Vývoj UHPLC-MS/MS metody pro separaci alkaloidů čeledi Amaryllidaceae

Abstract

Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of analytical chemistry Candidate: Kristýna Burianová Supervisor: prof. PharmDr. Lucie Nováková, Ph.D. Consultant: PharmDr. Veronika Pilařová, Ph.D. Title of Doctoral Thesis: Development of UHPLC-MS/MS method for separation of Amaryllidaceae alkaloids In this master thesis, a method for the separation of 30 alkaloids of the Amaryllidaceae family was developed using ultra-high performance liquid chromatography coupled with tandem mass detection (UHPLC-MS/MS). Chromatographic conditions and mass spectrometer parameters were optimized. In the first step of optimization, screening of 10 analytical columns was performed on an ultra-high performance liquid chromatography coupled with photodiode array detection (UHPLC-PDA). Those tested columns were Acquity UPLC BEH C18, Acquity UPLC CSH C18, Acquity UPLC BEH Phenyl, Acquity UPLC CSH Phenyl-hexyl, Arion Plus C18, Acquity UPLC BEH Shield RP18, Ascentis Express RP-Amide, ACE Excel C18-PFP, Kinetex PFP, and Kinetex F5. Acquity UPLC BEH Phenyl, Acquity UPLC CSH Phenyl-hexyl, and Acquity UPLC CSH C18 as they provided the highest separation score for the analyzed isomeric groups of analytes were selected for further optimization. The optimization of MS conditions was performed by...

Univerzita Karlova v Praze, Farmaceutická fakulta v Hradci Králové Katedra analytické chemie Kandidát: Kristýna Burianová Školitel: prof. PharmDr. Lucie Nováková, Ph.D. Konzultant: PharmDr. Veronika Pilařová, Ph.D. Název diplomové práce: Vývoj UHPLC-MS/MS metody pro separaci alkaloidů čeledi Amaryllidaceae V této diplomové práci byla vyvíjena metoda pro separaci 30 alkaloidů čeledi Amaryllidaceae pomocí ultra-vysokoúčinné kapalinové chromatografie ve spojení s tandemovou hmotnostní detekcí (UHPLC-MS/MS). Byly optimalizovány chromatografické podmínky a parametry hmotnostního spektrometru. V první fázi optimalizace byl proveden screening 10 analytických kolon na systému ultra-vysokoúčinné kapalinové chromatografie ve spojení s detektorem fotodiodového pole (UHPLC-PDA). Jednalo se o kolony Acquity UPLC BEH C18, Acquity UPLC CSH C18, Acquity UPLC BEH Phenyl, Acquity UPLC CSH Phenyl-hexyl, Arion Plus C18, Acquity UPLC BEH Shield RP18, Ascentis Express RP-Amide, ACE Excel C18-PFP, Kinetex PFP a Kinetex F5. K další optimalizaci byly vybrány kolony Acquity UPLC BEH Phenyl, Acquity UPLC CSH Phenyl-hexyl a Acquity UPLC CSH C18, protože poskytly nejvyšší skóre separace analyzovaných izomerních skupin analytů. Optimalizace MS podmínek byla provedena nastavením parametrů iontového zdroje a nastavením SRM...