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Development of a UHPLC-MS/MS method suitable for the determination of cabozantinib in blood serum
Kleinová, Jana ; Kozlík, Petr (advisor) ; Hraníček, Jakub (referee)
The aim of this Bachelor thesis was to develop a UHPLC-MS/MS method for the determination of Cabozantinib in blood serum. This method was used to monitor the release of the active substance in a model organism - the rat. First, the mass spectrometer setup was optimized, where MRM transitions for Cabozantinib and isotope-labelled Cabozantinib-D4 were found. For Cabozantinib, an MRM transition of 502.2 → 323.1 was found with optimal energy levels of Q1 = -26.0 V, CE = -40.0 V and Q2 = -22.0 V. For Cabozantinib-D4, the MRM transition of 506.2 → 327.0 was found with optimal energy levels being Q1 = -26.0 V, CE = -39.0 V and Q2 = -22 V. The method setup parameters were as follows: The chromatography column used was a Poroshell 120 EC-C18, 2.1x50 mm, 1.9 um, (Agilent Technologies). The mobile phase consisted of acetonitrile (B) and formic acid (A), the flow rate of the mobile phase was 0.350 ml/min, the analysis time was 5.5 min and the injection volume was 1 μl at gradient elution (time: 0; 0.5; 2.0; 3.0; 3.5; 4.0; 5.5 min, B: 10; 10; 60; 100; 10; 10 %). The method was linear (weighted regression 1/x2 ) with a regression coefficient equal to 0.9978, showing an excellent linearity of the method. The precision (relative standard deviation) of the method was within 14 %. The accuracy (relative error) was...
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Comparison of different tandem mass detection scans in the analysis of cannabidiol
Tichá, Tereza ; Kozlík, Petr (advisor) ; Kubíčková, Anna (referee)
The aim of this bachelor thesis was to compare different tandem mass spectrometer scans in order to develop an LC-MS/MS method suitable for the determination of cannabidiol in rat serum. In this bachelor thesis the mass spectrometer and high-performance liquid chromatography method were optimized. The optimum parameters were as follows: chromatographic column Acquity UPLC BEH C18 50 × 2,1 mm, 1,7 μm from Waters (Wexford, Ireland). The mobile phase consisted of methanol and water of LC-MS purity. The flow rate of the mobile phase was 0,4 ml/min, the column temperature 40 řC, the temperature in an autosampler 15 řC, the sample injection volume was 1 μl and the analysis time was 7 minutes under gradient elution conditions. The MRM transitions of highest intensity were monitored. For CBD, the MRM transition in positive mode was 315,2 → 193,1 (Q1 = -10 V, CE = -24 V, Q3 = -20 V), for isotopically labeled cannabidiol D3 was chosen 318,4 → 196,0 (Q1 = -10 V, CE = -23 V, Q3 = -20 V). The calibrations in solvent and rat serum were performed in MRM and SIM mode. No calibration was constructed for the SCAN mode due to its low selectivity, which made it able to detect cannabidiol only from concentration of 312,5 ng/ml. The highly selective MRM was able to operate over the entire observed concentration range of...
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Development of a UHPLC-MS/MS method for determination of rivaroxaban in rat serum
Plášilová, Denisa ; Kozlík, Petr (advisor) ; Hraníček, Jakub (referee)
This bachelor thesis is focused on development of a specific UHPLC-MS/MS method for the determination of rivaroxaban in rat serum. At first, the setting of the mass spectrometer was optimized. Suitable MRM transitions were found for rivaroxaban and its isotopically labeled internal standard. For rivaroxaban was found the MRM transition 436.1 → 145.05 with optimal energy levels Q1 = -12.0 V; CE = -30.0 V; Q2 = -27.0 V. For rivaroxaban D4 was found the MRM transition 440.1 → 145.0 with optimal energy levels Q1 = -22.0 V; CE = -31.0 V; Q2 = -25.0 V. The ion source setting parameters were as follows: nebulizing gas flow 3 l/min; heating gas flow 10 l/min; interface temperature 300 řC; desolvation temperature 526 řC; DL temperature 250 řC; heat block temperature 400 řC; drying gas flow 10 l/min. The optimal chromatographic method was as follows: A Poroshell 120 SB AQ, 100 × 2.1 mm, 2.6 µm (Agilent) chromatographic column; the mobile phase consisted of acetonitrile with the addition of 0.1% formic acid (A) and distilled water with the addition of 0.1% formic acid (B); flow rate of the mobile phase 0.5 ml/min; gradient elution (time: 0-1-2-3.5-4-6.5 min; A: 20-20-80-80-20-20 % v/v); autosampler temperature 15 řC; column temperature 40 řC; time of analysis 6.5 minutes; injection volume 2 µl. The optimized...
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DEVELOPMENT OF A UHPLC-MS/MS METHOD FOR NILOTINIB DETERMINATION IN RAT SERUM
Černá, Kateřina ; Kozlík, Petr (advisor) ; Křížek, Tomáš (referee)
This bachelor thesis aimed to develop a UHPLC-MS/MS method for the determination of nilotinib in rat serum. The developed UHPLC-MS/MS method was used to monitor the pharmacokinetic release of the active substance in a rat model organism in a project focused on the formulation of a tablet containing nilotinib with a slower release than before. The optimal conditions of the method were as follows. Chromatographic column Acquity UPLC BEH PHENYL 100x2.1 mm, 1.7 μm from Waters. The mobile phase consisted of methanol and distilled water, both with the addition of 0.1% formic acid using gradient elution. The flow rate of the mobile phase was 0.3 mL/min, the temperature in an autosampler 15 řC, the column temperature 40 řC, the analysis time 6.5 minutes, and the injection volume 2 μl. The MRM transition monitored for nilotinib was: 530.2 → 289.10 (Q1 = -26 V; CE = -31 V; Q3 = -20 V) and for nilotinib D6: 536.2 → 295.15 (Q1 = -26 V; CE = -31 V; Q3 = -14 V). The setting of the ion source was as follows: nebulizing gas flow 3 L/min; drying gas flow 10 L/min; source temperature 300 řC; desolvation capillary temperature 250 řC. The method was partially validated. The coefficient of determination 1.0000 shows the excellent linearity of the method. The accuracy, expressed as a relative error, was up to 20 %. The...
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