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Analysis, separation and physico-chemical characterization of peptide hormones by capillary electrophoresis
Šolínová, Veronika ; Jelínek, Ivan (advisor) ; Feltl, Ladislav (referee) ; Coufal, Pavel (referee) ; Samcová, Eva (referee)
I 11 CoNcr,usroNs The work bas demonstrated that capinary zone elechophoresis and micelrar electokinetic cbromatography are high-perfomrance and high-sensitive methods for qualitative and quantitative anďysis and for physico-chemicď characÚerization ofpeptide hormones. using the strategy for the rational selection of experimental conditions for cE analysis and separations ofpeptides, ionogenic peptides were separated as cations in the classical and isoelectric buffers in the acidic region, pH 2- 2'5, and as anions in amphoteric buffer in weakry arkaline conditions, pH = 8'1. Non-ionogenic peptides were analyzed by miceilar electrokinetic chromatography with micelrar phase consisting of anionic detergent sodium dodecylsulphate in alkaline conditions, pH 8.8. The experimental conditions were selected with the aim to avoid the adsorption of peptides on the capillary wall, which was particularly reached with suppression of dissociation of si'anol gÍoups on the imer capillary wall in the acidic pH region and with electrostatic repursion negatively charged peptides from negatively charged dissociation silanol groups on the inner capillary wall in the alkaline pH region. The pH values of background electrolýes were chosen in the region, where the biggest differences in calculated specific charges of analyzed...
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High-performance chromatographic methods for determination of proteins
Vařilová, Tereza ; Pacáková, Věra (advisor) ; Feltl, Ladislav (referee) ; Coufal, Pavel (referee) ; Kučerová, Zdenka (referee)
Conclusions This Thesis gives an overview of high-performance riquid chromatography in various separation modes for a characterization of selected proteins. various affinity stationary phases were prepared for analysis of pepsin, seminal plasma proteins and selected glycoproteins' choice of the affinity system suitable for a particular purpose is always a comprex probrem in which number of aspects must be considered. properties of the proteins to be separated and ligands used had to be taken into an account. The separation system should not influence the biorogicai activity ofproteins separated. From this point of view it is clear that for any each individuar system is necessary to find specific conditions for I) immobilizationof ligand, and II) separation conditions. Stationary phases with immob'ized 3,5-diiodo-L-tyrosine exhibited sufÍicient selectivity, proved to be useful for analysis ofporcine pepsin A and are appricable to practical sampres of human pepsin' Reversed-phase crs corumn is suitable for anarysis ofpepsin fragments obtained after a-chymotrypsin digestion. A study oť interaction of various forms of pepsin and pepsinogen are essentiar because of their high importance as crinicar diagnostic markers and as stomach enzyme. "lA IŤ Aťfinity chromatography with heparin immobilized to Toyopearl...
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Polymethacrylate monolithic columns in capillary liquid chromatography
Grafnetter, Jan ; Coufal, Pavel (advisor) ; Pacáková, Věra (referee) ; Ventura, Karel (referee) ; Sýkora, David (referee)
3 Conclusions This Thesis gives chromatographic and structural characteristics of the butyl methacrylate monolithic columns prepared for the capillary liquid chromatography. Results of this Thesis clearly demonstrate the possibility of preparation of capillary monolithic columns with a good repeatability of the principal chromatographic parameters. A completely new technique for preparation of the monolithic columns for use in the on-column detection mode was introduced. Different photometric detection modes were applied to monolithic capillary columns in CLC. The advantages and disadvantages of the on-column detection mode in comparison to the additional detection capillary were studied and described. It was shown that the best separation and the most sensitive quantification in the short analysis time could be reached by using the monolithic columns with detection window placed directry on the separation capillary, just after the stationary phase or within the monolith. The attempts leading to preparation of chiral monolithic stationary phases, which would allow enantioseparation of selected chiral analýes' were not totally successful. No time was left to find the suitable chromatographic conditions for enantioseparations. \ I
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Characterization of electrophoretic separation systems,utilization of system peaks for determination of surfactant properties and optimalization of complex separation systems by the LFER model
Lokajová, Jana ; Tesařová, Eva (advisor) ; Coufal, Pavel (referee) ; Ševčík, Juraj (referee)
Conclusions o We have developď new method for CMC determination that is based on the measurementsof sysrem peaks eigenmobilities in capiiluy "t*o"oo"r*tr. The proposed method is moreprecise and eliminates specific systematic errors .o.*'n* when using some s*ndaťdmethods. The proposed method has U".n opti*i""a "o^ia,sysrcm peaks. ..Ý9 .r4. uE'tl UP.lmzeo considering intensities (amplitudes) of o The implementation of mice description ", ",".o".#'ÍÍ.',11Tffi :: jlTl.Ti#'ff ffiTr,"JT::,l:systems. Simulation prognrms curÍently allow user. to cď"ulate system eigenmobilities andeven to optimize the micellar separation systen. We h;\ *:ffi J:ffi.;: oÍ extended simulation o.o,*,",.u".il.;:T"T1'ffi ' ffill: We have clarified the mobi strength, which "uu,", .pri..ll,l?.jln.fi.r::ř""".J:"# peak due to the BGES ionic additional peak has "oo.a,.o in the electrophe.oror, ,oo**"r' As a consequence an :::::::::'*":u.'*..in.*oo.". identifi cation ", i"*,,.,"'ť"i:TT. *i:ajff:oeen possible using rhe set of different derecrion waveilil:" capirrary zone electrophoresis has been used to revear a structure and charge of nanoparticresin the recentry synthesized bulky porymer micelres ,n-'ltt".*, pH regions and to t3 characterize the effect of the PvP fraction added to PS.PMA on the final structure. ouÍ results have confirmed that...
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