Table 3.
Reported liquid chromatographic (LC) and gas chromatographic (GC) methods for the quantification of acetazolamide in human urine, plasma or serum.
| Method | Matrix | Column | Derivati-zation | Mobile phase | Sample preparation | LOD; LOQ | Ref. | |
|---|---|---|---|---|---|---|---|---|
| Liquid chromatography | ||||||||
| LC-MS/MS | Plasma | Hypurity advance (50 mm × 4.6 mm) | none | MeCN + 0.1% FA | 100 μL plasma + IS + FA. SPE. Evaporation to dryness and reconstitution with 500 μL MeCN +0.1% FA | n.r.; 50 ng/mL | [6] | |
| HPLC-DAD | Urine | HP-Hypersil ODS-Cls (250 mm × 4.0 mm) | none | A: phosphate buffer; B: MeCN | 2 mL urine + 0.5 g sodium phosphate + 0.5 g NaCl. Extraction with 4 mL EA, addition of 5% lead acetate to the organic phase. Centrifugation, evaporation to dryness, reconstitution in 300 μL MeOH; 10 μL injection | 8 ng/mL | [7] | |
| HPLC-DAD | Urine | HP-LiChrospher 100 RP 18 (125 mm × 4.0 mm) | none | MeCN-H2O | native urine; column-switching technique | 10 ng/mL | [8] | |
| LC-MS/MS | Plasma (beagle) | Shimadzu VP-ODS C18 (150 mm × 2.0 mm) | none | A: H2O; B: MeCN | 100 μL plasma + IS. Protein precipitation with 600 μL MeCN. | n.r.; 200 ng/mL | [9] | |
| UHPLC-HRMS | Urine | Zorbax SB-C8 (2.1 mm × 50 mm) | none | A: H2O + 1 mM ammonium acetate + 0.001% AA; B: MeOH + 1 mM ammonium acetate + 0.001% AA |
100 μL urine + IS; centrifugation | 50 ng/mL; n.r. | [10] | |
| LC-MS/MS | Urine | Supelco Discovery HS-C18 (50 mm × 2.1 mm) |
none | A: H2O + 0.2% FA; B: MeOH + 0.2% FA | SPE. Evaporation to dryness and reconstitution with H2O + 0.2% FA. Filtration through a 0.45-μm membrane. | 25 ng/mL; n.r. | [11] | |
| UHPSFC-MS/MS | Urine | Acquity UPC2 BEH (100 mm × 3.0 mm) | none | CO2 + MeOH + 10 mM FA + 2% H2O | 100 μL urine and 10-fold dilution with H2O-MeCN | 0.15 ng/mL; n.r. | [12] | |
| Gas chromatography | ||||||||
| GC-ECD | Serum | Glass columns (150 cm × 0.18 cm) | CH3I | Nitrogen | 100 μL serum + IS + 1 mL tetrapentylammonium + 0.2 mL NaOH + H2O. Addition of 1 mL CH2Cl2 with 5% CH3I. Evaporation of the organic phase, reconstitution with 1 mL toluene washing with aqueous silver sulphate. | 500 ng/mL | [13] | |
| GC-MS | Urine | HP Ultra 1 (25 m × 0.20 mm) | CH3I | Helium | 1 mL urine + 25 μL NaOH 6 M + IS + 150 μL tetrahexylammonium hydrogensulphate + 5 mL CH3I. Extraction with toluene, centrifugation. Filtration through SM-7 resin, evaporation to dryness and reconstitution in 100 μL toluene. | 50 ng/mL; n.r. | [14] | |
| GC-MS | Urine | HP (25 m × 0.20 mm) | CH3I | Helium | 5 mL urine + IS, filtration through XAD-2 resin. Evaporation to dryness and reconstitution with 200 μL Me2CO. Addition of 20 μL CH3I and K2CO3. Derivatization for 3 h at 60 °C. | n.r. | [15] | |
| GC-MS | Urine | DB1 MS (5 m × 0.10 mm) | CH3I | Helium | 2 mL urine + 80 μL NaOH + 1 mL CH2Cl2-2-propanol. Reconstitution with 50 mg K2CO3 + 400 μL of Me2CO/CH3I. Derivatization using microwaves for 10 min at 900 W. Evaporation to dryness and reconstitution with 100 μL Me2CO. | 3 ng/mL | [16] | |
| GC-MS | Urine | VF-DA (12 m × 0.20 mm) | CH3I | Helium | 5 mL urine + IS + 200 mg Na2SO4 + NaOH + 5 mL organic solvent. Evaporation to dryness and reconstitution with CH3I. | 50 ng/mL; n.r. | [17] | |
| GC-MS | Urine | HP-1 (12 m × 0.2 mm) | CH3I | Helium | 2 mL urine + 0.02 THA + 6 mL CH3I in toluene. Derivatization for 30 min at 50 °C. SPE. Evaporation to dryness and reconstitution with 50 μL EA. | 10 ng/mL; n.r. | [18] | |
| GC-MS | Urine | HP5 (18 m × 0.2 mm) | CH3I | Helium | 5 mL urine + IS. SPE. Evaporation to dryness. Derivatization with CH3I in acetone +50 mg K2CO3 under microwave irradiation. | 50 ng/mL | [19] | |
| GC-MS | Urine | Agilent Ultra 1 (17 m × 0.2 mm) | MSTFA NH4I, PrSH | Helium | 2.5 mL urine + acetate buffer + IS + Na2CO3. Extraction with diethylether-2-propanol (5:1, v/v) + Na2SO4. Evaporation to dryness. Two-step silylation: 1) 50 μL MSTFA + MeCN (10 min, 80 °C); 2) 50 μL MSTFA/NH4I/PrSH (10 min, 80 °C) | n.r. | [21] | |
| GC-MS | Urine | Optima 17 (15 m × 0.25 mm) | PFB-Br | Helium | 50 μL urine + IS, evaporation to dryness. Addition of 100 μL EtOH, evaporation to dryness. Reconstitution with 100 μL MeCN +10 μL PFB-Br + 10 μL Hünig base, 60 min at 30 °C. Evaporation to dryness and reconstitution with 200 μL toluene. | 0.3 pmol; 25 μM | Present study | |
Abbreviations. AA, acetic acid; EA, ethyl acetate; EtOH, ethanol; FA, formic acid/formate; IS, internal standard; MeCN, acetonitrile; MeOH, methanol; Me2CO, acetone; n.r., not reported; SPE, solid-phase extraction.