Search results for "Reversed-phase chromatography"
showing 7 items of 187 documents
Peptide mapping by reversed-phase high-performance liquid chromatography employing silica rod monoliths.
2003
In this paper, a general procedure is described for the generation of peptide maps of proteins with monolithic silica-based columns. The peptide fragments were obtained by tryptic digestion of various cytochrome c species with purification of the tryptic fragments achieved by reversed-phase high-performance liquid chromatographic methods. Peak assignment of the various peptides was based on evaluation of the biophysical properties of the individual peptides and via mass spectrometric identification. The performance of several different monolithic sorbents prepared as columns of identical cross-sectional dimensions were investigated as part of these peptide mapping studies and the data evalu…
Solute retention in reversed-phase chromatography as a function of stationary phase properties: Effect of n-alkyl chain length and ligand density
1988
Two series of bonded phases were synthesized employing LiChrospher Si 100, 10 μm and n-alkyldimethylmonochlorosilanes as silanizing reagents. In series A the n-alkyl chain length, n, of the bonded phase was varied between 1 and 20 at a constant ligand density of 3.5±0.2μmol·m−2. In series B the ligand density, d, was gradually changed from 0 to 4.1μmol·m−2 on the C1, C4, C6, C8 and C18 bonded phases, respectively.
Super-high-speed liquid chromatography of proteins and peptides on non-porous Micra NPS-RP packings
1999
Abstract The new generation of non-porous silica RP packings commercially available from Micra Scientific was tested for separations of peptides and proteins by means of the gradient HPLC. Extremely high-speed separations were achieved using conventional chromatographic equipment: six proteins could be completely separated within six seconds. Tryptic digest peptides could be resolved in more then 40 components within 2–3 min. The effect of the experimental parameters such as temperature, flow rate etc. was investigated.
Comparative study on the column performance of microparticulate 5-μm C18-bonded and monolithic C18-bonded reversed-phase columns in high-performance …
1999
In this paper we report on the results of a comparative study on the performance of Purospher RP 18e, 5 μm, columns and prototypes of monolithic columns named SilicaROD from Merck, Darmstadt, Germany. The studies were performed on HPLC equipment with minimum extra column contribution. The plate height linear velocity dependency of the Purospher RP 18e column showed a minimum of H of about 10–15 μm at a linear velocity of 1 mm/s. The H versus u curves of the monolithic columns followed the same course. Yet, the curves remained flat up to a linear velocity of about 7 mm/s, where the Purospher RP 18e column could not be operated anymore due to the extremely high back-pressure. In conclusion th…
Determination of Homovanillic Acid in Human Plasma Using HPLC with Electrochemical Detection and Automated Solid Phase Extraction
1993
Abstract An isocratic HPLC method with electrochemical detection for the quantification of homovanillic acid (HVA) is described. The method included automated solid phase extraction on C-18-reversed phase material, followed by separation on a 3-μm Nucleosil 100 C18 column (250 mm × 4.6 mm I. D.) with a 100 mM citric acid solution (pH 6.6) containing 4% acetonitrile (v/v) as eluent at a flow rate of 0.6 ml/min. Isovanillinic acid served as internal standard. Extractability of both analytes was ca. 80 %. After extraction of 1 ml of plasma, coefficients of variation of replicate analyses were below 10 % in the naturally occuring concentration range.
Plasticizer extraction of Taxol infusion solution from various infusion devices.
1996
Taxol solution extracts the plasticizer DEHP (di(2-ethylhexyl)phthalate) from polyvinyl chloride (PVC) materials. In order to minimize patient exposure to DEHP, Taxol solutions should be prepared and administered in PVC-free materials. Particulate matter may form in Taxol infusion solution over time, so that in-line filtration with microporous membranes not greater than 0.22 microns is advisable. The purpose of this study was to evaluate the suitability of various administration- and in-line filter-sets for Taxol application. The extent of leached DEHP was determined using a Reversed Phase HPLC assay specific for DEHP. The four tested administration-sets, labeled as PVC-free, were all found…
Use of a three-factor interpretive optimisation strategy in the development of an isocratic chromatographic procedure for the screening of diuretics …
2000
Screening of diuretics in urine is feasible through direct injection of the samples into the chromatographic system and isocratic reversed-phase liquid chromatography (RPLC) with micellar-organic mobile phases of sodium dodecyl sulfate (SDS) and 1-propanol. The surfactant coverage of the chromatographic column makes the addition of organic competing amines less necessary than in conventional aqueous-organic RPLC to achieve well-shaped peaks. Also, the range of elution strengths of micellar mobile phases required to elute mixtures of hydrophobic and hydrophilic diuretics is smaller. This allows the isocratic separation of the diuretics within adequate analysis times. An interpretive methodol…