Table 1.
Details of the Bioanalytical Methods for the Evaluated Statins
| Method: | Atorvastatin/2‐OH‐atorvastatin | Pravastatin | Rosuvastatin | |
|---|---|---|---|---|
| Analyte | Atorvastatin | 2‐OH‐Atorvastatin | Pravastatin | Rosuvastatin |
| Internal standard | Atorvastatin‐d 5 | 2‐OH‐Atorvastatin‐d 5 | Pravastatin‐d 3 | Rosuvastatin‐d 6 |
| Extraction method | Liquid‐liquid | Liquid‐liquid | Liquid‐liquid | |
| HPLC column | Waters Acquity UPLC HSS T3, 1.8 μm, 2.1 x 50 mm | Waters Acquity UPLC BEH C18, 2.1 x 50 mm, 1.7 μm | Phenomenex Luna C18(2), 2 x 30 mm, 5 μm | |
|
Mobile phase (Gradient components A and B) |
A: Water:acetic acid at 100:0.1 (v:v) B: Acetonitrile: acetic acid at 100:0.1 (v:v) |
A: Water:formic acid at 100:0.1 (v:v) B: Acetonitrile:water: formic acid at 95:5:0.1 (v:v:v) |
A: Water:formic acid at 100:0.1 (v:v) B: Methanol:formic acid at 100:0.1 (v:v) |
|
| Retention time | 1.33 min | 1.22 min | 1.16 min | 2.6 min |
| Mass spectrometer settings | ||||
| Ionization | Positive Turbo Ion Spray® | Negative Turbo Ion Spray® | Positive Turbo Ion Spray® | |
| Mode | MRM | MRM | MRM | |
| m/z monitored | ||||
| Analyte | 559.3 → 440.3 | 575.3 → 440.3 | 423.2 → 321.2 | 482.2 → 258.2 |
| Internal Standard | 564.3 → 445.3 | 580.3 → 445.3 | 426.2 → 321.2 | 488.2 → 264.2 |
| LLOQ | 0.1 ng/mL | 0.1 ng/mL | 0.1 ng/mL | 0.05 ng/mL |
| Precision (%CV) | ||||
| Within‐day | ≤4.7 | ≤5.9 | ≤4.4 | ≤6.0 |
| Between‐day | ≤3.7 | ≤5.0 | ≤3.8 | ≤5.7 |
| Accuracy (%RE) | ||||
| Within‐day | −2.7 to 3.3 | −0.7 to 5.0 | −2.3 to 4.0 | −1.6 to 10.0 |
| Between‐day | −2.0 to 2.5 | 0.0 to 3.8 | −1.9 to 2.7 | 1.2 to 6.0 |
%CV, coefficient of variation, expressed as a percentage; %RE, relative error, expressed as a percentage; LLOQ, lower limit of quantitation; MRM, multiple reaction monitoring.