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PubMed1994

Determination of pharmaceuticals and related impurities by capillary electrophoresis.

In the first part of this work, the use of capillary electrophoresis (CE) for the separation of two groups of pharmaceuticals, namely a metabolite of tamoxifen and a basic drug substance, DS1 was investigated. The effects of pH and types of modifiers, e.g. surfactant, bile salt, gamma-cyclodextrin and hydroxypropyl-beta-cyclodextrin on selectivity, separation and peak shape were studied. Besides achieving complete separation of the compounds, the CE system was capable of providing separation with significant improvements in overall peak shape of the compounds compared with HPLC. In the case of the basic drug substance DS1, validation of the CE system developed in terms of linearity, selectivity, sensitivity and reproducibility was satisfactorily performed. At the same time, a study of the sample solvent matrix effects on the separation of this group of compounds was examined. The system was successfully applied to the analysis of laboratory-synthesized samples. Good correlation was observed between CE and HPLC, although higher efficiency and faster speed of separation were obtained using the CE system developed. For the tamoxifen metabolite, special emphasis was placed on the use of CE for the separation of the pair of isomers. This was readily achieved through the introduction of gamma-cyclodextrin in the electrolyte. Resolution of at least 1.5 was obtained for the isomers using the CE method.

2-Hydroxypropyl-beta-cyclodextrin↗
PubMed1994

Analysis of structural specificity in antibody-antigen reactions by capillary electrophoresis with laser-induced fluorescence detection.

A rapid and simple procedure for screening antibodies for binding to an antigen is proposed. A fluorescent hapten-dye conjugate was prepared by labeling the amino moiety of the hapten with a commercially available reactive cyanine dye, Cy5 (excitation maximum: 650 nm, emission maximum: 670 nm). A fixed amount of the Cy5-hapten was titrated with serial dilution of the antibody. Each of the titration mixture was analyzed by capillary electrophoresis (25 cm x 20 microns column) monitored by laser-induced fluorescence (laser: 10 mW helium-neon, 632.8 nm). Free and antibody-bound Cy5-hapten were analyzed simultaneously on the electropherogram. Competitive immunoassay of hapten was demonstrated with low-end sensitivity of 5.10(-8) M, about 10x more sensitive than the present drug screening methods. Using morphine as an example, the screening of various antibodies (from different vendors) and cross-reactivity of morphine analogues using the present procedure will be discussed.

Antibodies↗
PubMed1994

Mixed polymer networks in the direct analysis of pharmaceuticals in urine by capillary electrophoresis.

Two-component polymer mixtures of polyethylene oxide-polydextran have been investigated as unique separation media for capillary electrophoresis. The effects of concentration of the individual polymers and their mixtures on the electroosmotic velocity and electrophoretic mobility of small pharmaceutical compounds were investigated. The molecular masses of polymers, buffer concentrations and percentages of organic solvents and cyclodextrins were varied to explore their effects on the separation process. The plate height against field strength curves were also generated for a better understanding of the kinetic processes involved. The two-component polymer mixtures were found as stable and selective media for the analysis of an anti-ulcer drug famotidine directly in untreated urine.

Buffers↗
PubMed1995

Separation of beta-lactoglobulin A, B and C variants of bovine whey using capillary zone electrophoresis.

beta-Lactoglobulin is a whey protein that affects milk composition and product functionality and which can be present in up to eight genetic variant forms. A free zone capillary electrophoresis method has been developed to separate and identify the beta-lactoglobulin A, B and C variants. Three buffer systems [borate, 2-(N-morpholino)-ethanesulphonic acid (MES) and bis(2-hydroxyethyl)imino-tris(hydroxymethyl)methane (Bistris)] were examined over a range of pH values and with the addition of the separation buffer modifiers Tween 20 and/or ethanolamine. The most successful combination of these was 50 mM MES at pH 8.0 with the addition of 0.1% Tween 20 which clearly resolved the three variants from both each other and from the other whey proteins even though the MES buffer was acting well outside its pKa range (pH 5.3-7.3). The retention times and identification of the individual variants were verified by spiking with commercially purified beta-lactoglobulin A and B proteins and a beta-lactoglobulin AC whey. The method was then used to phenotype beta-lactoglobulin in a sample population of New Zealand Jersey cows.

Amino Acid Sequence↗
PubMed1995

Suppression of chiral recognition of 3-hydroxy-1,4-benzodiazepines during micellar electrokinetic capillary chromatography with bile salts.

During the development of a micellar electrokinetic chromatographic screening method for 1,4-benzodiazepines, peak splitting and broadening were observed for some 3-hydroxy-1,4-benzodiazepines (oxazepam, lorazepam, temazepam and lormetazepam). This phenomenon occurred when the micellar phase consisted of bile salts and can be ascribed to the chiral nature of these surfactants. As the bile salts were applied in order to reduce the capacity factors to an appropriate level, enantiomer separation was not an objective and even disturbing. By increasing the analysis temperature, the chiral recognition of these compounds could be suppressed.

Anti-Anxiety Agents↗
PubMed1995

Rapid determination of drugs in biofluids by capillary electrophoresis. Measurement of antipyrine in saliva for pharmacokinetic studies.

A micellar electrokinetic capillary chromatography method was developed that permitted the resolution of antipyrine from endogenous compounds and its quantitation in neat saliva in as little as 1 min. Final conditions were: SpectraPhoresis 1000, 30(23) cm x 50 microns silica capillary, 50 mM sodium phosphate pH 9.6, 50 mM SDS, 10 s hydrodynamic load, detection scanning 200-300 nm or 260 nm, run 25 kV. To overcome the effects of Joule heating the capillary was cooled to 15 degrees C. Sensitivity was < 10 microM and linearity extended to 350 microM. Comparison with an HPLC assay demonstrated that hydrodynamic injection gave a loading bias unless samples and standards were of equal viscosity. For 75 samples from five subjects the correlation of CE vs. HPLC was then r = 0.99.

Antipyrine↗
PubMed1995

Resolution of the enantiomers of oxamniquine by capillary electrophoresis and high-performance liquid chromatography with cyclodextrins and heparin as chiral selectors.

The methods of separation of the enantiomers of the chiral drug oxamniquine are compared, between HPLC with either cyclodextrins and their related derivatives as chiral selectors in the mobile phase or immobilised in a chiral stationary phase (as Cyclobond I and II) and between capillary zone electrophoresis (CZE) where the cyclodextrins are added to the buffer solution. The HPLC experiments, which included structured method optimisation were largely unsuccessful in resolving the enantiomers, with the exception of when a Chiral-AGP protein stationary phase was introduced into the programme. However although this chiral stationary phase provided baseline resolution of the enantiomers the stability of the method was suspect to small changes in the pH (0.2 units). In contrast the CZE method developed for both cyclodextrins and their derivatives gave good resolution of the enantiomers and method stability (R.S.D. < 1%, n = 10 on precision). The basis of the interaction mechanism between selector and selectand was shown as a 1:2 relationship of cyclodextrin to analyte by NMR. In addition the polysaccharide, heparin was investigated as a chiral additive and excellent resolution of the oxamniquine was achieved with 3 mM heparin in 50 mM sodium dihydrogenphosphate (pH 3.0) as buffer in CZE, which also gave a stable procedure. This method allowed the detection of each of the enantiomers in the presence of the other down to 0.23% (m/m). The overall composition of the heparin material from different sources can however be slightly variable and this can result in small differences in resolution capability.

Capillary Action↗
PubMed1995

Comparison of capillary electrophoresis with traditional methods to analyse bovine whey proteins.

The separation of the four major whey proteins by sodium dodecyl sulphate (SDS)-capillary gel electrophoresis (CGE) is described. Whilst commercially purified whey proteins could be analysed using the recommended protocol, the more complex nature of an acid whey and a reconstituted whey protein concentrate (WPC) powder necessitated considerable refinement of the CGE sample buffer. Individual whey proteins in the acid whey and WPC samples were then also separated and quantitated using capillary zone electrophoresis, polyacrylamide gel electrophoresis (PAGE) and HPLC methods and the results were compared. The values obtained for alpha-lactalbumin (alpha-Lac) and beta-lactoglobulin (beta-Lg) were consistent throughout the various methods, although size-exclusion HPLC, SDS-PAGE and SDS-CGE could not separate the two beta-Lg variants or the glycosylated form of alpha-Lac from the beta-Lg. There was considerable variation in the values for the bovine serum albumin and immunoglobulin determined by the different methods and it was concluded that none of the methods could satisfactorily quantitate all four whey proteins.

Animals↗
PubMed1995

Separation and detection of 4-hexadecylaniline maltooligosaccharide derivatives with packed capillary liquid chromatography-frit fast atom bombardment-mass spectrometry.

A LC-MS method is under development for the separation and detection of mixtures of native glycolipids and of oligosaccharide derivatives. The LC system is based on slurry-packed capillary columns. Frit fast atom bombardment (frit-FAB) is used as the LC-MS interface and ionisation technique and the column is connected to the frit via a 50 microns I.D. fused-silica capillary liner. Post column addition of matrix is achieved using a 50 microns I.D. fused-silica capillary liner with 2.5% (v/v) matrix solution. The two liners are joined through a septum and end side by side against the frit. The detection limit was found to be less than 1 pmole in the negative ion mode. A mixture of tetra to deca maltooligosaccharides reductively aminated with 4-hexadecylaniline (M4-10-HDA) was separated on a straight phase silica column using gradient elution.

Aniline Compounds↗
PubMed1995

Analysis of trichloroacetic acid in the urine of workers occupationally exposed to trichloroethylene by capillary gas chromatography.

A gas chromatographic procedure is described for the determination of trichloroacetic acid in urine, the major metabolite of trichloroethylene exposure. Trichloroacetic acid was derivatised to its methyl ester with BF3/methanol reagent and then extracted into toluene and analysed by capillary gas chromatography using electron-capture detection. The response was linear in the range 0.4-100 mg/l of trichloroacetic acid in urine and showed a relative recovery of 99.6%. The procedure is suitable for monitoring occupational exposure to trichloroethylene.

Absorption↗
PubMed1995

On-line coupling of micellar electrokinetic chromatography to electrospray mass spectrometry.

The possibility of selectivity enhancement in capillary electrophoresis-mass spectrometry (CE-MS) by hyphenating micellar electrokinetic chromatography (MEKC) and electrospray mass spectrometry (MS) is described for two quaternary ammonium compounds. Direct coupling of MEKC to MS is hazardous because of the contamination of the ion source due to presence of an excess of micelle forming agent in the MEKC buffer. Therefore, a coupled-capillary setup with the possibilities of voltage switching and buffer renewal has been designed. Such a system allows on-line heartcutting of the zones of interest in the MEKC capillary with subsequent transfer via a second capillary to the mass spectrometer.

Buffers↗
PubMed1989

Complete denture retention. Part I: Physical analysis of the mechanism. Hysteresis of the solid-liquid contact angle.

Analysis of physical forces involved in the mechanism of denture retention clearly shows that the hysteresis of the liquid-solid contact angle governs the viscoelastic response of the system dislodgment. This result is in contradiction to the most commonly admitted view according to which perfect wettability is necessary to obtain good retention and implies the existence of the high advancing contact angle of saliva on a prosthetic material.

Capillary Action↗
PubMed1990

A lining system for composite resin fillings.

Microleakage at the margins is a major problem with composite resin fillings. This study was performed to determine whether a liner that expands on contact with water could prevent leakage. A microfilled composite resin that has a high rate of contraction (taking place while curing) was used in class V cavities of extracted teeth. An experimental liner containing aluminosilicate crystals was painted on the dentin walls before insertion of the filling material. The specimens were thermocycled and sectioned. Class V fillings were also placed in the teeth of beagles. The teeth were extracted after 14 weeks. The depth to which the dye penetrated was measured microscopically. Results showed that the expanding liner almost completely prevented penetration of the dye in both in vitro and animal experiments.

Adolescent↗