Octadecylsilica Particles Research Articles

Systematic Evaluation of Different Coating Chemistries Used in Thin-Film Microextraction.

A systematic evaluation of eight different coatings made of solid phase extraction (SPE) and carbon-based sorbents immobilized with polyacrylonitrile in the thin-film microextraction (TFME) format using LC-MS/MS was described. The investigated coatings included graphene, graphene oxide, multi-walled carbon nanotubes (MWCNTs), carboxylated MWCNTs, as carbon-based coatings, and polystyrene-divinylbenzene (PS-DVB), octadecyl-silica particles (C18), hydrophilic–hydrophobic balance particles (HLB) and phenyl-boronic acid modified particles (PBA), as SPE-based coatings. A total of 24 compounds of diverse moieties and of a wide range of polarities (log P from −2.99 to 6.98) were selected as probes. The investigated coatings were characterized based on their extraction performance toward the selected probes at different pH values and at optimized desorption conditions. In the case of SPE-based coatings, PS-DVB and HLB exhibited a balanced extraction for compounds within a wide range of polarities, and C18 showed superior extraction recoveries for non-polar analytes. Carbon-based coatings showed high affinity for non-polar compounds given that their main driving force for extraction is hydrophobic interactions. Interestingly, among the studied carbon-based coatings, graphene oxide showed the best extraction capabilities toward polar compounds owing to its oxygen-containing groups. Overall, this work provided important insights about the extraction mechanisms and properties of the investigated coatings, facilitating the coating selection when developing new TFME applications.

Fused-silica capillary internally modified with nanostructured octadecyl silica for dynamic in-tube solid-phase microextraction of polycyclic aromatic hydrocarbons from aqueous media

The internal surface of a fused-silica capillary was made much more porous, more adsorptive and resistant to chemical and mechanical stresses by chemical coating using nanostructured octadecyl silica particles. The modification process was conducted using a simple and green nucleosynthesis procedure. The internally modified capillary was used as an in-tube solid-phase microextraction (IT-SPME) device for the extraction and preconcentration of ultratrace levels of polycyclic aromatic hydrocarbons (PAHs) in aqueous samples, followed by determination using gas chromatography-flame ionization detection (GC-FID) system. Morphology and structure of the sorbent was characterized by FT-IR, SEM and energy dispersive X-ray techniques. The effects of a range of experimental variables on the efficiency of the IT-SPME-GC-FID method were evaluated and optimized. Under the optimal conditions, good linearity (R2 > 0.99) was obtained for the calibration graphs over the range of 1–4000 ng mL−1. Detection limits were 0.22–0.47 ng mL−1 and relative standard deviations were obtained in the range 4.4–10.3%. Recoveries of the spiked samples were found to be 78.7–103.5%. The proposed IT-SPME-GC-FID strategy was applied successfully for the analysis of PAHs in real water samples. The results demonstrated good analytical performances, compared with those reported elsewhere for the sampling and quantification of PAHs in aqueous media.

Rapid enantiomeric separation and simultaneous determination of phenethylamines by ultra high performance liquid chromatography with fluorescence and mass spectrometric detection: application to the analysis of illicit drugs distributed in the Japanese market and biological samples

A rapid enantiomeric separation and simultaneous determination method based on ultra high performance liquid chromatography (UHPLC) was developed for phenethylamine-type abused drugs using (R)-(-)-4-(N,N-dimethylaminosulfonyl)-7-(3-isothiocyanatopyrrolidin-1-yl)-2,1,3-benzoxadiazole ((R)-(-)-DBD-Py-NCS) as the chiral fluorescent derivatization reagent. The derivatives were rapidly enantiomerically separated by reversed-phase UHPLC using a column of 2.3-µm octadecylsilica (ODS) particles by isocratic elution with water-methanol or water-acetonitrile systems as the mobile phase. The proposed method was applied to the analysis of products containing illicit drugs distributed in the Japanese market. Among the products, 1-(3,4-methylenedioxyphenyl)butan-2-amine (BDB) and 1-(2-methoxy4,5-methylenedioxyphenyl)propan-2-amine (MMDA-2) were detected in racemic form. Furthermore, the method was successfully applied to the analysis of hair specimens from rats that were continuously dosed with diphenyl(pyrrolidin-2-yl)methanol (D2PM). Using UHPLC-fluorescence (FL) detection, (R)- and (S)-D2PM from hair specimens were enantiomerically separated and detected with high sensitivity. The detection limits of (R)- and (S)-D2PM were 0.12 and 0.21 ng/mg hair, respectively (signal-to-noise ratio (S/N) = 3).

Fast CEC-MS using poly(dimethylsiloxane) microinjector, short packed column, and low-sheath-flow interface

A fast CEC-MS approach based on a microinjector and a short CEC column was developed. Poly(dimethylsiloxane) was used as the substrate for microinjector fabrication. A short capillary column (∼5 cm) packed with 5 μm octadecyl silica particles was inserted into the microinjector. The microinjector CEC device was interfaced to ESI-MS using a low-flow sheath liquid interface. The device delivers the advantages of sample introduction, pre-concentration, elution, and fast analysis as in chip-CEC yet avoids the difficulty of packing stationary material into the chip. The online pre-concentration and CEC-MS analysis capabilities of this device were demonstrated by analysis of a six-triazine mixture. A signal enhancement of 20-99-fold was achieved with a sample loading time of 180 s.

Flow-Through Fourier Transform Infrared Sensor for Total Hydrocarbons Determination in Water

A new flow-through Fourier transform infrared (FT-IR) sensor for oil in water analysis based on solid-phase spectroscopy on octadecyl (C18) silica particles has been developed. The C18 non-polar sorbent is placed inside the sensor and is able to retain hydrocarbons from water samples. The system does not require the use of chlorinated solvents, reducing the environmental impact, and the minimal sample handling stages serve to ensure sample integrity whilst reducing exposure of the analyst to any toxic hydrocarbons present within the samples. Fourier transform infrared (FT-IR) spectra were recorded by co-adding 32 scans at a resolution of 4 cm(-1) and the band located at 1462 cm(-1) due to the CH(2) bending was integrated from 1475 to 1450 cm(-1) using a baseline correction established between 1485 and 1440 cm(-1) using the areas as analytical signal. The technique, which provides a limit of detection (LOD) of 22 mg L(-1) and a precision expressed as relative standard deviation (RSD) lower than 5%, is considerably rapid and allows for a high level of automation.

Comprehensive supercritical fluid chromatography × reversed phase liquid chromatography for the analysis of the fatty acids in fish oil

The separation of the phenacyl esters of the fatty acids originating from a fish oil extract by means of a comprehensive analysis using silver-ion (SI) supercritical fluid chromatography (SFC) and reversed phase liquid chromatography (RP-LC) in the first and second dimensions, respectively, is described. The combination ensured a high orthogonality and peak capacity, particularly when compared to the comprehensive RP-LC × 2RP-LC separation achieved by using a configuration with two columns in parallel in the second dimension. The construction of the SI-SFC × RP-LC interface consists of two two-position/ten-port switching valves, of which one is equipped with two loops packed with octadecyl silica (ODS) particles. Compared to the SFC × RP-LC configuration described in an earlier publication, the peak capacity in the second dimension was increased. Water was not only added as make-up fluid to the SFC effluent to ensure analyte focusing, but also as rinsing medium of the loops prior to the transfer of the fractions to the second dimension. In the SFC dimension, high efficiency and loadability were obtained by coupling two wide-bore columns (4.6 mm ID) in series. Evaporative light scattering (ELSD) and ultraviolet (UV) detection with standard and high-pressure flow cells were evaluated in terms of data acquisition speed and suppression of signal interferences originating from the supercritical carbon dioxide (CO 2) expansion.

Microchip reversed-phase liquid chromatography with packed column and electrochemical flow cell using polystyrene/poly(dimethylsiloxane)

A microchip pressure-driven liquid chromatography (LC) with a packed column and an electrochemical flow cell has been developed by using polystyrene (PS) and poly(dimethylsiloxane) (PDMS). The cylindrical separation column with packed octadecyl silica particles was fabricated in the PS substrate. The three electrode system (working, reference, and counter electrode) for amperometric detection was fabricated onto the PS substrate, using the Au deposition, photolithography, and chemical etching. The detector flow cell was formed by sealing the electrode system with a PDMS chip containing a channel. In this flow cell, the effect of working electrode width (in the direction of flow) on chromatographic parameters, such as peak width and peak resolution were studied in electrode width ranging 50–5000 μ m. The effect of electrode width on sensitivity (current intensity, current density, and S/N ratio) was also examined. The sensitivity was discussed by simulating the concentration profile generated around the working electrode. The effects of the column packing size and the column size on the separation efficiency were examined. In this study, a good separation of three catechins was successfully achieved and the detection limits for (+)-catechin, epicatechin, and epigallocatechin gallate were 350, 450, and 160 nM, respectively.

Drug-Membrane Interactions Studied in Phospholipid Monolayers Adsorbed on Nonporous Alkylated Microspheres

Characterization of interactions with phospholipids is an integral part of the in vitro profiling of drug candidates because of the roles the interactions play in tissue accumulation and passive diffusion. Currently used test systems may inadequately emulate the bilayer core solvation properties (immobilized artificial membranes [IAM]), suffer from potentially slow transport of some chemicals (liposomes in free or immobilized forms), and require a tedious separation (if used for free liposomes). Here the authors introduce a well-defined system overcoming these drawbacks: nonporous octadecylsilica particles coated with a self-assembled phospholipid monolayer. The coating mimics the structure of the headgroup region, as well as the thickness and properties of the hydrocarbon core, more closely than IAM. The monolayer has a similar transition temperature pattern as the corresponding bilayer. The particles can be separated by filtration or a mild centrifugation. The partitioning equilibria of 81 tested chemicals were dissected into the headgroup and core contributions, the latter using the alkane/water partition coefficients. The deconvolution allowed a successful prediction of the bilayer/water partition coefficients with the standard deviation of 0.26 log units. The plate-friendly assay is suitable for high-throughput profiling of drug candidates without sacrificing the quality of analysis or details of the drug-phospholipid interactions.

Microstructure and Rheology of Thermoreversible Nanoparticle Gels

Naïve mode coupling theory is applied to particles interacting with short-range Yukawa attractions. Model results for the location of the gel line and the modulus of the resulting gels are reduced to algebraic equations capturing the effects of the range and strength of attraction. This model is then applied to thermo reversible gels composed of octadecyl silica particles suspended in decalin. The application of the model to the experimental system requires linking the experimental variable controlling strength of attraction, temperature, to the model strength of attraction. With this link, the model predicts temperature and volume fraction dependencies of gelation and modulus with five parameters: particle size, particle volume fraction, overlap volume of surface hairs, and theta temperature. In comparing model predictions with experimental results, we first observe that in these thermal gels there is no evidence of clustering as has been reported in depletion gels. One consequence of this observation is that there are no additional adjustable parameters required to make quantitative comparisons between experimental results and model predictions. Our results indicate that the naïve mode coupling approach taken here in conjunction with a model linking temperature to strength of attraction provides a robust approach for making quantitative predictions of gel mechanical properties. Extension of model predictions to additional experimental systems requires linking experimental variables to the Yukawa strength and range of attraction.

On-line monitoring of methanol in n-hexane by an organic-phase alcohol biosensor

An organic-phase alcohol biosensor has been developed by co-entrapping alcohol oxidase and horseradish peroxidase within an ionotropy polymer hydrogel matrix fabricated from silica gel particles, hydroxyethyl carboxymethylcellulose, an adduct of 3-methoxy-4-ethoxybenzaldehyde and 4-tert-butylpyridinium acetohydrazone, and octadecylsilica particles. The viability of the immobilised enzymes for the biocatalytic reaction of methanol in n-hexane was comparatively studied by using a bulk cell or a volume-changeable flow-through cell coupled with an oxygen optical transducer. It was found that the microenvironment around the enzyme, the deterioration property of the enzyme, the substrate throughput and the mass transfer process of the reactant in the bioreactor were the crucial parameters affecting the performance of the alcohol organic-phase biosensor. Our optimal biosensor was constructed from a flow-through cell packed with small particles of immobilised enzymes and it could maintain the biocatalytic reaction at high and stable rate for on-line detection of methanol in n-hexane under flow operation mode. The biosensor had an analytical working range of 2.3-90 mM methanol in n-hexane. The response times (t95) were 4.5 and 7.5 min for 60 and 10 mM methanol, respectively. The operational lifetime of the biosensor was more than 45 assays and the shelf lifetime was longer than 2 weeks. The biosensor has been successfully applied to determine the methanol content in a commercial gasoline-methanol blend sample with good recovery.

An efficient slurry packing procedure for the preparation of columns applicable in capillary electrochromatography and capillary electrochromatography‐electrospray‐mass spectrometry

AbstractA procedure is described for the slurry packing of 50‐μm ID fused silica capillaries with 3‐μm octadecyl silica (ODS) particles for capillary electrochromatography (CEC) and its hyphenation with electrospray ionisation mass spectrometry (ESI/MS). A homogeneous packed bed is obtained by using a slow packing process in an upward direction with a balanced density slurry solvent and MeOH as packing solvent. Special attention was paid to the in‐ and outlet frit preparation in order to avoid gas bubble formation which renders CEC‐ESI/MS problematic. Frits were made out of the packed bed itself, sintered in water, by using a perforated heating ribbon; they were not longer than 1 mm. In CEC‐UV, column efficiencies up to 300,000 plates per meter were obtained. Absence of gas bubbles was ascertained by the straightforward coupling to ESI/MS. A make‐up flow of 3 μL/min H2O/MeOH containing 0.1% HCOOH was used in the sheath flow interface. Steroids and carbamates were analysed with a 0.1% triethylamine‐acetic acid buffer (pH 8.9) containing varying amounts of acetonitrile. In CE‐ESI/MS, efficiencies dropped by ca. 20% but spectral data were excellent.

Large-bore particle-entrapped monolithic precolumns prepared by a sol–gel method for on-line peptides trapping and preconcentration in multidimensional liquid chromatography system for proteome analysis

The present report describes the preparation and characterization of large-bore particle-entrapped monolithic precolumns, which are suitable for incorporation into a two-dimensional liquid chromatography (2D-LC) system for proteome analysis. The fritless precolumns with different inner diameter (i.d.) (320 and 530 μm) were rapidly and successfully prepared by entrapping octadecylsilica (ODS) particles (5 μm, 300 Ǻ) prepacked into fused silica capillaries with a sol–gel network, which was formed by hydrolysis and polycondensation of methyltriethoxysilane (MTES). By optimizing the composition of the sol solution, the resulting large-bore monolithic precolumns of 5 mm length allow a flow rate of 20 μL/min loading buffer at a reasonable low back pressure of 25 bar or less and are capable of withstanding up to 300 bar inlet pressure. Scanning electron micrograms of the precolumns profile showed that the evolving sol–gel network joined particles to each other and onto the column wall, and no cracking or shrinkage of the column bed was observed even in 530 μm-i.d. capillary. The performance of the particle-entrapped monolithic precolumns used for preconcentration and desalting of proteolytic digest was evaluated by on-line coupling the large-bore precolumns with a capillary reversed-phase liquid chromatographic (RPLC) column followed by UV detection. The laboratory-made monolithic precolumns with 320 and 530 μm i.d. were characterized by using BSA tryptic digest or peptide standards as the analytes with respect to sample loading capacity, linearity, recovery and reproducibility, etc. The results indicate that the large-bore and short precolumns (5 mm × 320 μm i.d. or 5 mm × 530 μm i.d.) allow sample fast loading at a flow rate of 30 or 60 μL/min. The precolumns also have a mass loading capacity for BSA peptides of about 70 μg and for standard peptides of about 80 μg. Good linear calibration curves ( R 2 > 0.99) were obtained and the limits of detection (signal-to-noise ratio, S/N = 3) were improved by more than 60-fold and were between 0.53 and 1.32 ng/μL even with a UV absorbance detector. The total recovery was found to be approximately 90–100% for BSA digest and standard peptides. The day-to-day relative standard deviation (RSD) values for recoveries of BSA peptides on a single precolumn ranged from 4.66 to 7.56% and 2.68 to 3.05% for precolumn back pressure, while the column-to-column RSD values were 3.51–6.13% and 1.22–1.26% for recoveries of BSA peptides and precolumn back pressure, respectively. With good precolumn reproducibility, no significant degradation or decrease in precolumn performance was showed even after ∼150 preconcentration/desorption cycles. The precolumns also proved to be resistant to salt buffer with high concentration and low-pH mobile phase. The large-bore particle-entrapped monolithic precolumns will be further used in a high-throughput 2D-LC array system coupled with tandem matrix assisted laser desorption/ionization-time of flight–time of flight–mass spectrometry (MALDI-TOF–TOF–MS) detection for proteome analysis.

Isolation of phenolic antioxidant compounds by SFC

Specially designed chromatographic columns were evaluated for antioxidant compounds separation by supercritical fluid chromatography (SFC) with pure CO 2. Columns studied involved the use of commercial octadecylsilica particles (ODS) and silica particles coated with a stationary phase commonly used in gas chromatography, such as CW20M (polyethylene glycol) of high polarity. In order to select the appropriate chromatographic conditions to elute antioxidant compounds of phenolic structure, carnosic acid was selected as a model molecule and a theoretical study of the solubility of carnosic acid was performed. Carnosic acid was eluted using CO 2 as mobile phase (without modifiers) with a pressure programming up to 37 MPa (and at different temperatures) but only when coated particles were used. No elution was possible when ODS particles were tested, probably due to the strong interactions of the acid group of the molecule with the active sites remaining in the column. A separation of a complex mixture of a rosemary antioxidant extract (obtained by supercritical fluid extraction) using the columns designed in the present work is shown.

Influence of moderate Joule heating on electroosmotic flow velocity, retention, and efficiency in capillary electrochromatography

The influence of Joule heating on electroosmotic flow velocity, the retention factor of neutral analytes, and separation efficiency in capillary electrochromatography was investigated theoretically and experimentally. A plot of electrical current against the applied electrical field strength was used to evaluate the Joule heating effect. When the mobile phase concentration of Tris buffer exceeded 5.0 mM in the studied capillary electrochromatography systems using particulate and monolithic columns (with an accompanying power level of heat dissipation higher than 0.35 W/m), the Joule heating effect became clearly noticeable. Theoretical models for describing the variation of electroosmotic flow velocity with increasing applied field strength and the change of retention factors for neutral analytes with electrical field strength at higher Tris buffer concentrations were analyzed to explain consequences of Joule heating in capillary electrochromatography. Qualitative agreement between experimental data and implications of the theoretical model analysis was observed. The decrease of separation efficiency in capillary electrochromatography with macroporous octadecylsilica particles at high buffer concentration can be also attributed to Joule heating mainly via the increased axial diffusion of the analyte molecules and dispersion of solute bands by a nonuniform electroosmotic flow profile over the column cross-section. However, within a moderate temperature range, the contribution of the macroscopic velocity profile in the column arising from radial temperature gradients is insignificant.

Spongiform immobilization architecture of ionotropy polymer hydrogel coentrapping alcohol oxidase and horseradish peroxidase with octadecylsilica for optical biosensing alcohol in organic solvent.

An organic-phase optical alcohol biosensor consisting of alcohol oxidase and horseradish peroxidase coimmobilized in a spongiform hydrogel matrix of hydroxethyl carboxymethyl cellulose, an adduct of 3-methoxy-4-ethoxy benzaldehyde, 4-tert-butylpyridinium acetohydrazone, silica gel particles, and octadecylsilica particles in conjunction with an optical oxygen transducer has been successfully fabricated. The novel enzyme entrapment structure was mainly characterized with desirable solvent permeability, high efficiency of mass transfer for reactants, and good accessibility and stability of the immobilized enzymes. The biosensor could work in water-miscible solvent such as a solvent mixture of acetonitrile and phosphate aqueous buffer, as well as hydrophobic organic solvent such as n-hexane. The biosensor had the highest sensitivity to methanol in both solvent systems. Under the stop-flow mode, the biosensor had the analytical working ranges from 80 microM to 90 mM methanol in n-hexane and 0.10 to 90 mM methanol in acetonitrile/buffer. When the biosensor functioned in n-hexane, it could take benzaldehyde as an alcohol substrate and was free from any pH disturbance. In the presence of coimmobilized horseradish peroxidase, the operational life of the biosensor was 60 assays and the shelf life was longer than two weeks. The biosensor has been satisfactorily applied to the determination of methanol in commercial gasoline-methanol blend samples.

An organic-phase optical phenol biosensor coupling enzymatic oxidation with chemical reduction

This paper presents a recycle amplification optical biosensor to monitor phenol in hydrophobic organic solvents. Tyrosinase was first immobilised by entrapping it in a copolymer membrane of poly(vinyl alcohol)-hydroxyethyl carboxymethylcellulose doped with octadecylsilica particles. The biosensor was then constructed by co-mixing small particles of the immobilised tyrosinase with the adduct of L-ascorbic acid-poly(vinyl alcohol) (AsA-PVA) in conjunction with an optical oxygen transducer. The biosensor was characterised by its amplifying response to phenol, stable biocatalytic activity of entrapped-tyrosinase, free of interference from o-quinone polymerisation, and large water buffer capacity in hydrophobic organic solvents. The working range of the biosensor to phenol was 0.08-40 mmol dm(-3) in the flow mode. The response times (95%) of the biosensor were 4-7 min for phenol. The operational lifetime was more than 40 assays and the shelf lifetime of the biosensor was longer than 3 months. The biosensor has been successfully applied to quantify the phenol contents in some commercial ointment samples.

Hydrogel network entrapping cholesterol oxidase and octadecylsilica for optical biosensing in hydrophobic organic or aqueous micelle solvents.

Two optical cholesterol biosensors have been fabricated by immobilizing cholesterol oxidase (ChOx) and octadecylsilica (ODS) particles in hydrogel network matrixes of copolymer of poly(vinyl alcohol) (PVA)/hydroxyethyl carboxymethyl cellulose (HECMC), and sol-gel, respectively. In conjunction with an optical oxygen transducer, the immobilized ChOx in the sol-gel/ODS matrix was assembled as an optical cholesterol biosensor to continuously detect free cholesterol in aqueous micelle solution, while the immobilized ChOx in the PVA/HECMC/ODS matrix was constructed as an organic-phase optical cholesterol biosensor for the continuous analysis of free cholesterol in hydrophobic organic solvent. The compositions and properties of the immobilization matrixes, the effects of solvents and the analytical features were studied in detail. Both biosensors showed stable and reliable responses toward free cholesterol. For the aqueous micelle cholesterol biosensor, the analytical working range was from 0.05 to 8.0 mM cholesterol, the response time was 7-12 min, the operation life was more than 35 assays, and the shelf life was approximately 4 months. For the organic-phase cholesterol biosensor, the analytical working range was from 0.07 to 18.0 mM cholesterol, the response time was 4-8 min, the operation life was more than 120 assays, and the shelf life was longer than 5 months. The organic-phase cholesterol biosensor has been successfully applied to determine the free cholesterol content in commercial butter samples.

Synthesis of acylglucuronides of drugs using immobilized dog liver microsomes octadecylsilica particles coated with phospholipid

Immobilized dog liver microsome octadecylsilica (ODS) particles coated with phospholipid were developed for the synthesis of acylglucuronides of drugs. The phospholipid-coated ODS particles were readily prepared by stirring a solution containing l-α-dipalmitoylphosphatidylcholine with the ODS particles, in which the phospholipid was absorbed on the ODS surfaces by hydrophobic interaction between the acyl group of phospholipid and the otcadecyl group of the ODS particles. Similarly, the microsome-immobilized particles were readily prepared by stirring a buffer solution containing dog liver microsomes with the phospholipid-coated ODS particles, in which the microsomes were immobilized on the phospholipid-coated ODS particles by hydrophobic binding. The microsome-immobilized particles exhibited UDP-glucuronosyltransferase activity which catalyzed the glucuronidation of ketoprofen and a nonpeptide endothelin receptor antagonist, S-1255 ([ R]-[+]-2-[benzo(1,3)dioxol-5-yl]-6-isopropyl-4-[4-methoxyphenyl]-2 H-chromene-3-carboxylic acid), to the corresponding acylglucuronide in the presence of uridine 5 ′-diphosphate (UDP)-glucuronic acid, and two acylglucuronides of ketoprofen and S-1255 were synthesized using the microsome-immobilized particles. These acylglucuronides were synthesized by simply shaking the microsome-immobilized particles adsorbed on the substrate in a buffer solution containing UDP-glucuronic acid with a thermostated mixer. The molecular weights and chemical structures of the synthesized acylglucuronides were identified by mass spectrometry and nuclear magnetic resonance, respectively. The productivity of S-1255 acylglucuronide using microsome-immobilized particles was approximately threefold higher than that observed with free microsomes, whereas the ketoprofen acylglucuronide productivity was slightly lower than that observed with free microsomes. The present method should be very useful for the synthesis of acylglucuronides of drugs, which are slightly soluble aqueous solutions in the drug development stage.

Separation of selected humic degradation compounds by capillary electrochromatography with monolithic and packed columns

The separation of selected lignin/humic substance (HS) degradation compounds by capillary electrochromatography (CEC) with a methacrylate-based monolithic column and a conventional column packed with 5 microm octadecyl silica (ODS) particles is presented. The effects of organic modifier concentration, pH of the mobile phase, ionic strength, applied voltage, and temperature on the separation were investigated to determine the optimal separation conditions. With the increase of pH in the mobile phase, some of analytes start to ionize and both chromatographic partition and electrophoresis can play roles in separation simultaneously. Accordingly, different selectivity from high-performance liquid chromatography (HPLC) and capillary electrophoresis (CE) could be achieved. The performances of both kinds of columns were compared. The results showed that the peaks of compounds obtained on the former column were much wider than those on the latter one, although good separation efficiency of alkylbenzenes could be readily achieved; the most probable reasons for this behavior and method to solve this problem were briefly discussed. The CEC of a soil fulvic acid with a monolithic column produced partly resolved broad bands; by means of nuclear magnetic resonance (NMR) analysis a wide range of oxygen derived aromatic substitution patterns was found with prominent contributions from phenolic and carboxylic groups.

Immunoaffinity screening with capillary electrochromatography.

Highly efficient capillary electrochromatographic separations of cardiac glycosides and other steroids are presented. Employing butyl-derivatized silica particles as stationary phase resulted in a nearly three times faster electroosmotic flow (EOF) compared to capillary electrochromatography (CEC) with octadecyl silica particles. On-column focusing with a preconcentration factor of 180 was performed and separation efficiencies of up to 240,000 plates per meter were obtained. Using label-free standard UV absorbance, detection limits of 10-80 nM were reached for all steroids tested. For screening of cardiac glycosides, e.g., digoxin and digitoxin in mixtures of steroids, CEC was combined with immunoaffinity extraction using immobilized polyclonal anti-digoxigenin antibodies and F(ab) fragments. Simply adding small amounts of antibody carrying particles to the samples and comparing chromatograms before and after antibody addition allowed screening for high affinity antigens in mixtures with moderate numbers of compounds. Under conditions of competing antigens, affinity fingerprints of immobilized anti-digoxigenin and anti-digitoxin antibodies were obtained, reflecting the cross-reactivity of eleven steroids. The method provides high selectivity due to the combination of bioaffinity interaction with highly efficient CEC separation and UV detection at several wavelengths in parallel. This selectivity was exploited for the detection of four cardiac glycosides in submicromolar concentrations in an untreated urine sample.