Simultaneous quantification of four first line antitubercular drugs and metabolites in human plasma by hydrophilic interaction chromatography and tandem mass spectrometry.
J Chromatogr B Analyt Technol Biomed Life Sci
; 1105: 129-135, 2019 Jan 15.
Article
em En
| MEDLINE
| ID: mdl-30584977
ABSTRACT
Co-infection of tuberculosis in HIV-patients is a major health concern worldwide and especially so in Sub-Saharan Africa. To enhance the study of potential drug-drug interactions when simultaneously treating the two infections, a liquid chromatography tandem mass spectrometry method was developed for the quantitation of the four first line anti-tuberculosis drugs isoniazid, rifampicin, pyrazinamide, ethambutol and four of their major metabolites in human plasma. Analytes were extracted from 200⯵L of plasma using a sequential liquid-liquid extraction with ethyl acetate at neutral and acidic pH. The combined extracts were analyzed by liquid chromatography with mass spectrometric detection in a multiple reaction monitoring mode. The chromatographic separation was performed on a hydrophilic interaction column using a stepwise gradient with two mobile phases consisting of water with 0.3% formic acid and methanol with 0.3% formic acid, respectively. The total run time of each analysis was 4â¯min. The lower limit of quantification applied was 40â¯ng/mL for ethambutol, acetylisoniazid and 25-desacetylrifampicin, 60â¯ng/mL for 5-hydroxypyrazinamide, 80â¯ng/mL for isoniazid and isonicotinic acid, 200â¯ng/mL for rifampicin and 320â¯ng/mL for pyrazinamide. The method was validated according to US Food and Drug Administration guidance. The method exhibited adequate accuracy (87.1-114.9%), precision (CVâ¯<â¯12.8%) and specificity. Recovery and matrix effect were consistent (CVâ¯<â¯11.9%). The extracted samples were stable in the autosampler at 8⯰C for up to 24â¯h as well as after three freeze-thaw cycles (recoveryâ¯>â¯86.3%). The method has been shown to be robust for the analysis of the stated drugs and metabolites in human plasma obtained from 73 patients receiving these four first line anti-tuberculosis drugs.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Cromatografia Líquida
/
Espectrometria de Massas em Tandem
/
Antituberculosos
Tipo de estudo:
Prognostic_studies
Limite:
Humans
Idioma:
En
Revista:
J Chromatogr B Analyt Technol Biomed Life Sci
Assunto da revista:
ENGENHARIA BIOMEDICA
Ano de publicação:
2019
Tipo de documento:
Article
País de afiliação:
Suécia