Metabolism and excretion of 1-hydroxymethylpyrene, the proximate metabolite of the carcinogen 1-methylpyrene, in rats.

1-Methylpyrene, an alkylated polycyclic aromatic hydrocarbon and environmental carcinogen, is activated by side-chain hydroxylation to 1-hydroxymethylpyrene (1-HMP) and subsequent sulfo conjugation to the DNA-reactive 1-sulfooxymethylpyrene. In addition to the bioactivation, processes of metabolic detoxification and transport greatly influence the genotoxicity of 1-methylpyrene. For a better understanding of 1-HMP detoxification in vivo we studied urinary and fecal metabolites in rats following intraperitoneal doses of 19.3mg 1-HMP/kg body weight (5 rats) or the same dose containing 200µCi [(14)C]1-HMP/kg body weight (2 rats). After 48h, 48.0% (rat 1) and 29.1% (rat 2) of the radioactivity was recovered as 1-HMP in the feces. Six major metabolites were observed by UV and on-line radioactivity detection in urine samples and feces after HPLC separation. The compounds were characterized by mass spectrometry, (1)H NMR and (1)H-(1)H COSY NMR spectroscopy, which allowed assigning tentative molecular structures. Two prominent metabolites, 1-pyrene carboxylic acid (M-6) and the acyl glucuronide of 1-pyrene carboxylic acid (M-5) accounted for 17.7% (rat 1) and 25.2% (rat 2) of the overall radioactive dose. Further, we detected the acyl glucuronide of 6-hydroxy-1-pyrene carboxylic acid (M-1) and 8-sulfooxy-1-pyrene carboxylic acid (M-3) together with two regioisomers of M-3 (M-2 and M-4) differing in position of the sulfate group at the pyrene ring. In urine samples, the radioactivity of 1-pyrene carboxylic acid and its five derivatives amounted to 32.4% (rat 1) or 45.5% (rat 2) of the total [(14)C]1-HMP dose.

Insect Adhesion Secretions: Similarities and Dissimilarities in Hydrocarbon Profiles of Tarsi and Corresponding Tibiae.

Spatially controlled in vivo sampling by contact solid phase microextraction with a non-coated silica fiber combined with gas chromatography-mass spectrometry (GC-MS) was utilized for hydrocarbon profiling in tarsal adhesion secretions of four insect species (Nicrophorus vespilloides, Nicrophorus nepalensis, Sagra femorata, and Gromphadorhina portentosa) by using distinct adhesion systems, viz. hairy or smooth tarsi. For comparison, corresponding samples from tibiae, representing the general cuticular hydrocarbon profile, were analyzed to enable the statistical inference of active molecular adhesion principles in tarsal secretions possibly contributed by specific hydrocarbons. n-Alkanes, monomethyl and dimethyl alkanes, alkenes, alkadienes, and one aldehyde were detected. Multivariate statistical analysis (principal component and orthogonal partial least square discriminant analyses) gave insights into distinctive molecular features among the various insect species and between tarsus and tibia samples. In general, corresponding hydrocarbon profiles in tarsus and tibia samples largely resembled each other, both qualitatively and in relative abundances as well. However, several specific hydrocarbons showed significantly different relative abundances between corresponding tarsus and tibia samples, thus indicating that such differences of specific hydrocarbons in the complex mixtures might constitute a delicate mechanism for fine-tuning the reversible attachment performances in tarsal adhesive fluids that are composed of substances originating from the same pool as cuticular hydrocarbons. Caused by melting point depression, the multicomponent tarsal adhesion secretion, made up of straight chain alkanes, methyl alkanes, and alkenes will have a semi-solid, grease-like consistency, which might provide the basis for a good reversible attachment performance.

Contact solid-phase microextraction with uncoated glass and polydimethylsiloxane-coated fibers versus solvent sampling for the determination of hydrocarbons in adhesion secretions of Madagascar hissing cockroaches Gromphadorrhina portentosa (Blattodea) by gas chromatography-mass spectrometry.

Molecular profiles of adhesion secretions of Gromphadorrhina portentosa (Madagascar hissing cockroach, Blattodea) were investigated by gas chromatography mass spectrometry with particular focus on a comprehensive analysis of linear and branched hydrocarbons. For this purpose, secretions from the tarsi (feet), possibly contributing to adhesion on smooth surfaces, and control samples taken from the tibiae (lower legs), which contain general cuticular hydrocarbons that are supposed to be not involved in the biological adhesion function, were analyzed and their molecular fingerprints compared. A major analytical difficulty in such a study constitutes the representative, spatially controlled, precise and reproducible sampling from a living insect as well as the minute quantities of insect secretions on both tarsi and tibiae. Thus, three different in vivo sampling methods were compared in terms of sampling reproducibility and extraction efficiency by replicate measurement of samples from tarsi and tibiae. While contact solid-phase microextraction (SPME) with a polydimethylsiloxane (PDMS) fiber showed higher peak intensities, a self-made uncoated glass fiber had the best repeatability in contact-SPME sampling. Chromatographic profiles of these two contact-SPME sampling methods were statistically not significantly different. Inter-individual variances were larger than potentially existing minor differences in molecular patterns of distinct sampling methods. Sampling by solvent extraction was time consuming, showed lower sensitivities and was less reproducible. In general, sampling by contact-SPME with a cheap glass fiber turned out to be a viable alternative to PDMS-SPME sampling. Hydrocarbon patterns of the tarsal adhesion secretions were qualitatively similar to those of epicuticular hydrocarbon profiles of the tibiae. However, hydrocarbons were in general less abundant in tarsal secretions than secretions from tibiae.

Analysis of chemical profiles of insect adhesion secretions by gas chromatography-mass spectrometry.

This article reports on the chemical analysis of molecular profiles of tarsal secretions of the desert locust Schistocerca gregaria (Forsskål, 1775) by gas chromatography hyphenated with quadrupol mass spectrometry (GC-MS) as well as (1)H-nuclear magnetic resonance ((1)H NMR) spectroscopy. Special focus of this study was to elaborate on sampling methods which enable selective microscale extraction of insect secretions in a spatially controlled manner, in particular tarsal adhesive secretions and secretions located on cuticle surfaces at the tibia. Various solvent sampling procedures and contact solid-phase microextraction (SPME) methods were compared in terms of comprehensiveness and extraction efficiencies as measured by signal intensities in GC-MS. Solvent sampling with water as extraction solvent gave access to the elucidation of chemical profiles of polar compound classes such as amino acids and carbohydrates, but is extremely tedious. Contact SPME on the other hand can be regarded as a simplified and more elegant alternative, in particular for the lipophilic compound fraction. Many proteinogenic amino acids and ornithine as well as carbohydrate monomers arabinose, xylose, glucose, and galactose were detected in tarsal secretions after acid hydrolysis of aqueous extracts. Qualitatively similar but quantitatively significantly different molecular profiles were found for the lipid fraction which contained mainly n-alkanes and internally branched monomethyl-, dimethyl-, and trimethyl-alkanes in the C23-C49 range as well as long chain fatty acids and aldehydes. Especially, hydrocarbons with >C40 carbon numbers have previously been rarely reported for insect secretions. The results suggest that the investigated insect secretions are complex emulsions which allow the attachment of tarsi on various otherwise incompatible materials of smooth and rough surfaces. The solid consistence of the established alkanes at ambient temperatures might contribute to a semi-solid consistence of the adhesive, amalgamating partly opposing functions such as slip resistance, tarsal release, desiccation resistance, and mechanical compliance. The methods developed can be extended to other similar applications of studying compositions of insect secretions of other species.

Four new flavonol glycosides from the leaves of Brugmansia suaveolens.

Four new flavonol glycosides were isolated from the leaves of Brugmansia suaveolens: kaempferol 3-O-ß-D-glucopyranosyl-(1'''→2'')-O-α-L-arabinopyranoside (1), kaempferol 3-O-ß-D-glucopyranosyl-(1'''→2'')-O-α-L-arabinopyranoside-7-O-i-D-gluco-pyranoside (2), kaempferol 3-O-ß-D-[6'''-O-(E-caffeoyl)]-glucopyranosyl-(1'''→2'')-O-α-l-arabinopyranoside-7-O-ß-D-glucopyranoside (3), and kaempferol 3-O-ß-D-[2'''-O-(E-caffeoyl)]-glucopyranosyl-(1'''→2'')-O-α-l-arabinopyranoside-7-O-ß-D-glucopyranoside (4). The structure elucidation was performed by MS, 1D and 2D NMR analyses.

Spectroscopic and chromatographic characterisation of a pentafluorophenylpropyl silica phase end-capped in supercritical carbon dioxide as a reaction solvent.

This research uses solid-state nuclear magnetic resonance (NMR) spectroscopy to characterise the nature and amount of different surface species, and chromatography to evaluate phase properties of a pentafluorophenylpropyl (PFPP) bonded silica phase prepared and end-capped using supercritical carbon dioxide (sc-CO2) as a reaction solvent. Under sc-CO2 reaction conditions (at temperature of 100 °C and pressure of 414 bar), a PFPP silica phase was prepared using 3-[(pentafluorophenyl)propyldimethylchlorosilane] within 1h. The bonded PFPP phase was subsequently end-capped with bis-N,O-trimethylsilylacetamide (BSA), hexamethyldisilazane (HMDS) and trimethylchlorosilane (TMCS) within 1h under the same sc-CO2 reaction conditions (100 °C/4141 bar). Elemental microanalysis, thermogravimetric analysis (TGA), and scanning electron microscopy (SEM) were used to provide support data to solid-state NMR and chromatographic evaluation. Results revealed a surface coverage of 2.2 µmol/m(2) for the non-end-capped PFPP silica phase while the PFPP phase end-capped with BSA gave a higher surface coverage (3.9 µmol/m(2)) compared to HMDS (2.9 µmol/m(2)) and TMCS (2.8 µmol/m(2)). (29)Si CP/MAS NMR analysis of the PFPP end-capped with BSA shows a significant decrease in the amount of Q(3) (free silanols) and Q(4) (siloxane groups) species, coupled with the absence of the most reactive Q(2) (geminal silanols) in addition to increased amount of a single resonance peak centred at +13 ppm (MH) corresponding to -Si-O-*Si-CH3 bond. (13)C CP/MAS NMR shows the resonance corresponding to the propyl linkage (CH3CH2CH2-) and methyl groups (Si(CH3)n) confirming successful silanisation and endcapping reactions in sc-CO2. Chromatographic evaluation of the BSA end-capped PFPP phase with Neue text mixture revealed improved chromatographic separation as evidenced in the enhanced retention of hydrophobic markers and decreased retention for basic solutes. Moreover, chromatography revealed a change in column selectivity for the BSA end-capped PFPP phase with dipropylphthalate eluting before naphthalene, indicating decreased silanol groups and increased hydrophobicity. The extend of BSA end-capping as measured by the increase in column efficiency (67,260 N/m vs. 60,480 N/m) on a 2.1 i.d.×50 mm column, methylene group selectivity (α(CH(2)) = 2.27 vs. 2.14) and decreased silanophilic interactions (S=3.7 vs. 4.10) indicate that the increase in carbon loading (3.9 µmol/m(2) vs. 2.2 µmol/m(2)) and improvement in chromatography in good peak shape and symmetry is attributed to end-capping with trimethylsilyl groups.

Synthesis, characterisation and chromatographic evaluation of pentafluorophenyl and phenyl bonded silica phases prepared using supercritical carbon dioxide as a reaction solvent.

Pentafluorophenyl and phenyl silica stationary phases offer alternative selectivity compared to alkyl bonded C18 and C8 stationary phases, through other interactions such as π-π interactions, dipole-dipole and hydrogen bond interactions. Pentafluorophenyl and phenyl silica bonded stationary phases were efficiently prepared in sc-CO2 specifically pentafluorophenyl propyl (PFPP), pentafluorophenyl (PFP), phenyl propyl (PP) and phenyl (P) silica stationary phases. The bonded phases were characterised by elemental analysis, thermogravimetric analysis (TGA), BET, and by solid-state NMR spectroscopy. Chromatographic performance of the supercritical fluid generated phases was also investigated using the Neue test. The authors present results which demonstrate that pentafluorophenyl and phenyl stationary phases can be prepared successfully under supercritical conditions of 100 °C, 414 bar in a reaction time of 1h with surface coverage comparable to traditional organic solvent based methods. Chromatographic results reveal that the pentafluorophenyl propyl (PFPP) phase provides superior separation performance for Neue test solutes despite having a lower ligand density (C: 5.67%, 2.2 µmol/m²) compared to the phenyl propyl (PP) analogue having the highest ligand density (C: 6.67%, 2.5 µmol/m²). The difference chromatographic performance is attributed to the polarity of the CF bond in PFPP phase. Moreover, as the alkyl chain length decreases, the hydrophobic interaction also decreases, and the PFPP phase (with a propyl linkage) provides better separation compared to the PFP phase.

Simulation of the chromatographic separation process in HPLC employing suspended-state NMR spectroscopy - comparison of interaction behavior for monomeric and hydride-modified C18 stationary phases.

The interactions of different analytes with monomeric and hydride-modified stationary phases have been investigated employing suspended-state NMR spectroscopy. The suspended-state high-resolution/magic-angle-spinning (1)H-NMR spectrum of an analyte in the presence of C(18) SP material shows a splitting into two sets of signals for the analyte molecule. One state reflects a closer interaction between analyte and C(18) -modified surface that results in an upfield shift and broader signal half-widths. This phenomenon suggests that the analyte exists in two environments. We report a systematic approach upon the investigation on the interaction in the interface of analyte, mobile phase, and modified silica through synthesis of differently modified silica with a gradual increase in surface coverage. The determination of the signal half-widths and chemical shifts revealed a relationship between the modification technique of the C(18) SPs and the chromatographic and NMR spectroscopic behavior. Increasing ligand density results in higher shielding of the NMR signals for the analyte in the "adsorbed" state. The measurement of spin-lattice relaxation times T(1) of the analyte molecule correlate NMR parameter together with separation behavior in HPLC. Furthermore, suspended-state and solid-state NMR measurements revealed different alkyl chain mobilities for the monomeric and hydride-modified SPs.

Monitoring the on-line titration of enantiomeric omeprazole employing continuous-flow capillary microcoil 1H NMR spectroscopy.

The titration of the (S)-enantiomer of omeprazole with the (R)-enantiomer in chloroform-d(1) is monitored by continuous-flow capillary microcoil (1)H NMR spectroscopy employing a microcoil with a detection volume of 1.5 µl. The observed changes of the (1)H NMR chemical shifts indicate the formation of a heterochiral (R,S) dimer of omeprazole via its sulfinyl group and the NH group of the benzimidazole ring.

Detection of DNA adducts originating from 1-methoxy-3-indolylmethyl glucosinolate using isotope-dilution UPLC-ESI-MS/MS.

1-Methoxy-3-indolylmethyl (1-MIM) glucosinolate, present at substantial levels in several food crops (e.g., broccoli and cabbage), forms DNA adducts in vitro and is mutagenic to bacterial and mammalian cells after activation by the plant enzyme myrosinase. Moreover, a breakdown product, 1-MIM alcohol, is metabolized to a secondary reactive intermediate by some mammalian sulfotransferases (SULTs). First, we incubated herring-sperm DNA with 1-MIM glucosinolate in the presence of myrosinase. We identified and synthesized the predominant adducts, N(2)-(1-MIM)-dG and N(6)-(1-MIM)-dA, and developed an UPLC-ESI-MS/MS method for their specific detection using isotopic dilution. Second, we demonstrated both DNA adducts in target cells (Salmonella typhimurium TA100 and Chinese hamster V79) of standard mutagenicity tests treated with 1-MIM glucosinolate/myrosinase as well as in 1-MIM alcohol-treated Salmonella and V79 cells engineered for expression of human SULT1A1. Similar excesses of N(2)-(1-MIM)-dG over N(6)-(1-MIM)-dA adducts were found in all cellular models independent of the test compound (1-MIM glucosinolate or alcohol), whereas dA adducts predominated in the cell-free system. Finally, we detected both DNA adducts in colon tissue of a mouse orally treated with 1-MIM glucosinolate. We are going to use this specific and sensitive method for investigating genotoxic risks of food-borne exposure to 1-MIM glucosinolate in animal and human studies.

Time-dependent column performance of cholesterol-based stationary phases for HPLC by LC characterization and solid-state NMR spectroscopy.

Three different cholesterol-based stationary phases were investigated with respect to their time-dependent separation behavior. The examined stationary phases differ in the used spacer molecule and the synthesis route and were used under routine laboratory conditions over a period of two years. The chromatographic behavior of the three phases was determined by using a standard reference material in addition to a separation of a steroid mixture. The surface chemistry and the modification of these with the chemically bonded moiety were investigated with nuclear magnetic resonance (NMR) spectroscopy and elemental analysis. Through applying different techniques we determined changes in retention and selectivity; solid-state NMR spectra showed changes in the surface chemistry dependent on the synthesis route. Superior long-term stability was observed for the undecanoate-cholesterol (UDC-Chol) column in terms of hydrophobic retentiveness and selectivity.

Preparation and characterization of bonded silica hydride intermediate from triethoxysilane and dimethylmethoxysilane using supercritical carbon dioxide and dioxane as reaction medium.

This research examines bonding methodology, surface coverage and silanol conversion efficiencies on the preparation of silica hydride (SiH) intermediate from triethoxysilane (TES) and dimethylmethoxysilane (DMMS) using sc-CO(2) and dioxane as reaction solvent. Under sc-CO(2) reaction conditions (at temperature and pressure of 100 °C, 414 bar, respectively and 3h reaction time), the surface coverages of SiH (evaluated from %C obtained from elemental analysis) prepared with DMMS (3.39 µmol/m(2)) and TES (4.46 µmol/m(2)) increased by 2- and 4-folds respectively, when compared to reaction performed in dioxane (2.66 µmol/m(2), SiH, DMMS and 0.69 µmol/m(2), SiH, TES). The relatively higher surface coverage of SiH from TES over DMMS generated in sc-CO(2) is due to the inherent trialkoxy moiety of the TES that favours siloxane crosslinkage, forming polymeric surface attachments to yield a higher ligand density than the monomeric DMMS ligand. A conversion efficiency of ∼84.4% of SiH prepared from TES in sc-CO(2) estimated from (29)Si CP/MAS NMR analysis is comparable to TES silanization in dioxane or toluene. Moreover, silica hydride (SiH) conversion efficiency of ca. 42.4% achieved for the hydride intermediate prepared from DMMS in sc-CO(2) is more superior to 33.3% efficiency obtained in dioxane. The differences in conversion efficiencies is attributed to the ability of sc-CO(2) being able to access silica pores that are inaccessible in organic solvents. Bonded silica hydride from TES, DMMS prepared in sc-CO(2) were characterized using elemental analysis, thermogravimetric analysis (TGA), BET surface area, Fourier transform infrared (FI-IR) and solid state NMR spectroscopy. Silica hydride technology/chemical functionalization of silica in sc-CO(2) avoid extended purification steps (i.e. filtration and washing), generation of waste organic solvent and the need of costly or energy consuming drying processing with improved modification efficiency.

From single to multiple microcoil flow probe NMR and related capillary techniques: a review.

Nuclear magnetic resonance (NMR) spectroscopy is one of the most important and powerful instrumental analytical techniques for structural elucidation of unknown small and large (complex) isolated and synthesized compounds in organic and inorganic chemistry. X-ray crystallography, neutron scattering (neutron diffraction), and NMR spectroscopy are the only suitable methods for three-dimensional structure determination at atomic resolution. Moreover, these methods are complementary. However, by means of NMR spectroscopy, reaction dynamics and interaction processes can also be investigated. Unfortunately, this technique is very insensitive in comparison with other spectrometric (e.g., mass spectrometry) and spectroscopic (e.g., infrared spectroscopy) methods. Mainly through the development of stronger magnets and more sensitive solenoidal microcoil flow probes, this drawback has been successfully counteracted. Capillary NMR spectroscopy increases the mass-based sensitivity of the NMR spectroscopic analysis up to 100-fold compared with conventional 5-mm NMR probes, and thus can be coupled online and off-line with other microseparation and detection techniques. It offers not only higher sensitivity, but in many cases provides better quality spectra than traditional methods. Owing to the immense number of compounds (e.g., of natural product extracts and compound libraries) to be examined, single microcoil flow probe NMR spectroscopy will soon be far from being sufficiently effective as a screening method. For this reason, an inevitable trend towards coupled microseparation-multiple microcoil flow probe NMR techniques, which allow simultaneous online and off-line detection of several compounds, will occur. In this review we describe the current status and possible future developments of single and multiple microcoil capillary flow probe NMR spectroscopy and its application as a high-throughput tool for the analysis of a large number of mass-limited samples. The advantages and drawbacks of different coupled microseparation-capillary NMR spectroscopy techniques, such as capillary high-performance liquid chromatography-NMR spectroscopy, capillary electrophoresis-NMR spectroscopy, and capillary gas chromatography-NMR spectroscopy, are discussed and demonstrated by specific applications. Another subject of discussion is the progress in parallel NMR detection techniques. Furthermore, the applicability and mixing capability of tiny reactor systems, termed "microreactors" or "micromixers," implemented in NMR probes is demonstrated by carbamate- and imine-forming reactions.

Synthesis and characterisation of bonded mercaptopropyl silica intermediate stationary phases prepared using multifunctional alkoxysilanes in supercritical carbon dioxide as a reaction solvent.

This research employed (29)Si and (13)C Cross-Polarisation/Magic Angle Spinning (CP/MAS) NMR spectroscopy to characterise the nature and amount of surface species of di-and trifunctional mercaptopropylsilane (MPS) bonded silica using supercritical carbon dioxide (sc-CO(2)) as a reaction solvent without additives (co-solvent) or catalysts. The MPS stationary phases were prepared within 1h at a temperature and pressure of 70°C and 414 bar, respectively. Complementary analysis including elemental analysis, thermogravimetric analysis (TGA), DRIFT spectroscopy and BET surface area measurements were employed to characterise the bonded MPS intermediate stationary phases in support of data obtained from solid-state NMR analysis. The results revealed that modification of silica with a trimethoxymercaptopropylsilane (MPTMS) results in ligand surface coverage that is larger than when dimethoxymethylmercaptopropysilane (MPDMMS) is employed as a silanisation reagent. This observation is attributed to greater reactivity and cross-linkage of trifunctional silane. Reaction in sc-CO(2) in comparison to reflux in organic solvents, is rapid, reducing product recovery procedures.

Enlightening the past: analytical proof for the use of Pistacia exudates in ancient Egyptian embalming resins.

Mastic, the resinous exudate of the evergreen shrub Pistacia lentiscus, is frequently discussed as one of the ingredients used for embalming in ancient Egypt. We show the identification of mastic in ancient Egyptian embalming resins by an unambiguous assignment of the mastic triterpenoid fingerprint consisting of moronic acid, oleanonic acid, isomasticadienonic and masticadienonic acid through the consolidation of NMR and GC/MS analysis. Differences in the observed triterpenoid fingerprints between mummy specimens suggest that more than one plant species served as the triterpenoid resin source. Analysis of the triterpenoid acids of ancient embalming resin samples in the form of their methyl- and trimethylsilyl esters is compared. In addition we show a simple way to differentiate between residues of mastic from its use as incense during embalming or from direct mastic application in the embalming resin.

Retention and selectivity effects caused by bonding of a polar urea-type ligand to silica: a study on mixed-mode retention mechanisms and the pivotal role of solute-silanol interactions in the hydrophilic interaction chromatography elution mode.

The separation properties of five silica packings bonded with 1-[3-(trimethoxysilyl)propyl]urea in the range of 0-3.67 µmol m⁻² were investigated in the hydrophilic interaction chromatography (HILIC) elution mode. An increase of the ligand surface density promoted retention of non-charged polar compounds and even more so for acids. An opposite trend was observed for bases, while the amphoteric compound tyrosine exhibited a U-shaped response profile. An overall partitioning retention mechanism was incompatible with these observations; rather, the substantial involvement of adsorptive interactions was implicated. Support for the latter was provided by column-specific changes in analyte retention and concomitant selectivity effects due to variations of salt concentration, type of salt, pH value, organic modifier content, and column temperature. Silica was more selective for separating compounds differing in charge state (e.g. tyramine vs. 4-hydroxybenzoic acid), while in cases where structural differences of solutes resided in non-charged polar groups (e.g. tyramine vs. 5-hydroxydopamine, nucleoside vs. nucleobase) more selective separations were obtained on bonded phases. Hierarchical cluster analysis of the home-made urea-type and three commercial amide-type bonded packings evinced considerable differences in separation properties. The present data emphasise that the role of the packing material under HILIC elution conditions is hardly just the polar support for a dynamic coating with a water-enriched layer. Three major retention mechanisms are claimed to be relevant on bare silica and the urea-type bonded packings: (i) HILIC-type partitioning, (ii) HILIC-type weak adsorption such as hydrogen bonding between solutes and ligands or solutes and silanols (potentially influenced by individual degrees of solvation, salt bridging, etc.), (iii) strong electrostatic (ionic) solute-silanol interactions (attractive/repulsive). Even when non-charged polar bonded phases are used, solute-silanol interactions should not be discounted, which makes them a prime parameter to be characterised by HILIC column tests. Multi/mixed-mode type separations seem to be common under HILIC elution conditions, associated with a great deal of selectivity increments. They are accessible and controllable by a careful choice of the type of packing, the mobile phase composition, and the temperature.

Online coupling of enantioselective capillary gas chromatography with proton nuclear magnetic resonance spectroscopy.

The hyphenation of enantioselective capillary gas chromatography and mass spectrometry is not always sufficient to distinguish between structural isomers, thus requiring peak identification by NMR spectroscopy. Here the first online coupling of enantioselective capillary gas chromatography with proton nuclear resonance spectroscopy is described for the unfunctionalized chiral alkane 2,4-dimethylhexane resolved on octakis(6-O-methyl-2,3-di-O-pentyl)-gamma-cyclodextrin at 60 degrees C. NMR allows constitutional and configurational isomers (diastereomers and enantiomers) to be distinguished. Enantiomers display identical spectra at different retention times, which enable an indirect identification of these unfunctionalized alkanes. The presented method is still at an early development stage, and will require instrumental optimization in the future.

Retention and selectivity properties of carbamate pesticides on novel polar-embedded stationary phases.

This study describes the use of stationary phases with polar functionality suitable for the chemical analysis of carbamates pesticides and comparing with conventional alkyl C8 and C18 phases. The emphasis of this study was to compare the selectivity and retention of the pesticides on different stationary phases, bonded onto 1.7 microm partially porous silica particles under isocratic separation condition. Four stationary phases including: phenylaminopropyl (PAP) phase, bidentate propylurea-C18 (BPUC(18)), C8 and C18, were successfully bonded on the partially porous silica spheres as evidenced by (29)Si and (13)C solid-state NMR analysis. The phenylaminopropyl phase exhibited smaller retentivity and enhanced selectivity compared to the alkyl C8 phase; the analysis time to run separation of the six carbamate pesticides (i.e., methomyl, propoxur, carbofuran, carbaryl, isoprocarb, and promecarb) on the PAP phase was threefold faster than alkyl C8 phase. In a similar manner, the BPUC(18) phase shows similar selectivity to that of the PAP phase, but with longer retentivity; although the BPUC(18) phase is characterized with a lesser degree of retentivity for the carbamate pesticides than the conventional alkyl C18 phase. We propose that pi-pi and weak polar interactions between the carbamate pesticides and the PAP phase dominates the separation mechanism and providing a superior selectivity; faster separation time was also achieved as a result of smaller retentivity. Whereas the C8 and C18 bonded phases exhibits only hydrophobic interactions with the pesticides, leading to larger retentivity. The BPUC(18) phase is shown to interact via polar-polar interactions in addition to hydrophobic interactions with the pesticides, providing similar selectivity with the PAP phase but with larger retentivity.

Characterization of binding affinities in a chromatographic system by suspended state HR/MAS NMR spectroscopy.

In the current work a racemate of (R)- and (S)-benzylmandelate was separated with a stereoselective polysaccharide-based chiral stationary phase by HPLC. To elucidate the occurring chiral molecular recognition processes in the heterogeneous system used, NMR spectroscopy was chosen under high resolution/magic angle spinning (HR/MAS) NMR conditions in the suspended state. Therefore, and as a proof of concept, a combination of several NMR methods such as spin-lattice relaxation time (T(1)) measurements (T(1)), the saturation transfer difference, and the 2D experiment of the transferred nuclear overhauser enhancement spectroscopy technique were applied. With HR/MAS NMR it is feasible to combine NMR and chromatography to achieve further insights into the separation process.

Thermally induced radical hydrosilylation for synthesis of C18 HPLC phases from highly condensed SiH terminated silica surfaces.

Silicon hydride terminated silica surfaces were prepared at high temperatures by a chlorination-reduction sequence. SiH groups are desired for further surface modification as an alternative to the native silanol groups which are unfavorable for RPLC applications. Only few silanol groups remain in these materials and mostly SiH moieties with the highest degree of cross-linking are obtained. The retention properties of basic analytes on the SiH terminated material confirm that the surfaces is mostly free of silanols and that therefore the remaining SiOH groups are bulk species. A reagentless, radical initiated hydrosilylation reaction is introduced for the functionalization of the hydride terminated surface with 1-octadecene. (13)C CP/MAS NMR and DRIFT spectroscopy demonstrate the reaction of the carbon-carbon double bond and the SiH group as well as the linkage of C18 groups to the silica surface. These novel C18 materials show promising performance in RPLC separation, especially for the separation of organic bases.