Preparation of Various Bonded Phases for HPLC Using Monochlorosilanes

Abstract

Monochlorosilanes have been prepared with yields of about 80% through a catalytic hydrosilylation of terminal olefins. Subsequently, the silanes are chemically bonded to silica to obtain: (i) n-octyldimethylsilyl bonded phases with reproducible surface coverage ranging from 0.8 to 3.5 μmol/m², (ii) propyldimethylsilyl bonded phases with different functional groups at the γ-position, all showing a nearly equal surface coverage of some 3.3 μmol/m², and (iii) n-alkyldimethylsilyl bonded phases with chainlengths ranging from 1 to 22 carbon atoms and with surface coverages ranging from 3.9 μmol/m² for RP-1 to 3.0 μmol/m² for the RP-22 bonded phase. A simplified model based on the pore structure of silica allows an explanation and estimation of the maximum surface coverage as a function of the chainlength of the bonded phase.     

Preparation and Chromatographic Properties of Some Chemically Bonded Phases for Reversed-Phase Liquid Chromatography

Abstract

Four different monomeric chemically bonded phases suitable for reversed-phase liquid chromatography have been prepared. Duplicate preparations, elemental analysis, post-treatment with TMS and chromatographic properties all indicate that maximum coverage has been obtained. The chromatographic activity of the silica surface is discussed on the basis of TMS-phases with various surface coverages. It is observed that logarithmic retention data for homologous series and different RP-phases-partially taken from the literature—form a set of straight lines with a common intersection point.     

Chemical Modification of Silica Gels After Preliminary Thermal Treatment

Abstract

Chemically bonded phases have been prepared using thermally treated silicagels as basic material. Four bonding reactions with trimethylchlorosilane (TMCS) were successively performed on each silicagel in order to obtain the maximum coverage.

The effect of the chemical modification on the surface structure of packings and on their retention behaviour in reversed-phase chromatography was investigated. The maximum coverage with trimethylsiloxy groups was observed with silicagels previously treated at 300–400°C while the minimum concentration of hydroxyl groups was calculated for silicagels previously treated at 600°C.

Significant differences in retention were observed for these phases in reversed-phase chromatography. Silicagels with maximum coverage were found to be the most convenient for the analysis of apolar solutes while silicagels with minimum hydroxyl group concentration were the most convenient for the analysis of polar solutes.     

Coverage homogenity of chemically bonded C18 phases for HPLC. Investigation of the changes in the free surface energy of the packings

Summary Packings containing chemically bonded C₁₈ groups with different coverage density were prepared using monochlorooctadecylsilane. The method permitting the determination of the free surface energy on the basis of adsorption measurements of two different liquids, noctane and water, was utilized to evaluate the coverage homogeneity of the silica gel surface by the alkylsilyl ligands. It was found that the packing is characterized by a coverage greater than 3.8 mol/m², it is homogenous and the support surface is fully screened.     

Reaction of Silica Gel with Trimethylsilyl Donors Using Conditions Useful for End-Capping HPLC Bonded Phase Packings

Abstract

Several trimethylsilyl (TMS) donors including bis-N, O-trimethylsilylacetamide (BSA), bis-N, O-trimethylsilyltrifluoro-acetamide (BSTFA), hexamethyldisilane (HMDS), trimethylchlorosilane (TMCS) with and without pyridine, 2-trimethylsiloxypent-2-en-A-one (TMSOP), trimethylsilyldimethylamine (TMSDMA), and trimethylsilylimidazole (TMSIM) were reacted with 60 Å silica. TMSIM at 60°C with reaction times of one hour or less gave coverage of silica gel equivalent to that provided by refluxing TMCS and pyridine in toluene for 67 hrs. Reactivity of the TMS donors studied may be ranked in the following order: TMSIM > TMSDMA > BSTFA > BSA > TMCS > TMSOP > HMDS. TMSIM appears to be the reagent of choice for end-capping of alkyl bonded HPLC phases.     

Characterization of Amino-, Cyano-alkyl Bonded Silica Columns in Normal-Phase Liquid Chromatography by Using Steroids

Abstract

In order to characterize the chemically bonded phases in HPLC analysis, the retention behavior of fifteen steroids including estrogen, androgen, progestogen and corticoid were systematically examined using dioxan as the stronger component in an n-hexane-binary system. A linear relationship between the logarithm of the capacity ratio and logarithm of the molar concentration of the binary solvent was confirmed for amino- and cyano-type bonded as well as non-bonded silica gel columns. Based on the retention indices of these phases, the retentivity of the packing materials was determined as follows: the amino-type is similar to and the cyano-type is weaker (0.7 times) than bare silica gel when using dioxan as the stronger component. The specific retentivity of an amino column for polar steroids containing phenolic and alcoholic hydroxyl groups suggests a molecular interaction associated with hydrogen bonding between the polar packing surface and solute compounds. The selectivity of amino packing was found to be larger than cyano packing whose retention selectivity is similar to a bare silica gel.     

Microcalorimetric Study of Methanol Adsorption on Initial Silica and Silica Modified with Polyfluoroalkyl Groups

The adsorption of methanol on initial silica and modified silica samples containing large mesopores is studied by the adsorption–calorimetric method. The grafted tridecylfluoroalkyl groups have a tilted orientation on the silica and physically screen the part of the surface OH groups that have not been involved in the reaction with a modifier. Adsorbed methanol makes the modifying layer looser, thus facilitating the accessibility of methanol molecules to these hydrophilic adsorption sites. Concentrations of OH groups involved in the chemical interaction with molecules of the modifier, OH groups physically screened by its organofluoric radicals, and OH groups located on the surface areas free of the modifier are quantitatively estimated. An additional silanization of the modified silica leads to coverage of silica surface areas that are free of organofluoric modifier with trimethylsilyl radicals. The heat of interaction between the methanol molecules and silica surface hydroxyl groups is determined; it is equal to 60 kJ/mol. The structure of the modifying organofluoric layer and changes in this structure that resulted from additional silanization of the surface and from the methanol adsorption are discussed.     

Chemical end-grafting of homogeneous polystyrene monolayers on mica and silica surfaces

Homogeneous polystyrene monolayers covalently end-attached on mica and silica surfaces were obtained using a "graft to" methodology. The grafting was achieved via nucleophilic substitution between silanol groups (Si-OH) containing surface and monochlorosilyl terminated polystyrene (PS). Different parameters, such as surface activation, grafting reaction time, polymer concentration, nature of solvent, and presence of catalyst, were investigated to determine the optimal conditions for creating very homogeneous and stable polymer monolayers. Ellipsometry, atomic force microscopy (AFM), surface forces apparatus (SFA), and contact angle measurements were used to characterize the polymer-grafted layers. An efficient plasma activation procedure was established to create a maximum number of silanol groups on mica surfaces without increasing the surface roughness. Surface reactivity was investigated by grafting trimethylchlorosilane (TMS) on OH-activated mica and silica. The maximum TMS surface coverage on activated mica is similar to that observed for silica. The stability of covalently attached TMS and PS layers in toluene and water were investigated. Both grafted layers (TMS and PS) partially detached from the mica and silica surfaces when immersed in water. Hydrolysis of the siloxane bond between the monochlorosilyl groups and the surface is the most probable cause of layer degrafting. The degrafting was much slower with the long PS polymer chains, compared to the small TMS molecules, which may act as a protective layer against hydrolysis.     

Influence of the Amount of Bonded Non-Polar Phase and of the Length of Attached Alkyl Chains on Retention Characteristics of Silica-Based Sorbents for Reversed-Phase High Performance Liquid Chromatography

Abstract

The retention characteristics of two sets of chemically bonded non-polar silica packings with a high surface concentration of functional groups have been compared in the reversed-phase mode of liquid chromatography: (i) the conventional packings prepared by chemical modification with trimethylsilyl, heptyl, dodecyl and octadecyl groups, and (ii) mixed-phase materials where differences in the amount of organic bonded phase were achieved by bonding octadecyl and trimethylsilyl groups in different proportions. The retention data of two homologous series of solutes show that these two classes of packings are distinctly different; in particular, the capacity factors and selectivities are always higher on the mixed-phase bonded silica packings at the same percentage of carbon.     

Herstellung oberflächenmodifizierter Adsorbentien

Zusammenfassung Die Reaktion von Phenyltrichlorsilan mit Silicagel wurde an drei Pr?paraten (eng-, mittel- und grobporig) mit bekannter Hydroxylgruppenkonzentration im Temperaturbereich von 20–350 °C untersucht. Die Analyse der modifizierten Produkte ergab bei jeder Reaktionstemperatur eine Phenyl-monochlorsilyl-Gruppierung als neue Oberfl?chenschicht. Daraus folgt, da? eine Phenyltrichlorsilanmolekel selektiv mit zwei oberfl?chenst?ndigen Hydroxylgruppen reagiert. Dieses Ergebnis wurde best?tigt durch Bestimmungen der Hydroxylgruppenkonzentration der Silicagele vor und nach der Reaktion. Es konnten maximal 75% der vorhandenen Hydroxylgruppen umgesetzt werden. Dies entspricht einer Belegung von 3,6μmol Phenylchlorsilylgruppen pro m². Nach der Hydrolyse des modifizierten Gels fiel ein weitgehend hydrophobes Produkt an. Bei dem mittelund grobporigen Pr?parat wurde die spezifische Oberfl?che und die mittlere Porenweite durch die Umsetzung nicht ver?ndert. Bei dem engporigen Pr?parat führte die Umsetzung mit Phenyltrichlorsilan zu einem nichtpor?sen Produkt. Die phenylierten Silicagele k?nnen als Gerüstsubstanzen in der Gas-Fest-Chromatographie und in der Gelpermeationschromatographie eingesetzt werden. Sie dienen weiterhin als Ausgangsprodukte zur Darstellung synthetischer Kationenaustauscher.

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Summary The surface reaction of phenyltrichlorsilane with three different species of silica gel of definite pore structure and known concentration of hydroxyl groups on the surface was studied in the temperature range of 20–350 °C. On the basis of the analytical results of the modified silica gel species it is suggested that phenyltrichlorosilane reacts selectively with two hydroxyl groups forming a phenyl-monochloro-silyl surface layer at all reaction temperatures under investigation. This result was confirmed by the determination of the concentration of the hydroxyl groups on the surface before and after the reaction. The maximum yield of the surface coverage attained under the best reaction conditions was 3,6 μmole phenyl-monochloro-silyl groups m⁻² indicating that 75% of the total amount of the hydroxyl groups had reacted. Hydrolysis of the modified silica gel yielded a white hydrophobic adsorbens. For the wide-pored silical gel and for the species with pores in the transitional range (20<d<400 ?) the value of the specific surface and the mean pore diameter remained unchanged by the modification. The silica gel with narrow pores gave a non porous adsorbent by the modification. The phenylated silica gels were tested as packing materials for GSC- and GPC columns. Furthermore, they were employed for the preparation of synthetic cationexchangers.

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Wir danken Herrn Prof.H. W. Kohlschütter für die F?rderung dieser Arbeit sowie für wertvolle Diskussionen.     

Preparation and Evaluation of P-tert-Butyl-calix[8]arene-bonded Silica Stationary Phase for High Performance Liquid Chromatography

ABSTRACT

A new p-fert-butyl-calix[8]arene-bonded silica gel stationary phase was synthesized through heterogeneous functionalisation of suspended porous silica. A characterization of its structure was carried out by using elemental analysis, FTIR and ¹³C solid state NMR spectroscopy. Chromatographic performance of the new packing material was investigated by employing polycyclic aromatic hydrocarbons (PAHs) as probes and using methanol-water as mobile phase. The investigations show that the new stationary phase behaves as a reversed phase stationary phase. The liquid chromatographic separation of PAHs solutes on the new bonded phase was compared with that on a p-tert-butyl-calix[4]arene-bonded silica stationary phase. The new p-tert-butyl-calix[8]arene-bonded phase exhibited higher retention and better separation selectivity, although the carbon content and coverage of the new packing material was lower than that of the p-tert-butyl-calix[4]arene bonded silica stationary phase. A possible retention mechanism for the new packing material was also proposed.     

Inaccessible hydroxyl groups on silica are accessible in supercritical CO2

The three main types of hydroxyl groups on a silica surface are classified as isolated, hydrogen bonded, and inaccessible. The isolated and hydrogen bonded groups are the most important as these readily exchange with D(2)O and thus are exposed to reactant molecules. However, it has generally been accepted that the inaccessible groups do not participate in surface reactions as only a small fraction of these groups exchange with D(2)O. It is shown that the inaccessible hydroxyl groups on nonporous fumed silica and mesoporous MCM-48 silica powders and films fully exchange with D(2)O and are reactive with octadecylydimethylchlorosilane when supercritical CO(2) is used as the solvent. Furthermore, it is found that the CO(2) penetrating the regions containing the inaccessible groups is not removed by simple evacuation but rather slowly diffuses from the silica over periods of months.     

Evaluation of phenyldimethylethoxysilane treated high-performance thin-layer chromatographic plates. Application to analysis of flavonoids inScutellariae radix

Summary The retention and selectivity of flavonoids (baicalin, baicalein, wogonin, oroxylin A) inScutellariae radix have been studies by high-performance thin-layer chromatography on phenyldimethylethoxysilane-treated silica plates. The silica plates treated with phenyl groups were used for physical and chemical analysis. From elemental carbon analysis, the maximum number of bonded phenyl surface groups per gram was calculated to be 0.467×10²¹ (Oginal silica plate: Merck Art. 15109, Silica gel 100 F₂₅₄). With methanol-1/15 M phosphate buffer (pH 6.2) mixtures as mobile phase, baicalin, baicalein, wogonin, and oroxylin A inScutellariae radix were separated. It has been shown that phenyl-treated plates are more suitable for selective separation of baicalin, baicalein, wogonin, and oroxylin A than octadecyl-treated plates.     

Characterization of Column Packings in Normal-Phase Liquid Chromatography. II. Retention Behavior of Fat-Soluble Vitamins in Amino- and Cyano-Propyl Bonded Silica Gel and Binary Solvent Systems

Abstract

As continuation of our study on the characterization of column packings in normal-phase HPLC analysis, the retention indices of ten fat-soluble vitamins on aminopropyl and cyanopropyl bonded silica columns were systematically estimated using binary solvents containing ethyl acetate (EtOAc), tetrahydrofuran (THF) or 2-propanol (PrOH) in n-hexane. A linear relationship between the logarithm of the capacity ratio and that of the solvent composition was confirmed. The retentivity and selectivity for these chemically bonded packing columns were determined as follows: the amino-type column has stronger and cyano-type column weaker retentivity than the bare silica column for n-hexane-EtOAc or THF binary systems. Specific adsorption of tocopherol derivatives containing phenolic hydroxyl groups on the amino column was observed. To obtain high efficiency in the separation of fat soluble vitamins, peak sharpness and asymmetry factors were measured using three columns and three binary solvents. The bare silica and PrOH binary solvent generally gave superior peak shape for all vitamin samples.     

Effect of the surface coverage of endcapped C18-silica on the excess adsorption isotherms of commonly used organic solvents from water in reversed phase liquid chromatography

The excess adsorption isotherms of methanol, ethanol, 2-propanol, acetonitrile, and tetrahydrofuran from water were measured on five different silica-based packing materials by the minor disturbance method. These materials were prepared with the same lot of 5-microm particles (average pore size 90 A), all endcapped with trimethylchlorosilane (TMS), and bonded to octadecyl chains with different surface coverages (0, 0.42, 1.01, 2.03, and 3.15 micromol/m2). The relative adsorption of one eluent by respect to a second one informs on the heterogeneity of the material (alkyl-bonded and bare silica regions) and on the accessibility of the unreacted silanol groups to the mobile phase. It is shown that the total surface area of the adsorbent decreases with increasing degree of surface coverage with octadecyl chains and that the relative surface area of the regions occupied by accessible silanol groups to the regions occupied by alkyl-bonded groups decreases. For the five columns, an average of 10% of the adsorbent surface area is covered of bare silica accessible to the liquid phase, with a minimum of 5% with tetrahydrofuran and a maximum of 12% with ethanol or 2-propanol. Increasing the surface coverage by the C18 chains causes a significant increase of the attraction potential of the hydrophobic surface toward the organic solvent. This result is confirmed by the increase of the number of adsorbate monolayers with increasing bonding density of the octadecyl chains. This number is twice larger for the 315C18 column than for the C1 column.     

Epoxide‐derived mixed‐mode chromatographic stationary phases for separation of active substances in fixed‐dose combination drugs

A method for the preparation of novel mixed‐mode reversed‐phase/strong cation exchange stationary phase for the separation of fixed‐dose combination drugs has been developed. An epoxysilane bonded silica prepared by vapor phase deposition was used as a starting material to produce diol, octadecyl, sulfonate, and mixed octadecyl/sulfonate groups bonded silica phases. The chemical structure and surface coverage of the functional groups on these synthesized phases were confirmed by fourier‐transform infrared and solid‐state ¹³C NMR spectroscopy and elemental analysis. Alkylbenzene homologs, basic drugs, nucleobases and alkylaniline homologs were used as probes to demonstrate the reversed‐phase, ion exchange, hydrophilic interaction and mixed‐mode retention behaviors of these stationary phases. The octadecyl/sulfonate bonded silica exhibits pronounced mixed‐mode retention behavior and superior retentivity and selectivity for alkylaniline homologs. The mixed‐mode retention is affected by either ionic or solvent strength in the mobile phase, permiting optimization of a separation by fine tuning these parameters. The mixed‐mode stationary phase was applied to separate two fixed‐dose combination drugs: compound reserpine tablets and compound methoxyphenamine capsules. The results show that simultaneous separation of multiple substances in the compound dosage can be achieved on the mixed‐mode phase, which makes multi‐cycles of analysis for multiple components obsolete.     

Experimental Study of Model Bonded Stationary Phases for Liquid Chromatography I. Silica/Polyethyleneoxide

Abstract

Model bonded phases have been prepared by reaction of polyethyleneoxide of various molecular weights (200 < M < 5.10⁶) on silica.

The retention behavior of solutes on these bonded stationary phases for liquid chromatography depends on silica loadings, grafted molecule length, solute size and solvent nature. Different mechanisms such as dissolution effect in the grafted phase, adsorption on mineral support and steric exclusion due to the residual porosity of silica, are involved in the observed separations. An expression of elution volumes in relation with these mechanisms is proposed.

Chromatographic data and thermodynamic predictions are in good agreement.     

Analysis of silica hydride and surface hydrosilation reactions by solid-state NMR in the preparation of<Emphasis Type="Italic"> p</Emphasis>-chlorobenzamide bonded silica phase

²⁹Si and ¹³C CP-MAS NMR spectroscopy was used to follow the conversion of native silica to a p-chlorobenzamide bonded silica material. The benzamide bonded phase was prepared via a hydrosilation reaction of a hydride silica intermediate with p-chloro-N-allylbenzamide. Solid-state NMR was used to show the disappearance of reactive surface hydride species (M_H) and to identify newly formed bonded chemical species on the silica surface. DRIFT spectroscopy, elemental analysis, and specific surface-area determinations (BET) of the prepared phases are also reported.     

Ionic liquid-modified silica as a new stationary phase for chromatographic separation

A new stationary phase based on silica modified with 1-methyl-3-propylimidazolium chloride was synthesized and characterized in this paper. A derivative of 1-methyl-3-propylimidazolium chloride was used to chemically modify the surface of silica particles to act as the stationary phase for HPLC. The modified particles were characterized by Fourier Transform Infrared (FT-IR), ¹³C NMR spectroscopy and thermogravimetric analysis (TGA). The surface modification procedure rendered particles with a surface coverage of 0.89 μmol/m² of alkylimidazolium chloride. Columns packed with the modified silica and blank silica particles were tested under HPLC conditions. Preliminary evaluation of the stationary phase for HPLC was performed using aromatic compounds as model compounds. The separation mechanism appears to involve multiple interactions including ion exchange, hydrophobic and electrostatic interactions.     

Properties and diversity of C18 bonded phases

Summary The effect of the properties of the silica gel matrix, and of the octadecylsilane used, on the chromatographic selectivity of C18 bonded phases is described. Preparation of bonded phases with monochlorodimethyloctadecylsilane is easier because absolute water exclusion is not required in this case. The surface coverage (mol/m²) is smaller than with dichloromethyl- or trichlorooctadecylsilanes, but in these cases a secondary silanization with HMDS is essential to remove newly formed silanol groups. Bonded phases on Si 60 give the largest amount of hydrocarbon per unit column volume, large k values and good selectivity, however, surface coverage and efficiency of these packed columns is less than with Si 100 silica. The chemical properties of the silica surface also influence bonded phase selectivity. Bonded phases should not be compared with only a single eluent, because properties of bonded phases differ with test solutes and test conditions, i.e. eluent composition.Presented at the 14th International Symposium on Chromatography London, September, 1982In Part from Ph. D. Thesis H. Schmidt, Saarbrücken 1978