0000000000204443
AUTHOR
P. Roumeliotis
Separation of unmodified α-amino acid enantiomers by reverse phase HPLC
A novel chiral phase system is presented for the resolution of unmodified α-amino acid racemates which is composed of a reverse phase packing coated with N-alkyl-L-hydroxyproline (where alkyl is n-C7H15-, n-C10H21- and n-C16H33-) and a hydro-organic eluent containing copper(II)acetate. The factors controlling retention and enantioselectivity such as concentration of Cu(II)ion and pH of the eluent, addition of NH4Ac to the eluent, type and content of organic solvent of hydro-organic eluent and column temperature were examined. The extremely high enantioselectivity observed (α up to 16) is assumed to be caused by a three site sorbate-sorbent interaction involving bidentate coordination of two…
Optimization of the phase system in the analysis of polynuclear aromatics (PNA) from diesel engine exhaust by high performance liquid chromatography (HPLC)
Polynuclear aromatics (PNA) were collected and enriched from diesel engine exhausts by means of a probe and an absorber system. Group separation of PNA from other constituents of exhaust was accomplished through elution with n-heptane on a silica column. A variety of phase systems ranging from unpolar to strong polar packings and eluents, respectively, were tested to resolve PNA into single species. Of these a n-octadecyl bonded silica packing and methanol/water was found to be the most optimal with regard to resolution for real PNA mixtures. Separation was completed within 2–3 h at isocratic conditions.
Preparative separation of proteins and enzymes in the mean molecular-weight range of 10,000–100,000 LiChrosorb diol® packing by high-performance size-exclusion chromatography
Abstract LiChrossorb Diol® packing has been to be well-suited for the separation of proteins and enzymes according to a size-exclusion mechanism in a mean molecular-weight (MW) range between 10,000 and 100,000. Loadability of a small bore column of 6 mm I.D. (A) and a large-bore column of 23.5 mm I.D. (B), both of 250 mm in length, were examined. Defining a 20% decrease of the number of theoretical plates as loadability limit the volume load at constant mass of chymotrypsinogen as representative test solute was ≈ 100 μl for column (A) and 1500 μl for column B at 0.78 · 10-5 g/g of packing for column A and 0.78 · 10-6 g/g of packing for column B, respectively. Mass load to constant injection…
On-line high-pressure extraction-high-performance liquid chromatography
Abstract A coupling unit is described that allows on-line operation of a high-pressure extraction apparatus with a high-performance liquid chromatograph. The unit consists of two high-pressure sample-injection valves, connected in series. The first valve operates as a switching valve to the loop and controls the release of pressure along a packed microbore column. Two short, packed microbore columns are positioned between the first and second valve, sampling the extracted compounds over a desired period and simultaneously operating as sample loop for the second valve; the latter is the injector for the high-performance liquid chromatographic column. The unit was tested by continuous extract…
Assessment and optimization of system parameters in size exclusion separation of proteins on diol-modified silica columns
Abstract On diol-modified silica columns the retention of proteins is governed by a size exclusion effect, but superimposed on this are some secondary effects, i.e. , ionic and diol-ligand interactions which can be controlled and adjusted reproducibly by varying the eluent composition. The eluent composition also affects the column efficiency and peak shape. Both dependences can be employed to obtain a better resolution of proteins than can be expected from size exclusion alone.
Determination of Phenols from Automobile Exhaust by Means of High-Performance Liquid Chromatography (HPLC)
Abstract The paper describes a procedure to determine ten representative phenols (phenol, m-, p-, o-cresol, 2,4-, 2,6-, 3,4-, 3,5-xylenol, 2,3,6-, 2,4,6-trimethylphenol) in engine exhaust. Sampling was performed by a conventional Grimmer device. A sample of the water condensate collected was extracted with ether. After concentration of the ether extract phenols were isolated from other interfering compounds by stepwise elution on a Lobar column packed with LiChroprep Si 60 under controlled conditions. Relative recovery of the majority of reference phenols ranged from 90-100%. Separation of phenols into individual components was performed on polar packings as well as on Reverse Phase packing…
Effect of the hydrophobic spacer in bonded [Cu(l-hydroxyprolyl)alkyl]+ silicas on retention and enantioselectivity of α-amino acids in high-performance liquid chromatography
Abstract The following chiral ligands were bonded to silica: [Cu( l -hydroxyprolyl)methyl) + on LiChrosorb Si 60 (1), [Cu( l -hydroxyprolyl)methyl] + on LiChrosorb Si 100 (2) and [Cu( l -hydroxyprolyl) n -octyl] + on LiChrosorb Si 100 (3). The packings contained residual iodomethyl- and ω-iodooctyl groups at the surface. Studies on packing 1 and 2 under comparable conditions in eluents containing 10 -4 M copper acetate showed a higher retention on 1 than on 2 but a much better enantioselectivity in the latter case. The retention of enantiomers on all packings examined was found to be governed by the eluent pH and methanol content as well as by the concentration of ammonium acetate. The vari…
Study on the efficiency of assembled packed microbore columns in HPLC
Stainless steel columns (internally mirror-finished, 125 or 250nm in length, of bore 1.0 or 1.6mm) were slurry-packed with 5µm and 4µm reversed phase silicas (Hypersil ODS, LiChrosorb RP-8 and RP-18 and Superspher RP-8. The HPLC equipment consisted of a pump LC5A (Shimadzu) or a pump 2150 (LKB Instruments), a rheodyne valve 7413 with loops of 0.5, 1.0 and 5.0mm3 and a Jasco-Uvidec 100-II UV detector, variously with one of three specially constructed cells of 0.2, 0.4 and 1.3mm3 volume. Columns were assembled using two types of coupling device employing a stainless steel capillary of 0.12mm bore. The effect of sample volume, design of coupling device in assembled column, detector cell volume…
Structure and properties ofn-alkyldimethylsilyl bonded silica reversed-phase packings
Summary The effects of the modifier functionality and the chain length ofn-alkylchlorosilanes on the surface structure of packings and on their retention behaviour in reversed-phase chromatography were investigated. Comparative retention studies on three silica packings that had been treated to maximum conversion withn-octyltrichlorosilane (I),n-octylmethyldichlorosilane (II) andn-octyldimethylmonochlorosilane (III) showed that the most pronounced reversed-phase character is obtained by using III as the modifying reagent. A series ofn-alkyldimethylsilyl bonded silica packings were prepared with widely differingn-alkyl chain lengths. Although the packings are very hydrophobic, the maximum su…
Novel porous carbon packings in reversed-phase high-performance liquid chromatography
Abstract Porous carbon packings of particle size 5–20 μm were prepared by calcination of purified active carbons and cokes having narrow size ranges. Highly efficient columns packed with 10-μm particles were obtained by means of the high-viscosity slurry technique, and exhibited about 3000 theoretical plates at 100 mm length and optimal eluent flow-rate. The loading of columns, θ0.1, was determined to be 25–50 μg of solute per gram of packing. Eluotropic series of solvents were found to be similar to those on pyrocarbon-modified carbons and silicas. The log k′ values for homologous compounds increased linearly with the hydrocarbonaceous surface area at constant eluent composition. Studies w…
Anwendungsmöglichkeiten der HPLC bei der Trennung, Isolierung, Identifizierung und quantitativen Bestimmung von Substanzen aus komplexen Stoffgemischen—am Beispiel von Carbonsäuren in Natriumaluminatlaugen aus dem Bayerprozeß der Aluminiumoxidgewinnung
Es wird die Anwendung der Hochdruckflussigchromatographie (HPLC) in Bezug auf die Identifizierung und quantitative Bestimmung von Carbonsauren in Aluminatlaugen aus dem Bayerprozes beschrieben. Die Vorgehensweise besteht in folgenden Teilschritten: Optimierung von HPLC Phasensystemen zur Trennung von synthetischen Gemischen von Carbonsauren, Aufarbeitung der Aluminatlauge, semi-praparative Isolierung von Substanzen, Identifizierung auf flussigchromatographischem Wege und durch Massenspektrometrie, quantitative Bestimmung durch Peakhohenauswertung mit Hilfe externer Standards.