Chromatographic and liquid chromatography-tandem-mass spectrometry determination of first-line anti-tuberculosis drugs in human plasma

Autor: S. A. Soloveva, E. A. Bessonova, Liudmila Kartsova
Rok vydání: 2016
Předmět:
Zdroj: Аналитика и контроль. 20:161-167
ISSN: 2073-1450
2073-1442
Popis: In the current study the possibility of the simultaneous determination of anti-tuberculosis ( anti-TB ) drugs (isoniazid, pyrazinamide, ethambutol and rifampicin) using the reversed-phase HPLC (RP-HPLC) and ion-pair chromatography was investigated. A selective and highly sensitive liquid chromatography/tandem mass spectrometry method for the simultaneous determination of four anti-TB drugs (isoniazid, pyrazinamide, ethambutol and rifampicin) in the human plasma was developed. The detection was carried out using the multiple reaction-monitoring (MRM) modes with positive polarity. The fragmentation conditions for each drug were optimized, and the conditions for the blood plasma preparation for HPLC/MS analysis, including the protein precipitation with ACN with a ratio of 3:1 (by volume), were chosen. The matrix effects on the separation and ionization of anti-TB drugs were estimated by the post-extraction additives method. It was shown that the 10-fold dilution of plasma extracts was sufficient to decrease the influence of the sample matrix. The stability of anti-TB drugs during the analysis (in autosampler at 1 and 12 hours) and at storage conditions (3 cycles of freeze-thaw) was studied. The method for increasing the stability of rifampicin using ascorbic acid (1 mg/ml) as antioxidant was provided. The LOD with UV detection were 2 – 20 µg/ml, in SIM-mode – 2 – 15 ng/ml and in MRM-mode – 0.5 – 10 ng/ml respectively. The possibility of the determination of four anti-TB drugs in real plasma samples of patients with tuberculosis undergoing drug therapy in optimized conditions HPLC/MC was shown. Key words: anti-tuberculosis drugs, RP HPLC, LC/MS/MS, matrix effect (Russian) DOI: http://dx.doi.org/10.15826/analitika.2016.20.2.007 E.A. Bessonova, L.A. Kartsova, S.A. Soloveva Saint-Petersburg State University, 7/9 Universitetskaya nab., St. Petersburg, 199034, Russian Federation
Databáze: OpenAIRE
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