | 4.70% | 0.46% | 0.25% |
| Inter-assay CV | 3.12% | 2.25% | 1.44% |
| Accuracya | |||
| Intra-assay CV | 4.73% | −1.30% | −1.74% |
| Inter-assay CV | 5.80% | −1.15% | −1.80% |
| Stability | |||
| At −20°C (stable until 49 days) | −7.86% to 2.77% | – | −4.79% to 1.84% |
| At room temperature (stable until 6 hours) | −1.34% to 2.77% | – | −2.96% to 2.51% |
| Freeze and thaw (stable until 3 cycles) | −5.00% to 6.11% | – | −3.00% to 0.17% |
| Linearity | |||
| The linearity of the standard calibration curves was obtained (r of 1.00 on day 1, 1.00 on day 2, and 0.99 on day 3) | |||
| LLOQ | |||
| The LLOQ has been established at 10.00 ng/mL | |||
| Selectivity | |||
| The % difference of analyte interference ranged from 0.00% to 10.07%, while the % difference of internal standard interference ranged from 0.00% to 0.13%. From the result, it can be concluded that there was no interferences of the analyte and internal standard compounds | |||
| Range | |||
| The range of quantification has been established from 10.00 to 6,000.49 ng/mL | |||
Note:
a
Shown by the difference of the measured values to actual values (% difference).
Abbreviations: CV, coefficient of variation; LLOQ, lower limit of quantification; UPLC-MS/MS, ultra-performance liquid chromatography with tandem mass spectrometry.