Session Type: ePosters
Session Title: ePosters
Authors(s): A. Luf (1), V. Al Jalali (2), T. Stimpfl (1), M. Zeitlinger (2)
Authors Affiliations(s): (1) Department of Laboratory Medicine, Medical University of Vienna, Austria, (2) Department of Clinical Pharmacology, Medical University of Vienna, Austria
Background:
Gepotidacin is a novel, first in class triazaacenaphthylene antibiotic that inhibits bacterial DNA replication by a distinct mechanism of action. Distribution into peripheral tissues in human is unkown for gepotidacin and microdialysis (MD) is an ideal method to measure unbound drug concentrations in human plasma and tissue. The aim of this study was to evaluate if MD can be used with gepotidacin, investigate suitable internal standards (IS) for MD calibration and develop and validate a bioanalytical method for the quantitation of gepotidacin and the selected IS.
Methods:In-vitro MD experiments were designed as forward and reverse dialysis experiments. Perfusion solution consisted of an aqueous solution of gepotidacin (2µg/mL) and one of four different IS and the immersion solution was physiological saline (forward dialysis) or vice versa (reverse dialysis). MD samples were collected for two 30min and two 60min intervals.
The quantitative analysis of gepotidacin and the IS was performed on an AB SCIEX QTRAP 5500 mass spectrometer with positive ionization in MRM mode. Chromatographic separation was achieved by reversed phase chromatography (Phenomenex Luna Omega PS C18 100x2,1mm;1.6µm) with a Dionex Ultimate 3000 HPLC system. The bioanalytical HPLC-MS method, ranging from 5 to 2100ng for gepotidacin and 30 to 1100ng for the IS, was validated based on a validation plan complying with assessment criteria of the latest EMA guideline for bioanalytical method development.
Results:This in-vitro study demonstrated that recovery rates for gepotidacin (45.4%) and IS (47.3%) were constant over time and equal in both directions (forward/reverse). Additionally, all assessed substances proved to be suitable to serve as IS for MD calibration. Substance D was selected to serve as IS for future MD experiments, due to its comparable physiochemical properties. No interference was observed between substance D and gepotidacin during analysis.
Conclusions:A bioanalytical method for gepotidacin quantitation was successfully established and validated. Furthermore, the present study demonstrated that MD with gepotidacin is feasible and provides reliable results. The implementation of an IS in the MD experiments allows for calibration of the experiments without the need for an additional calibration period. These findings serve as a basis for future MD experiments in human tissue.
Keyword(s): gepotidacin, tissue pharmacokinetics, microdialysisSession Type: ePosters
Session Title: ePosters
Authors(s): A. Luf (1), V. Al Jalali (2), T. Stimpfl (1), M. Zeitlinger (2)
Authors Affiliations(s): (1) Department of Laboratory Medicine, Medical University of Vienna, Austria, (2) Department of Clinical Pharmacology, Medical University of Vienna, Austria
Background:
Gepotidacin is a novel, first in class triazaacenaphthylene antibiotic that inhibits bacterial DNA replication by a distinct mechanism of action. Distribution into peripheral tissues in human is unkown for gepotidacin and microdialysis (MD) is an ideal method to measure unbound drug concentrations in human plasma and tissue. The aim of this study was to evaluate if MD can be used with gepotidacin, investigate suitable internal standards (IS) for MD calibration and develop and validate a bioanalytical method for the quantitation of gepotidacin and the selected IS.
Methods:In-vitro MD experiments were designed as forward and reverse dialysis experiments. Perfusion solution consisted of an aqueous solution of gepotidacin (2µg/mL) and one of four different IS and the immersion solution was physiological saline (forward dialysis) or vice versa (reverse dialysis). MD samples were collected for two 30min and two 60min intervals.
The quantitative analysis of gepotidacin and the IS was performed on an AB SCIEX QTRAP 5500 mass spectrometer with positive ionization in MRM mode. Chromatographic separation was achieved by reversed phase chromatography (Phenomenex Luna Omega PS C18 100x2,1mm;1.6µm) with a Dionex Ultimate 3000 HPLC system. The bioanalytical HPLC-MS method, ranging from 5 to 2100ng for gepotidacin and 30 to 1100ng for the IS, was validated based on a validation plan complying with assessment criteria of the latest EMA guideline for bioanalytical method development.
Results:This in-vitro study demonstrated that recovery rates for gepotidacin (45.4%) and IS (47.3%) were constant over time and equal in both directions (forward/reverse). Additionally, all assessed substances proved to be suitable to serve as IS for MD calibration. Substance D was selected to serve as IS for future MD experiments, due to its comparable physiochemical properties. No interference was observed between substance D and gepotidacin during analysis.
Conclusions:A bioanalytical method for gepotidacin quantitation was successfully established and validated. Furthermore, the present study demonstrated that MD with gepotidacin is feasible and provides reliable results. The implementation of an IS in the MD experiments allows for calibration of the experiments without the need for an additional calibration period. These findings serve as a basis for future MD experiments in human tissue.
Keyword(s): gepotidacin, tissue pharmacokinetics, microdialysis
Medical University of Vienna, Department of Clinical Pharmacology
Session Type: ePosters
Session Title: ePosters
Authors(s): A. Luf (1), V. Al Jalali (2), T. Stimpfl (1), M. Zeitlinger (2)
Authors Affiliations(s): (1) Department of Laboratory Medicine, Medical University of Vienna, Austria, (2) Department of Clinical Pharmacology, Medical University of Vienna, Austria
Background:
Gepotidacin is a novel, first in class triazaacenaphthylene antibiotic that inhibits bacterial DNA replication by a distinct mechanism of action. Distribution into peripheral tissues in human is unkown for gepotidacin and microdialysis (MD) is an ideal method to measure unbound drug concentrations in human plasma and tissue. The aim of this study was to evaluate if MD can be used with gepotidacin, investigate suitable internal standards (IS) for MD calibration and develop and validate a bioanalytical method for the quantitation of gepotidacin and the selected IS.
Methods:In-vitro MD experiments were designed as forward and reverse dialysis experiments. Perfusion solution consisted of an aqueous solution of gepotidacin (2µg/mL) and one of four different IS and the immersion solution was physiological saline (forward dialysis) or vice versa (reverse dialysis). MD samples were collected for two 30min and two 60min intervals.
The quantitative analysis of gepotidacin and the IS was performed on an AB SCIEX QTRAP 5500 mass spectrometer with positive ionization in MRM mode. Chromatographic separation was achieved by reversed phase chromatography (Phenomenex Luna Omega PS C18 100x2,1mm;1.6µm) with a Dionex Ultimate 3000 HPLC system. The bioanalytical HPLC-MS method, ranging from 5 to 2100ng for gepotidacin and 30 to 1100ng for the IS, was validated based on a validation plan complying with assessment criteria of the latest EMA guideline for bioanalytical method development.
Results:This in-vitro study demonstrated that recovery rates for gepotidacin (45.4%) and IS (47.3%) were constant over time and equal in both directions (forward/reverse). Additionally, all assessed substances proved to be suitable to serve as IS for MD calibration. Substance D was selected to serve as IS for future MD experiments, due to its comparable physiochemical properties. No interference was observed between substance D and gepotidacin during analysis.
Conclusions:A bioanalytical method for gepotidacin quantitation was successfully established and validated. Furthermore, the present study demonstrated that MD with gepotidacin is feasible and provides reliable results. The implementation of an IS in the MD experiments allows for calibration of the experiments without the need for an additional calibration period. These findings serve as a basis for future MD experiments in human tissue.
Keyword(s): gepotidacin, tissue pharmacokinetics, microdialysisSession Type: ePosters
Session Title: ePosters
Authors(s): A. Luf (1), V. Al Jalali (2), T. Stimpfl (1), M. Zeitlinger (2)
Authors Affiliations(s): (1) Department of Laboratory Medicine, Medical University of Vienna, Austria, (2) Department of Clinical Pharmacology, Medical University of Vienna, Austria
Background:
Gepotidacin is a novel, first in class triazaacenaphthylene antibiotic that inhibits bacterial DNA replication by a distinct mechanism of action. Distribution into peripheral tissues in human is unkown for gepotidacin and microdialysis (MD) is an ideal method to measure unbound drug concentrations in human plasma and tissue. The aim of this study was to evaluate if MD can be used with gepotidacin, investigate suitable internal standards (IS) for MD calibration and develop and validate a bioanalytical method for the quantitation of gepotidacin and the selected IS.
Methods:In-vitro MD experiments were designed as forward and reverse dialysis experiments. Perfusion solution consisted of an aqueous solution of gepotidacin (2µg/mL) and one of four different IS and the immersion solution was physiological saline (forward dialysis) or vice versa (reverse dialysis). MD samples were collected for two 30min and two 60min intervals.
The quantitative analysis of gepotidacin and the IS was performed on an AB SCIEX QTRAP 5500 mass spectrometer with positive ionization in MRM mode. Chromatographic separation was achieved by reversed phase chromatography (Phenomenex Luna Omega PS C18 100x2,1mm;1.6µm) with a Dionex Ultimate 3000 HPLC system. The bioanalytical HPLC-MS method, ranging from 5 to 2100ng for gepotidacin and 30 to 1100ng for the IS, was validated based on a validation plan complying with assessment criteria of the latest EMA guideline for bioanalytical method development.
Results:This in-vitro study demonstrated that recovery rates for gepotidacin (45.4%) and IS (47.3%) were constant over time and equal in both directions (forward/reverse). Additionally, all assessed substances proved to be suitable to serve as IS for MD calibration. Substance D was selected to serve as IS for future MD experiments, due to its comparable physiochemical properties. No interference was observed between substance D and gepotidacin during analysis.
Conclusions:A bioanalytical method for gepotidacin quantitation was successfully established and validated. Furthermore, the present study demonstrated that MD with gepotidacin is feasible and provides reliable results. The implementation of an IS in the MD experiments allows for calibration of the experiments without the need for an additional calibration period. These findings serve as a basis for future MD experiments in human tissue.
Keyword(s): gepotidacin, tissue pharmacokinetics, microdialysis