TY - JOUR
T1 - A validated liquid chromatography tandem mass spectrometry assay for the analysis of pretomanid in plasma samples from pulmonary tuberculosis patients
AU - Malo, Andisiwe
AU - Kellermann, Tracy
AU - Ignatius, Elisa H.
AU - Dooley, Kelly E.
AU - Dawson, Rodney
AU - Joubert, Anton
AU - Norman, Jennifer
AU - Castel, Sandra
AU - Wiesner, Lubbe
N1 - Funding Information:
The development, validation and sample analysis were supported in part by the National Institute of Allergy and Infectious Diseases of the National Institutes of Health ( UM1 AI068634 , UM1 AI068636 and UM1 AI106701 ) and the FDA orphan drugs grant ( R01FD004794 ), and the National Institute of Mental Health ( AI068632 ). Kelly E. Dooley is supported by K24AI150349, and Elisa H. Ignatius is supported by T32 GM066691. The Global Alliance for TB Drug Development (TB Alliance) provided reference standards for pretomanid and PA-824-d5 for this study. The work is also based on the research supported in part by the National Research Foundation of South Africa ( HBG171031277340 ).
Publisher Copyright:
© 2020 Elsevier B.V.
PY - 2021/2/20
Y1 - 2021/2/20
N2 - A method for the extraction and quantification of pretomanid in 40 μL of human plasma, by high performance liquid chromatography with tandem mass spectrometry (LC–MS/MS) detection was developed and validated. Samples were prepared using liquid-liquid extraction and chromatographic separation was achieved on an Agilent Poroshell C18 column using an isocratic elution at a flow rate of 400 μL/min. Electrospray ionization with mass detection at unit resolution in the multiple reaction monitoring (MRM) mode on an AB Sciex API 3200 mass spectrometer was used. Over the validation period, accuracy, precision, selectivity, sensitivity, recovery and stability were assessed. The calibration range was 10 – 10 000 ng/mL. Inter- and intra-day precision, expressed as the coefficient of variation (%CV), was shown to be lower than 9% at all concentrations tested with accuracies between 95.2 and 110 %. The recovery was 72.4 % overall and reproducible at the low, medium and high end of the calibration range. The method was shown to be specific for pretomanid with no significant matrix effects observed. The validated method facilitated the analysis of pretomanid in plasma collected from adults with pulmonary TB as part of a clinical pharmacokinetic study.
AB - A method for the extraction and quantification of pretomanid in 40 μL of human plasma, by high performance liquid chromatography with tandem mass spectrometry (LC–MS/MS) detection was developed and validated. Samples were prepared using liquid-liquid extraction and chromatographic separation was achieved on an Agilent Poroshell C18 column using an isocratic elution at a flow rate of 400 μL/min. Electrospray ionization with mass detection at unit resolution in the multiple reaction monitoring (MRM) mode on an AB Sciex API 3200 mass spectrometer was used. Over the validation period, accuracy, precision, selectivity, sensitivity, recovery and stability were assessed. The calibration range was 10 – 10 000 ng/mL. Inter- and intra-day precision, expressed as the coefficient of variation (%CV), was shown to be lower than 9% at all concentrations tested with accuracies between 95.2 and 110 %. The recovery was 72.4 % overall and reproducible at the low, medium and high end of the calibration range. The method was shown to be specific for pretomanid with no significant matrix effects observed. The validated method facilitated the analysis of pretomanid in plasma collected from adults with pulmonary TB as part of a clinical pharmacokinetic study.
KW - LC–MS/MS
KW - Liquid-liquid extraction
KW - Pretomanid
KW - Tuberculosis (TB)
KW - Validation
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U2 - 10.1016/j.jpba.2020.113885
DO - 10.1016/j.jpba.2020.113885
M3 - Article
C2 - 33406472
AN - SCOPUS:85098667715
SN - 0731-7085
VL - 195
JO - Journal of Pharmaceutical and Biomedical Analysis
JF - Journal of Pharmaceutical and Biomedical Analysis
M1 - 113885
ER -