Biomedical subjects

H Lingeman

Publications and source records attributed to H Lingeman.

At least 19 recordsLinked to original sources

PubMed1999

Coupling of biological sample handling and capillary electrophoresis.

The analysis of biological samples (e.g., blood, urine, saliva, tissue homogenates) by capillary electrophoresis (CE) requires efficient sample preparation (i.e., concentration and clean-up) procedures to remove interfering solutes (endogenous/exogenous and/or low-/high-molecular-mass), (in)organic salts and particulate matter. The sample preparation modules can be coupled with CE either off-line (manual), at-line (robotic interface), on-line (coupling via a transfer line) or in-line (complete integration between sample preparation and separation system). Sample preparation systems reported in the literature are based on chromatographic, electrophoretic or membrane-based procedures. The combination of automated sample preparation and CE is especially useful if complex samples have to be analyzed and helps to improve both selectivity and sensitivity. In this review, the different modes of solid-phase (micro-) extraction will be discussed and an overview of the potential of chromatographic, electrophoretic (e.g., isotachophoresis, sample stacking) and membrane-based procedures will be given.

Electrophoresis, Capillary↗

PubMed1999

Comparison between transient isotachophoretic capillary zone electrophoresis and reversed-phase liquid chromatography for the determination of peptides in plasma.

Low levels of peptide drugs in human plasma can be determined employing off-line solid-phase extraction, followed by capillary zone electrophoresis with UV detection. A bioanalytical procedure is presented, using gonadorelin and angiotensin II in human plasma as model compounds. The solid-phase extraction method, based on a weak cation exchange mechanism, is able to remove interfering endogenous components from the plasma sample, extract the model peptides quantitatively, and give a possibility of concentrating the sample at the same time. Transient isotachophoretic conditions were kept to increase the sample loadability by about two orders of magnitude. Up to about 70% of the capillary was filled with the reconstituted extract, whereafter the peptides were selectively concentrated during the first 15 min. Subsequently, the concentrated sample zones were separated under capillary zone electrophoresis conditions, showing the technique's high resolution. For the model cationic peptides (gonadorelin, angiotensin II) good linearity and reproducibility was observed in the 20-100 ng/mL concentration range. A more extensive washing procedure permits quantitation of gonadorelin at the 5 ng/mL level. In comparison with a liquid chromatography analysis, superior mass sensitivity and separation are obtained with the transient isotachophoretic capillary zone electrophoresis method. Moreover, in this case equivalent sensitivity is achieved when it is directly compared with a liquid chromatography method with UV detection, keeping in mind that 60 times more sample is needed for the latter method. A further gain in sensitivity can be obtained when the analysis is combined with native fluorescence detection, as is demonstrated by combining liquid chromatography separation with fluorescence detection.

Chromatography, Liquid↗

PubMed1999

Determination of roquefortine C in blue cheese using on-line column-switching liquid chromatography.

A method is described for the determination of roquefortine C in (blue) cheese. After liquid-liquid extraction with a mixture of hydrochloric acid and methanol, and filtration, an aliquot is analysed using column-switching reversed-phase liquid chromatography. The recovery of roquefortine C in Fetta cheese is about 85%, the calibration curve is linear from 10 to 2500 ng g(-1) (r2 = 0.998), and the detection limit is about 10 ng g(-1). In different batches of Danish Blue concentrations of 1000-2000 ng g(-1) of roquefortine C are found. As regards the stability of roquefortine C its half-life in diffuse daylight is ca. 50 min, while after irradiation with ultraviolet light, it is about 10 min.

Acetates↗

PubMed1999

Capillary electrophoresis as a versatile tool for the bioanalysis of drugs--a review.

This review article presents an overview of current research on the use of capillary electrophoretic techniques for the analysis of drugs in biological matrices. The principles of capillary electrophoresis and its various separation and detection modes are briefly discussed. Sample pretreatment methods which have been used for clean-up and concentration are discussed. Finally, an extensive overview of bioanalytical applications is presented. The bioanalyses of more than 200 drugs have been summarised, including the applied sample pretreatment methods and the achieved detection limits.

Chromatography, Micellar Electrokinetic Capillary↗

PubMed1999

On-line dialysis-SPE-CE of acidic drugs in biological samples.

A fully automated method is presented for the determination of acidic drugs in urine and serum using on-line dialysis-solid-phase extraction (SPE)-capillary electrophoresis (CE) with UV detection. With non-steroidal anti-inflammatory drugs (NSAIDs) as test compounds, detection limits in the biological samples were 0.05-1.0 microgram ml-1. Calibration plots were linear over two orders of magnitude and the within-day and between-day repeatability were better than 10%. The CE capillary and SPE column were used for over 500 analyses; the dialysis membrane was replaced after 250 analyses. A general protocol for dialysis-SPE-CE which can be used for amphoteric and acidic drugs was devised. The present results show that this protocol has general validity and can be recommended for future work on other classes of drugs.

Acids↗

PubMed1998

At-line solid-phase extraction for capillary electrophoresis: application to negatively charged solutes.

The analysis of complex biological samples with capillary electrophoresis (CE) requires proper sample pretreatment. In this paper the applicability of solid-phase extraction (SPE) coupled at-line with CE is studied, by using a laboratory-made interface. A fresh (disposable) SPE cartridge is used for each sample to prevent carry-over effects. The sample handling procedure is performed parallel with the analysis of the previous sample, to improve sample throughput. Using this set-up, negatively charged test compounds (some non-steroid anti-inflammatory drugs) can be determined in serum and urine. The method is linear over at least two decades and detection limits are around 40 microg/l. A single capillary, flushed only once a week with a sodium hydroxide solution, was used without problems for the analysis of ca. 900 samples during 1 year. The robustness of the system was very good: no blocking of loop, interface or capillary was found during this period. Furthermore, the system was successfully used for overnight runs.

Anti-Inflammatory Agents, Non-Steroidal↗

PubMed1998

Determination of phenprocoumon in plasma and urine using at-line solid-phase extraction-capillary electrophoresis.

The use of capillary electrophoresis (CE) for the analysis of biological samples is rather problematic because of the large number of interferences present in the matrix. One of the possibilities to solve such problems is to couple solid-phase extraction (SPE) at-line with CE, a technique developed in our laboratory. In this study at-line SPE-CE is performed for the determination of the anticoagulant phenprocoumon in biological fluids. Plasma samples are injected after the addition of 1 vol.% of formic acid to release the drug from binding proteins, while urine samples can be directly injected. The procedure is linear between 0.2 and 30 microg ml(-1) with a correlation coefficient, r2, of 0.9996. The detection limit in plasma is 0.1 microg ml(-1), which is fully adequate in view of the concentrations, that have to be dealt with in practice. The phenprocoumon concentration in a plasma sample of a patient treated with the anticoagulant was 3.8 microg ml(-1).

Electrophoresis, Capillary↗

PubMed1998

Determination of roquefortine C in blue cheese using on-line column-switching liquid chromatography.

A method is described for the determination of roquefortine C in (blue) cheese. After liquid liquid extraction with a mixture of hydrochloric acid and methanol, and filtration, an aliquot is analysed using column-switching reversed-phase liquid chromatography. The recovery of roquefortine C in Fetta cheese is about 85%, the calibration curve is linear from 10 to 2500 ng g(-1) (r2 = 0.998), and the detection limit is about 10 ng g(-1). In different batches of Danish Blue concentrations of 1000-2000 ng g(-1) of roquefortine C are found. As regards the stability of roquelfortine C its half-life in diffuse daylight is ca. 50 min, while after irradiation with ultraviolet light, it is about 10 min.

Cheese↗

PubMed1998

Derivatization in capillary electrophoresis.

In recent years capillary electrophoresis (CE) has been developed into a versatile separation technique, next to gas and liquid chromatography (LC), well suited for the determination of a wide variety of e.g., pharmaceutical, biomedical and environmental samples. The main advantages of CE over chromatographic separation techniques are its simplicity and efficiency. It is well recognized, however, that the sensitivity and selectivity of the detection are relatively weak points of CE. One way to overcome these limitations is the conversion (derivatization) of the analytes into product(s) with more favourable detection characteristics. Although, in principle, almost any detection mode can be combined with a derivatization procedure, in practice, fluorescence monitoring is favoured in most cases. This paper aims to give a short overview on the various reagents that can be used for pre-, post- and on-column derivatization in CE. First, a short introduction is given on CE as an analytical technique, followed by a discussion of the pros and cons of the various modes of derivatization, a comparison of derivatizations in CE with derivatizations in LC, the principles of fluorescence and prerequisites for a good fluorophore and the potential of using diode lasers in combination with a labelling procedure. With respect to the derivatization reagents the emphasis is on the labelling of amino, aldehyde, keto, carboxyl, hydroxyl and sulfhydryl groups.

Amines↗

PubMed1998

On-line dialysis solid-phase extraction coupled to capillary electrophoresis.

A fully automated dialysis solid-phase extraction (SPE) sample preparation procedure is coupled on-line to capillary electrophoresis (CE) for the first time. The system is used to determine sulfonamides in serum and urine. The dialysis unit serves to remove proteins and particulate matter. Reconcentration of the analytes is performed with a small SPE column while (in)organic salts and other interferences are removed simultaneously. Finally, the analytes are desorbed and injected, via a homemade interface, into the CE system. Limits of detection (LOD) of 0.05-0.1 and 0.05-0.3 microg/mL are obtained in urine and serum, respectively. The within-day and between-day precisions are in the range of 2-6% and 3-8%, respectively, for a concentration of five times the LOD. The dialysis SPE-CE system was used over a period of six months for the analysis of over 500 serum and urine samples without problems such as clogging of the CE capillary or SPE column.

Body Fluids↗

PubMed1998

Development and validation of transient isotachophoretic capillary zone electrophoresis for determination of peptides.

Although capillary zone electrophoresis (CZE) is known for its high resolution power and low mass detection limits, the concentration detection limits are rather poor when ultraviolet absorbance detection is used. To overcome this limitation, several on-column transient isotachophoresis (tITP) protocols have been developed and validated for the determination of both cationic and anionic model peptides, separately. Using this preconcentration method, up to 72% of the capillary can be filled with sample solution, without any loss in resolution. Thus, without any modification of the hardware set-up, the sensitivity is increased about two orders of magnitude. For the model cationic peptides (gonadorelin, angiotensin II) good linearity and reproducibility is observed in the 20 to 100 ng/mL concentration range. For the anionic peptides (N-t-Boc-Pentagastrin and two related peptides), a tITP method was developed using a dynamically coated capillary. The coating was prepared by adding Fluorad FC-135 to the leading electrolyte buffer. In this way a positively charged bilayer was formed on the inside of the capillary, producing an electroosmotic flow towards the outlet using reversed polarity conditions. In this way, acceptable analysis times were achieved. Using the developed tITP method, up to 72% of the capillary can be filled with sample solution as well. The anionic peptides are separated even better than when using CZE conditions. Linearity and reproducibility in the 20-100 ng/mL range proved to be excellent.

Angiotensin II↗

PubMed1997

Evaluation of phytic acid as a buffer additive for the separation of proteins in capillary electrophoresis.

The use of phytic acid to improve protein analysis by capillary electrophoresis (CE) is becoming more and more popular. Due to its size and number of negative charges (up to 12) it provides a high ionic strength combined with a low conductance resulting in an efficient decrease of wall adsorption for proteins. Because of its twelve acidic groups, phytic acid can be used as a buffer over a wide pH range (pH 2-11). The limited wall adsorption of proteins using phytic acid-containing buffers is observed for buffers with a pH of 5.5 and higher. With a monoprotic buffer, most of the investigated proteins show wall adsorption at the pH values studied. In case of a phytic acid buffer, wall adsorption is reduced by a factor of 2-4. The use of phytic acid both as a modifier and as a pH buffer results in more pronounced differences between the various protein mobilities compared with the use of monoprotic buffers. As a result this feature can be used to improve resolution in protein separations.

Buffers↗

PubMed1997

Particle-loaded membranes for sample concentration and/or clean-up in bioanalysis.

Solid-phase extraction nowadays is a major sample preparation tool. The latest development in this area is the introduction of particle-loaded membranes (membrane-extraction disks). The potential of these extraction membranes in bioanalysis is discussed with respect to recoveries, reproducibility, sensitivity and speed. A comparison is made between liquid-liquid extraction and solid-phase extraction using traditional sorbents and extraction disks, and off-line and on-line techniques. Particle-loaded membranes are available in disks with diameters of 4-90 mm. The 25-90 mm disks are mainly used for off-line extractions of mainly environmental samples, while the 4 mm disks are available in the so-called drug tubes that can be used in the same way as conventional extraction cartridges for the extraction of drugs from biological fluids. The main advantage of using drug tubes is the smaller desorption volume and, therefore, the increased sensitivity. Cutting smaller disks, from the commercially available disks, allows the use of on-line extractions in column-switching systems. The main conclusion is that in many cases particle-loaded membranes are more efficient than packed solid-phase extraction cartridges.

Chemical Fractionation↗

PubMed1997

Pre-, on- and post-column derivatization in capillary electrophoresis.

This survey gives a short overview of the various reagents and procedures that can be used for pre-, post- and on-column derivatization in capillary electrophoresis. First there is an introduction about capillary electrophoresis as an analytical technique; this is followed by a discussion of the pros and cons of the various modes of derivatization and a comparison with liquid chromatography. In the following paragraphs the reagents for a number of functional groups are discussed. The emphasis is on derivatization of the amino group. Most of the information on the reagents and derivatization procedures is listed in tables together with information on the detection mode, analytes, sensitivity and samples. In addition to the amino group, information is given on labeling of aldehyde, keto, carboxyl, hydroxyl and sulfhydryl groups.

Amino Acids↗

PubMed1996

Solid-phase extraction of polar pesticides from environmental water samples on graphitized carbon and Empore-activated carbon disks and on-line coupling to octadecyl-bonded silica analytical columns.

The suitability of Empore-activated carbon disks (EACD), Envi-Carb graphitized carbon black (GCB) and CPP-50 graphitized carbon for the trace enrichment of polar pesticides from water samples was studied by means of off-line and on-line solid-phase extraction (SPE). In the off-line procedure, 0.5-2 l samples spiked with a test mixture of oxamyl, methomyl and aldicarb sulfoxide were enriched on EnviCarb SPE cartridges or 47 mm diameter EACD and eluted with dichloromethane-methanol. After evaporation, a sample was injected onto a C18-bonded silica column and analysed by liquid chromatography with ultraviolet (LC-UV) detection. EACD performed better than EnviCarb cartridges in terms of breakthrough volumes (> 2 l for all test analytes), reproducibility (R.S.D. of recoveries, 4-8%, n = 3) and sampling speed (100 ml/min); detection limits in drinking water were 0.05-0.16 microgram/l. In the on-line experiments, 4.6 mm diameter pieces cut from original EACD and stacked onto each other in a 9 mm long precolumn, and EnviCarb and CPP-50 packed in 10 x 2.0 mm I.D. precolumn, were tested, and 50-200 ml spiked water samples were preconcentrated. Because of the peak broadening caused by the strong sorption of the analytes on carbon, the carbon-packed precolumns were eluted by a separate stream of 0.1 ml/min acetonitrile which was mixed with the gradient LC eluent in front of the C18 analytical column. The final on-line procedure was also applied for the less polar propoxur, carbaryl and methiocarb. EnviCarb could not be used due to its poor pressure resistance. CPP-50 provided less peak broadening than EACD: peak widths were 0.1-0.3 min and R.S.D. of peak heights 4-14% (n = 3). In terms of analyte trapping efficiency on-line SPE-LC-UV with a CPP-50 precolumn also showed better performance than when Bondesil C18/OH or polymeric PLRP-S was used, but chromatographic resolution was similar. With the CPP-50-based system, detection limits of the test compounds were 0.05-1 microgram/l in surface water.

Adsorption↗

PubMed1995

Determination of the anticancer drug metabolite WR1065 using pre-column derivatization and diode laser induced fluorescence detection.

A liquid chromatographic (LC) procedure using alumina as stationary phase in both the pre- and the analytical column, is reported for the determination of WR1065, the active metabolite of the amino- and thiol-containing anticancer drug WR2721. After pre-column derivatization of the thiol group, the analyte is determined by LC with diode laser induced fluorescence detection in the near-infrared. Selective removal of excess label is achieved by means of column switching; it allows the detection of 5 x 10(-9) M WR1065 in water and 10-fold diluted, deproteinated plasma samples. The detection limit is determined by the derivatization reaction and not by the fluorescence detection of the labelled analyte. Endogeneous thiols do not interfere.

Antineoplastic Agents↗

PubMed1994

Automated column liquid chromatographic determination of amoxicillin and cefadroxil in bovine serum and muscle tissue using on-line dialysis for sample preparation.

A fully automated method is described for the determination of amoxicillin and cefadroxil in bovine serum and muscle tissue. The method is based on the on-line combination of dialysis and solid-phase extraction for sample preparation, and column liquid chromatography with ultraviolet detection. In order to enhance the UV detectability of the analytes, post-column addition of 0.1 M sodium hydroxide is performed. The method shows good linearity and repeatability for both analytes in serum as well as in muscle tissue; the limits of detection in these samples are 0.05 microgram/ml and 0.2 microgram/g, respectively. The method has a sample throughput of 30 samples per 24 h.

Amoxicillin↗

PubMed1994

General strategies and selection of derivatization reactions for liquid chromatography and capillary electrophoresis.

The general strategies, reasons and the different possibilities for the derivatization of biomedically important compounds are reviewed. Different approaches apply for small versus large analyte molecules, different advantages and disadvantages are visualized with pre- and post-column arrangements. Particular interest is focused upon solid-phase derivatization reagents.

Chromatography, High Pressure Liquid↗