Table 2.
Multi-analyte assays for first- and second-line anti-tuberculosis drugs and single -analyte assays for second-line drugs using LC-MS/MS.
| Drug | Internal standard | Prep. time* (min) | Sample volume (µl), matrix | Protein precipitation solvent | Chromatographic column | Range (µg/ml) | Mobile phase Solvent A, B and C, pH | Run time (min) | Retention time (min) | Mass transitions |
|---|---|---|---|---|---|---|---|---|---|---|
| R, H, E, Z [44] | Rifabutin (R), 6-aminonicotinic acid (H, E, Z) | >10 | 50, serum | methanol | YMC Co, Hydrosphere C18 (50 × 2.0 mm, 3 μm) | R 1.1–320, H 0.07–19, E 0.07–18, Z 0.4–290 | A: 0.3% formic acid in 100% methanol B: 0.3% formic acid in water. pH: NA | 4 | R 3.43, H 1.27, E 0.89, Z 134 | R 824–792, H 138–121, E 205–116, Z 124–81 |
| R, H, E, Z, S [45] | Rifaxim (R), Phenformin hydrochloride (other) | >12 | 100, plasma | methanol, formic acid, ammonium acetate | GL Sciences, Interstil HILIC (75 × 2.1 mm, 3 μm) | R, H, Z 0.004–4, E 0.005–0.5, S 0.01–16 | A: methanol B: water with 0.1% formic acid and 5 mM ammonium acetate** pH 3.0. | 2 | R 1.42, H 1.41, E 1.22, Z 1.64, S 1.11 | R 823.7–791.6, H 138.1–121.1, E 205.3–116.2 Z 124.1–81.1, S 582.5–263.3 |
| R, H, E, Z [46], [43] | 6,7-dimethyl-2,3-di(2-pyridyl)quioxaline (R), Thymidine (H, E, Z) | >10 | 200, plasma | acetonitrile | Waters, Acquity HSS T3 (150 × 2.1 mm, 1.8 μm) | R 0.47–60, H, E 0.12–15 Z 0.55–70 | A: water with 0.05% formic acid B: acetonitrile with 0.05% formic acid pH: NA | 6 | R ca 4.4, H ca 1.6, E ca 0.8, Z ca 3.1 | R 824–792, H 138–79, E 205–116, Z 124–81 |
| R, H, E, Z, S [47] | Phenacetin | >18 | 100, plasma | methanol, formic acid | Agilent, Zorbax SB-C18 (100 × 2.1 mm, 3.5 μm) | R, Z 0.2–4, H 0.08–2, E 0.0002–1, S 0.002–0.2 | A: 0.02% heptaflourobutyric acid, 0.2% formic acid in methanol B: 0.02% heptaflourobutyric acid, 0.2% formic acid in water pH: NA | 8.5 | R ca 7.2, H ca 1.3, E ca 1.6, Z ca 1.7, S ca 1.5 | R 823.4–791.2, H 138.1–120.9, E 205.3–116.2 Z 124.1–79.1, S 582.3–262.6 |
| R, H, E, Z [50] | D3-Rifampicin (R), D4-isoniazid (H) 15 N,D3-pyrazinamide (Z), D4-ethambutol dihydrochloride (E) | >82 | 50, plasma | methanol, acetonitrile | Merck, Chromolith Flash RP-18e (25 × 4.6 mm, 2 μm) | R 0.75–30, H 0.5–10, E 0.25–10, Z 4–80 | A: 5 mM ammonium acetate, 5% methanol and 0.45% formic acid in water. B: 5 mM ammonium acetate, 95% methanol and 0.45% formic acid in water C: 5 mM ammonium acetate in water. pH: NA | 10.2 | R ca 5.5, H ca 1.1, E ca 0.9, S ca 1.5 | R 824.0–792.5, H 138.1–121.1, E 205.1–116.1, Z 124.1–81.1 |
| R, H, E, Z, Lfx, Mfx, Am, Km, S, Eto, Pto, Cs, Lzd, Cfz, PAS, Amx [48] | Apramycin (Z, Am, Km, S), D4-Moxifloxacin (other) | >10 | 100 (Z, Am, Km, S) 50 (other), plasma | acetonitrile | Waters, Atlantis HILIC (150 × 2.1 mm, 3 μm) (Z, Am, Km, S) Waters, Atlantis dC18 (150 × 2.0 mm, 3 μm) (other) | R, Lfx, Mfx, Pto 0.2–10, H, E, Eto 0.1–5.0, Z 2.0–100, Am, Km, S, PAS 1.0–50, Cs 0.8–40, Cfz 0.04–2.0, Lzd, Amx 0.4–20 | A: 0.1% formic acid solution B: 0.1% formic acid in acetonitrile pH: NA | Z, Am, Km, S 9 Other 13 | R ca 7.3, H ca 2.1, E ca 1.8, Z ca 2.8, Lfx ca 6, Mfx ca 6.5, Am ca 2.6, Km ca 2.6, S ca 2.6, Eto ca 3.3, Pto ca 4.7, Cs ca 2, Lzd ca 6.7, Cfz ca 7.5, PAS ca 6.3, Amx ca 2.8 | R 823.4–791.1, H 138.0–121.0, E 167.1–107.0, Z 124.0–81.0, Lfx 362.0–318.0, Mfx 402.0–384.0, Am 586.2–425.3, Km 485.5–163.3, S 582.2–263.3, Eto 167.1–107.0, Pto 181.0–154.3, Cs 103.0–58.0, Lzd 338.0–296.0, Cfz 473.2–431.2, PAS 154.0–119.0, Amx 366.0–114.0 |
| Lfx, Mfx, Km, S, Pto, Cs, Lzd, PAS [52] | Gm (Km), Dihydrostreptomycin (S), Lomefloxacin (Lfx, Mfx, Lzd), Ethionamide (Pto), Muscimol (Cs), 4-aminobenzoic acid (PAS) | >8 | 50 (Lfx, Mfx, PAS), 10 (other), serum | acidified methanol | Waters, Acquity HSS T3 (50 × 2.1 mm, 1.8 µm) | Lfx, Mfx, Lzd 1.0–20, Km, S, Cs, 5–100, Pto 0.5–10, PAS 0–100 | A: 10 mM ammonium formate in 0.1% formic acid B: acetonitrile in 0.1% formic acid pH: NA | 3 | Lfx 1.37, Mfx 1.90, Km 0.51, S 0.56, Pto 1.88, Cs 0.63, Lzd 1.96, PAS 1.42 | Lfx 362.3–318.2, Mfx 402.4–384.3, Km 485.5–163.1, S 582.6–263.2, Pto 181.0–154.1, Cs 103.0–75.1, Lzd 338.3–296.2, PAS 154.1–119.0 |
| Bdq [55] | D6-bedaquiline | >6 | 10, serum | acetonitrile, methanol | Thermo Scientific, Hypurity C18 (50 × 2.1 mm, 3 μm) | 0.05–6 | A: 5 g/l ammonium acetate in 100% acetic acid 25 ml/L and 100% triflouroacetic acid 2 ml/L B: water C: acetonitrile pH: NA | 2.6 | 1.9 | 555.1–58.4 |
| Dlm [62] | OPC-14714 | >7 | 50, plasma | acetonitrile | Waters, UPLC BEH C18 (50 × 2.1 mm, 1.7 μm) | 0.001–0.5 | A: 5 mM ammonium bicarbonate and 0.2% ammonium hydroxide in water B: 0.2% ammonium hydroxide in methanol pH: NA | 4 | 1.9 | 535.1–352.2 |
| Mpm, Ipm [64] | D6-Meropenem | >10 | 100, plasma | methanol | Thermo Fischer Scientific, Accucore C18 (100 × 2.1 mm, 2.6 μm) | 0.5–32 | A: 2 mM ammonium formate and 0.1% formic acid in water B: acetonitrile with 0.1% formic acid | 9 | Mpm ca 3.4, Ipm 3.04 + 3.09 | Mpm 384.2, Ipm 300.1 |
| Mpm [65] | D6-Meropenem | >12 | 50, plasma | acetonitrile | Waters, Acquity UPLC BEH C18 (100 × 2.1 mm, 1.7 μm) | 0.27–150 | A: 0.1% formic acid in water B: 0.1% formic acid in acetonitrile pH: NA | 3 | 1.05 | 384.2–141.0 |
| Clv**** [66] | NA | >5 | 1000, serum | acetonitrile | Waters, Acquity UPLC BEH C18 (100 × 2.1 mm, 1.7 μm) | 0.1–5 | A: methanol B: 0.01% formic acid in water pH: NA | 5.8 | 1.05–1.35 | 198.1–136.2 |
All studies used protein precipitation as extraction method, gradient elution for the mobile phase and positive electrospray ionization mode unless specified. *Times specified in each manuscript for different sample preparation steps (e.g. vortexing and centrifugation), **isocratic elution, ***solid phase extraction, ****negative electrospray ionization mode. LC-MS/MS = liquid chromatography-tandem mass spectrometry, Prep. time = sample preparation time, R = rifampicin, H = isoniazid, E = ethambutol, Z = pyrazinamide, Lfx = levofloxacin, Mfx = moxifloxacin, Am = amikacin, Km = kanamycin, S = streptomycin, Eto = ethionamide, Pto = prothionamide, Cs = cycloserine, Lzd = linezolid, Cfz = clofazimine, Bdq = bedaquiline, Dlm = delamanid, PAS = p-aminosalicylic acid, Ipm = imipenem, Mpm = meropenem, Amx = amoxicillin, Clv = clavulanate, Gm = Gentamycin, NA = not available.