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Study of phenolic compounds in some biologic materials using method LC/MS
Měřínská, Radana ; Zemanová, Jana (referee) ; Kočí, Radka (advisor)
Presented bachelor thesis was focused on suitable methods for analysis of phenolics, especially modern instrumental methods were studied. In experimental part some standard phenolic compounds were analysed using combined technique HPLC/ESI-MS with off-line and/or on-line detection. Mass spectra of individual stadards and/or mixtures were determined in MS full scan and/or MS/MS full scan mode. Ionisation of chosen standard compounds was done in pozitive and/or negative mode. Parent ions of followed compounds were identified: reserpine (PI 609.9, NI 607.7), acid chlorogenic (NI 353), quercetin (PI 303.5, NI 301.5), rutin (PI 611, NI 609), morin (PI 303.1, NI 301.1). Typical fragments of studied standards were produced using MS/MS full scan mode. By fragmentation characteristic product ions were identified: acid chlorogenic (NI 191), quercetin (PI: 229.1 and 257.1; NI: 179.0 and 151.0), rutin (in MS/MS NI: 301; in MS3 NI: 179, 151.1), morin (PI: 285.1 a 257.1; NI: 283.2, 273.2, 257.1, 229.1). Optimal conditions of phenolic HPLC separation were determined including gradient elution and Restek column use. In conclusion on-line HPLC/MS detection using negative ionization was observed as the best configuration for phenolic analysis.
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Imunomagnetic separation of lactic acid bacteria using magnetic microparticles functionalised by antibodies
Vaňásek, Jakub ; Španová, Alena (referee) ; Trachtová, Štěpánka (advisor)
Immunomagnetic separation is based on binding of antibody with antigen, where antibody is bound to magnetic particle. In this thesis there were used particles of magnetic pearl cellulose with antiLactobacillus and antiBifidobacterium antibodies. Immunomagnetic separation method was optimalized and verified for its efficiency and specifity with bacterial and yeast cells. This cells were identified by polymerase chain reaction. Efficiency of immunomagnetic separation was verified on probiotic meat product, where Lactobacillus cells were isolated. With DNA from isolated Lactobacillus cells the high resolution melting was performed. The results show presence of several bacterial strains of Lactobacillus species.
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Supercritical fluid chromatography
Planeta, Josef
The principle of supecritical fluid chromatography, current state and instrumentation possibilities, hyphenation with MS or MALDI-TOF are described. The examples of separation are presented, and the possibility of using SFC for obtaining physico-chemical data are discussed.
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The Result of High Dose Therapy with Peripheral Blood Stemm Cells (PBCs) Support at Patients with Multiple Myeloma
Mecl, Jan ; Špička, Ivan (referee) ; Kořen, Jan (advisor)
The submitted thesis is based on comparison of therapeutical effectivity of two different methods using in multiple myeloma patients for therapy. The best therapy seems to be single or tandem autologous stem cell transplantation ASCT with stimulation and separation of periferal blood stem cells (PBSCs). The stimulation could be done by CFA (cyclofosfamid) or cytostatik combinaňtion, e.g. IVE (ifosfamide, vepeside, epirubicine) in combination with G - CSF (granulocyte - colony - stimulating factor). The question which is not answered is which one of those methods is more effective. And which one of single or tandem ASCT is more profitable if any. The goal of the study is to compare two different methods of stimulation and separation of PBSCs (CFA and IVE). I compare the effectivity from a viewpoint of breeding PBSCs and also time consuming of the separation meňthods. Nobody has done this comparison before us. To separate PBSCs was used COBE Spectra apheresis systém. Spectra is efficient to separate the whole blood into its components. The group for my study were 167 patients with Multiple Myeloma. Part was stimulated with CFA the other part with IVE. In every one of those groups we compare the number of PBSCs represent by CD34+ cells, type and period of several separation methods and also differences between...
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