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  • Column liquid chromatography  (349)
  • Industrial Chemistry
  • Inorganic Chemistry
  • Seismology
  • ddc:330
  • Springer  (353)
  • 1985-1989  (351)
  • 1950-1954  (2)
  • 101
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    Chromatographia 28 (1989), S. 412-414 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Dinitrobenzene isomers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary An efficient, reproducible and rapid high-performance liquid chromatographic method, in normal phase mode, for the analysis of the three dinitrobenzene isomers is described. The method affords good linearity for each isomer in the range 10–160 μg ml−1. The total analysis time is only 10 minutes, and the method shows an accuracy of ±1.25% with a coefficient of variation from 0.30% to 2.85% for different levels of the dinitrobenzene isomers.
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  • 102
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    Chromatographia 28 (1989), S. 493-496 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Nucleic acid components ; Buffer salt concentration ; Variation of retention
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary In the reversed-phase high-performance liquid chromatographic studies of nucleotides, nucleosides and bases, phosphate and acetate are commonly used as buffer materials. The concentration of these buffer salts in the mobile phase exerts noticeable influence both upon the retardation and upon the separation of the nucleic acid components. This can be of certain usage in the fine adjustment of the elution order of individual components. The possible reasons for the observed data are discussed.
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  • 103
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Optical resolution ; Polymeric adsorbents ; Penicillin sulfoxide ; Radical polymerization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Polymethacrylamide carrying penicillin sulfoxide skeleton, poly(benzyl 1-oxo-6-methacrylaminopenicillanate) was synthesized in various solvents. Optical resolution efficiency of the high-performance liquid chromatography columns comprising the polymers was investigated. It was found that the radical polymerization solvent influenced the optical resolution capability of the resultant polymer, and that less hydrogen-bonding solvent was preferred for effective resolution.
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  • 104
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    Chromatographia 25 (1988), S. 31-36 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Radioactivity detector ; Solid scintillator ; Fibre-packed flow cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A new type of flow-cell radiation detector for use in liquid chromatography which is packed with aligned scintillator fibers is described. A primary advantage of the fiber packed cell is that light generated by the scintillator is absorbed to a much less extent by the fibers than by the powder scintillator used in conventional flow cells. A detection efficiency of 55% has been obtained for carbon-14 using 0.1-mm diameter hand-pulled glass fibers. Computer modeling has shown that even better results can be obtained by using smaller and more uniform diameter fibers which will allow better packing. The fiber cell also demonstrates back pressures which are a factor of 50 less than the conventional cell and much less susceptibility to absorption of compounds because of its lower surface area.
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  • 105
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    Chromatographia 25 (1988), S. 854-860 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Polystyrene ; Gradient elution ; Turbidimetry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Polystyrene samples of narrow molecular-weight distribution have been eluted according to their molecular weight from columns packed with bare silica Si50, phenyl, or C18 bonded phase by gradients of methanol and tetrahydrofuran (THF) or ofiso-octane and THF. Among the six combinations investigated,iso-octane/THF with a silica column formed a proper normal-phase system whereas methanol/THF with a C18 column formed a proper reversed-phase system. The combinations of C18 column andiso-octane/THF or of Si50 column and methanol/THF gradient did not correspond to the approved polarity rules in high-performance liquid chromatography but were nevertheless effective in separating polystyrene mixtures by molecular weight. Methanol andiso-octane are nonsolvents for polystyrene whereas THF is a solvent. The solubility of polystyrene as a function of molecular weight and concentration was determined by means of turbidimetric titration of solutions in THF with the nonsolvents used in the gradients. The solubility and elution characteristics were almost identical on C18 columns or in methanol/THF combinations. The elution from phenyl bonded phase and Si50 columns usingiso-octane/THF gradients required more THF than the solubility experiments. Information is also given on the occurrence of multimodal elution patterns.
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  • 106
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Polycyclic aromatic compounds ; Alkylated PAC ; Retention characteristics ; Normal phase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Retention characteristics of series of polymethyl and mono-n-alkyl derivatives of benzene and pyrene, and also of parent polycyclic aromatic hydrocarbons (PAH), were studied using silica and aminopropyl- and cyanopropyl-modified silica. Differences in the selectivities for the studied compound groups were found between the three phases. Deviations from linear behaviour in plots of log (k′)vs. carbon number were observed for the methyl series. These are explained in terms of differences in π-electron delocalisation within the aromatic ring systems. Further, the effect of methyl substitution on selectivity decreased with an increasing number of aromatic rings. Results were obtained which indicated that the primary adsorption site in a cyano column used in normal phase mode, at least for PAH molecules, is the cyano group.
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  • 107
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    Chromatographia 25 (1988), S. 43-50 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Reversed phase ; Void volume ; Adsorption isotherms
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary In reversed-phase liquid chromatography with n-alkyl bonded silica, the dead volume (V0) of the column is theoretically indeterminate owing to adsorption of organic modifier on n-alkyl chains and of water on silanol groups. With binary mobile phases, retention volumes of the mobile phase components and of their deuterated species are relaeed to the adsorption isotherms and V0 by equations which can be solved with some assumptions on the adsorbed layer composition. Methanol-water and acetonitrile-water systems are studied. As the experimental excess isotherm shows a linear part in the concentration range 50–80% in organic modifier, the hypothesis of an adsorbed layer of constant composition in this range is possible. When increasing the water content of the mobile phase, adsorption of water occurs up to saturation of silanol groups. Then the assumption of a constant water content for a mobile phase having more than 50% of water is applied. With the hypothesis of a constant adsorbed content of organic modifier when the eluent has more than 80% of organic modifier, V0 and the absolute isotherms are calculated over the entire range of mobile phase composition. Experimental retention behavior of the mobile phase components are totally explained by these V0 determinations. The retention times of commonly used V0 markers are compared with V0 values. It is shown that, when buffering the eluent, no visible effect on the distribution equilibrium is observed, so that injection of concentrated potassium nitrate is a convenient method to measure V0. With a few solutes with are UV detectable it is possible to measure V0 whatever the mobile phase composition in methanol-water and acetonitrile-water systems.
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  • 108
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Stationary phase solubility parameters ; Cyano-silica ; Amino-silica ; Diol-silica
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Extended solubility parameters have been determined for aminopropyl, cyanopropyl and 1,2-dihydroxypropyl propyl ether (diol) normal bonded-phase HPLC columns. Parameters were calculated from both the retention data of solutes (partition model) and empirically determined solvent strengths (adsorption-displacement model). The use of solvent strengths to calculate solubility parameters for these silica-based bonded stationary phases appears to be superior, since this technique avoids many of the problems that arise from their inherent heterogeneity. Normalized solubility parameters were also used to position these columns on a stationary phase selectivity triangle. The amino and cyano phases appear in regions of the tringle expected based on the properties of the pure liquids, but an ether linkage in the diol apparently neutralizes some of the acidity expected from this phase.
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  • 109
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Macrolide antibiotic azithromycin ; Intermediates of azithromycin ; Optimization of separation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary HPLC is the most convenient method for the assay of the azithromycin (10-dihydro-10-deoxo-11-methyl-11-azaerythromycin A), a semisynthetic macrolide antibiotic and its reaction intermediates. Data concerning the effects of pH, temperature and column type are presented. The analytical procedures enabling the reliable assay of azithromycin and its intermediates, as well as other impurities are defined. Use of a reversed-phase octadecyl column, a pH of 9.3–9.5, and isocratic mode at ambient temperature are the best conditions of analysis.
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  • 110
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Metabolic disorders ; Xanthine and 2,8-dihydroxyadenine ; Urine analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary 2,8-Dihydroxyadeninuria and xanthinuria are inborn enzyme disorders which must be under lifelong therapeutic control. Quantitative determination of 2,8-DHA and xanthine using reversed phase HPLC was performed. A new application for the separation of purines shortens analysis-time. Examples of the determination of 2,8-DHA and xanthine before and under certain therapies are shown. Long term monitoring of the patients offers the possibility of a reliable prophylaxis against stone recurrence.
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  • 111
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Indirect detection of non-electrolytes ; Simulation of induced peaks ; RP systems
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Induced peaks observed in indirect detection of nonelectrolytes in high-performance liquid chromatography are simulated. The equation representing the signal intensity of the induced peaks is derived, and it is verified by the experimental results. The peak area is proportional to (K b ′ +1) k a ′ /| a ′ —k b ′ |, where k a ′ ad k b ′ are the capacity factors of the visualization agent and the analyte, respectively.
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  • 112
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    Chromatographia 25 (1988), S. 162-166 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Ion chromatography ; Analysis of metal ions ; Post column derivatisation ; Amperometric detection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A method for determination of metal ions subsequent to separation by ion chromatography has previously been developed. The method is based on Indirect Amperometric Detection whereby the decrease in the oxidation current, due to a dithiocarbamate ligand added post column, is monitored. Upon elution from the chromatographic column the metal ions are complexed by the ligand. As the complexes formed are electroinactive at the applied potential, the background current decreases according to the metal ion concentration. The method developed in this work involves addition to the reagent of Zn(II) as a auxiliary metal ion to displace the analyte metals from the chromatographic eluent ligand complexes after separation. Sodium bis(2-hydroxy-ethyl) dithiocarbamate was used as the postcolumn derivatising reagent. The addition of Zn(II) to the reagent causes some unforeseen behaviour in the chromatographic system.
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  • 113
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    Chromatographia 25 (1988), S. 199-204 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Column switching ; Catecholamines ; Urine ; Serum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary We developed an automated, two-column HPLC-method that can be used routinely to quantify the catecholamines norepinephrine and epinephrine in body fluids. The method is based on a new, laboratory-prepared SEC-HPAC column material, on the application of a microprocessor-controlled column-switching technique, and on the use of an integrated reaction system for postcolumn derivatization and trihydroxyindol-fluorescence monitoring. It allows the direct injection (upto 500 μl) of an appropriate biological fluid and distinguishes on its integrated sample-processing mode, its practicability, its chemoselectivity and-specivity, its detection limit (2 pg), its within- and between-run precision and its speed of analysis.
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  • 114
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Pheophytinatonicke(II) ; Chlorophyll ; Pheophytin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary HPLC determination of pheophytinatonicke(II) (Pheo-Ni) prepared by the replacement of magnesium(II) in chlorophyll with nicke(II) is described. The good separation of PheoNi was obtained by using chemically bonded C18 as the stationary phase and acetone-methanol (50∶50, vol/vol) as the mobile phase. Conventional spectrophotometric method was also used for the determination of PheoNi. For the synthetic samples prepared by mixing (pheophytinato a) nicke(II) [(Pheo-a) Ni] and (pheophytinato b) nicke(II) [(Pheo-b) Ni], analytical values obtained by the spectrophotometric method were very high compared to those obtained by HPLC. In the proposed HPLC method, (Pheo-a) Ni and (Pheo-b). Ni could be determined in the concentration range of 0.028–30μg/ml and 0.038–30μg/ml with relative standard deviations (n=10) of 3.1% and 0.8%, respectively.
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  • 115
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Flavonoid compounds ; Structure-retention relationships ; Reversed-phase systems
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Reversed-phase HPLC retention behaviour of different flavonoid compounds in different columns and with a range of methanol-buffer eluents was examined. Repeatibility and reproducibility of logk′ and Δlogk′ values were analyzed. Sharp group contributions, expressed as Δlogk′, were observed. These did not depend significantly on acid modifier type, column packing characteristics (C8 and C18) and dimensions (5,7 and 10μm) or the percentage of methanol in the mobile phase. A list of twelve group contributions is presented which is in agreement with literature data.
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  • 116
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    Chromatographia 25 (1988), S. 443-446 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Large scale separations ; Dissolution of silica ; Metallic contaminants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Possible contamination of purified product by the silica matrix is of considerable importance when considering process scale high-performance liquid chromatography (HPLC) as a separation methodology. The stability of silica supports in normal phase, reversed phase and affinity chromatography modes has been examined with organic solvents and water. Both inorganic and organic components were found, contributed by the support, and the chromatographic and distillation equipment. In all cases, based on assumed values of production, they were found to contribute less than 0.01% of impurities. In critical cases crystallisation or dialysis may be considered as a last stage of purification, with re-chromatography to maximise yields.
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  • 117
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Fast atom bombardment mass spectrometry ; Nonionic detergents
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Microcolumn liquid chromatography (LC) combined with fast-atom-bombardment mass spectrometry (FABMS) was applied to the analysis of nonionic detergents. The porous silver filter attached to the top of the capillary fused-silica tubing worked as the interface for coupling of micro LC and FABMS.
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  • 118
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    Chromatographia 25 (1988), S. 577-581 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Process scale ; Examples of separations
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The flexibility of process scale high performance liquid chromatography is demonstrated by three examples of reversed-phase separations (a) a standard purification (b) isolation of trace compounds, and (c) recovery from crystallisation mother liquors. It was shown that a material, thought to be acceptably pure, contained a toxic impurity, and a remaining impurity, a previously unseen component of potential therapeutic interest. Favourable cost data were indicated for example (3).
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  • 119
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    Chromatographia 25 (1988), S. 717-720 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Reversed-phase chromtography ; Antifungla agent ; Pharmacokinetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A high-performance liqid chromatographic method is reported for the measurement of miconazole in systemic, fungal infectious patients. Pharmacokinetic data are presented for a single patient receiving miconazole therapy. Sample preparation involves protein precipitation by acetonitrile (1:1, vol/vol). Analyses are carried out on a reversed-phae chromatographic system using octadecylsilane stationary phase: a mobile phase consisting of 0.05 M acetate buffer (pH 7.4)acetonitrile (20:80, vol/vol) is used to elaute miconazole is quantified on the basis of ultraviolet absorption at 220 nm. The precision of the method ranged from 3.21% at 0.5 mg/L to 0.85% at 2.0 mg/L. The limit of quantification was established as 0.1 mg/L. Interference from other drugs that are co-adimistered such as amphotericin B, 5-fluorocytosine of ketoconazole and most other comonly encountered drugs was not observed.
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  • 120
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Fractional factorial experimental designs ; Ruggedness test ; Method validation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Recently much attention has been directed to the validation of analytical methods. The level of validation will depend on the method application. If a method is developed for general use, for instance by different analysts, instruments or laboratories, then the effect of these changes need evaluation. This is normally referred to as ruggedness testing, where the limitations of the method to changes in the specified conditions are examined. This paper examines the use of Plackett-Burman fractional factorial experiment designs for ruggedness testing in high performance liquid chromatography (HPLC). The ability of these designs to identify confounding effects is applied to the stability indicating analysis of Salbutamol and its major degradation product, AH4045. The results show that confounding effects, proving extremely pertinent to HPLC, can be identified. The assay proved rugged to most tested changes although not to the use of column packing material from different manufacturers.
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  • 121
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Alkyl-bonded phases ; Retention behaviour ; Ligand density ; n-alkyl chain length
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary 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. The capacity factors of benzoic acid esters and anilines as solutes were found to increase linearly with the n-alkyl chain length of packings at constant eluent composition (series A) up to a so-called critical chain length, ncrit, where the capacity factor remained constant. ncrit was in the range from 11 to 14. The same pattern was observed when plotting the solute capacity factor against the ligand density of the packing at constant n and constant eluent composition (series B). The critical ligand density, αcrit, varied between 2.3 and 3.2 μmol·m−2 depending on n and the solute. Furthermore, solute retention was slightly higher on RP packings with an even number of carbon atoms in the alkyl chain than on those with an odd number. These findings are consistent with the results of Berendsen and de Galan (J. Chromatogr., 196, 21 (1980)), Dill (J. Phys. Chem., 91, 1980 (1987)) and Simpson and Lau (to be published). The observed phenomena reflect the dynamic structure of RP silicas which are related to the mobility of solvated n-alkyl chains. Due to the lack of a precise conformational analysis of the surface of RP silicas, a semiquantitative model was applied to interprete the described dependencies.
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  • 122
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    Chromatographia 25 (1988), S. 265-271 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Chiral phases ; Polymer coating ; Quinine substituted polysiloxanes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Stationary phases for chiral separations have been synthesized by chemical modification of porous small particle silica using new procedures of fixation of the chiral moiety. So called pre-polymers of the methylpolysiloxane polysiloxane type are immobilized on silica surfaces by different procedures. These pre-polymers are substituted by chiral groups. Their synthesis is done externally i.e. not in-situ on the support surface. The immobilization on the silica surface is achieved by crosslinking and/or by chemical bonding. Anchor groups within the pre-polymer (e.g. SiH) as well as on the silica surface (SiOH) give rise to the chemical bonding therewith. Chiral phases with quinine as the chiral moiety were obtained which show high separation efficiency as well as chemical stability, in addition to the enantiomeric selectivity required for the separation of certain types of aliphatic and aromatic alcohols.
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  • 123
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    Chromatographia 25 (1988), S. 288-294 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Mobile phase volume ; Ionic solutes ; Eluent electrolyte
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A new method for the determination of the mobile phase volume (V m) in liquid chromatography is presented based on the model regarding the retention of ionic solutes in the presence of eluent electrolytes. TheV m value can be determined by measuring the retention volumes of two ions that have the same charge in two eluent electrolyte systems. Compared with the methods using isotopically labelled eluent components or inorganic salts asV m markers, the method presented is proved to give more reasonableV m values for both normal and reversed-phase liquid chromatography. As well as in binary mixed solvent systems, theV m values in single solvent systems can be determined by this method.
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  • 124
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    Chromatographia 25 (1988), S. 339-342 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Gradient elution ; Prediction of retention
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A general equation for the final retention of a solute chromatographed under conditions of stepwise gradient elution has been derived. The elution process and the distances travelled by solutes as a function of eluent volume were simulated by computer for the optimization of stepwise gradient prorams from isocratic HPLC data. The validity of the equations was experimentally veritied.
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  • 125
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    Chromatographia 25 (1988), S. 272-278 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Polyaromatic hydrocarbons ; Molecular parameters ; Retention prediction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The present research studies the possibility of using the correlation dependence between molecular parameters of unsubstituted polyaromatic hydrocarbons (PAH) and their retention in reversed-phase liquid chromatography to optimize the conditions for the separation and identification of unknown peaks on the chromatograms of multicomponent mixtures. A linear correlation equation, that takes the number and environment of the carbon atom in the PAH molecule into account as well as the differences in the specific interactions of isomeric molecules with polar eluent, has been proposed. The adequacy of the proposed PAH retention model was verified by comparing the calculated retention values with the experimental data. The possibility of identifying unsubstituted PAH according to the number of carbon atoms of various types and according to the values of the molecules lengths (calculated on the basis of the retention of these substances under different eluent compositions) was exemplified by various chromatographic systems (reversed phase-eluent-PAH molecules).
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  • 126
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    Chromatographia 25 (1988), S. 327-331 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Olefin group selectivity ; Methylene group selectivity ; Non-aqueous reversed-phase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The effects of mobile phase composition upon olefin group selectivity (the ratio of the retention factor of a n-alkane to 1-olefin of equal carbon number) has been examined for non-aqueous reversed-phase liquid chromatorphy. Under time-normalized conditions, large variations in olefin group selectivities were noted as the mobile phase constitutents were changes. However, methylene group selectivities were found to be insensitive to the nature of the mobile phase under these conditions. Mobile phases containing alcohols demonstrated low olefin group selectivities compared to those containing acetonitrile as weak solvent. The results of this study explain variations previously observed in the LC separation of olive oil triglycerides that differ in the number of methylene groups and double bonds.
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  • 127
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Cyano bonded phases ; Residual silanol groups ; Diethyldithiocarbamates
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary When a cyano bonded phase is used together with a nonaqueous eluent, it is universal that the silanol groups which remain at the surface of the silica gel after bonding affect the retention of solutes. With solutes containing such atoms as N, S and O, hydrogen bond may form between the solute and the residual silanol group, leading to dual retention mechanism. Based on the understanding of retention mechanism, methods were developed to separate metal-diethyldithiocarbamate (DDTC) chelates and crown ethers on cyano bonded phase, the mobile phase being a nonaqueous solvent containing triethylamine (TEA).
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  • 128
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    Chromatographia 25 (1988), S. 409-412 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Separation of proteins ; Biological activity after separation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The proteins of the large ribosomal subunit fromEscherichia coli have been separated by size-exclusion, ion-exchange and reversed-phase high-performance-liquid chromatography (HPLC) using various buffer systems. The biological activity of the isolated proteins was tested via their ability to assemble into active 50S subunits (total reconstitution). The activity of the reconstituted subunits was measured with poly(U)-dependent poly-(Phe) synthesis. Reversed-phase HPLC techniques yielded active proteins (80–100%) by application of 2-propanol or acetonitrile. Proteins prepared by size-exclusion chromatography employing ammonium acetate as buffer also gave highly active proteins (70%). On the other hand, separation of the proteins on ion-exchange columns, using urea containing buffers, resulted in reduced activity (up to 50%).
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  • 129
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Phospholipids ; Amniotic fluid ; Fetal lung maturity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A new quantitative analytical method for the determination of phospholipids in amniotic fluid by high performance liquid chromatography (HPLC) is described. In addition to the main compounds, phosphatidylcholine (lecithin) and sphingomyelin, the so-called minor phospholipids, phosphatidylglycerol, phosphatidylinositol and phosphatidylethanolamine can also be determined. Separation is achieved using a guard-column of Lichrosorb Si 60 and an analytical column of Lichrosorb DIOL. Acetonitrile/water is used as mobile phase at an elevated temperature. By determining the recovery rates, the within-run and the between-run precision, it was shown that sufficient accuracy and precision could be achieved for all the parameters examined. The method is highly sensitive, the detection limit for sphingomyelin is 0.2 μg and 0.1 μg for all the other components. A single determination of 5 phospholipids in an amniotic fluid sample takes about two hours. By performing simultaneous extractions it is possible to analyse 5 samples per day.
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  • 130
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Reversed-phase diphenyl and naphtylethyl bonded phases ; Retention/molecular polarizability relationships ; Peropyrene aromatics
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    Topics: Chemistry and Pharmacology
    Notes: Summary The retention behavior of 15 peropyrene-type polycyclic aromatic hydrocarbons was investigated on various bonded stationary phases in reversed-phase liquid chromatography. On diphenyl and naphthylethyl bonded phases, high correlations were obtained between the molecular polarizability of solutes and their retention. However, very low or no correlations were found on various octadecyl bonded phases. These facts are discussed by using the electrostatic interaction concept between the solutes and the stationary phase. We conclude that these observations are due to two reasons: the difference in the degree of planarity of polycyclic aromatic hydrocarbons and the high ability of planarity recognition of octadecyl bonded phases.
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  • 131
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    Chromatographia 25 (1988), S. 504-506 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Porous polymeric sorbent ; Solid-phase extraction ; Chlorophenols
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A new porous polymer obtained by the copolymerization of two cross-linking agents: 1,4-di(methacryloyloxymethyl) naphthalene and divinylbenzene was used for the off-line preconcentration of chlorophenols from water solutions by solid-phase extraction. In order to establish its applicability to preconcentration the recoveries and breakthrough volumes of phenol, 2-; 3-chlorophenols; 2,3-; 2,4-dichlorophenols and 2,4,6-trichlorophenols were studied. The recoveries of the studied compounds were compared with those obtained for chemically bonded phases containing hexyl and octadecyl groups.
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  • 132
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    Chromatographia 25 (1988), S. 515-519 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Retention/structure relationships ; Retention prediction ; Substituted phenols
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    Topics: Chemistry and Pharmacology
    Notes: Summary A retention prediction system (RPS) for substituted phenols in reversed-phase HPLC using a ternary mobile phase was investigated. The RPS was used to predict the retention times of the phenols, and then evaluated by comparing measured and predicted retention data. Excellent agreement between both values was obtained. In addition, the retention mechanism of the phenols was investigated by means of a correlation analysis relating the parameters of the RPS to the physicochemical properties of the phenols. The three properties investigated (partition coefficient, hydrophobic substituent constant and Hammett's constant) were used to describe quantitatively the structure-retention relationship. Significant correlating equations between these descriptors and the retention data were obtained.
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  • 133
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; NAD analogs ; NAD glycohydrolase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Pig brain NAD glycohydrolase immobilized on Affi-Gel 10 or nylon 6 was used for the conversion of NAD into 3-acetylpyridine adenine dinucleotide (APAD) or 3-aminopyridine adenine dinucleotide (AAD). A reversed-phase chromatographic system consisting of a C18 Resolve column and phosphate buffer (pH 6.2)-methanol as the mobile phase was used to monitor the production of APAD and AAD.
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  • 134
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    Chromatographia 25 (1988), S. 603-608 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Ion interaction reagent HPLC ; Separation of amines
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Ion interaction reagent RP-HPLC has been employed for the separation of some typical aliphatic and aromatic amines. The effects on retention of the alkyl chain length of the eluent, and of the eluent flow-rate have been studied. The use of solutions of hexylaminium-, octylaminium-and decylaminium-salicylate has been tested and compared, employing both conductometric and spectrophotometric detection. On the basis of the results obtained, the optimal experimental conditions can be chosen for each separation. Aliphatic and aromatic mono and diamines can be separated and detected, with an average sensitivity of the order of 40 ng without any pretreatment or derivatization. The retention data obtained for amines, compared with those obtained with the same ion interaction reagents for anions, help in the interpretation of the mechanism involved in the technique.
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  • 135
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    Chromatographia 25 (1988), S. 609-612 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Gas chromatography ; Triglyceride composition of grapes ; Identification system
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The triglyceride composition of the Tempranillo grape (Vitis vinifera) has been examined by a combination of HPLC and GLC. To identify the triglycerides, equations were applied relating log k′ with the molecular variables: equivalent carbon number, chain length and number of double bonds in each of the fatty acids in the glycerides. Ten triglycerides were found, the principal ones being trilinolein (35.75%), dilinoleyl-olein (21.03%) and dilinoleyl-palmitin (17.02%).
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  • 136
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Collagenase ; Enzyme activity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The activity of bacterial collagenase Clostridiopeptidase A was estimated using a labelled synthetic peptide, 4-phenylazobenzyloxycarbonyl-L-Pro-L-Leu-Gly-L-Pro-D-Arg, as substrate. The N-protected dipeptide obtained after enzymatic hydrolysis of Leu-Gly peptide bond was quantified by reversed-phase, high-performance liquid chromatography using 4-phenylazobenzyloxycarbonyl-L-Pro-L-Phe as internal standard. The time dependence of the appearance of the hydrolysis product and the dependence of rates of hydrolysis on collagenase concentration were linear. Kinetic parameters for collagenase were determined to test the suitability of the described procedure.
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  • 137
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Racemic resolution ; Effect of temperature
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The effect of temperature on the resolution of racemates was investigated with respect to the resolution of racemic mixtures of aminoacid esters and 2,2,2-trifluoro-1-(9 anthryl) ethanol. Generally the capacity factor (k′) was found to increase with decreasing temperature. The values of the difference in the standard change of molar Gibbs energy, (ΔΔG), between the more retained and the less retained enantiomer do not appear to vary in a particular fashion. The sign of the enthalpic (ΔΔH) and the entropic, (ΔΔS) quantities for all the mixtures also depend upon the nature of the solute mixture.
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  • 138
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    Chromatographia 25 (1988), S. 791-796 
    ISSN: 1612-1112
    Keywords: Gel permeation ; Column liquid chromatography ; Column switching ; Additives in cellulose acetate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The capability of elution-elution multi-dimensional liquid chromatography was investigated. A “column scaling” approach was evaluated for the quantification of low-molecular-weight additives in cellulose acetate. A small-bore (1-mm i.d.) gel-permeation column was used to separate the higher-molecular-weight polymer from the lower-molecular-weight components. Once separated these additives were transferred to a C18 reversed-phase column via a switching valve. The reversed-phase system successfully separated and quantified individual additives. Analysis time for an ultraviolet inhibitor, Tinuvin®P, in cellulose acetate, including re-equilibration, was approximately 30 minutes. Both accuracy and precision were good. Precision over a three day period was about 1.5%.
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  • 139
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    Chromatographia 25 (1988), S. 811-814 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Separation of trimedlure isomers ; Medfly ; Ceratitis capitata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Semi-preparative high-performance liquid chromatographic procedures on 5 μm silica were developed for the isolation of gram quantities of eight trimedlure isomers (trans: A, B1, B2, and C;cis: V, W, X, and Y) for comparative biological evaluation and NMR studies. Isolations were made from an eight-component 2∶1cis: trans-trimedlure mixture, a four-componentcis-trimedlure mixture, trimedlure-B2:X and trimedlure-C:W epimerization merization mixtures, a trimedlure-B1:Y:B2 mixture, and a trimedlure-A concentrate.
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  • 140
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    Chromatographia 26 (1988), S. 133-138 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Electrochemical detection ; Flavonoids ; Metal-complexes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The electrochemical and chromatographic behaviour of flavonoid standards and of flavonoids extracted from food (green tea, black tea and onions) is investigated with respect to metalbinding properties. It is shown that metals such as iron, copper or aluminium are complexed by flavonoids, preferrably by those having an aromatic o-dihydroxy structure. This is confirmed by cyclic voltammetry on HPLC fractions (stopped flow) and by AAS measurement of metals. As the complexing sites of flavonoids are closely related to electrochemical properties, this is used for an indirect detection of metal species at low oxidation potentials. For iron species in particular a sensitive and selective detection is possible. For copper reductive detection can also be used.
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  • 141
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Electrochemical detection ; Morphine ; Polymeric column
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A simple, sensitive, accurate and reliable method for morphine determination in biological samples has been developed. It uses reverse-phase HPLC on a polymeric column with an eluent (0.05 mol/L dibasic sodium phosphate: acetonitrile, 85∶15) at pH 9.5, allowing both the suppression of ionization of morphine amine and the promotion of oxidation of the phenolic group. Amperometric detection at mild oxidizing potential (350 mV) proves very selective, and, therefore, only a simple and rapid one-step liquid-liquid sample preparation is required. Under these conditions “clean” chromatograms are obtained even with complex biological matrices such as cadaveric blood, urine and hair. Minimum detectable amount of morphine is about 200 pg injected. A calibration line with a correlation coefficient of 0.99996 has been produced over the range 7.5–250 ng/mL. Precision results comparable to other HPLC methods.
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  • 142
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Electrochemical detector ; Phenolics ; Lignocellulosic materials
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The application of HPLC with an electrochemical detector for the determination of phenolics in lignocellulosic materials is reported. The separation of phenolic acids and aldehydes (gallic acid, p-hydroxybenzoic acid, vanillic acid, p-coumaric acid, syringic acid, ferulic acid, vanillin, syringaldehyde and p-hydroxybenzaldehyde) on two different columns (reversed phase C6 and styrene-divinylbenzene PLRP-S) is shown. Chromatograms of phenolic compounds in neutral, basic and oxidative extracts of wheat straw treated with NaOH and white rot fungusStropharia rugosoannulata are reported along with quantitative results.
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  • 143
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    Chromatographia 26 (1988), S. 234-236 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Preparative-scale separations ; Teicoplanin ; Structure determination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Teicoplanin, a lipoglycopeptide antibiotic active against gram-positive bacteria, is produced as a complex mixture consisting of six major components and four chemically related minor cmmponents. Preparative HPLC was used to isolate small amounts of the pure minor components, in order to determine their structures. In the present paper the isolation procedures are presented, as well as the analytical HPLC conditions. The retention times of the minor components with respect to those of the more abundant compounds, permittedus to make hypotheses on their structures, which were then confirmed by NMR and FAB-MS investigations.
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  • 144
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Phase-transfer catalysis ; On-line derivatization ; Phenolic steroids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Commercially available equipment from two manufacturers served to set up an automated system for the precolumn phase-transfer-catalyzed dansylation of phenolic steroids, using ethynyl estradiol (EE) and estradiol (E) as model compounds. Using different mixing techniques, the on-line determination of EE and E in 200μl untreated urine samples was achieved at a rate of 3–8 analyses per hour. Detection limits were calculated to be 3–5 ng/ml. Calibration curves in urine were linear over two orders of magnitude with r=0.999 (n=5) for EE and r=0.999 (n=6) for E. The repeatability of the determination of EE in urine (1μg/ml) was 3.9% (RSD; n=20) and of E (1.5μl/ml) 3.8% (RDS; n=10). The use of plasma instead of urine in the on-line procedures was not possible due to rapid formation of emulsions, but E and EE were determined in 100μl plasma samples using a mild off-line mixing procedure in 10min. Detection limits were calculated to be ca 10ng/ml. A reaction detector, based on a solvent-segmented system, was developed for the on-line post-column dansylation of phenols and was coupled with a reversed-phase LC system. The highly selective system showed excellent linearity over at least two orders of magnitude with r=0.9999 (n=6) for both phenol and 2,5-dimethylphenol. The reproducibility was good with RSD values of around 2%. Detection limits for loop injections from standard solutions were calculated to be between 4 and 11ng.
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  • 145
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Enantiomeric purity ; Chiral HPLC ; Oxyphenonium ; Post-column extraction detection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A method is described for the determination of the enantiomeric purity (enantiomeric excess) of the anticholinergic drug oxyphenonium. The method for this quaternary ammonium compound is based on the direct HPLC analysis with a chiral stationary phase. Two kinds of α1-acid glycoprotein-bonded phases were used. For the detection a post-column extraction with fluorescence detection of the ion-pair counter ion dimethoxyantracene sulphonate was used.
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  • 146
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    Chromatographia 26 (1988), S. 297-299 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Electrochemical detection ; 2-Mercaptobenzothiazole ; River water
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A method is proposed for the determination of 2-mercaptobenzothiazole in river waters using multi-electrode electrochemical detection HPLC. 2-mercaptobenzothiazole determination is unsatisfactory by gas chromatography as it degrades readily on the column. Multielectrode electrochemical detection HPLC combines sensitivity and the ability to ‘screen out’ other electrochemically active species. The development work leading up to the proposed method is discussed. The method has a limit of detection of 0.798μgl−1 2-mercaptobenzothiazole and a total standard deviation of 2.06μgl−1 2-mercaptobenzothiazole at a concentration of 7.97μgl−1 2-mercaptobenzothiazole in river water.
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  • 147
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    Chromatographia 26 (1988), S. 369-371 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Urinary trypsin inhibitors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Human urine contains a family of trypsin inhibitors. Procedures for their purification and characterization involve laborious techniques and the conclusions are different in the identification of the separated compounds. We report results obtained by applying our RP-HPLC method to some procedures adopted by different authors.
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  • 148
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Determination of S-Carboxymethyl-L-cysteine ; Precolumn derivatization ; HPLC on-line clean-up
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary In order to follow levels of S-Carboxymethyl-L-cysteine in biological fluids for a period as long as three half-lives after drug administration during pharmacokinetic studies, an improved method for its determination had to be developed. Like the previous one, this method uses a protein precipitation step followed by an O-Phthalaldehyde derivatization step and then an HPLC on-line clean-up. This latter was obtained by means of a switching valve system, including a Nucleosil CN 5 μm (3 cm × 4.6 mm i.d.) precolumn and a Spherisorb ODS 5 μm (15 cm×4.6 mm i.d.) analytical column. The sensitivity limit was improved to 0.1 μg/ml in plasma samples and 0.2 μg/ml in urine samples. This method was applied in studies comparing single (0.75 g) and repeated (0.75 g tid) oral administration of the drug to 30 elderly patients and 20 healthy volunteers. Results showed that the half-life was 40% longer in elderly patients than in healthy volunteers, and that area under the plasma concentration versus time curve (AUC) values in elderly patients were twice those obtained with young subjects.
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  • 149
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    Chromatographia 25 (1988), S. 895-898 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Ozone ; Formaldehyde ; Annular denuder ; Air monitoring
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A denuder sampling method combined with HPLC analysis for the simultaneous determination of formaldehyde and ozone in ambient air is described. It is based on the reactions of CH2O and O3 with 2,4-dinitrophenylhydrazine (DNPH) and 4-allyl-2-methoxyphenol (eugenol)_respectively, both acting as coatings of two annular denuders connected in series. Formaldehyde released from the ozonolysis of eugenol is quantitatively collected on a third downstream DNPH-coated denuder. The two DNPH denuders are then extracted and analyzed as hydrazone derivative by HPLC with UV absorbance detection. The stoichiometric factor of the eugenol-ozone reaction was found to be 2.0±0.1 moles of O3 per mole of CH2O. The limits of detection are 0.8μgm−3 CH2O and 3μgm−3 O3 for 100l air sampled, corresponding to 1-h sampling at 1.7l min−1.
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  • 150
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Silica modifiers ; Aromatic hydrocarbons
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The retention parameters of benzene, naphthalene; anthracene and phenanthrene on untreated silica gel were determined in the presence of four modifiers: 1-methyl-2-pyrrolidone (MP), 4-formyl morpholine (FM), γ-butyrolactone (BL) and sulfolane (SF). Better column performance was found in the case of FM and MP only.
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  • 151
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    Chromatographia 25 (1988), S. 870-874 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Gel-permeation ; Pectinase activity ; Aspergillus Niger andRhizopus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A rapid liquid chromatographic (HPLC) method for the determination of pectinolytic activity of enzymes produced by Aspergillus Niger and Rhizopus species is reported. Compared with more conventional methods, HPLC is more reliable and has a much improved maximum sensitivity. The limit of quantitation of galacturonic acid is 0.1μg.
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  • 152
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    Chromatographia 25 (1988), S. 887-890 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Thiopental ; Pharmacokinetics ; Serum and plasma samples
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A high-performance liquid chromatographic method for the analysis of thiopental in 100μl of human serum or plasma is described. The procedure involves protein precipitation with acetonitrile. The supernatant is directly injected into a chromatograph containing a reversed-phase CLC-ODS (Shimadzu) column. A 50∶50 (v/v) mixture of water-acetonitrile, at a flow-rate of 1.0ml/min is used as the mobile phase. Detection is carried out ata wavelength of 280nm. Total analysis time per sample is 10min. The assay was found to be linear in the range of 0.1 to 120μg/ml. Reproducibility was good, with intra-assay coefficients of variation from 1.780 to 3.208% and inter-assay coefficients of variation from 3.241 to 4.860%. The absolute recoveries were 97.4 to 101,4%. Other drugs were tested for potential interference with the assay, but none was found.
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  • 153
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Chiral separations ; Chymotrypsin on silica ; Amino acids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A new HPLC stationary phase was synthesized by the covalent immobilization of α-chymotrypsin on silica. This increased the stability of the enzyme, without decreasing its activity. The initial chromatographic studies show that this phase can be used for chiral separations of enantiomeric solutes. The stereochemical resolutions of amino acids and amino acid derivatives are reported.
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  • 154
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    Chromatographia 25 (1988), S. 1034-1038 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Determination of propafenone ; 5-OH propafenone ; Reversed-phase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary This paper presents a method for the simultaneous detection of propafenone and 5-hydroxypropafenone (5-OH propafenone) using HPLC. The method is sensitive, selective, and reproducible. The chromatographic separation is based on a 25×0.4 cm 5 μm ODS column, a mobil phase of 0.1 M potassium phosphate (K2HPO4) buffer (pH 2.5) and acetonitrile (63∶37), and UV absorbance detection. There is excellent intra- and interassay variation. Retention times of the internal standard, propafenone, and 5-OH propafenone are 4.3, 6.0 and 2.9 minutes respectively. This method shows linearity over the therapeutic range for propafenone and 5-OH propafenone (0.15 to 3.0 μg/ml and 0.075 to 1.5 μg/ml respectively). No interference has been found from other commonly administered drugs, including several antiarrhythmic drugs, at therapeutic levels.
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  • 155
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Soyabean leaf extracts ; Kaempferol glycosides ; Kaempferol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary An HPLC method is described for the determination of kaempferol glycosides in soybean leaf extracts. The method is rapid and can provide qualitative and relative quantitative results for 9 kaempferol glycosides. The flavonol glycosides are extracted from soybean leaves by shaking the samples in 50% methanol; the extracts are filtered, evaporated to dryness and reconstituted in methanol before further purification through a C-18 Sep-Pak column. The samples are injected onto a C-18 HPLC column, separated by gradient elution with a 1% phosphoric acid: methanol mixture and detected on a UV/VIS diode-array detector. Flavonols were monitored at 265 and 348 nm and spectra from 180 to 400 nm were stored and used as an aid in identification. Relative quantities of the kaempferol glycosides among soybean cultivars were calculated from their proportion of peak area in the chromatograms. Total kaempferol concentration of the extracts was calculated after acid hydrolysis of the kaempferol glycosides to the aglycone and comparison of peak areas to kaempferol standards.
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  • 156
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Gel permeation ; Human serum IgG ; Fluorescent immune complex ; Fluoresence detection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A high-performance liquid gel-permeation chromatographic method is described for the determination of human serum immunoglobulin G (IgG) by separating the fluorescent immuno complex from the free fluorescence-labeled antibody. Fluorescence-labeled antibody used in this study was fluorescein isothiocyanate (FITC)-labeled Fab fragment goat anti-human IgG (anti-IgG Fab). Immuno complexes and antibody of different molecular sizes can be separated. FITC-labeled anti-IgG Fab was added to the serum and the mixture is passed through the column. An immuno complex separates as well-delineated peak in the column void volume, and was measured by the fluorescence of the column eluate (Ex=490nm, Em=520nm). The total analysis time for a serum sample was approximately 15min. The minimum detection limit was 25 mg/dl. The relative standard deviation was below 2% (peak area). The results of the HPL-GPC analysis correlate well with those obtained by laser nephelometric assay (r=0.992).
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  • 157
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    Chromatographia 25 (1988), S. 1067-1071 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Retention behaviour of 1,n-halo(alkylthio)alkanes ; Effect of the halogen atom ; Effect of the number of methylene groups
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The RPLC retention of 1,n-halo(alkylthio)alkanes on an ODS phase with methanol/water (95%/5%) has been investigated. Replacement of a methyl group in a dialkyl sulphide R-S-R by a halogen atom X, to yield the title compounds X-(CH2)n-S-R, generally results in a retention decrease. This retention loss results either from the halogen atom proper, or from its immediate vicinity, or from a reduced retention contribution of the alkyl units. The methylene groups (n) between halogen and sulphur produce somewhat greater retention than the methylene groups in the alkyl groups R.
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  • 158
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Aliphatic amines ; Separation factors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary An analytical method is presented for the identification and separation of aliphatic monoamines, diamines and polyamines. Dansyl chloride as the derivatizing agent and fluorimetric detection were employed. The behaviour of different column packings and eluent compositions was tested and compared. The interaction mechanisms of the dansylated compounds with stationary and mobile phases are discussed on the basis of the analysis of the capacity factors evaluated in the homologous series of monoamines CH3(CH2)nNH2, with n varying up to 9 and diamines NH2(CH2)nNH2, with n varying up to 12. An application of this method to a real sample is presented.
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  • 159
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Detection, chiroptical, polarimetric, circular dichroic ; Enantiomers ; Sorbents, optically active
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Polarimetric and/or circular dichroic detection of enantiomers after liquid chromatography on optically active sorberts has been used for the following purposes: determination of enantiomeric purity in spite of peak overlap, investigation of enantiomerization during chromatography, monitoring of racemizations and recording of circular dichrograms without preparative enrichment of enantiomers. Novel examples for all applications are give. The general prospects for chiroptical detection are discussed.
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  • 160
    ISSN: 1612-1112
    Keywords: Size-exclusion chromatography ; Column liquid chromatography ; Peptides ; Hydrolysates
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Gel filtration (size-exclusion) and high-performance liquid chromatography have been used to separate peptic peptides from haemoglobin hydrolysate. Elution profiles on Sephadex G-25 displayed nine fractions with molecular weights lower than 6500 daltons. Each fraction was analysed for total amino acid content and showed less than 1% free amino acids. Reversed phase HPLC, using ammonium acetate buffer and acetonitrile as solvent, was applied to each fraction in order to obtain pure peptide peaks. The importance of acquiring a better knowledge of such an hydrolysate is discussed. Various potential applications of this type of hydrolysate, some of them already being undertaken, are envisaged.
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  • 161
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Charge transfer mechanisms ; Aromatic esters ; Structure/retention relationships
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    Topics: Chemistry and Pharmacology
    Notes: Summary The separation of aromatic esters has been studied on acceptor bonded phases. It is likely that donor-acceptor complex (DAC) formation occurs between the acceptor phase and the electrons of the carbonyl groups. The influence of substituents attached to the aromatic rings and the role of the alkyl chains of phthalates on retention are discussed.
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  • 162
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Gradient elution ; Prediction of retention ; Aldehydes and amines
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    Topics: Chemistry and Pharmacology
    Notes: Summary A general chromatographic model has been set up starting from a set of equations based on the concept of the velocity of a solute along the column. The composition of the mobile phase is taken into account solely as a numerical factor entering into suitable equations and totally independent of the chemical-properties of the constituents. A few isocratic experimental runs are necessary as input data, and subsequently a small amount of computational effort is sufficient to make predictions of retention times under gradient elution conditions for solutes of whatever chemical structure. The prediction errors are dependent on the steepness of the linear gradient chosen but are, in any case, acceptably low.
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  • 163
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    Chromatographia 25 (1988), S. 437-442 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Large scale separations ; Optimisation procedures
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Scale up of analytical High-Performance Liquid Chromatography (HPLC) to the process scale offers promise as a preferred separation method for many high value fine chemicals, both for development purposes and continuous production. It's early adoption is curtailed by the widely held perception that preparative HPLC is a low efficiency high cost technique of limited application. This view has largely beeen generated by the generalised use of large particle size, low quality angular silica supports, and difficulties with packing methodologies. A systematic study of a new silica, in new axially compressed equipment using Plackett-Burman statistics, has demonstrated that very high efficiences, leading to high capacities, can be attained.
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  • 164
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Racemic resolution ; Aminoacid esters ; Mobile phase composition
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The work in this paper demonstrates the effect of the mobile phase composition on interactions involving racemic esters of aminoacids and a chiral stationary phase derived from N-formylisoleucine. Aprotic solvents have been employed to test the resolution of these racemates. An increase in the alcohol concentration of the mobile phase results in the deterioration of selectivity. These solvents seem to favour high separation factors. The solvent 1,1,2-trichlorotri fluoroethane is a suitable mobile phase modifier for normal phase liquid chromatographic resolution of enantiomers of aminoacid esters.
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  • 165
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    Chromatographia 25 (1988), S. 697-700 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Micro columns ; Fluorometric detection ; Packed flow cell
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    Topics: Chemistry and Pharmacology
    Notes: Summary A packed flow cell was used for fluorometric detection in micro high-performance liquid chromatography. The flow cell consisted of fused-silica tubing packed with the same material as the separation column. A focusing effect of the stationary phase on the signal intensity was observed, leading to an improvement of the mass detection limit, as achieved by on-column detection.
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  • 166
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    Chromatographia 25 (1988), S. 815-816 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; On-column detection ; Fused silica microcolumn ; Fibre optics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Modification of the UV absorbance detector for LC is described. It is based on a simple combination of a commercial photometer with optical fibres to conduct the light. The performance of the detector is illustrated by the analysis of polyaromatic hydrocarbons on a packed fused-silica column. The minimum detectable concentration for naphthalene was measured as 6·10−6M.
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  • 167
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Inductive effects ; Resonance effects ; Methylquinoline derivatives
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    Topics: Chemistry and Pharmacology
    Notes: Summary Hydrophobicity of methylquinolines was compared with their retention data measured on two stationary phases by reversed-phase HPLC, and with the experimental partition coefficients (log P). The influence of the substituent position on the hydrophobicity is discussed and a two-parameter correlation equation is proposed permitting the prediction of the log k′ values with help of the inductive and resonance effects.
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  • 168
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Ion-pair chromatography ; Valve switching ; Fluorouracil
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Fluorouracil has been analyzed with a reversed-phase ion-pair liquid chromatographic system applying PRP-1 as the stationary phase. A valve switching technique was applied for the separation of the analyte and some other chemotherapeutic agents. The described procedure gives a rapid screening for the analyte in urine and plasma samples without interferences from late eluting solutes. Biological samples are pretreated by means of a liquid-liquid extraction resulting in a recovery of about 52% for plasma and urine with minimum detectable amounts of about 5ng/ml. The sensitivity and the selectivity of the procedure allows the monitoring of fluorouracil in body fluids and subsequent pharmaco-kinetic studies of the fluoropyrimidine.
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  • 169
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    Chromatographia 26 (1988), S. 120-124 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Chlorophylls ; Pheophytins ; Fluorimetric detection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A HPLC method for the separation and estimation of chlorophylls and pheophytins in fresh and frozen peas was assessed with the aim of following the colour changes of the products during storage under frozen conditions. The method involves the acetone extraction of the pigments, followed by HPLC on RP18 with isocratic elution by acetone: ethanol:water, (70:17:13), detecting and estimating the separated compounds by fluorimetry. The mobile phase composition was selected by monitoring the separation efficiency of several mixtures of solvents by HPLC. The reliability and the accuracy of the method were checked.
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  • 170
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Modified polystyrene-divinylbenzene
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    Topics: Chemistry and Pharmacology
    Notes: Summary One possible method of characterizing chemicall-modified stationary phases is to describe their chromatographic properties. In this study we investigated several chemically-modifiedpolystyrene-divinylbenzene packings for the separation of nitroarenes, polycyclic aromatic hydrocarbons and aminosubstituted aromatic hydrocarbons. Chromatography was carried out on commercially available polymer stationary phases, for example C-18 or vinylpyridine modified polystyrene. In addition, a chemically-immobilized polymer packing was prepared by introducing nitrogroups, which were established by elemental analysis and infrared spectroscopy.
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  • 171
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Polyamine and nucleotide metabolism ; Ornithine-utilizing key-enzymes ; Enzyme-targeted chemotherapy ; DL-α-difluoromethylornithine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Ion-exchange and reversed-phase high performance liquid chromatographic methods have been developed and applied to the analysis of metabolites related to the biosynthesis of natural polyamines some amino acids and nucleotides in P388 leukemia cells. Key-enzyme activities concerning the ornithine utilization were measured by quantitating dansylated putrescine and citrulline. Thus the efficacy of enzyme-targeted chemotherapy using DL-α-difluoromethylornithine as inhibitor of ornithine-utilizing enzymes could be monitored. A marked increase of uridine-5′ and cytidine-5′ phosphates induced by DL-α-difluoromethylornithine treatment was demonstrated in P388 leukemia cells using ion-pair and ion-exchange liquid chromatographic methods.
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  • 172
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Analgesic peptides ; Opioids ; Glycopeptides ; Glycosylenkephalins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary N 1.5-Glycosyl enkephalins are potent analgesics. Two enkephalin-related glycosyl peptides: [DMet2, Pro5] enkephalin (N 1.5 β-glucopyranosyl] amide and [DMet2, Pro5] enkephalin [N 1.5 β-galactosyl] amide have been prepared following the liquid phase procedure. The chromatographic behavior of both the final glycosyl enkephalins and their synthetic intermediates on reversed-phase columns has been tested. Semipreparative, reversed-phase, HPLC conditions have been developed in order to obtain a simple and rapid procedure for the purification of the two synthetic pentapeptides. Thus, using isocratic elution, injections of 10mg of crude material after gel filtration gave homogeneous (99.9%) glycosyl enkephalins.
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  • 173
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Acrylic acid ; Polyacrylic gelling agents ; Superabsorbent polymers
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    Topics: Chemistry and Pharmacology
    Notes: Summary High-performance liquid chromatographic methods (reversed-phase and ion exclusion) for the determination of monomeric acrylic acid in polyacrylic gelling agents are described. Acrylic acid is extracted by two different methods. The methods were applied to samples containing acrylic acid from 1–1000 mg/kg.
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  • 174
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    Chromatographia 25 (1988), S. 919-922 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Nifedipine ; Normal phase ; Plasma ; Liquid-liquid extraction
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    Topics: Chemistry and Pharmacology
    Notes: Summary For the determination of nifedipine in plasma a sensitive and selective method is required. The use of ‘on-line’ pre-column enrichment, followed by ‘reversed phase’ separation and UV-detection at 350 nm proved to be too susceptible. Therefore, detection was carried out after a post-column ‘on-line’ reduction and photoreaction step using a fluorescence detector. Although this method proved to be extremely sensitive (limit of detection 0.1 ng/ml plasma) and selective, a number of problems cropped up caused by the reduction agent which finally prevented the procedure being used in routine analysis. As a consequence, the following method was developed. After liquid-liquid extraction of nifedipine a 1/3 of the extract was chromatographed on a normal phase (diOH) system and detected at 235 nm, because detection at 350 nm was not sensitive enough. This method has a limit of detection of 1 ng nifedipine/ml plasma and the calibration curve is linear up to 320 ng/ml. The recovery lies around 82% and the standard deviation for the range 6–320 ng/ml is less than 5%. So far about 2000 plasma samples have been analysed by this method.
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  • 175
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    Chromatographia 25 (1988), S. 957-960 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Electrochemical Detection ; Ethinyloestradiol
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    Topics: Chemistry and Pharmacology
    Notes: Summary The analysis of ethinyloestradiol in tablets has been carried out by reversed phase high performance liquid chromatography (HPLC) using electrochemical detection. The drug was eluted with aqueous acetonitrile containing sodium acetate as background electrolyte. The mobile phase was used as extracting solvent, and 4-tertpentylphenol was used as internal standard.
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  • 176
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    Chromatographia 25 (1988), S. 969-973 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Carboxylic acid phase ; Addition reactions ; Synthesis of new bonded phases
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    Topics: Chemistry and Pharmacology
    Notes: Summary The allyl bonded phase is used as an intermediate in the synthesis of a weak cation exchange material. The double bond is first converted to an aldehyde by an ozonation reaction and then subsequent oxidation to the carboxylate group is accomplished with dichromate. Diffuse reflectance Fourier transform infrared spectroscopy (DRIFT) is used to monitor the conversion process. The presence of the aldehyde functional group is confirmed by reaction with 2,4-dinitrophenylhydrazine. ESCA spectra reveal no residual chromium on the silica surface. The product is tested chromatographically by the retention of alkylarylketones, the ion-exchange behavior of both monovalent and divalent cations, the retention of nucleosides and bases, and the separation of a mixture of hemoglobins.
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  • 177
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; UV detection ; Solubility ; Polystyrene ; Sample precipitation ; Gradient elution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Gradient elution of polystyrene standards on reversed phase C18 columns by methanol/tetrahydrofuran or methanol/dichloromethane mixtures yielded a strange effect of the molecular weight of the sample on the specific peak are (mAUs per μg sample injected). The effect did not occur when pure dichloromethane was used as an eluent. Further to this, dependence of specific peak area on flow rate was observed in gradient elution with methanol/tetrahydrofuran mixtures. It was found that these effects were due to polymer elution at the verge of precipitation. Depending on the dwell time of the sample in the column, opalescence occurred that added to the UV signal used for monitoring the elution.
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  • 178
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Reversed-phase ; Ion-pair ; Red blood cells ; Tumour cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The determination of nucleotides, nucleosides, and nucleobases was carried out in cells of different metabolic complexity: in mature and immature red blood cells, in Ehrlich ascites tumour cells from different proliferation stages, and in other tumour cells. The maturation of reticulocytes to erythrocytes is accompanied by loss of organelles and energy-requiring processes as well as the switch from aerobic to anaerobic ATP production. The profile of purine nucleotides, nucleosides, bases, and pyridine dinucleotides, by reversed-phae HPLC, shows large concentration changes during the maturation of red blood cells. The concentrations of purine mono and triphosphates are two to four times greater in reticulocytes in comparison with erythrocytes; the difference in the concentrations of nucleosides and nucleobases between reticulocytes and erythrocytes is even greater. Application of ion-pair HPLC showed that the Ehrlich ascites cells loose major portions of purine mono-, diand triphosphates between the 7th and 11th day after inoculation. Fast growing solid sarcoma tumours of rats (MV 202 Ner) contain higher amounts of nucleotides than slowly growing tumours of identical cell type.
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  • 179
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Micro enzyme-immobilized columns ; Separation of aldehydes ; Derivatization and fluorometric detection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Aldehydes in liquors were analyzed by micro high-performance liquid chromatography with post-column derivatization on the enzyme-immobilized glass beads. β-nicotinamide adenine dinucleotide (NAD) was converted to its reduced form (NADH) on the enzyme-immobilized column in the presence of an aldehyde and a thiol, followed by fluorometric detection. A detection limit of 2.5 ppm (50 pg) was achieved for acetaldehyde.
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  • 180
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    Chromatographia 25 (1988), S. 526-530 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Doxycycline and Demeclocycline ; Plasma and urine samples ; Solid phase extraction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A method for the selective quantification of doxycycline in human plasma and urine has been developed using HPLC with UV-detection at 350 nm. Analyte and internal standard were extracted from plasma by extraction columns filled with octadecylsilica. Urines were only mixed with diluted acid prio to injection. The influence of pH on peak shapes is discussed as well as comparative investigations on selectivity and peak symmetry on commerical octadecylsilica. The method was successfully applied to the samples of a clinical study with an oral single dose of 100 mg. Precision and accuracydata of the assay and calculated pharmacokinetic parameters are presented.
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  • 181
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Microcolumn ion chromatography ; Indirect photometric detection ; Monovalent cations
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    Topics: Chemistry and Pharmacology
    Notes: Summary Monovalent cations were indirectly detected via postcolumn suppressor ion replacement in microcolumn ion chromatography. The ion-repalcement column loaded by chromophoric ions was connected to the suppressor column. The eluent, nitric acid, was converted to water through the suppressor and anion-replacement columns, while the analyte cations were coeluted with the chromophoric anions. The analyte cations were indirectly detected by measuring UV absorptions of the chromophoric anions.
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  • 182
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    Chromatographia 25 (1988), S. 593-597 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Solid phase extraction ; Amiodarone in serum ; Addition of ammonia to eluent
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary An improved liquid-solid extraction procedure for the purification and determination of amiodarone and desethylamiodarone in biological fluids is proposed. The sample is passed through a silica-C18 column, and after washing, the analytes are eluted with methanol. The determination is then accomplished by RP HPLC using an octadecyl silica column and a mobile phase of methanol containing 0.0015% of ammonium hydroxide. The effect of ammonia concentration on the capacity factors of the analytes has been used for estimating the acid dissociation constants of the investigated secondary and tertiary amines in methanol.
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  • 183
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Indirect photometric detection ; Signal intensity evaluation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary We have experimentally evaluated the equation representing the peak area of induced peaks in indirect photometric detection of nonelectrolytes as a function of the amount and concentration of the analyte and the visualizing agent and their capacity factor, the phase ratio of the column and the absorptivity and the pathlength of detection. Aliphatic alcohols and hydrocarbons were used as the test analytes. Most of the analytes behaved according to the relationship expressed by the equation.
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  • 184
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    Chromatographia 25 (1988), S. 685-690 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Adsorption on solid support ; Polar solutes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The properties of chloroform as a liquid stationary phase in reversed-phase liquid chromatography have been investigated. Stable systems and complete pore filling of the support were obtained. Uncharged acids were slightly adsorbed by the support. Amines in the cationic form showed tailing, which can be attributed to adsorption on the support material. The influence of several reversed-phase supports on retention and peak symmetry has been studied. Little difference was found for uncharged acids, but retention and peak shape of the cations was dependent on the specific support. Also on the poly(styrene-divinylbenzene) copolymer PRP-1 cations gave tailing peaks. The results indicate that the surface of reversed-phase supports is heterogeneous. From this and other studies it can be concluded that only certain reversed-phase silicas, such as μ-Bondapak, are suitable supports for cationic solutes. In liquid-liquid chromatography polar solutes only obey a liquid-liquid distribution model if the liquid stationary phase molecules contains a hydrophobic group, as well as a polar function.
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  • 185
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Ligand exchange ; On-Line preconcentration ; Opiate pentapeptide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary In order to study the occurrence of the opiate pentapeptide β-[D-Ala2,4, Tyr5] CM-5-NH2 in the 10−9–10−5 M concentration range, we have developed a new method consisting of extraction of the peptide from its buffered aqueous medium onto a reversed phase cartridge of 50 μL void volume, followed by on-line injection onto a Cu(II) modified silica gel column. The acetonitrile-rich mobile phase allows detection of the natural fluorescence of the peptide at the picomole level.
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  • 186
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Expert systems ; Optimization of operating parameters
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Given the mobile and the stationary phase and values for the physical parameters such as temperature and pH, a separation can be optimized by varying the so-calledchromatographic parameters. These include the column dimensions, particle size, operating conditions (e.g. flow rate, attenuation) and instrumentation (e.g. detector cell, time constant). Optimization of the chromatographic parameters implies finding the best possible set of values, which we define as yielding (i) sufficient separation and (ii) sufficient sensitivity in (iii) the shortest possible time. Finding the best possible conditions (the global optimum) is very difficult for chromatographers in practice. An expert system is described that allows chromatographic optimization to be performed for isocratic separations. An initial chromatogram is required to consult the system. In return, the system provides a complete set of chromatographic parameters, which represents the global optimum within the limits set by the required resolution and signal-to-noise ratio specified by the user. The tolerated flow and pressure ranges, the volume of the available detector cells and the time constant of the detection system are constraints during the optimization. A separate module of the system concerns the sample preparation for pharmaceutical formulations in solid dosages and aqueous solutions. Prototype expert systems have been successfully implemented in the expert-system shell Knowledge Craft on a MicroVAX workstation.
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  • 187
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Expert systems ; Selectivity optimization ; Optimization criteria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Systematic procedures for the optimization of chromatographic selectivity require objective criteria to characterize the quality of separation in a chromatogram. Numerous criteria have been suggested. Different criteria yield different results and the choice will depend on a large number of factors. It is genuinely difficult to select the most suitable criterion in a particular situation. For these reasons, an expert system has been developed to assist chromatographers in the selection of optimization criteria. A structured representation of the required knowledge and its implementation in an expert-system shell are presented in this paper.
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  • 188
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    Chromatographia 26 (1988), S. 300-304 
    ISSN: 1612-1112
    Keywords: Gas chromatography ; Column liquid chromatography ; Polycyclic aromatic hydrocarbons ; Phenolic compounds ; Air pollution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary An improved procedure for the determination of the less volatile air pollutants of recycled closed atmospheres by activated carbon-Soxhlet extraction is proposed. After total treatment of desorbate, PAH determination is then accomplished by RP-HPLC using both adsorptiometric and fluorimetric UV detection; phenolic compounds are determined by GC-MS. This technique is suitable for very low levels of PAH and phenolic compounds.
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  • 189
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Isotachophoresis ; Sample pretreatment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary This study focusses attention on the possibilities of preparative isotachophoresis (ITP) as a sample pretreatment technique prior to liquid chromatographic (HPLC) analysis. The increased demand for accurate and less time consuming analysis necessitates that sample pretreatment procedures, should be develop in parallel with other improvements (e. g. in detection and separation) which can be observed. The preparation isotachophoresis was performed on gel slabs and the zones of interest were subsequently cut out, desorbed and the desorbates analyzed by HPLC. In this study satisfactory recoveries of between 85–90% with a standard deviation of 1–5% were observed for blank experiments. For spiked serum and urine samples the recoveries in general decreased with decreasing spiked drug concentrations. These observations are discussed in this paper.
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  • 190
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Electrochemical detection ; β-Naphthoquinone sulfonate ; Labelling reagent ; Direct labelling in body fluids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A new sensitive, high-performance liquid chromatography (HPLC) method using electrochemical detection (ECD) is reported for the determination of 21 amphetamine-like compounds such as amphetamine, methamphetamine, methoxyphenamine, phentermine, ephedrine, mescaline, STP and methylenedioxyamphetamine, in biological fluids. Primary and secondary amines in biological specimens were easily labelled with β-naphtho-quinone-4-sulfonate (BNQS) at 60°C for 30 min. Hydrodynamic voltamograms of BNQ-derivatives showed a maximum response at an applied potential of around zero V (against Ag/AgCl) in reducing model. BNQ derivatives of amines were analyzed by HPLC using a reducing ECD at zero V (Ag/AgCl). Separation was on a μ Bondasphere C18 with mobile phase: acetonitrile-methanol-0.01 M H2SO4. Good separations of twenty one amphetamine-analogs were obtained. Only 50 μl of biological fluid containing those drugs at a concentration of over 1 μg ml−1 were needed direct for determination with good accuracy. When extraction was carried out prior to the labelling reaction, the detection limits were much improved to less than 10 ng ml−1. This method was successfully applied to determination of methamphetamine in hair.
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    Chromatographia 26 (1988), S. 13-18 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Hydrophobic interaction chromatography ; Proteins ; Salt effects ; Effect of MgCl2
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary It has been noted in the literature that certain salts enter into specific interaction with proteins. As a result of this, they may act as salting-in agents. We have investigated the effect of magnesium chloride which is known to possess such unusual properties on the retention of proteins in hydrophobic-interaction chromatography. First the retention behaviour of amino acids and small peptides having a wide polarity range was studied on reversed-phase columns using eluents containing (NH4)2SO4, MgSO4 or MgCl2, in wide the concentration ranges. For less polar eluites plots of the logarithmic retention factors against the salt concentration were found to be linear, whereas the more polar species showed irregular behavior. The retention of a wide range of proteins was measured on a TSK Phenyl-5-PW column using eluents containing (NH4)2SO4, MgSO4 or MgCl2 at different concentrations. The salt-mediated retention was regular with (NH4)2 SO4 and MgSO4 although MgSO4 showed a lesser effect than that predicted by the surface-tension increment. The effect of MgCl2 was quite irregular: the retention factors either increased or decreased or remained unchanged depending on the protein. These results corroborate earlier observations regarding the particular effect of MgCl2 and suggest the modulation of selectivity in hyrophobic-interaction chromatography by the addition of MgCl2 to the eluent.
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    Chromatographia 26 (1988), S. 417-422 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Oligomeric series ; Separation selectivity ; Oligoethylene glycols ; Ethoxylated nonylphenols
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The simultaneous dependence of the retention in oligomeric series on the number of repeat structural units and on the mobile phase composition may be described by very similar equations for reversed-phase and for normal-phase systems. In reversed-phase systems, the separation selectivity of the individual oligomers is determined mainly by the size and by the polarity of the repeat structural unit, but the influence of a bulky and polar structural residue may also become important so that even reversed order of elution may be observed for oligomeric series with the same oligomeric units but significantly different end groups. For example, oligoethylene glycols are eluted in the order of increasing size of the oligomers, whereas ethoxylated nonylphenols are eluted in the order of decreasing size. In normal-phase systems, the separation selectivity in oligomeric series depends on the adsorption energy and on the adsorbed area of the oligomeric unit. If the oligomeric unit is small, the concentration of the polar solvent in the binary organic mobile phase has only a minor effect on retention and selectivity, which may be controlled by taking account of the nature of the adsorbent and of the polar solvent or by varying the proportion of two polar solvents in a ternary mobile phase.
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  • 193
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    Chromatographia 26 (1988), S. 19-28 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Chiral LC and SFC phases ; Polymer coating ; Chemical bonding ; Polysiloxane pre-polymers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Chirally substituted Si−H-containing polysiloxanes were synthesized, which can be immobilized on small particle silica gel as well as on the smooth surfaces of fused silica capillaries. Immobilization is achieved either by crosslinking or by chemical bonding to the surfaces via silanol groups; both reactions can only be performed by addition of H2PtCl6, which acts as catalyst for hydrosilylation and as stoichiometric reagent for crosslinking. Chiral substituents of systematically varied chemical structure were introduced into the polysiloxanes by hydrosilylation. The mechanism of immobilization was investigated by spectroscopic methods, notably29Si-NMR. Homogenous stationary-phase coatings of variable film thickness and corresponding retentivity can easily be achieved. The enantioselectivity of the phase systems was characterized in dependency on the chemical structure of the chiral selectors attached to the polysiloxane chain of the chiral stationary phases and also in terms of the functional groups introduced into the solutes by derivatisation.
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  • 194
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    Chromatographia 23 (1987), S. 796-802 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Steroids ; Rat tissue ; Purification by extraction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A high-performance liquid chromatographic method for analysis of steroids in the tissue of laboratory animals is described. A two-stage solid extraction procedure, utilizing Sep-pak C18 and graphitized carbon black, gives a purified product. Steroids in this extract were separated by multistep gradient elution with a reversedphase liquid chromatography system, using acetonitrilewater as solvent. Several columns with different geometry and phase loading were investigated. Optimum resolution was obtained with a medium covered and spherical particle. The applicability of this method was successfully demonstrated in the analysis of the testis and adrenal gland of rat.
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  • 195
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    Chromatographia 23 (1987), S. 814-817 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Thin layer chromatography ; Ammonium tungstophosphate ; Aminoacids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The chromatographic behaviour of aminoacids on columns of silica coated with different percentages of ammonium tungstophosphate at several buffer phosphate concentrations is reported. The stationary phase shows a high selectivity towards aminobutyric acids and methyl derivatives of tryptophan due to the distance between the carboxylic group and the −NH 3 + group involved in the cation-exchange process and/or to the steric hindrance by a methyl substituent. A relationship between HPLC and HPTLC data has been verified and applied to the prediction of the separation of six aminoacids.
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  • 196
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    Chromatographia 23 (1987), S. 934-938 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Calibration ; Confidence interval ; Limit of determination ; Limit of calibration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary In HPLC calibration the expressions lowest calibration limit and determination limit are defined in statistical terms. The lowest calibration limit is the minimum mass in the measured series of calibration points. It is calculated from the confidence interval of the inverse of the calibration function as the lowest mass limit that may be differentiated from zero mass with a preset probability of error. If the calculated lowest calibration limit is lower than the actual data, points at lower concentration may be measured. The determination limit is the smallest concentration of an analysis that is differentiated from the concentration zero or an apparent blind value in the calibration curve with a given probability of error. Using two different UV-detectors (variable wavelength and photodiode-array) the lowest calibration limit is experimentally evaluated and compared with specific data for the detectors.
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  • 197
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Pre-column derivatization ; Ultraviolet detector sensitivity ; Dihydroartemisinin ; Diacetyldihydrofluorescein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The use of diacetyldihydrofluorescein (DADF) for derivatization of dihydroartemisinin (dihydroqinghaosu, DHQHS) is proposed. The reaction between DHQHS and this reagent in the presence of 4-dimethylaminopyridine (DMAP) and N,N′-dicyclohexylcarbodiimide (DCC) was complete in 8 hours at room temperature giving about 80 per cent theoretical yield. The derivative showed intense UV absorption, thus providing a sensitivity of 0.1 nanogram by UV detection after column separation. The influences of the ratio of the reagents, reaction temperature, chromatographic conditions and the extent of detection linearity were investigated. The reaction gave consistent results and chromatographic separation was not affected by an excess of the reagent or side products.
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  • 198
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    Chromatographia 23 (1987), S. 395-400 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Photokinetic data ; Reaction mechanism ; (E)-1-phenylpropene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A combined method of high-performance liquid chromatography and UV-absorption spectroscopy is described for use in dynamic systems. It provides a way of obtaining detailed mechanistic information as well as quantitative data (rate constants, quantum yields), even though neither mechanism nor photoproducts are known. The kinetic treatment of the photoreaction of (E)-1-phenylpropene was successful, showing a photo-isomerization and a parallel step. Information on this additional step is obtained by using HPLC to observe the changes in the concentrations of the reactants' selectivity. The necessity of process control is demonstrated.
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  • 199
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    Chromatographia 23 (1987), S. 419-422 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Fatty acid esters ; Particulate material in seawater
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A method for the determination of fatty acids of the molecular weight range of propionic to erucic acid is reported. The acids are converted to either p-bromophenacyl-or p-phenylphenacylesters and the esters are separated by HPLC and detected by UV-absorption. Twentytwo esters, including saturated, monoolefinic and polyunsaturated ones were separated within one HPLC-run. The detection limit is about 5 pmol per fatty acid. The application of the method to particulate material is described.
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  • 200
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Anion-exchange stationary phases ; Polymer coating ; Cross-linking
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary PHEB=POLY(2-hydroxy, 3N-ethylenediamino)butadiene has been synthesized. This unsaturated pre-polymer can be immobilized on different, preferably inorganic supports of suitable porosity such as silica or alumina by cross-linking. Weak anion-exchange phase are obtained. Such phases do not have the disadvantages of organic polymeric phases, regarding pressure stability and swelling by solvent influence. The ion-exchange capacity and the retentivity of IE-separations can deliberately be varied via the thickness of the cross-linked polymer layer. Alumina can also be coated with PHEB applying the same cross-linking procedure. Such phases exhibit an exceptional chemical stability even when operated with mobile phases at very high pH-values. Separations of excellent selectivity are achieved for inorganic anions, as well as with mixtures of aliphatic and aromatic free acids also including hydroxy- (mono- or poly)carboxylic acids.
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