Characterization of the acidity of residual silanol groups in microparticulate and monolithic reversed-phase columns

The residual silanol acidity and activity of several microparticulate and monolithic C18 columns has been measured from the retention of LiNO3 in the columns with a methanol/buffer (1 mM in Na+) (60:40 v/v) mobile phase buffered to different pH values. For Luna C18 (2) and LiChrospher RP-18 columns, at least two different types of silanols with different acidity for each packing, were observed. Purospher RP-18e and Chromolith RP-18e packings present evidence of some active silanols only at pH values close to their basic pH stability limit or higher. The results obtained have been compared with those obtained previously for Resolve C18, Resolve Silica, Symmetry C18, Symmetry Silica, XTerra MSC18 and Underivatized XTerra. A modification of an equation previously proposed has been applied to all columns studied and the results obtained have been used to classify the columns according to their silanol acidity and activity. The method allows the prediction of the extent of the silanol activity of the columns studied at a particular mobile phase pH.     

Comparison of the acidity of residual silanol groups in several liquid chromatography columns

The silanol acidity of Waters Resolve C18, Waters Resolve silica, Waters Symmetry C18, Waters Symmetry silica, Waters XTerra MS C18 and underivatized XTerra columns has been measured from the retention of LiNO3 with a methanol/water (60:40) mobile phase buffered to different pH values. The Li+ cation is retained by cationic exchange with the background cation of the mobile phase (Na+) through the ionized silanols. The number of active silanols increases in the order: XTerra MS C18 < Symmetry C18 < underivatized XTerra < Resolve C18 < Resolve silica approximately equal to Symmetry silica. XTerra MS C18 does not present any residual silanol acidity up to s(s)pH 10.0 (pH in 60% methanol) as measured by LiNO3. The underivatized XTerra packing and Symmetry C18 present active silanols only at s(s)pH values higher than 7.0. For the other three columns, two different types of silanols with different acidity (s(s)pKa values about 3.5-4.6 and 6.2-6.8, respectively) have been observed. Symmetry C18 shows evidence of the presence of active basic sites that retain NO3- by anionic exchange.     

Evaluation of the influence of residual silanol groups on the total retention in reversed-phase chromatography

It was demonstrated that the characteristics of stationary phases for reversed-phase chromatography can be compared by analyzing the retention of several compounds belonging to a homologous series during elution with one and the same composition of mobile phase or the retention of a nonpolar compound during elution with several mobile phases of different compositions. The slope of the corresponding linear correlations can be used to characterize the hydrophobicity of the columns (stationary phases) under study. To analyze the polar interactions between the sorbate and the stationary phase it is advantageous to study the retention of a polar substance in several (at least in two) mobile phases of different compositions. The degree of displacement of the linear correlation relative to that for nonpolar substances can be considered a measure of the influence of residual silanol groups on the total retention characteristics.     

A new method for assessing the effect of residual silanol groups on total retention in reversed-phase HPLC

A new approach is proposed to assess the effect of residual silanol groups (hydrophilicity) on the retention of polar substances in reversed-phase high-performance liquid chromatography (HPLC). It is shown that the numerical characteristic of the relative position of trend lines in the coordinates logk(B) vs. logk(A) of the relative retention parameters (at different mobile phase compositions) for any stationary phase relative to the stationary phase selected as a standard is preferable to the separation selectivity of the same pair of substances. Full information can be obtained by determining the retention of corresponding adsorbates with two different mobile phase compositions for each column under study. It was shown that the pair of adsorbates p-toluidine-p-cresol exhibits higher sensitivity to a change in the hydrophilicity of stationary phases as compared to the pair aniline-phenol. However, the use of a nonpolar substance as a reference compound is more informative.     

Reactivity of silanol groups on zeolite surfaces

 Alkyl group introduction into zeolite surfaces was attempted utilizing the reactivity of surface silanol groups as lattice defects resulting from hydrothermal or acid treatment of zeolites. The reaction was carried out using organosilane and alcohols, and traced by in situ IR measurement as well as by elemental analysis. In situ IR measurement demonstrated high reactivity of terminal silanol groups. A good correlation was noted between the amount of reacted groups and the amount of terminal silanol groups, indicating high reactivity of terminal silanol groups and very low reactivity of silanol groups that form hydroxy nests. Treated zeolite exhibited high hydrophobicity, such that it floats on water. Received: 17 February 1998 Accepted: 26 May 1998     

Method for estimating the residual silanol activity of reversed-phase chromatographic adsorbents

The retention of some organic bases on reversed-phase adsorbents was studied as a function of the concentration of KH₂PO₄ in the mobile phase. It was shown that, for some adsorbents, retention decreases with an increase in the salt concentration, while an opposite dependence was observed for the other adsorbents. Such behavior of adsorbates can be explained by the prevailing silanophilic interactions in the former case and hydrophobic interactions in the latter case. The observed dependence can be used to change the chromatographic selectivity in the analysis of complex mixtures containing organic bases and to assess the properties of reversed-phase columns. Diphenhydramine hydrochloride was proposed as a test substance to assess the properties of the columns in the analysis of multicomponent pharmaceutical preparations. It was used in the comparative study of the properties of some adsorbents.     

Interaction of trimethylchlorosilane with silanol groups silica surface in the presence of amines

Experimental and quantum chemical studies of ClSi(CH₃)₃ interactions with SiOH groups catalyzed by amines were carried out using aerosil samples. The catalytic effects are due to the enthalpy factor of rate constants through transition state stabilization by NR₃.     

Density of silanol groups on the surface of silica precipitated from a hydrothermal solution

The physicochemical properties of amorphous silica precipitated from a hydrothermal solution were studied. Low-temperature nitrogen adsorption in conjunction with the BET method was used to determine the specific surface area of this silica. Based on thermogravimetry data, the total content of water was estimated. A comparison of the thermogravimetry data with the Zhuravlev physicochemical constants made it possible to determine the temperature dependences of the concentration of surface and internal silanols over a temperature range of from 200 to 1200°C. A new type of amorphous silica with enhanced internal water content was revealed. The distinctions between the mechanisms of the removal of surface and internal water were established.     

IR spectroscopic investigation of internal silanol groups in different zeolites with pentasil structure

Various isomorphically substituted high-silica zeolites, including ZSM-5, beta, ferrisilicates, gallosilicates and borosilicates, were investigated by diffuse reflectance IR spectroscopy. The narrow band at 3735 cm–1 and the broad band at 3500 cm–1 were assigned to internal silanol groups. These defect sites in the as-synthesized Na-forms of zeolites arise during the template decomposition.     

Reducing residual silanol interactions in reversed-phase liquid chromatography. Thermal treatment of silica before derivatization

This study describes the thermal pretreatment of a silica gel between 150 and 800 degrees C before derivatization with dimethyloctadecylchlorosilane as a means of reducing residual silanol activity in HPLC bonded stationary phases. A time study was done from 12 to 48 h to find the optimum time needed for dehydroxylation. With increasing pretreatment temperatures, the number of reactive silanols is reduced from 8 micromol/m2 to essentially zero at 1000 degrees C (where sintering occurs). The effects of the thermal pretreatments were observed with diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) and solid state cross polarization magic angle spinning (CP-MAS) 29Si NMR. Following derivatization, residual silanol activity and pH stability were tested by packing columns with the derivatized silica and carrying out a series of reversed-phase liquid chromatography (RPLC) experiments. Residual silanol activity was greatly reduced when the silica was pretreated at 800 degrees C, leading to less peak tailing for basic solutes. In a strongly basic mobile phase (pH 11.5) the pretreated silica was surprisingly stable, although bond cleavage of C18 groups from the surface was observed.     

Characterization of immobilized artificial membrane (IAM) and XTerra columns by means of chromatographic models

Immobilized artificial membranes (IAMs) prepared from phosphatidylcholine analogs are used as stationary phases in liquid chromatography systems to model drug partitioning between an aqueous phase (mobile phase) and a cell membrane (IAM column). Two different chromatographic models, which describe retention as a function of solute and column-mobile phase properties, have been applied to characterization of an IAM and two reversed phase C18 columns (Waters XTerra MSC18 and XTerra RP18) with acetonitrile-water mobile phases. The comparison of the results shows that the phosphatidylcholine group makes IAM column more polar than both XTerra columns, specially in terms of hydrogen-bond acceptor ability. XTerra RP18 is slightly more polar than XTerra MSC18 because of the presence of the embedded carbamate polar group.     

Characterization of sol-gel immobilized lipases

The immobilization of lipases within a chemically inert hydrophobic sol-gel support, which is prepared by polycondensation of hydrolyzed tetramethoxysilane (TMOS) and methyltrimethoxysilane (MTMS) or iso-butyltrimethoxysilane (iso-BTMS), results in heterocatalysts. The heterocatalysts so prepared showed a dramatically enhanced catalytic activity and stability as measured by the hydrolysis and transesterification of soybean oil. The lipase/sol-gel materials were characterized by nitrogen adsorption to determine their specific surface area. Solid state NMR was used to reveal the degree of cross-linking of the sol-gel materials. Scanning electron microscopy and atomic force microscopy were used to observe the morphology of the biocatalysts. Transmission electron microscopy and confocal microscopy were used to investigate the enzyme distribution within the sol-gel materials. The characterization studies showed that the most active lipase-containing sol-gel was a non-porous amorphous material with enzyme randomly distributed throughout the sol-gel material. The activity of the immobilized enzyme did not correlate to the degree of cross-linking or the specific surface area of the sol-gel materials. The highly retained activity of the immobilized enzyme was more likely attributed to the conformational changes of the enzyme during the immobilization, which result in enzyme's fixation in a more favorable conformation and to the lipophilic environment of the hybrid matrix structure which facilitates the transport of the hydrophobic substrate to the active sites.     

Multiquantum processes in the enzyme molecules immobilized on biological membranes

A physical model of an oxidation–reduction reaction for immobilized enzyme is considered. The influence of the electric-field intensity of a biological membrane on the enzyme reaction rate is analyzed. It is shown that the low-frequency dipole-active vibration of the oppostite charge groups of the substrate and enzyme relative to each other leads to multiquantum excitation of the system over the conformational degree of freedom. This excitation provides an abrupt increasing of the tunnel decay rate of the substrate–enzyme complex on a free product and a free enzyme. This way of the reaction is more probable in comparison with the usual overbarrier process. Some consequences for the immobilized cytochrome P-450 are discussed. © 1998 John Wiley & Sons, Inc. Int J Quant Chem 66 : 255–260, 1998     

Relative importance of hydrogen bonding and coordinating groups in modulating the zinc-water acidity

The presence of second-sphere -NH(2) groups in the proximity of a zinc(ii)-bound water molecule enhances its acidity by ca. 2 pK(a) units.     

Quantitative 1H NMR analysis of reacted silanol groups in silica nanoparticles chemically modified with monochlorosilanes

Quantitative analysis of reacted silanol groups in silica nanoparticles modified chemically with monochlorosilanes was performed by ¹H NMR after treatment with cesium fluoride. Silica nanoparticles were modified chemically by the reaction between the silanol groups and monochlorosilanes, and the structure of the organic moiety anchored onto the silica surface was confirmed with solid‐state ¹³C NMR. As monochlorosilanes react with silanol groups at 1:1 ratio unlike di‐ or trichlorosilanes, the number of the silanes introduced into silica nanoparticles equals that of reacted silanol groups. Organically modified silica nanoparticles were dissolved using cesium fluoride, and the amount of the soluble organic compounds originated from the introduced silanes was determined by a ¹H NMR internal standard method using pyrene as the reference. Those values determined by ¹H NMR were in good agreement with those determined by elemental analysis. Thus, the number of reacted silanol groups per one particle was calculated on the basis of the results obtained by the ¹H NMR method, and the values were highly dependent on the steric structure of the introduced silanes. Copyright © 2010 John Wiley & Sons, Ltd.     

Statistical optimization of the silylation reaction of a mercaptosilane with silanol groups on the surface of silica gel

Thiol-modified silica is often used as an intermediate product for further synthesis of modified stationary phases for chromatography or purification processes. Different conditions were used to synthesize such thiol-modified particles, but systematic optimizations remained scarce. In this study the reaction conditions for the synthesis of mercaptopropyl-modified silica were optimized. The general synthetic method consists in slurrying the silica gel in toluene before adding 3-mercaptopropyldimethoxymethylsilane together with a tertiary amine as catalyst (here dimethylaminopyridine). Reaction time and temperature were optimized using a full factorial design of experiment (DoE) from 3 to 25 h with temperature varying between 45 and 105 °C. The surface coverage of the silica with mercaptopropyl-groups was analyzed by two different ways (elemental analysis and chemical surface reaction with 2,2′-dipyridyl disulfide followed by HPLC-UV analysis of stoichiometrically liberated pyridyl-2-thione). We obtained a three-dimensional (3D) plot of the surface coverage as a function of reaction time and temperature. The arch-shaped hyperplane allowed us to determine an optimum with regard to time and temperature, which yields to the highest surface coverage possible. We also verified that the increase of the surface coverage does not lead to a decrease of the stability of the surface modification by subjecting the gels to treatment with high temperature and acidic conditions. The stability was monitored by different chromatographic methods. Moreover, ²⁹Si cross-polarization-magic angle spinning (CP-MAS) NMR spectra of materials prepared by different conditions allowed to confirm that the Si species on the surface were essentially the same, while there was only a minute difference in signal intensities for the individual Si species for materials obtained by distinct temperatures.