Table 1.
Assay | Mobile Phase | Gradient Profile | Detection Method | Retention Times |
---|---|---|---|---|
Curcuminoids | 10 mM ammonium acetate (pH 4.2) | 40% CH3CN (0–20 min) → 75% CH3CN (25 min) →40% CH3CN (26–28 min) | Curcuminoids: 425 nm Phenacetin: 254 nm | Curcumin: 25.2 min Demethoxy: 24.6 min Bisdemethoxy: 23.6 min Phenacetin (IS): 3.6 min |
Triazolam | 10 mM phosphate buffer (pH 7.0) | 25% CH3CN/ 15% CH3OH (0–17 min) → 32% CH3CN/ 20% CH3OH (18 min) → 45% CH3CN/ 20% CH3OH (25 min) → 25% CH3CN/ 15% CH3OH (27–32 min) | 220 nm | 1′-OH-triazolam: 15.2 min 4-OH-triazolam:16.7 min Diazepam (IS): 26.5 min |
Flurbiprofen | 20 mM phosphate buffer (pH 2.2) | 35% CH3CN (0 min) → 50% CH3CN (5 min) → 75% CH3CN (15 min) → 35%CH3CN (17–20 min) | 4′-OH-flurbiprofen: ex 260 nm, em 320 nm Phenacetin: 220 nm | 4′-OH-flurbiprofen: 8.9 min Phenacetin (IS): 6.5 min |
S-mephenytoin | 0.5% formic acid | 10% CH3OH (0 min) → 80% CH3OH (4 min) → 10% CH3OH (4.1–12 min) | 4-OH-mephenytoin (MS-MS): 235.5 (width 5) → 150.2 Diazepam (MS): 285.3 | 4-OH-mephenytoin: 3.7 min Diazepam (IS): 5.4 min |
Dextromethorphan | 20 mM phosphate buffer (pH 2.2) | 20% CH3CN (0–5 min) → 60% CH3CN(14 min) →20% CH3CN (15–18 min) | ex 280 nm, em 310 nm | Dextrorphan: 7.8 min Pronethalol (IS): 12.1 min |
Phenacetin | 0.5% formic acid | 0% CH3CN (0 min) → 50% CH3CN (8 min) → 0% CH3CN (8.1–12 min) | APAP (+, MS-MS): 152.1 (1.0 width) → 110.0 Sulfaphenazole (+): 315.3 | APAP: 2.9 min Sulfaphenazole (IS): 6.6 min |
Chlorzoxazone | 20 mM phosphate buffer (pH 2.2) | 18% CH3CN (0 min) → 50% CH3CN (15 min) → 18% CH3CN (16–20 min) | 295 nm | 6-OH-chlorzoxazone: 8.8 min Phenacetin (IS): 12.8 min |
Bupropion | 0.02% TFA | 5% CH3CN (0 min) → 60% CH3CN (5 min)→5% CH3CN (5.2–10 min) | Hydroxybupropion (+, MS- MS): 256.0 (2.0 width) → 238.0 Trazodone (+): 372.6 | Hydroxybupropion: 2.1 min Trazodone (IS): 4.1 min |
Acetaminophen (+ Mode) | 10 mM Ammonium Acetate (pH 4.2) | 0% CH3CN (0–4 min) → 50% CH3CN(14 min) →0% CH3CN (14.1–23 min) | APAP-glucuronide (MS-MS): 345.2 (10 width) → 152.2 Sulfaphenazole: 315.3 | APAP-glucuronide: 1.4 min Sulfaphenazole (IS): 10.7 min |
Acetaminophen (− Mode) | 10 mM Ammonium Acetate (pH 4.2) | 0% CH3CN (0 min) → 50% CH3CN (10 min) → 0% (10.1–15 min) | APAP-sulfate (MS-MS): 230.2 (10 width) → 150.2 Sulfaphenazole: 313.3 | APAP-sulfate: 2.9 min Sulfaphenazole (IS): 7.1 min |
IS: internal standard, ex: excitation; em: emission