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F M Everaerts

Publications and source records attributed to F M Everaerts.

At least 19 recordsLinked to original sources

Chiral interactions in capillary zone electrophoresis: computer simulation and comparison with experiment.

Chiral interaction in capillary electrophoresis can be modeled using pK values, mobilities of analytes, and their formation constants with the chiral selector. An existing steady-state simulation program for CE (HPCESIM) was recently extended with a chiral submenu involving the chiral parameters listed above. These were experimentally determined in both our laboratories for mandelic acid and terbutaline using hydroxypropylated beta-cyclodextrin as chiral selector. A comparison was made between both sets of parameters and between experimental electropherograms and those obtained from simulation. Error analysis of the results indicate the sensitivity of the obtained results.

Computer Simulation↗

Quality control of histamine and methacholine in diagnostic solutions with capillary electrophoresis.

Solutions of histamine and methacholine bromide in different matrices for diagnostic purposes were analyzed for stability and quality control using capillary electrophoresis. Histamine (2,[4-imidazolyl]ethylamine) [CAS No. 51-45-6] was determined using a 0.1 M Tris-borate buffer of pH 8.3 with 5.10(-5) M cetyltrimethylammonium bromide (CTAB) and 0.005% poly(vinyl alcohol) (PVA) and detected at 214 nm using clenbuterol [4-amino-alpha-(tert.-butylaminomethyl)-3,5-dichlorobenzyl alcohol] [37148-27-9] as an internal standard. Metacholine bromide (acetyl-beta-methacholine bromide) [333-31-3] was determined with a 0.01 M creatinine-chloride buffer of pH 4.85 and detected with indirect UV at 230 nm using potassium as an internal standard. Histamine solutions were stable for a prolonged period of time, whereas under enforced degradation conditions methacholine was hydrolyzed, yielding acetic acid and (tentatively) beta-methylcholine as reaction products.

Drug Stability↗

Training software for electrophoresis.

Computer programs, simulating electrophoretic separations, were evaluated and discussed with respect to their suitability for training purposes. Quite a number of them, mainly those dealing with steady-state phenomena, are sufficiently fast and user-friendly for the purpose of visualization of electrophoretic principles. Transient-state or dynamic models, however, are more suitable for the advanced user, mainly because of their inherent complexity and long calculation times.

Electrophoresis↗

Detection of point mutations in DNA using capillary electrophoresis in a polymer network.

The use of capillary electrophoresis (CE) in a polymer network for single-strand conformation polymorphism (SSCP) is investigated. SSCP is a method to detect DNA point mutations, essential in the diagnosis of several diseases. The PCR (polymerase chain reaction) amplified p53 gene, a tumour suppressor gene known to be frequently mutated in malignant cells, was subjected to CE analysis. Two single-strand DNA fragments of 372 bp in length differing in only one nucleotide could be separated. We conclude that SSCP using CE in a polymer network is a powerful method for the detection of point mutations in DNA sequences.

Base Sequence↗

Determination of aminoglycoside antibiotics in pharmaceuticals by capillary zone electrophoresis with indirect UV detection coupled with micellar electrokinetic capillary chromatography.

Aminoglycoside antibiotics can be determined by capillary zone electrophoresis (CZE) with indirect UV detection in the anionic mode with a reversed electroosmotic flow (EOF) by addition of FC 135 to the background electrolyte. The effective mobilities of thirteen aminoglycoside antibiotics were determined as a function of pH. Applying CZE with indirect UV detection in the anionic mode and reversed EOF coupled with micellar electrokinetic capillary chromatography with the cationic surfactant cetyltrimethylammonium bromide, both neutral and charged antibiotics can be determined in combined pharmaceuticals. As an example, neomycin and hydrocortisone were determined in Otosporin eardrops.

Aminoglycosides↗

Determination of sulphonamides in pork meat extracts by capillary zone electrophoresis.

For sixteen sulphonamides the effective mobility was measured as a function of pH and from the effective mobilities determined in two different electrolyte systems the pK value and mobility at infinite dilution were calculated. A pH of 7.0 was found to be the optimum pH for the separation for both standard mixtures and mixtures of sulphonamides dissolved in pork meat extracts. For the determination of the sulphonamides in pork meat only a very simple pretreatment consisting of extraction with acetonitrile and centrifugation is suitable, as the matrix effects at pH 7.0 do not affect the separation. Calibration graphs for five sulphonamides were constructed, and regression coefficients of at least 0.999 were obtained. The limit of detection for the method varies from 2 to 9 ppm for a pressure injection time of 10 s (injection volume ca. 18 nl) using a Polymicro Technology capillary of length 116.45 cm, distance between injection and detection 109.75 cm and I.D. 50 microns.

Animals↗

Comparison of isotachophoresis, capillary zone electrophoresis and high-performance liquid chromatography for the determination of salbutamol, terbutaline sulphate and fenoterol hydrobromide in pharmaceutical dosage forms.

Reversed-phase high-performance liquid chromatography (RP-HPLC), isotachophoresis (ITP) and capillary zone electrophoresis (CZE) were applied to the determination of salbutamol, terbutaline sulphate and fenoterol hydrobromide in commercially available pharmaceutical dosage forms. The comparison showed that especially with the use of ITP, high concentrations of other charged sample components can disturb the separation process. If special attention is paid to ensure a complete separation, all methods give comparable results. For the regression lines of the calibration graphs, regression coefficients of at least ca. 0.999 and nearly zero intercepts are obtained with relative standard deviations of ca. 1-2% for peak area or zone lengths. By applying the different techniques, often different components of the sample matrix can be detected, i.e., a more complete impression of the sample composition can be obtained by using all the three techniques.

Albuterol↗

Capillary electrophoresis of peptides. Analysis of adrenocorticotropic hormone-related fragments.

Capillary electrophoresis can be used successfully to analyse small peptides to give additional information to that obtained using high-performance liquid chromatography (HPLC). The separation of a modified adrenocorticotropic hormone (4-9) fragment (Org 2766) and several of its fragments was investigated using capillary zone electrophoresis. Prediction of migration in aqueous systems using pKa-related data and the migration behaviour using sodium dodecyl sulphate in the buffer are discussed, as is the choice of buffer systems. The electrophoretic patterns are compared with the HPLC separation.

Adrenocorticotropic Hormone↗

Ultraviolet-absorbing organic anions in uremic serum separated by capillary zone electrophoresis, and quantification of hippuric acid.

Organic anions accumulated in blood serum of patients with chronic renal failure were separated by a novel technique: closed-system capillary zone electrophoresis (CZE) in a pH6 carrier-electrolyte system. Hippuric acid (HA), p-hydroxyhippuric acid, and uric acid were identified by their co-elution with standards prepared in ultrafiltered normal serum and by comparison with the corresponding ultraviolet-detected peaks positively identified in the HPLC analyses. Analysis time for the entire profile is 8 min. Repeatabilities (CVs) of CZE migration times and peak areas of the three acids in serum samples were about 0.7% and 6%, respectively. We quantified HA in 10 ultrafiltered uremic serum samples and compared results with those by a previously described HPLC procedure. The very good agreement further supports the identification of hippuric acid. Accuracy and precision of the CZE method were similar to those for the HPLC gradient-elution method, but analysis time for HA (8 min) is much less than by HPLC (90 min). Our technique is very suitable for selective, rapid analysis for (ultraviolet-absorbing) anionic constituents in ultrafiltered uremic serum, without any sample pretreatment.

Anions↗

Concept of response factor in capillary isotachophoresis. Determination of drugs in solution for intravenous injection.

Twenty-six drugs in solutions for intravenous injection were determined by capillary isotachophoresis with only one calibration point for each component. The maximum deviation of the labelled concentration was 2%. A new calibration constant is introduced, viz., the response factor (RF, dimensionless), which is independent of the diameter of the capillary, the construction of the universal detector and the driving current used during detection. It is shown that the RF can be used on different equipment, using different currents during detection. It appears that the RF is usable for routine analysis when a deviation of 5% is acceptable. Daily one-point recalibration, however, improves this value to 1%.

Electrophoresis↗

A first screening for hemocompatibility of a universal support for selective and specific hemoperfusion.

A modified filmadsorber is presented, intended to be used for selective and specific hemoperfusion. It consists of a spirally wound cellulose nitrate film, onto which--after chemical activation with sodium periodate--albumin is bound, stabilized, sterilized and activated with glutaraldehyde. Various bioactive ligands containing amino groups can be coupled to this support. A subsequent treatment with dimethylamino borane stabilizes the bonds between cellulose nitrate, albumin, glutaraldehyde and ligand. Columns in which a second layer of albumin is bound to the support as a model for a bioactive ligand were first screened for hemocompatibility using rabbits. Leukocyte, thrombocyte and hematocrit behaviour during hemoperfusion showed that hemocompatibility of the support was good.

Adsorption↗

Analysis of the aqueous phase of human cervical mucus by reversed-phase high-performance liquid chromatography and capillary isotachophoresis.

The aqueous phase of human cervical mucus was analysed by reversed-phase high-performance liquid chromatography (HPLC) and capillary isotachophoresis (ITP). With HPLC, seventeen ultraviolet-absorbing and eight fluorescent components and with ITP five anionic and four cationic components could be determined. The sample pre-treatment consisted of a simple ultrafiltration. Ten samples from fertile women and eleven samples from infertile women were analysed. In six samples from the infertile group higher median concentrations of several components were found. This may be an indication of disturbances in the biochemical processes of the cervical mucus of woman with fertility problems.

Cervix Mucus↗

Isotachophoresis as a candidate reference method in analytical chemistry. Determination of sodium in serum.

Isotachophoresis seems a likely candidate for a reference method, as it is more accurate and precise than common routine methods. The response is highly linear and depends on a well defined transport number of the leading ion, stability of the driving current, mobility of the separand, which is well controlled by the leading electrolyte and the use of a high-resolution detector. The suitability of isotachophoresis as a reference method was investigated for the determination of sodium in human serum. The operational conditions were 0.01 M K+/citrate (leading electrolyte) at pH 5.5 and 0.01 M creatinine X HCl (terminating electrolyte). Both n-butylamine and ammediol could be used as internal standards. The calibration graph constructed from standard solutions, diluted by weight with the internal standard, yielded a correlation coefficient of 0.99994 (n = 50) in the working range. The method seems especially useful for the determination of any ionic solute in, e.g., clinical samples (lithium, calcium, creatinine or drugs).

Butylamines↗

Isotachophoresis of allergenic extracts.

One aspect of the isotachophoretic determination of protein patterns in biological samples of interest is the characterization of allergens. This group of (glyco) proteins, causing allergic reactions, is used both for diagnosis and in the treatment of allergy. The aim of this investigation was to obtain a maximum amount of information, within one run, on the (glyco)protein composition of a number of allergenic extracts (e.g., from pollen or house dust mites). Commercially available extracts were dialysed prior to analysis to remove disturbing buffer constituents. A high-pH system was chosen in order to obtain a maximum amount of information from the samples (1-2 microliter). The leading electrolyte was 0.01 M C1-, buffered with Tris (pH 8.2), containing 0.2% w/v hydroxyethylcellulose, and the terminating electrolyte was beta-alanine, buffered to pH 10 with Ba(OH)2. The total analysis time was 15-20 min using a PTFE capillary (0.2 mm I.D.). The pre-separation current was 30 microA and the current during detection was 15 microA. UV absorption was measured at 280 nm. For optimal discrimination of the compounds of interest, an ampholyte mixture was used for spacing. The analytical procedure yielded highly reproducible UV patterns. Significant differences between various allergenic extracts were observed. It was concluded that isotachophoresis is a powerful method for the physico-chemical characterization of individual allergenic extracts, e.g., with respect to manufacturing and quality control.

Allergens↗

Determination of quinine in beverages, pharmaceutical preparations and urine by isotachophoresis.

The suitability of isotachophoresis for the determination of quinine in different samples was investigated. The operational conditions were 0.01 M potassium-morpholinoethanesulphonic acid (MES) (pH 6.0) with 0.05% Mowiol as the leading electrolyte and ca. 0.005 M creatinine-MES as the terminating electrolyte. The analyses were carried out at 25 microA in a 0.2 mm I.D. PTFE capillary with UV and conductivity detection. Quinine-containing beverages were degassed by sonification and directly injected. The limit of detection was 5 mg/l with a 4 microliter injection volume. The allowed concentrations could be determined with sufficient accuracy. Analgesic preparations were dissolved in a solution of 5 X 10(-3) M MES with sonification. The quinine levels found agreed well with the declared values. The other constituents of the pharmaceuticals did not interfere with the analysis. Urine samples from volunteers were analysed after consumption of tonic. The samples were extracted with dichloromethane-isopropanol (95:5), vortexed, centrifuged, evaporated to dryness, the residue dissolved in 5 X 10(-3) M MES and analysed. At a concentration factor of 33, the limit of detection was ca. 60 micrograms in 48-h urine: 2-15% of the quinine consumed was excreted as the parent compound in the first 48 h after consumption. The combination of the extraction procedure and the operational system makes the method suitable for the determination of a number of other alkaloids in physiological samples.

Analgesics↗

Determination of heavy metals by isotachophoresis.

The suitability of isotachophoresis for the analysis of metals in, e.g., environmental samples was studied. In a cationic operational system the heavy metals Fe, Cu, Ni, Cd, Co, Zn, Pb and Mn were simultaneously determined. The separation was achieved through complex formation with one of the counter ions, hydroxyisobutyric acid. The other counter ion was acetic acid, the leading ion was 0.02 M potassium or sodium (pH 4.1) and the terminator was H+. The analysis time was 15 min at 60 microA in a 0.2 mm I.D. capillary. Aqueous samples containing ppm and ppb amounts were enriched on a cation exchanger with an extremely low affinity for sodium (Chelex 100). Good recovery, linearity, precision and accuracy were obtained even down to the ppb range. Although the sensitivity of the method is not greater than that of some of the more established methods for the individual metals, a great advantage of isotachophoresis is the simultaneous determination of the metals, with equal response factors. An example is given of the determination of metals, including aluminium, in serum.

Cation Exchange Resins↗

Determination of conjugated bile acids in human bile by isotachophoresis in a non-aqueous solvent using a.c. conductivity and UV detection.

A method for the determination of conjugated bile acids in human bile using isotachophoresis in 95% methanol is described. The leading ion is 0.01 M chloride, the counter ion is hydroxylamine at its pK value and the terminating ion is N-2-hydroxyethylpiperazine-N'-2-ethanesulphonic acid (HEPES). The sample preparation consists of C18-silica cartridge adsorption. Microlitre amounts of the methanol eluate are injected and analysed within 20 min in a 0.2 mm I.D. PTFE capillary. The sensitivity of the method is better than 50 ng of each of the conjugated bile acids using a.c. conductivity detection.

Bile↗