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Analýza,separace a fyzikálně-chemická charakterizace peptidových hormonů kapilární elektroforézou
Šolínová, Veronika ; Jelínek, Ivan (vedoucí práce) ; Feltl, Ladislav (oponent) ; Coufal, Pavel (oponent) ; Samcová, Eva (oponent)
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 (vedoucí práce) ; Feltl, Ladislav (oponent) ; Coufal, Pavel (oponent) ; Kučerová, Zdenka (oponent)
High-performance chromatographic methods for determination of proteins Ab strakt disertačnípráce Praha,2006 Autor: Mgr. Tereza Vařilová Školitelé: Prof. RNDr.Věra Pacáková, CSc. (Katedra analýické chemie, Karlova Univerzita, Praha) Prof. Dr. Hans-Gerd Janssen (Unilever Research & Development, Nizozemí) Ana|ýza biologicky aktivních látek je rychle se rozvíjejícíobor, kteý tvoří důležitou část aplikací v analýické chemii. Mezi látkami jako jsou peptidy, hormony, léky a drogy tvoří proteiny důleŽitou skupinu bíologicky aktivních látek. Naročným úkolem je izo|ace, separace a identifikace iednotlivých složek složitýchsměsí (např. krevní sénrm). Pro t1to f{9|1z p2ií vysokoúčinnéchromatografické metody svénezastupitelné místo. Proteomika je dynamicky se rozvijejícía dtúežitávědní disciplína. Jejípokrok by nebyl možný bez r{rvoje v metodách analytických a preparativních separací. Komplexnost biologický.ch systémůklade velképoŽadavky na pouŽité metody. V současnosti se běžné vytlŽivá šest ruzných módů HPLC pro ana|ýzu proteinů: iontově qiměrurá chromatografie (IEC)' vylučovací(SEC), afinitní (ALC), chromatografie s hydrofobní interakcí (HIC), chronratograÍie na normální fazi (i.IPLC) a chromatografie na reverzni fázi (RPLC). Dva z těchto módů, konkrétně afinitní chromatograťte a vylučovacíchromatografie, byly...
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Polymethakrylátové monolitické kolony v kapilární kapalinové chromatografii
Grafnetter, Jan ; Coufal, Pavel (vedoucí práce) ; Pacáková, Věra (oponent) ; Ventura, Karel (oponent) ; Sýkora, David (oponent)
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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Popis elektroforetických separačních systémů, využití systémových píků k charakterizaci vlastností surfaktantů a optimalizace složitějších systémů pomocí modelu LFER
Lokajová, Jana ; Tesařová, Eva (vedoucí práce) ; Coufal, Pavel (oponent) ; Ševčík, Juraj (oponent)
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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