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Supercritical fluid adsorption and desorption of lipids on various adsorbents
Mojca Škerget, Željko Knez, 2007, izvirni znanstveni članek

Opis: In present work the feasibility of using supercritical fluid chromatography (SFC) for separation of free fatty acids (FFA) and triglycerides has been examined. Lipids such as oleic acid and sunflower oil (containing 96% of triglycerides) were used; adsorption and desorption on adsorbents such as silica gel and neusilin using supercritical $CO_2$ as mobile phase was investigated. The experiments were performed at 40 °C and in the pressure range from 13 to 30 MPa. Adsorption kinetic lines were determined by analysingbreakthrough curves. Several parameters such as rate of adsorption and desorption, loading, desorbed quantity and diffusion coefficients in solid phase were calculated.
Ključne besede: supercritical fluid chromatography, lipids, oleic acid, triglycerides, adsorption, desorption
Objavljeno: 31.05.2012; Ogledov: 860; Prenosov: 12
.pdf Polno besedilo (167,88 KB)

Multivariate data analysis of erythrocyte membrane phospholipid fatty acid profiles in the discrimination between normal blood tissue and various disease states
Zdenka Cencič-Kodba, Darinka Brodnjak-Vončina, Marjana Novič, Uroš Potočnik, 2010, izvirni znanstveni članek

Opis: The investigation presented here aims to compare the fatty acid composition of red blood cells of patients with different disease states and to test the hypothesis that the changes in fatty acid profiles derived from erythrocyte phospholipids might be relevant to various diseases. The study sample consisted of 342 blood donors, among them 135 with inflammatory bowel disease, 53 with uterine leiomyoma, 14 with verified absence of uterine leiomyoma, 52 with asthma, 18 with colon adenomas, and 70 blood samples without any of mentioned diseases that was used as a control group. After the isolation of erythrocytes from blood samples, total extracted lipids were separated by solid-phase extraction (SPE) into non polar lipids and polar phospholipids. After the saponification of phospholipid fraction, the esterification process followed with boron trifluoride-methanol reagent. The fatty acid methyl ester (FAME) composition of the total red blood cell phospholipid fraction was analyzed by gas chromatography (GC) with flame ionization detector (FID). Additionally two fatty aldehyde dimethyl acetals (hexadecanal and octadecanal dimethyl acetals; 16:0 DMA and 18:0 DMA) derived from erythrocyte membrane plasmalogen phospholipids were also determined. The resulting fatty acid and plasmalogen linked fatty acid composition was evaluated by the principal component analysis (PCA). We demonstrated decreased levels of omega-3 polyunsaturated fatty acids (n-3 PUFAs) in red blood cell membrane of patients with colon adenomas. Also, a large negative correlation was observed among all samples between the quantity of saturated acids and arachidonic (20:4n6) acid as well as saturated acids and adrenic (22:4n6) acid. In PCA score plot a group of female donors is distinguished mainly by the content of linoleic (18:2n6) acid; a small subgroup shows its concentration highly above the average value. At the same time, the same subgroup has both dimethyl acetals below the average concentrations. The study demonstrates feasibility of multivariate data analysis in discrimination of patients with different diseases according to fatty acid profile and suggests considerable differences in membrane fatty acid profiles in patients with various disease states.
Ključne besede: erythrocyte phospholipids, cell membrane, fatty acid profiles, differentiation, disease state, gas chromatography, principal component analysis
Objavljeno: 31.05.2012; Ogledov: 1228; Prenosov: 11
.pdf Polno besedilo (280,87 KB)

Chiral discrimination in mobile phases for HPLC
Renato Lukač, Andrew J. Clark, Syma Khalid, Alison Rodger, Alan Snedden, P. Mark Rodger, 2002, izvirni znanstveni članek

Opis: In this paper we present an algorithm and some preliminary results of a computer simulation study designed to elucidate the molecular interaction mechanisms associated with chiral discrimination in chiral high performance liquid chromatography (HPLC). Molecular dynamics simulations have been performed on a novel active stationary phase constituent based on disaccharides and a model analyte in a typical solvent used as mobile phase in chiral HPLC. The results are interpreted in terms of typical binding geometries and energies found from the simulations. This paper provides basic algorithms for predicting enantiometric selectivity and for investigating the implications for choice of parameters such as solvent polarity and temperature for optimising chiral HPLC separations
Ključne besede: physical chemistry, liquid crystals, liquid chromatography, computer simulations, spectroscopy
Objavljeno: 07.06.2012; Ogledov: 498; Prenosov: 1
URL Polno besedilo (0,00 KB)

Determination of major phenolic acids, phenolic diterpenes and triterpenes in Rosemary (Rosmarinus Officinalis L.) by gas chromatography and mass spectrometry
Maša Islamčević Razboršek, Darinka Brodnjak-Vončina, Valter Doleček, Ernest Vončina, 2007, izvirni znanstveni članek

Opis: A gas chromatographic-mass spectrometric (GC-MS) method for the simultaneous identification and quantification of seven major phenolic and terpenic compounds in Rosmarinus officinalis L. was developed. The compounds were identified as trimethylsilyl (TMS) derivatives of phenolic acids (caffeic and rosmarinic acid), phenolic diterpene (carnosic acid), and pentacyclic triterpenes (ursolic, oleanolic, betulinic acid and betulin). These compounds have been identified by retention time and comparison of mass spectra. The procedure involves ultrasonic extraction using solvent mixture of tetrahydrofuran and ethanol. Extracts were fractionated by size exclusion chromatography (SEC) after purifying on graphitised carbon. The fraction with phenolic and terpenic compounds was derivatised prior to GC-MS analysis using N-methyl-N-trimethylsilyl trifluoroacetamide (MSTFA) as a derivatisation reagent. The derivatisation process was optimized regarding temperature and reaction time. The linearity of the method was tested in concentration range 4-25 mg L$^{–1}$. The correlation coefficients ($r^2$) were in the range of 0.997 to 0.999. The average recoveries for all compounds ranged from 80 to 82%. The GC-MS technique is specific and sensitive, and can be used for simultaneous identification and determination of a wide range of phenolic and terpenic compounds in different plants even at trace levels.
Ključne besede: Rosmarinus officinalis, diterpenes, triterpenes, size exclusion chromatography, gas chromatography, mass spectrometry
Objavljeno: 21.12.2015; Ogledov: 355; Prenosov: 5
.pdf Polno besedilo (227,83 KB)

Supercritical fluid chromatography and scale up study
Miha Oman, Petra Kotnik, Mojca Škerget, Željko Knez, 2014, izvirni znanstveni članek

Opis: The influence of process parameters on supercritical fluid chromatography (SFC) and scale-up on to the preparative scale was investigated. In this scope, dependency of pressure, temperature, type and concentration of modifier, and type of stationary phase on the separation were examined separately. Experiments were performed on silica stationary phases in the range of pressures from 100 bar to 250 bar and temperatures from 30°C to 65°C, with use of ethanol and methanol as modifier. Results obtained on analytical scale were used for scale up on preparative (to production) scale. The aim of this study is presentation of the state-of-the-art of SFC through separate effects of process parameters on the separation and investigation of development, benefits and drawbacks of scaling-up process of the SFC.
Ključne besede: supercritical fluids, supercritical fluid chromatograpy, chromatography, scale up study, SFC
Objavljeno: 21.12.2015; Ogledov: 359; Prenosov: 7
.pdf Polno besedilo (842,20 KB)

Gut microbiota patterns associated with colonization of different clostridium difficile ribotypes
Jure Škraban, Sašo Džeroski, Bernard Ženko, Domen Mongus, Simon Gangl, Maja Rupnik, 2013, izvirni znanstveni članek

Opis: Abstract Introduction Materials and Methods Results Discussion Supporting Information Acknowledgments Author Contributions References Reader Comments (0) Figures Abstract C. difficile infection is associated with disturbed gut microbiota and changes in relative frequencies and abundance of individual bacterial taxons have been described. In this study we have analysed bacterial, fungal and archaeal microbiota by denaturing high pressure liquid chromatography (DHPLC) and with machine learning methods in 208 faecal samplesfrom healthy volunteers and in routine samples with requested C. difficile testing. The latter were further divided according to stool consistency, C. difficile presence or absence and C. difficile ribotype (027 or non-027). Lower microbiota diversity was a common trait of all routine samples and not necessarily connected only to C. difficile colonisation. Differences between the healthy donors and C. difficile positive routine samples were detected in bacterial, fungal and archaeal components. Bifidobacterium longum was the single most important species associated with C. difficile negative samples. However, by machine learning approaches we haveidentified patterns of microbiota composition predictive for C. difficile colonization. Those patterns also differed between samples with C. difficile ribotype 027 and other C. difficile ribotypes. The results indicate that not only the presence of a single species/group is important but that certain combinations of gut microbes are associated with C. difficile carriage and that some ribotypes (027) might be associated with more disturbed microbiota than the others.
Ključne besede: microbiota, clostridium difficile, chromatography
Objavljeno: 19.06.2017; Ogledov: 26; Prenosov: 2
.pdf Polno besedilo (1,01 MB)

Determination of topiramate in human plasma using liquid chromatography tandem mass spectrometry
Tanja Vnučec, Lea Cvitkovič-Maričič, Helena Prosen, Darinka Brodnjak-Vončina, 2013, izvirni znanstveni članek

Opis: The LC-MS/MS method for determination of the anti-epileptic drug topiramate (TPM) in human plasma was developed and validated for pharmacokinetic and bioequivalence study purposes. For quantitative determination of TPM values the method with deuterated internal standard (topiramate-d12) and liquid chromatography with tandem mass spectrometry was used. TPM was extracted from the human plasma using the solid-phase extraction procedure on a Strata X extraction column. Negative ions were monitored in the selected reaction monitoring mode (SRM) and transitions m/z 338.2 > 78.2 and m/z 350.3 > 78.2 were used for the quantitative evaluation of TPM and the internal standard, respectively. The results obtained from validation were statistically evaluated according to the requirements of European Medicines Agency (EMA) and Food and Drug Administration (FDA) regulatory guidelines. The linearity of the method was checked within a concentration range from 10 to 2000 ng/mL. Successful validation confirmed that this method is precise, accurate, sensitive and therefore suitable for determination of topiramate plasma levels in pharmacokinetic and bioequivalence studies.
Ključne besede: topiramate, liquid chromatography tandem mass spectroscopy, human plasma, bioequivalence study
Objavljeno: 18.08.2017; Ogledov: 18; Prenosov: 0
.pdf Polno besedilo (189,19 KB)

Comparison of methods for determination of polyphenols in wine by HPLC-UV/VIS, LC/MS/MS and spectrophotometry
Vesna Bukovac, Matija Strlič, Drago Kočar, 2009, izvirni znanstveni članek

Opis: Phenolic antioxidants are usually grouped into flavonoids and non-flavonoids, according to their structure. With regard to the tannic character, phenolic antioxidants are further subdivided to tannic phenols and non-tannic phenols. Collectively, these compounds contribute to the high antioxidant capacity of wine. In this work, we compare determination of gallic acid, catechin, epicatechin, resveratrol, quercetin, dihydrobenzoic acid, sinapic acid, vanillic acid, caffeic acid, chlorogenic acid, ferullic acid, ellagic acid, p-coumaric acid and caftaric acid in 141 wine samples using two liquid chromatographic methods and detection systems, i.e. with UV detection and mass-spectrometric detection. In addition, we applied the conventional Folin-Ciocalteu spectrophotometric method for determination of the total phenolic content in wine samples and compared the results with those obtained using the chromatographic methods. Despite satisfactory correlations statistically significant differences between HPLC-UV/VIS and LC/MS/MS were established, which could be related to coelution not detectable with UV/VIS detectors. The correlations between results of the spectrophotometric method and sum of LC/MS/MS determinations are not satisfactory and are different for white, red, and rosé wines.
Ključne besede: food analysis, wine, antioxidants, chromatography
Objavljeno: 17.08.2017; Ogledov: 41; Prenosov: 0
.pdf Polno besedilo (185,67 KB)

Determination of candesartan in human plasma with liquid chromatography - tandem mass spectrometry
Vanja Forjan, Lea Cvitkovič-Maričič, Helena Prosen, Darinka Brodnjak-Vončina, 2016, izvirni znanstveni članek

Opis: A sensitive, specific and rapid liquid chromatography - tandem mass spectrometry method was developed and validated for the determination of candesartan in human plasma. Analyte was separated from endogenous components present in plasma by solid phase extraction. Chromatographic separation was performed on Gemini C18 analytical column using mobile phase acetonitrile – 5 mM ammonium formate pH 2 (90:10, v/v) at flow rate of 0.3 mL/min. For detection, tandem mass spectrometry in SRM mode with positive electrospray ionization was used. The mass transitions m/z 441.1 > 263.1 and 445.1 > 267.1 were used to determine candesartan by using candesartan-d4 as an internal standard. After development, the method was validated according to the requirements of EMA regulatory guidelines in the concentration range 1 - 400 ng/ml in human plasma. Limit of quantification (LLOQ) was 1 ng/ml. The developed and validated method proved to be very fast and reproducible and was therefore successfully implemented in pharmacokinetic and bioequivalence studies with large number of study samples.
Ključne besede: candesartan, liquid chromatography, tandem mass spectrometry, human plasma
Objavljeno: 17.08.2017; Ogledov: 38; Prenosov: 0
.pdf Polno besedilo (215,54 KB)

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