Protein precipitation method for determination of clobazam and N-desmethylclobazam in human plasma by LC-MS/MS.
Biomed Chromatogr
; 34(1): e4710, 2020 Jan.
Article
em En
| MEDLINE
| ID: mdl-31630425
ABSTRACT
A protein precipitation method for the determination of clobazam (CLB) and its major active metabolite N-desmethylclobazam (N-CLB) in human plasma by liquid chromatography tandem mass spectrometry (LC-MS/MS) was established. CLB and N-CLB were extracted from human plasma samples by protein precipitation with methanol. Analyte separation was done using a Phenomenex Kinetex™ Biphenyl (50 × 2.1 mm, 1.7 µm) column using isocratic elution with a mobile phase of 5 mm ammonium formate with 0.01% ammonium hydroxide (40%) and methanol (60%) at a flow rate of 0.4 mL/min and an injection volume of 10 µL. The detection was performed on a triple quadrupole mass spectrometer in multiple reaction monitoring mode to monitor the precursor-to-product ion transitions of m/z 301.1 â 259.0, 306.0 â 263.9 for CLB and CLB-D5 and 287.0 â 245.0, 292.0 â 250.0 for N-CLB and N-CLB-D5 in positive electrospray ionization mode, respectively. The method was validated over a concentration range of 2.0-750 ng/mL for CLB and 0.7-200 ng/mL for N-CLB on SCIEX Triple Quad 4500 MS System. Total run time was 5 min. This method has been designed for bioequivalence study for formulations containing 20 mg of CLB.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Benzodiazepinas
/
Cromatografia Líquida
/
Espectrometria de Massas em Tandem
/
Clobazam
Tipo de estudo:
Diagnostic_studies
/
Prognostic_studies
Limite:
Humans
Idioma:
En
Revista:
Biomed Chromatogr
Ano de publicação:
2020
Tipo de documento:
Article
País de afiliação:
Armênia