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Biomedical subjects

J Crommen

Publications and source records attributed to J Crommen.

At least 73 records · Page 4Linked to original sources
PubMed1994

Knowledge-based system for the automated solid-phase extraction of basic drugs from plasma coupled with their liquid chromatographic determination. Application to the biodetermination of beta-receptor blocking agents.

Techniques for the preparation of biological samples are often based nowadays on solid-phase extraction (SPE). The different SPE steps can be performed automatically on disposable extraction cartridges (DECs) by means of a sample processor. A knowledge-based system was developed to facilitate the development of fully automated methods for the solid-phase extraction of relatively hydrophobic basic drugs from plasma, coupled with their determination by high-performance liquid chromatography (HPLC). The DEC filled with 50 mg of cyanopropyl-bonded silica phase is first conditioned with methanol and buffer solution (pH 7.4). After sample application, the DEC sorbent is washed with the same buffer. The analytes are then desorbed with an appropriate eluent and the eluate is finally diluted with the same buffer as used in the HPLC mobile phase before injection. Under these conditions, only three variables are still to be optimized: the composition and volume of the elution solvent and the volume of buffer to be added to the eluate. On the basis of this general strategy, a decision tree providing information about suggested starting conditions and guidelines for the optimization of the three variables was developed and implemented by use of a hypermedia software. This didactic expert system was evaluated using several beta-receptor blocking agents as model compounds and the operating conditions obtained for the automated SPE of these compounds are presented. A method for the determination of propranolol in plasma using the SPE conditions deduced from the knowledge-based system was validated. The absolute recovery of propranolol is ca. 93% and the limit of detection is 1.3 ng ml-1. The mean within-day and between-day reproducibilities are 2.3 and 3.6%, respectively.

Adrenergic beta-Antagonists↗
PubMed1994

Chiral separation of basic drugs by capillary zone electrophoresis with cyclodextrin additives.

The enantiomers of a series of basic drugs were separated in capillary zone electrophoresis (CZE) in phosphate buffers, pH 3, containing beta-cyclodextrin or one of its derivatives as chiral selectors and uncoated fused silica capillaries thermostated at 15 degrees C. The nature of the cationic component of the background electrolyte was found to have a significant influence on achiral resolution and peak symmetry. The best results were obtained with triethanolamine, which was then used to adjust the buffer pH in all further experiments. The effects on chiral resolution of the nature and concentration of cyclodextrin, of the addition of methanol, and of capillary temperature were studied. Maximum resolution was obtained at a particular cyclodextrin concentration for each analyte, depending on the affinity of the analyte for this cyclodextrin. On the basis of the results, the effects of methanol addition and temperature on enantiomeric resolution could be explained and predicted. Numerous chiral separations are presented and suggestions for the rapid optimization of CZE enantioseparations with cyclodextrin additives are given.

Alkalies↗
PubMed1994

Determination of benzodiazepines by micellar electrokinetic chromatography.

A method for the separation and determination of benzodiazepines by micellar electrokinetic chromatography (MEKC) has been developed. Separation buffers consisted of aqueous solutions of glycine and triethanolamine (pH 9.0), containing sodium dodecyl sulfate (SDS) as surfactant and methanol as organic modifier. The effect of the concentration of SDS, methanol, glycine and triethanolamine on migration times and resolution was studied. Ten benzodiazepines were baseline separated at a 25 mM SDS concentration and 20% v/v methanol in a 75 mM glycine-250 mM triethanolamine buffer. Under these conditions, the within-day reproducibilities were 0.3-0.5% for migration times and 1.7-1.9% for peak areas at a concentration of 10 micrograms/mL. The limits of detection and quantification for oxazepam were 0.2 and 0.7 micrograms/mL, respectively, using an injection time of 5 s.

Benzodiazepines↗
PubMed1993

Fully automated determination of sulfamethazine in ovine plasma using solid-phase extraction on disposable cartridges and liquid chromatography.

An automatic sample preparation procedure followed by on-line injection of the sample extract into a HPLC system has been developed for the quantitative analysis of sulfamethazine and its N4-acetyl metabolite in ovine plasma. The sample clean-up was performed by solid-phase extraction (SPE) on C18 disposable extraction cartridges (DECs). All the sample handling operations were effected by a robotic auto-sampler. The DEC was first conditioned with methanol and phosphate buffer pH 7.4. After loading 1.0 ml of plasma sample onto the DEC, the latter was washed with the same buffer. The elution step was performed with methanol (0.25 ml) and the eluate was then diluted by adding 0.75 ml volume of phosphate buffer pH 6.4. A 20-microliters volume of the resultant solution was injected onto an octadecyl silica column preceded by a short guard column. The HPLC mobile phase was methanol-phosphate buffer pH 6.4 (25:75, v/v). Sulfamethazine and N4-acetylsulfamethazine were determined photometrically at 262 nm. Under these conditions, linear calibration curves ranging from 2 to 250 micrograms ml-1 have been obtained for both compounds. Drug recoveries were higher than 90% and typical relative standard deviation values were 0.7% (within-day) and 2.0% (between-day) at a plasma concentration of 50 micrograms ml-1.

Animals↗
PubMed1991

Automatic determination of diltiazem and desacetyldiltiazem in human plasma using liquid-solid extraction on disposable cartridges coupled to HPLC--Part I: Optimization of the HPLC system and method validation.

A sensitive and automated method for the analysis of diltiazem and desacetyldiltiazem in plasma has been developed using liquid-solid extraction (LSE) on disposable extraction cartridges (DECs) in combination with HPLC. After isolation from plasma, the analytes are separated on a highly deactivated octyl silica column with a mobile phase of methanol-0.05 M phosphate buffer (pH 7.4) (62:38, v/v). The analytes are monitored photometrically at 238 nm. The complete preparation of the plasma sample as well as the injection of the final extract on to the analytical column are performed automatically by means of a sample processor equipped with a robotic arm to which is attached a needle dispensing the different liquids. The internal standard solution is first added to the plasma sample. The DEC is then conditioned successively with methanol and phosphate buffer (pH 7.4). A 1.0-ml volume of sample containing the internal standard solution is applied on an extraction cartridge filled with cyanopropyl silica (50 mg). After the DEC has been washed with the same buffer, the analytes are eluted with 0.16 ml of methanol. A 0.14-ml volume of buffer is then passed through the DEC and 0.25 ml of the final extract is injected onto the HPLC column. The absolute recoveries of the drugs are about 90% and the limit of detection for diltiazem is 0.8 ng ml-1. Relative standard deviations of 2.6% (within-day) and 3.7% (between-day) have been obtained for this compound at a plasma concentration of 50 ng ml-1.

Automation↗
PubMed1991

Automatic determination of diltiazem and desacetyldiltiazem in human plasma using liquid-solid extraction on disposable cartridges coupled to HPLC--Part II: Optimization of liquid-solid extraction.

An automatic liquid-solid extraction (LSE) procedure to be coupled to HPLC for the determination of diltiazem and desacetyldiltiazem in plasma has been developed. The LSE operations are performed on disposable extraction cartridges (DECs) by means of a sample processor equipped with a robotic arm holding a needle through which the different liquids are dispensed. The operating parameters of LSE have been optimized with respect to recovery, detectability and reproducibility by using, whenever possible, aqueous solutions of the analytes. Different kinds of DECs have been tested. For the compounds studied, DECs filled with 50 mg of cyanopropyl silica have been selected. The influence of the pH of the buffer used in the washing step has been studied, leading finally to the selection of the same phosphate buffer (pH 7.4) as in the HPLC mobile phase. The minimum volume of methanol which still gives a nearly complete elution of the analytes from the extraction cartridges has been determined. Under these conditions, a high sensitivity can be obtained without an evaporation step. Moreover, the volume of buffer to be added to the methanolic eluate before injection into the HPLC system has been optimized in such a way that a focusing effect is obtained at the top of the analytical column while the dilution of the extract is minimized.

Automation↗
PubMed1989

A fully automated high-performance liquid chromatographic method for the determination of indomethacin in plasma.

A fully automated method is described, which enables the determination of indomethacin in plasma by reversed-phase HPLC following on-line sample enrichment and clean-up on a short pre-column. The plasma sample is introduced directly into the column switching system. The pre-column, filled with a pellicular bonded phase, is first washed with phosphate buffer, pH 7.4. The compounds retained on the pre-column are then eluted in the fore-flush mode and separated on an octadecylsilica column with a methanolic phosphate buffer (pH 7.4) mobile phase. Indomethacin is determined spectrophotometrically at 254 or 260 nm. The effect of changes in the pH and flow rate of the washing eluent are studied. Under the conditions selected, memory effects can be avoided, the absolute recovery of the drug is 70% and the limit of detection 10 ng ml-1 for a 100 microliter injection of plasma. At a concentration of 100 ng ml-1, the relative standard deviations (RSD) are 1.7% (within-day) and 3.5% (between-day), respectively.

Autoanalysis↗
PubMed1984

Determination of thiamine and thiamine phosphates in excitable tissues as thiochrome derivatives by reversed-phase high-performance liquid chromatography on octadecyl silica.

The analysis of thiamine and thiamine phosphates by high-performance liquid chromatography owes its high sensitivity to the fluorescent derivatives or thiochromes obtained by chemical oxidation in alkaline medium. The possibility of performing precolumn oxidation with potassium ferricyanide instead of using the hazardous cyanogen bromide has been investigated. The derivatization step has been optimized with respect to the following parameters: concentration of alkali and oxidant, presence of methanol and stability of the thiochromes . A gradient separation with 25 mM phosphate buffer (pH 8.4) and methanol as mobile phase components and an octadecyl silica column as stationary phase has been set up. The analytical run takes 14 min with the following elution order: thiochrome triphosphate, thiochrome pyrophosphate, thiochrome monophosphate and thiochrome. The minimum detectable amount is 0.05 pmol. The method was found suitable for the determination of thiamine compounds in excitable tissues such as nerves and electric organs as well as in proteins extracted from membranes of these organs. It may be useful to study the role of thiamine in the electrical activity of these tissues at the molecular level.

Animals↗
PubMed1984

Indirect UV detection of hydrophilic ionized compounds in reversed-phase liquid chromatography by use of a UV-absorbing ion of the same charge.

Ion-pair reversed-phase liquid chromatographic systems have been developed that allow the detection and quantitation of low amounts of hydrophilic ionized substances, which otherwise display little or no capability for optical detection. The mobile phase contains a UV-absorbing ion of the same charge and hydrophobic character as the samples, most often together with a hydrophobic counter-ion to give the samples suitable retention; the stationary phase consists of octadecylsilica. Chromatograms featuring two system peaks are usually obtained, and these give rise to some deviations from the usual response pattern. In the present work the influence on retention and detection sensitivity of the non-absorbing counter-ion, the UV-absorbing ion and the uncharged organic modifier have been studied systematically. Principles for optimizing the detector response by changing the mobile phase composition have been deduced. Examples of applications of the indirect detection technique to the analysis of hydrophilic compounds of pharmaceutical and biological interest, such as amino acids, water-soluble vitamins and piperazine, are presented.

Journal Article↗
PubMed1983

Ion-pairing detection technique in reversed-phase high-performance liquid chromatography of drugs and related compounds.

The principles and practice of UV-detection methods based on the ion-pairing technique are discussed with respect to reversed-phase high-performance liquid chromatography. The detection of amino acids, aliphatic alcohols and vitamins is described together with the rationale for optimizing sensitivity of detection in using the ion-pairing technique for pharmaceutical analysis.

Journal Article↗
PubMedDate not supplied

Determination of verapamil and norverapamil in human plasma by liquid chromatography: comparison between a liquid-liquid extraction procedure and an automated liquid-solid extraction method for sample preparation.

A conventional liquid-liquid extraction (LLE) procedure with high-performance liquid chromatography (HPLC) has been developed for the determination of verapamil and its main metabolite, norverapamil, in plasma. After addition of the internal standard, plasma samples were basified with phosphate buffer (pH 9.0) and extracted with a mixture of cyclohexane-dichloromethane. After centrifugation, the organic layer was separated and the analytes were extracted back into a 0.1 N sulphuric acid solution containing 2-aminoheptane. An aliquot of this aqueous phase was then injected directly onto the HPLC column. This LLE procedure has been compared with an automated liquid-solid extraction (LSE) method that has been developed in parallel. Good linearity was obtained using both extraction methods. The absolute recoveries for the two analytes were ca 95% with the automated LSE procedure and slightly lower (ca 84%) for the LLE method. The automated method gives better results with respect to detectability and precision, but the LLE procedure is simpler to develop, requires much less expensive equipment, and remains a useful alternative when the number of samples to be analysed is limited.

Buffers↗
PubMedDate not supplied

Determination of meprobamate in pharmaceutical dosage forms also containing carbromal by liquid chromatography and indirect photometric detection.

In a pharmaceutical form also containing carbromal, meprobamate could not be quantified selectively by classical methods described in pharmacopoeias due to a significant interference from carbromal. Consequently, reversed-phase HPLC methods have been developed to separate the two active ingredients using indirect photometric detection to visualize and determine meprobamate which has very poor chromophoric properties. Different parameters influencing the sensitivity of the indirect response, such as the nature of the highly absorbing compound added to the mobile phase (the marker) as well as the methanol content and the pH of this phase, have been studied. Two chromatographic systems containing benzoic acid or cinnamic acid as the marker, have been optimized and validated. Good linearity and reproducibility have been obtained with both systems but the cinnamic acid method has the advantage that meprobamate and carbromal can be determined simultaneously at 273 nm.

Benzoates↗