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1.
Chromatographic behavior of epirubicin and its analogues on high-purity silica in hydrophilic interaction chromatography 总被引:1,自引:0,他引:1
Hydrophilic interaction chromatography has been applied for the separation of epirubicin and its analogues using high-purity silica column with aqueous-organic mobile phase. Parameters affecting the chromatographic behavior of the solutes such as organic modifier, buffer pH, ionic strength and sample size, have been investigated. Of utmost importance for successful separation of these analogues is the choice of organic modifier, since it impacts both the solvent selectivity and the ionization of silica silanols as well as buffer solution, and consequently the retention behavior of solutes. Acetonitrile was shown to offer superior separation of these analogues to methanol, isopropanol or tetrahydrofuran. Results of the effects of organic modifier, buffer pH and ion strength indicate that the retention mechanism is a mixed-mode of adsorption and ion exchange. In addition, an irreversible adsorption of these compounds was found on silica in the weakly acidic or neutral mobile phases, and the effect of various factors on irreversible adsorption was also preliminarily discussed. More significantly, these basic compounds have exhibited peaks with a slanted front and a sharp tail, a typical overloading peak profile belonging to the behavior of competitive anti-Langmuir isotherm by increasing the sample size at the experimental conditions. 相似文献
2.
In analysis of the temperature effect on chromatographic separations the influence of the adsorption of organic solvent on the retention properties of solute is generally not taken into account. In fact, adsorption behavior of solutes is strongly affected by competitive adsorption of organic solvents, which is temperature dependent. In this work changes of adsorption equilibrium of an organic solvent as well as a solute with temperature have been analyzed. Data of the excess adsorption of methanol from aqueous solutions on octadecyl-bonded silica have been acquired at different temperature. Experiments have been performed over a relatively narrow temperature range corresponding to typical chromatographic conditions, i.e., 10-50 degrees C. The competitive adsorption equilibria of model solutes (i.e., two homologous compounds: cyclopentanone and cyclohexanone) have been measured at different temperature and composition of the mobile phase. Temperature alterations to the retention properties were found to result from combined effects of changes in adsorption behavior of the organic solvent and of the solute. The influence of temperature on the separation selectivity has been considered. 相似文献
3.
Modelling retention in liquid chromatography as a function of solvent composition and pH of the mobile phase 总被引:2,自引:0,他引:2
The aim of this work was to develop a model that accurately describes retention in liquid chromatography (LC) as a function of pH and solvent composition throughout a large parameter space. The variation of retention as a function of the solvent composition, keeping other factors constants, has been extensively studied. The linear relationship established between retention factors of solutes and the polarity parameter of the mobile phase, E(N)T, has proved to predict accurately retention in LC as a function of the organic solvent content. Moreover, correlation between retention and the mobile phase pH, measured in the hydroorganic mixture, can be established allowing prediction of the chromatographic behavior as a function of the eluent pH. The combination of these relationships could be useful for modelling retention in LC as a function of solvent composition and pH. For that purpose, the retention behavior on an octadecyl silica column of a group of diuretic compounds covering a wide range of physico-chemical properties were studied using acetonitrile as organic modifier. The suggested model accurately describes retention of ionizable solutes as concomitant effects of variables included and is applicable to all solutes studied. We also aimed to establish an experimental design that allows to reproduce to a good approximation the real retention surface from a limited number of experiments, that is from a limited number of chromatograms. Ultimately, our intention is to use the model and experimental design for the simultaneous interpretive optimization of pH and proportion of organic solvent of the mobile phase to be used in the proposed separation. 相似文献
4.
The influence of the mobile phase on retention is studied in this paper for a group of over 70 compounds with a broad range of multiple functional groups. We varied the pH of the mobile phase (pH 3, 7, and 10) and the organic modifier (methanol, acetonitrile (ACN), and tetrahydrofuran (THF)), using 15 different stationary phases. In this paper, we describe the overall retention and selectivity changes observed with these variables. We focus on the primary effects of solvent choice and pH. For example, transfer rules for solvent composition resulting in equivalent retention depend on the packing as well as on the type of analyte. Based on the retention patterns, one can calculate selectivity difference values for different variables. The selectivity difference is a measure of the importance of the different variables involved in method development. Selectivity changes specific to the type of analyte are described. The largest selectivity differences are obtained with pH changes. 相似文献
5.
The effects of some experimental parameters, such as the volume fraction and type of organic modifier in the mobile phase, and the concentration, type and pH of the buffer on the electroosmotic flow velocity, the retention behavior of test solutes, and the column efficiency have been investigated in capillary electrochromatography (CEC) using an open-tubular column of 9.60 microm I.D. with a porous silica layer chemically modified with C18 as stationary phase. The retention of a group of polycyclic aromatic hydrocarbons (PAHs) used as a test mixture varied significantly by changing the organic modifier content in the hydroorganic mobile phase according to the reversed-phase-like selectivity of the stationary phase. In addition, an increase in the percentage of organic modifier resulted in a slight increase in the linear velocity of the EOF. On the other hand, when the phosphate buffer concentration was increased over the range 1-50 mM, the electroosmotic mobility fell dramatically, the retention of the solutes decreased steadily, and the plate height showed a significant increase. The results obtained with phosphate, trishydroxymethylaminomethane or 2-morpholinoethanesulfonic acid as buffers were similar when pH remained constant. Optimization in CEC was essential to achieve further enhancement of separation performance, because the analysis time and separation resolution are essentially affected when varying operating parameters. Separations of seven PAHs with more than 100000 plates are presented within 4 min analysis time. 相似文献
6.
The effect of organic modifier concentration on retention and selectivity of two piperazine diastereomers in a typical n-octadecyl-bonded silica (ODS) column was investigated at pH 6.4 and pH 3.0 using phosphate-buffered acetonitrile (MeCN/H2O) and methanol (MeOH/H2O) mobile phases. The results show the logarithmic retention factors decrease with increasing organic concentration in a less rectilinear fashion in the MeCN/H2O system than in the MeOH/H2O system at high organic concentrations at both pHs. At pH 6.4, the MeOH/H2O system provided significantly higher diastereomer selectivity than the MeCN/H2O system, which can be ascribed to the hydrogen bonding interaction of methanol (as a hydrogen donor) with the piperazine amine moiety of the solute (as a hydrogen acceptor). At pH 3.0, both mobile phases provided high selectivity, in which both acetonitrile and methanol acted as hydrogen acceptors, while the protonated amine acted as the hydrogen donor. The effect of temperature on retention and selectivity was also studied in the two mobile phase systems at both pHs. It was found that at pH 6.4 the retention and selectivity were enthalpically driven in the MeOH/H2O system, while entropically driven in the MeCN/H2O system. However, the retention was entropically driven and the selectivity enthalpically driven in both systems at pH 3.0. Locally preferential solvating and hydrogen bonding effects are proposed to explain the anomalous retention and selectivity behaviors. 相似文献
7.
In an effort to gain insight into the relationship between stationary phase solvation and selectivity, the use of short- and medium-chained-length alcohols (methanol, n-propanol, n-butanol, and n-pentanol) as mobile phase modifiers in reversed-phase liquid chromatography (RPLC) was investigated to determine their impact on chromatographic selectivity. A wide range of mobile phase compositions was evaluated because of the large effect exerted by solvent strength on selectivity. Employing a set of six vanillin compounds as retention probes, evidence is presented to support the view that an increase in the hydrophobicity of the organic modifier used in RPLC can increase the selectivity of the C18 alkyl bonded phase while simultaneously decreasing the retention time of the eluting solutes. Thus, we are presented with an interesting paradox: higher selectivity and shorter retention times, which can be attributed to changes in either solvent selectivity and/or stationary phase solvation by the organic modifier. 相似文献
8.
Poole CF Kiridena W DeKay C Koziol WW Rosencrans RD 《Journal of chromatography. A》2006,1115(1-2):133-141
Plots of the retention factor against mobile phase composition were used to organize a varied group of solutes into three categories according to their retention mechanism on an octadecylsiloxane-bonded silica stationary phase HyPURITY C18 with methanol-water and acetonitrile-water mobile phase compositions containing 10-70% (v/v) organic solvent. The solutes in category 1 could be fit to a general retention model, Eq. (2), and exhibited normal retention behavior for the full composition range. The solutes in category 2 exhibited normal retention behavior at high organic solvent composition with a discontinuity at low organic solvent compositions. The solutes in category 3 exhibited a pronounced step or plateau in the middle region of the retention plots with a retention mechanism similar to category 1 solutes at mobile phase compositions after the discontinuity and a different retention mechanism before the discontinuity. Selecting solutes and appropriate composition ranges from the three categories where a single retention mechanism was operative allowed modeling of the experimental retention factors using the solvation parameter model. These models were then used to predict retention factors for solutes not included in the models. The overwhelming number of residual values [log k (experimental) - log k (model predicted)] were negative and could be explained by contributions from steric repulsion, defined as the inability of the solute to insert itself fully into the stationary phase because of its bulkiness (i.e., volume and/or shape). Steric repulsion is shown to strongly depend on the mobile phase composition and was more significant for mobile phases with a low volume fraction of organic solvent in general and for mobile phases containing methanol rather than acetonitrile. For mobile phases containing less than about 20 % (v/v) organic solvent the mobile phase was unable to completely wet the stationary phase resulting in a significant change in the phase ratio and for acetonitrile (but less so methanol) changes in the solvation environment indicated by a discontinuity in the system maps. 相似文献
9.
Qiu H Wanigasekara E Zhang Y Tran T Armstrong DW 《Journal of chromatography. A》2011,1218(44):8075-8082
New zwitterionic stationary phases were synthesized by covalently bonding 3-P,P-diphenylphosphonium-propylsulfonate to silica gel. The resulting materials possess both a negatively charged sulfonate group and a positively charged quaternary phosphonium group, which means that there is no net charge over a wide pH range. The retention mechanism and chromatographic behavior of polar solutes under HILIC conditions were studied on these zwitterionic phases. Compared to the commercial ZIC-HILIC column and a bare silica gel stationary phase, the newly synthesized zwitterionic stationary phases provided greater retention, higher peak efficiency and better peak symmetry in the HILIC mode. The analytes examined included: β-blockers, nucleic acid bases and nucleosides, salicylic acid and its analogues, and water soluble vitamins. Factors, such as the type of organic modifiers, solvent composition, pH and the buffer concentration of the mobile phase, have been considered as potential variables for controlling the chromatographic retention of polar analytes. 相似文献
10.
Chromatographic effects of dedicated stationary and mobile phase variations in hydrophilic interaction chromatography (HILIC) were investigated using a set of nucleobases, nucleosides and deoxynucleosides as polar test solutes. Retention and selectivity profiles were comparatively mapped on four in-house developed silica materials modified with short alkyl chains (C4, C5) which carry hydroxyl functionalities (including diol motifs) as well as embedded sulphide or sulphoxide groups. These data were complemented by results obtained with two commercially available diol-type phases and a bare silica column. Besides elucidation of packing-related aspects this work concentrated specifically on extending aqueous HILIC (AQ-HILIC) to nonaqueous polar-organic elution conditions herein termed NA-HILIC. The exchange of the polar modifier water by various alcohols in ACN-rich mobile phases containing 5 mM ammonium acetate decreased the eluotropic strength of the resulting eluents. The gain in retention largely followed the order ethanol (EtOH)>methanol (MeOH)>1,2-ethanediol (Et(OH)2) and was accompanied by distinct effects on chromatographic selectivity. For example, on the most polar home-made packing the purine nucleoside selectivity guanosine/adenosine increased from 2.25 in the AQ-HILIC (kguanosine=8.3) to 7.33 (kguanosine=59) in the NA-HILIC mode when EtOH was employed as NA modifier while this value was 5.84 and 2.93 with MeOH and Et(OH)2, respectively (eluent: 5 mM ammonium acetate in ACN/modifier 90:10 v/v). Besides the type of protic modifier its percentage as well the retention and selectivity effects upon varying the ammonium acetate concentration and column temperature, respectively, were also investigated. Notable inter-column differences were found for all of these elution parameters. A mixed-mode retention model composed of partitioning and adsorption is proposed for both AQ- and NA-HILIC retention processes. The potential of (i) the implementation of novel polar bondings (such as ones containing sulphoxide functionalities) and (ii) the comprehensive exploitation of elution variables (type of protic modifiers, salt, etc.) for providing new selectivity increments to the separation of polar analytes in HILIC is emphasised. 相似文献
11.
The retention of ionogenic bases in liquid chromatography is strongly dependent upon the pH of the mobile phase. Chromatographic behavior of a series of substituted aniline and pyridine basic compounds has been studied on C18 bonded silica using acetonitrile-water (10:90) as the eluent with different pHs and at various concentrations of the acidic modifier counter anions. The effect of different acidic modifiers on solute retention over a pH range from 1.3 to 8.6 was studied. Ionized basic compounds showed increased retention with a decrease of the mobile phase pH. This increase in retention was attributed to the interaction with counter anions of the acidic modifiers. The increase in retention is dependent on the nature of the counter anion and its concentration in the mobile phase. It was shown that altering the concentration of counter anion of the acidic modifier allows the optimization of the selectivity between basic compounds as well as for neutral and acidic compounds. 相似文献
12.
13.
Small organic acids have shown significant retention on various stationary phases, such as amide, amino, aspartamide, silica
and sulfobetaine phase commonly used in hydrophilic interaction chromatography (HILIC). This study investigated the effect
of chromatographic conditions on the retention behavior of organic acids in HILIC using the tool of design of experiment (DOE).
The results of the DOE study indicated that both the content of organic solvent (i.e., acetonitrile) and salt concentration
in the mobile phase had significant effects on the retention of organic acids. Higher content of organic solvent in the mobile
phase led to a significant increase in retention on all types of stationary phases. Increasing salt concentration also resulted
in a moderate increase in retention; however, the effect of salt concentration varied with the type of stationary phase. The
study also revealed that column temperature had less impact on retention than organic solvent content and salt concentration
in HILIC. 相似文献
14.
J. G. M. Janssen P. J. Schoenmakers C. A. Cramers 《Journal of separation science》1989,12(10):645-651
The effects of organic modifiers on retention and peak shape in packed-column supercritical fluid chromatography were studied. The adsorption behavior of different modifiers was investigated on stationary phases consisting of hydrocarbon monolayers chemically bonded on silica. Adsorption isotherms for several modifiers were recorded using breakthrough measurements. The results were compared with those obtained by a simple method based on the injection of different sample sizes. Modifiers were selected to reflect various types of interactions with the silica support. Isotherms were found to be approximately Langmuirean. Experiments were performed to elucidate the influence of adsorbed molecules on the retention of selected test solutes. The introduction of a solvent modifier can lead to a substantial change in the mobile phase density, the effect of which is comparable with that of an increase in the density of pure carbon dioxide. Our results confirm that the effects of low concentrations of modifiers (between 0 and 2 percent) in packed-column SFC are largely due to deactivation of residual silanol groups on the silica support. The accessibility of the active sites was found to depend strongly on the size and structure of the modifier molecules. The decrease of retention due to the addition of low concentrations of modifiers could be described accurately by a model derived from Langmuir adsorption behavior. Some semi-quantitative rules for the selection of modifiers and the required concentrations for optimum deactivation of the support are discussed. 相似文献
15.
To understand the chromatographic process as a whole, whether it be for gas chromatography (GC), liquid chromatography (LC), or supercritical fluid chromatography (SFC), one needs to know the chemical and physical nature of the mobile and stationary phases and also the interactions that take place between analytes (solutes) and the two phases. An approach towards Investigating the ways that stationary and mobile phases contribute to chromatographic retention Involves exploring the effects of solvent polarity on the strength of the mobile phase. In SFC this could involve determining the polarity of several different modifier/carbon dioxide mobile phases. In this paper, the use of a solvatochromic indicator to learn more about the effects of SFC modifier/mobile phase polarity will be investigated and discussed using several different modifiers and a diolmodified silica column. 相似文献
16.
Joseph J. Pesek Maria T. Matyska Kavita V. Prajapati 《Journal of separation science》2010,33(19):2908-2916
Two novel silica hydride‐based fluorinated bonded phases have been synthesized using a hydrosilation procedure to test combined fluorine and hydride selectivity. The bonded moieties were characterized by elemental and spectral analysis. Chromatographic testing was done using hydrophilic analytes in the aqueous normal phase mode. At higher amounts of the nonpolar solvent in the mobile phase, there should be increased retention for solutes such as acids, bases and other polar compounds, whereas nonpolar solutes can be retained when water is increased as in RP chromatography. The synergistic effects of the fluorinated phase selectivity and aqueous normal phase retention on a hydride surface have been explored for small polar molecules. The stability and repeatability of the hydride‐based fluorinated stationary phases were evaluated. The use of acetone as the organic component in the mobile phase was also tested. 相似文献
17.
Summary The retention behavior of a set of polycyclic aromatic hydrocarbons in supercritical fluid chromatography have been studied
on a chemically bonded stationary phase based upon a side chain liquid crystalline polymer (LCP) with carbon dioxide-based
mobile phase. The effects of the mobile phase pressure, column temperature and amount of mobile phase organic modifier have
been investigated in order to detect a possible structural change in the liquid crystal polymer linked to the silica support.
The influence of these factors on the selectivity coefficients has also been studied. Two distinctive behaviors with temperature
are noted at low pressure on the one hand and at higher pressure on the other. This change in behavior is based on the density
of the supercritical CO2 and the PAH volatility rather than on any specific stationary phase structural change. Both lower mobile phase pressure and
amount of mobile phase modifier are required to obtain better selectivities. Better planarity recognition is observed in SFC
than in HPLC with these new bonded liquid crystal stationary phases. The bonded liquid crystal phase is only weakly affected
by the addition of organic modifier in the supercritical CO2. 相似文献
18.
Orentienė A Olšauskaitė V Vičkačkaitė V Padarauskas A 《Journal of chromatography. A》2011,1218(39):6884-6891
The potential of 1.7 μm ethylene bridged hybrid silica phase was investigated for the separation of twelve imidazolium-based ionic liquid cations. U-shaped retention profile was observed for all solutes with an increase in retention at both low and high acetonitrile content. Chromatographic behaviour of imidazolium cations in both hydrophilic interaction chromatography (HILIC) and per aqueous liquid chromatography (PALC) modes was studied by varying key parameters such as buffer concentration and pH, acid additive, organic modifier and column temperature. Experimental data provided some evidences that under PALC conditions cationic solutes are retained predominantly by mixed hydrophobic/ion-exchange interactions. In the HILIC mode, both partitioning and ion-exchange interactions are responsible for the retention of solutes. Compared to PALC, HILIC provided significantly higher efficiencies with less or even no peak tailing, better separation selectivity and greater resistance to overload. In PALC mode gradient elution was required to achieve adequate retentivity of all solutes but selectivity was not sufficient to distinguish between solutes with very similar hydrophobicity. In contrast, under HILIC conditions twelve solutes were almost completely resolved in less than 4 min by using isocratic elution. Summarizing, it could be concluded that ethylene bridged hybrid silica column providing a dual retention mechanism offers the possibility of selecting between the two retention modes with opposite separation selectivity, just by changing the composition of the mobile phase. 相似文献
19.
20.
Yuanzhong Yang Maria T. Matyska Reinhard I. Boysen Joseph J. Pesek Milton T. W. Hearn 《Journal of separation science》2013,36(7):1209-1216
In this study, the retention behavior of selected hydrophobic and polar bases on a minimally modified silica hydride phase was investigated. From these results and the associated retention plots, significant differences in the chromatographic dependencies of these two classes of basic compounds were evident. The polar bases exhibited strong retention with mobile phases of high organic solvent content, but displayed weak retention with mobile phases of high water content. In contrast, the hydrophobic bases showed “U‐shape” retention dependencies, indicative of the interplay of both RP and normal‐phase retention characteristics. These studies have demonstrated that hydrophobic and polar bases can be simultaneously separated on the same column either under typical RP‐like or aqueous normal‐phase‐like conditions, respectively, with distinctive selectivity. Finally, the effects of temperature on the RP and aqueous normal phase modality of separations with these analytes were investigated, where discrete changes in retention behavior were also observed. 相似文献