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Development of method for the determination of entecavir in biological materials using UHPLC-MS/MS
Gottvald, Tomáš ; Nováková, Lucie (advisor) ; Šatínský, Dalibor (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Tomáš Gottvald Supervisor: Doc. PharmDr. Lucie Nováková, Ph.D. Title of thesis: Development of method for the determination of entecavir in biological materials using UHPLC-MS/MS Entecavir is a synthetic guanosine nucleoside that plays an important role in treatment of chronical hepatitis B virus infection. This work was focused on development of highly sensitive method to determine entecavir concentrations in rat urine by ultra-high performance liquid chromatography-tandem mass spectrometry. Entecavir C2 13 N15 , stable isotopically labeled internal standard, was used for quantitation. Hydrophilic interaction chromatography seems to be a suitable technique for retention and separation of polar compounds present in analyzed sample. One of the main goals was to confirm this fact by comparison with commonly used reverse phase chromatography mode (RP-UHPLC) on BEH C18 stationary phase. Conditions for RP-UHPLC were optimized as follows: mobile phase composed of acetonitrile/0.01 % formic acid (4:96). The HILIC conditions on BEH Amide stationary phase were optimized using isocratic elution with mobile phase composed of acetonitrile/5 mM ammonium acetate pH 4.0 (75:25). HILIC method provided much...
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Development of method for the determination of entecavir in biological materials using UHPLC-MS/MS
Gottvald, Tomáš ; Nováková, Lucie (advisor) ; Šatínský, Dalibor (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Tomáš Gottvald Supervisor: Doc. PharmDr. Lucie Nováková, Ph.D. Title of thesis: Development of method for the determination of entecavir in biological materials using UHPLC-MS/MS Entecavir is a synthetic guanosine nucleoside that plays an important role in treatment of chronical hepatitis B virus infection. This work was focused on development of highly sensitive method to determine entecavir concentrations in rat urine by ultra-high performance liquid chromatography-tandem mass spectrometry. Entecavir C2 13 N15 , stable isotopically labeled internal standard, was used for quantitation. Hydrophilic interaction chromatography seems to be a suitable technique for retention and separation of polar compounds present in analyzed sample. One of the main goals was to confirm this fact by comparison with commonly used reverse phase chromatography mode (RP-UHPLC) on BEH C18 stationary phase. Conditions for RP-UHPLC were optimized as follows: mobile phase composed of acetonitrile/0.01 % formic acid (4:96). The HILIC conditions on BEH Amide stationary phase were optimized using isocratic elution with mobile phase composed of acetonitrile/5 mM ammonium acetate pH 4.0 (75:25). HILIC method provided much...
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Development of method for the determination of entecavir in biological materials using UHPLC-MS/MS
Gottvald, Tomáš ; Nováková, Lucie (advisor) ; Šatínský, Dalibor (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Tomáš Gottvald Supervisor: Doc. PharmDr. Lucie Nováková, Ph.D. Title of thesis: Development of method for the determination of entecavir in biological materials using UHPLC-MS/MS Entecavir is a synthetic guanosine nucleoside that plays an important role in treatment of chronical hepatitis B virus infection. This work was focused on development of highly sensitive method to determine entecavir concentrations in rat urine by ultra-high performance liquid chromatography-tandem mass spectrometry. Entecavir C2 13 N15 , stable isotopically labeled internal standard, was used for quantitation. Hydrophilic interaction chromatography seems to be a suitable technique for retention and separation of polar compounds present in analyzed sample. One of the main goals was to confirm this fact by comparison with commonly used reverse phase chromatography mode (RP-UHPLC) on BEH C18 stationary phase. Conditions for RP-UHPLC were optimized as follows: mobile phase composed of acetonitrile/0.01 % formic acid (4:96). The HILIC conditions on BEH Amide stationary phase were optimized using isocratic elution with mobile phase composed of acetonitrile/5 mM ammonium acetate pH 4.0 (75:25). HILIC method provided much...
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Development of method for the determination of entecavir in biological materials using UHPLC-MS/MS
Gottvald, Tomáš ; Nováková, Lucie (advisor) ; Šatínský, Dalibor (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Tomáš Gottvald Supervisor: Doc. PharmDr. Lucie Nováková, Ph.D. Title of thesis: Development of method for the determination of entecavir in biological materials using UHPLC-MS/MS Entecavir is a synthetic guanosine nucleoside that plays an important role in treatment of chronical hepatitis B virus infection. This work was focused on development of highly sensitive method to determine entecavir concentrations in rat urine by ultra-high performance liquid chromatography-tandem mass spectrometry. Entecavir C2 13 N15 , stable isotopically labeled internal standard, was used for quantitation. Hydrophilic interaction chromatography seems to be a suitable technique for retention and separation of polar compounds present in analyzed sample. One of the main goals was to confirm this fact by comparison with commonly used reverse phase chromatography mode (RP-UHPLC) on BEH C18 stationary phase. Conditions for RP-UHPLC were optimized as follows: mobile phase composed of acetonitrile/0.01 % formic acid (4:96). The HILIC conditions on BEH Amide stationary phase were optimized using isocratic elution with mobile phase composed of acetonitrile/5 mM ammonium acetate pH 4.0 (75:25). HILIC method provided much...
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