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PubMed2001

Determination of palladium in human urine by high-performance liquid chromatography and ultraviolet detection after ultraviolet photolysis and selective solid-phase extraction.

The high-performance liquid chromatographic method with UV detection described below permits the selective determination of traces of palladium in human urine. After UV photolysis, during which the complete organic matrix was destroyed, the palladium was selectively enriched by solid-phase extraction (SPE). The reversed-phase C18 SPE column material was loaded with the ligand N,N-diethyl-N'-benzoylthiourea (DEBT) which shows an excellent complexing capacity for palladium in acidic solutions and at room temperature. The Pd(DEBT)2 complex was eluted with ethanol. After isocratic separation on the analytical column (MeOH/H2O 98:2 (v/v)), the complex was detected at 274 nm. The detection limit was 10 ng Pd/l. The relative standard deviations (RSD) of the within-series imprecision were in the range between 11% (75 ng Pd/l) and 7% (180 ng Pd/l). The between-day imprecision was 11% (75 ng Pd/l) and 5% (180 ng Pd/l). The recovery rates ranged between 94 and 96%. Using this method, urine samples of 44 persons from the general population were analysed. Only in one urine sample could palladium be detected. For comparison, 10 persons with occupational palladium exposure were examined. The urinary concentrations ranged from <10 to 2,538 ng/l.

Calibration↗
PubMed2002

A study of the conditions of the supercritical fluid extraction in the analysis of selected anti-inflammatory drugs in plasma.

Supercritical fluid extraction (SFE) was employed to analyze selected anti-inflammatory drugs in plasma. Evaluation of selected drugs (ibuprofen, indomethacin, and flufenamic acid) was performed using the HPLC method on columns with the reverse phase C-18 and detection in the UV region of the spectrum. A study of the conditions of SFE carried out for 30 min at 50 degrees C investigated the magnitude of the pressure of carbon dioxide suitable for drug extraction, the selection of the collecting solvent, and the modification of CO2 with an organic solvent. The results of the study made it possible to determine the optimal procedure for SFE of ibuprofen, indomethacin, and flufenamic acid from plasma, which renders their HPLC quantification possible.

Anti-Inflammatory Agents, Non-Steroidal↗
PubMed2004

Solvent extraction of sr2+ and cs+ based on room-temperature ionic liquids containing monoaza-substituted crown ethers.

A series of N-alkyl aza-18-crown-6 ethers were synthesized and characterized by NMR spectroscopy and mass spectrometry. These monoaza-substituted crown ethers in ionic liquids were investigated as recyclable extractants for separation of Sr(2+) and Cs(+) from aqueous solutions. The pH-sensitive complexation capability of these ligands allows for a facile stripping process to be developed so that both macrocyclic ligands and ionic liquids can be reused. The extraction efficiencies and selectivities of these monoaza-substituted crown ethers for Na(+), K(+), Cs(+), and Sr(2+) were studied in comparison to those of dicyclohexano-18-crown-6 under the same conditions. The extraction selectivity order for dicyclohexano-18-crown-6 in the ionic liquids investigated here was K(+) >> Sr(2+) > Cs(+) > Na(+). The extraction selectivity order for N-alkyl aza-18-crown-6, in which the alkyl group is varied systematically from ethyl to n-dodecyl, was Sr(2+) >> K(+) > Cs(+) > Na(+) in 1-ethyl-3-methylimidazolium bis[(trifluoromethyl)sulfonyl]amide and 1-butyl-3-methylimidazolium bis[(trifluoromethyl)sulfonyl]amide and K(+) > Sr(2+) > Cs(+) > Na(+) in 1-hexyl-3-methylimidazolium bis[(trifluoromethyl)sulfonyl] amide and 1-octyl-3-methylimidazolium bis[(trifluoromethyl)sulfonyl]amide. The strong dependence of selectivity on the type of ionic liquid indicates an important role played by solvation in solvent extraction processes based on ionic liquids. The optimization of macrocyclic ligands and ionic liquids led to an extraction system that is highly selective toward Sr(2+).

Journal Article↗
PubMed2005

Calcium-induced contraction of sarcomeres changes the regulation of mitochondrial respiration in permeabilized cardiac cells.

The relationships between cardiac cell structure and the regulation of mitochondrial respiration were studied by applying fluorescent confocal microscopy and analysing the kinetics of mitochondrial ADP-stimulated respiration, during calcium-induced contraction in permeabilized cardiomyocytes and myocardial fibers, and in their 'ghost' preparations (after selective myosin extraction). Up to 3 microm free calcium, in the presence of ATP, induced strong contraction of permeabilized cardiomyocytes with intact sarcomeres, accompanied by alterations in mitochondrial arrangement and a significant decrease in the apparent K(m) for exogenous ADP and ATP in the kinetics of mitochondrial respiration. The V(max) of respiration showed a moderate (50%) increase, with an optimum at 0.4 microm free calcium and a decrease at higher calcium concentrations. At high free-calcium concentrations, the direct flux of ADP from ATPases to mitochondria was diminished compared to that at low calcium levels. All of these effects were unrelated either to mitochondrial calcium overload or to mitochondrial permeability transition and were not observed in 'ghost' preparations after the selective extraction of myosin. Our results suggest that the structural changes transmitted from contractile apparatus to mitochondria modify localized restrictions of the diffusion of adenine nucleotides and thus may actively participate in the regulation of mitochondrial function, in addition to the metabolic signalling via the creatine kinase system.

Adenosine Diphosphate↗
PubMed2003

An amobarbital molecularly imprinted microsphere for selective solid-phase extraction of phenobarbital from human urine and medicines and their determination by high-performance liquid chromatography.

A 40-60 microm amobarbital molecularly imprinted microsphere, used as a solid-phase selective sorbent for extracting phenobarbital from human urine and medicines, was prepared by a suspension polymerization method. A series of binding studies was performed in order to find optimal loading, washing and eluting conditions for solid-phase extraction. Under optimal conditions, good recoveries of phenobarbital in samples were obtained. Normally, molecularly imprinted polymers, prepared in bulk, require laborious work. Significant losses occur during the procedure of grinding and washing. In this work all molecularly imprinted polymers made into microsphere could be utilized, and the cost of the template was reduced too (the price of phenobarbital is twice that of amobarbital). As the phenobarbital to be extracted was different from the template molecule amobarbital, the interference caused by template leaking could be avoided in the assay.

Amobarbital↗
PubMed2005

The accuracy of maternal anthropometry measurements as predictor for spontaneous preterm birth--a systematic review.

OBJECTIVE: To assess the accuracy with which antenatal maternal anthropometric measurements predict the risk spontaneous preterm birth. STUDY DESIGN: (1) DATA SOURCES: Studies were identified without language restrictions from MEDLINE, EMBASE, PASCAL, BIOSIS, the Cochrane Library, MEDION, National Research Register, SCISEARCH and Conference Papers, and manual searching of bibliographies of known primary and review articles, and contact with authors. (2) STUDY SELECTION AND DATA EXTRACTION: Studies were selected if they used antenatal maternal anthropometric features (pre-pregnancy weight, maternal pregnancy weight gain and maternal height) to predict spontaneous preterm birth. Two reviewers independently selected studies and extracted data on their characteristics, quality and accuracy. Accuracy data were used to form 2 x 2 contingency tables of the maternal anthropometric test results with spontaneous preterm birth as the reference standard. (3) DATA SYNTHESIS: Heterogeneity assessments were carried out to aid the decision regarding pooling of the accuracy results. Likelihood ratios for positive (LR+) and negative (LR-) test results were calculated, and summary estimates were produced in absence of heterogeneity of the accuracy results. RESULTS: There were eight primary accuracy articles that met the selection criteria, which included a total of 122,647 asymptomatic women. There were six studies on pre-pregnancy weight where five measured the body mass index (BMI) and one used an arbitrary measure. There were four studies on the adequacy of pregnancy weight gain and two studies on maternal height as a predictor for the risk of preterm birth. One article contributed three studies, while two articles provided two studies each. The commonest reference standard used was birth before 37 weeks' gestation. None of the studies fulfilled the ideal test accuracy study criteria. There was heterogeneity in the accuracy results of pre-pregnancy BMI but not in the adequacy of weight gain. All three maternal anthropometric features were poor predictors of preterm labour. Pre-pregnancy BMI is a poor predictor of preterm birth before 37 weeks' gestation (LR+ that ranged from 0.96 (95% confidence interval (CI) 0.66-1.40) to 1.75 (95% CI 1.33-2.31)) as are the adequacy of pregnancy weight gain (summary LR+ of 1.81, 95% CI 1.45-2.30) and short maternal height (LR+ of 1.79 (95% CI 1.27-2.52). CONCLUSION: Routine antenatal maternal anthropometric measurements are not useful in predicting the risk of preterm birth before 37 weeks' gestation. Further studies should address their use in combination with other test but need to use a more clinically appropriate reference standard of preterm birth, such as birth before 32-34 weeks' gestation, and improve on the quality of study design.

Anthropometry↗
PubMed2002

High-performance liquid chromatographic assay for amiodarone N-deethylation activity in human liver microsomes using solid-phase extraction.

A selective and sensitive assay for amiodarone N-deethylation activity in human liver microsomes by high-performance liquid chromatography (HPLC) with UV detection is reported. The extraction of desethylamiodarone from incubation samples was performed by means of an original solid-phase extraction (SPE) procedure using a polymeric reversed-phase sorbent (Oasis HLB). The method was validated for the determination of desethylamiodarone with respect to specificity, linearity, precision, accuracy, recovery, limit of quantitation and stability. Amiodarone N-deethylation activity from low to high substrate concentrations using human liver microsomes was precisely determined without a concentration step. This method is applicable to the study in vitro of the metabolism of amiodarone.

Amiodarone↗
PubMed1999

[Determination of selected secondary metabolites and extracts of Philadelphus coronarius L].

The paper deals with the determination of selected secondary metabolites in the branches and leaves of Philadelphus coronarius L. Colorimetric methods were employed to determine the content of flavonoids (0.63% and 0.01%, respectively) and phenolic substances (3.25% and 0.70%, respectively) in leaves or branches. Also the content of extractive substances was determined in ethanolic, aqueous-ethanolic (85% and 60% ethanol), and aqueous extracts. This yielded 9.56%, 22.46%, 24.93%, and 33.15% of extractive substances in leaves and 2.20%, 6.04%, 8.84%, and 10.39% of extractive substances in branches.

Colorimetry↗
PubMed2006

Comparison of methods for detection of norovirus in oysters.

In the absence of culture methods for noroviruses, detection in foods relies on molecular techniques such as Reverse Transcription-Polymerase Chain Reaction (RT-PCR) on extracted viral RNA followed by PCR product confirmation by hybridisation and/or sequencing. However, in order to obtain a successful detection it is of great importance to remove the tissue inhibitors during the viral RNA extraction. To select the most efficient extraction procedure of oysters we have compared four protocols. A pool of digestive gland material from oyster samples was divided into 1.5 g portions and spiked with 10-fold dilutions of human faecal samples containing norovirus genogroup II. The samples were tested on three different occasions using four different sample treatment protocols. The protocols were assessed with regard to their ability to recover viral RNA and detect norovirus in spiked oysters and for their in-house reproducibility. One method using viral elution by a Mixer Mill Cell Disrupter resulted in a 10-fold better recovery than the other three protocols when an RT-seminested PCR (G2SKR/COG2F and G2SKR/G2SKF) detection approach was applied. Although less distinctive this was also the case when NoV was detected by a single round RT-PCR approach using the primers JV13i and JV12y. The second most efficient method was a method using chloroform extraction and polyethylene precipitation.

Animals↗
PubMed2006

Effect of pressure on the crystal structure of salicylaldoxime-I, and the structure of salicylaldoxime-II at 5.93 GPa.

The effect of pressure on the crystal structure of salicylaldoxime has been investigated. The ambient-pressure phase (salicylaldoxime-I) consists of pairs of molecules interacting through oximic OH...O hydrogen bonds; taken with phenolic OH...N intramolecular hydrogen bonds, these dimers form a pseudo-macrocycle bounded by an R4 4(10) motif. The dimers interact principally via pi...pi stacking contacts. Salicylaldoxime derivatives are used industrially as selective solvent extractants for copper; the selectivity reflects the compatibility of the metal ion with the pseudo-macrocycle cavity size. On increasing the pressure to 5.28 GPa the size of the cavity was found to decrease by an amount comparable to the difference in hole sizes in the structures of the Cu2+ salicylaldoximato complex and its Ni2+ equivalent. On increasing the pressure to 5.93 GPa a new polymorph, salicylaldoxime-II, was obtained in a single-crystal to single-crystal phase transition. PIXEL calculations show that the phase transition is driven in part by relief of intermolecular repulsions in the dimer-forming OH...O-bonded ring motif, and the ten-centre hydrogen-bonding ring motif of the phase I structure is replaced in phase II by a six-centre ring formed by oximic OH...N hydrogen bonds. The transition also relieves repulsions in the pi...pi stacking contacts. The intramolecular OH...N hydrogen bond of phase I is replaced in phase II by a intermolecular phenolic OH...O hydrogen bond, but the total interaction energy of the pairs of molecules connected by this new contact is very slightly repulsive because the electrostatic hydrogen-bond energy is cancelled by the repulsion term. The intra- to intermolecular hydrogen-bond conversion simply promotes efficient packing rather than contributing to the overall lattice energy.

Crystallization↗
PubMed1998

Propionic acid production by extractive fermentation. I. Solvent considerations.

Solvent selection for extractive fermentation for propionic acid was conducted with three systems: Alamine 304-1 (trilaurylamine) in 2-octanol, 1-dodecanol, and Witcohol 85 NF (oleyl alcohol). Among them, the solvent containing 2-octanol exhibited the highest partition coefficient in acid extraction, but it was also toxic to propionibacteria. The most solvent-resistant strain among five strains of the microorganism was selected. Solvent toxicity was eliminated via two strategies: entrapment of dissolved toxic solvent in the culture growth medium with vegetable oils such as corn, olive, or soybean oils; or replacement of the toxic 2-octanol with nontoxic Witcohol 85 NF. The complete recovery of acids from the Alamine 304-1/Witcohol 85 NF was also realized with vacuum distillation.

Fermentation↗
PubMed1994

Insulin resistance, hypersecretion of LH, and a dual-defect hypothesis for the pathogenesis of polycystic ovary syndrome.

OBJECTIVE: To review the literature dealing with the roles of insulin resistance and elevated LH levels in the development of the polycystic ovary syndrome and to outline a new hypothesis of the pathogenesis of this disorder. DATA SOURCES: We reviewed articles on the topics of insulin resistance, elevated LH levels, and polycystic ovary syndrome that were contained in the CD-PLUS MEDLINE system data base for years 1976-1994. METHODS OF STUDY SELECTION: Ninety-one original reports published in English-language, peer-reviewed biomedical journals were selected. DATA EXTRACTION AND SYNTHESIS: The selected studies were reviewed critically and their conclusions were evaluated. The available literature indicates that insulin resistance and increased LH secretion are frequent features of polycystic ovary syndrome and may be important in its pathogenesis. It appears that both the amplitude and the frequency of LH pulses are increased in this disorder. Although the causes of these abnormalities of LH secretion are unknown, they could be either primary (due to increased sensitivity of LH secretion to GnRH) or secondary (due to the effects of sex steroids on LH secretion). The cause of insulin resistance in polycystic ovary syndrome also is unknown. It most likely results from a post-binding defect in the insulin action pathway. There is both in vitro and in vivo evidence that elevated LH and hyperinsulinemia act synergistically to enhance ovarian growth, androgen secretion, and ovarian cyst formation. CONCLUSIONS: Based on the available literature, we propose a "dual-defect" hypothesis of polycystic ovary syndrome. We suggest that in a significant subset of patients, this disorder may be caused by a conjunction of two independent genetic defects: one that produces elevated LH secretion and another that produces insulin resistance. Thus, polycystic ovary syndrome develops as a result of the synergistic action of increased LH levels and hyperinsulinemia on the ovary. This working hypothesis may serve as a useful guide for further studies of the pathogenesis of polycystic ovary syndrome.

Female↗
PubMed2003

Quantitative determination of the HIV protease inhibitor atazanavir (BMS-232632) in human plasma by liquid chromatography-tandem mass spectrometry following automated solid-phase extraction.

A selective, accurate, and reproducible LC-MS-MS assay was developed for the determination of the HIV protease inhibitor atazanavir (BMS-232632) in human plasma samples. The method involved automated solid-phase extraction of atazanavir and a stable isotope analog internal standard (I.S.) using Oasis HLB 10 mg 96-well SPE plates. A portion of the reconstituted sample residue was injected onto a C(18) HDO analytical column which was configured with a triple quad mass spectrometer for analyte determination by positive ion electrospray. The assay was linear from 1.00 to 1,000 ng/ml with a lower limit of quantitation of 1.00 ng/ml. The inter- and intra-day coefficients of variation (C.V.) for the assay were <4%, and the accuracy was 99-102%. Atazanavir was stable in human plasma for at least 109 h at room temperature and for at least 1 year at -20 degrees C.

Atazanavir Sulfate↗
PubMed2003

High-performance liquid chromatographic determination of diltiazem in human plasma after solid-phase and liquid-liquid extraction.

A selective, sensitive, and accurate high-performance liquid chromatographic method for determination of diltiazem in plasma samples has been developed and validated. The effects of mobile phase composition, buffer concentration, mobile phase pH, and concentration of organic modifiers on retention of diltiazem and internal standard were investigated. Solid-phase and liquid-liquid extraction were examined and proposed for isolation of the drug and elimination of endogenous plasma interferences. A 5 microm Lichrocart Lichrospher 60 RP-select B chromatographic column was used; the mobile phase was acetonitrile-0.025 mol L(-1) KH(2)PO(4) (pH 5.5), 35:65 ( v/ v) at a flow-rate of 1.5 mL min(-1). The detection wavelength was 215 nm. The calibration plots were linear in the concentration range 20.0-500.0 ng mL(-1). The method has been implemented to monitor diltiazem levels in patient samples.

Calibration↗
PubMed2006

Determination of ofloxacin, norfloxacin, and ciprofloxacin in sewage by selective solid-phase extraction, liquid chromatography with fluorescence detection, and liquid chromatography--tandem mass spectrometry.

A solid-phase extraction (SPE) and liquid chromatographic (LC) method was developed for the determination of selected fluoroquinolone (FQ) drugs including ofloxacin, norfloxacin, and ciprofloxacin in municipal wastewater samples. Extraction of the FQs was carried out with a weak cation exchanger SPE cartridge, the Oasis WCX. The cartridge was washed with water and methanol as a cleanup before the FQs were eluted by a mixture of methanol, acetonitrile, and formic acid. Separation of the FQs was achieved by using a Zorbax SB-C8 column under isocratic condition at a flow rate of 0.2mL/min. Recoveries of the FQs in spiked final effluent samples were between 87 and 94% with a relative standard deviation of less than 6%. Several techniques have been evaluated for the detection of FQs in sewage extracts; they included fluorescence detection and electrospray ionization (ESI) mass spectrometry using either mass-selective detection or tandem mass spectrometry (MS/MS). When they were applied to sewage influent and effluent samples, the LC-MS/MS technique operating in the multiple reaction monitoring (MRM) mode proved to be best suited for the determination of FQs in sewage samples as it provided the highest sensitivity (limit of quantification 5ng/L) and selectivity. The observation of signal suppression (matrix effect) for some FQs in ESI LC-MS and LC-MS/MS is discussed and a solution is proposed. The three FQs were detected in all the sewage samples tested in this work, with median concentrations between 34 and 251ng/L.

Chromatography, Liquid↗
PubMed2004

Effects of an ethanolic salix extract on the release of selected inflammatory mediators in vitro.

Salix extracts are in current use for the treatment of pain and inflammation. In order to obtain an insight into the mechanism(s) of action of the ethanolic Salix extract 1520L--which is essentially similar to an extract for which clinical studies have demonstrated analgesic effectiveness--its effects were evaluated in an established in vitro assay test system using primary human monocytes. The IC50-values obtained for the inhibition of lipopolysaccharide (LPS)-induced release of prostaglandin E2 (PGE2) reflecting cyclooxygenase (COX)-2-mediated PGE2 release were 47 microg/ml and 0.6 microg/ml, for the Salix extract 1520L and rofecoxib-like research compound L745337, respectively. There was no effect on COX-1 and COX-2 activity. The Salix extract inhibited the LPS-induced release of tumor necrosis factor-alpha, interleukin-1beta and interleukin-6 with IC50-values of 180.0, 33.0 and 86.0 microg/ml, respectively. Both, salicin and salicylate, had no effect in any of the parameters. Our results indicate that Salix extract 1520L inhibits COX-2-mediated PGE2 release through compounds other than salicin or salicylate. Our data further suggest that the proprietary Salix extract is a weak inhibitor of proinflammatory cytokines.

Anti-Inflammatory Agents, Non-Steroidal↗
PubMedDate not supplied

Purification of glycoproteins by selective transport using concanavalin-mediated reverse micellar extraction.

A novel methodology for coupling liquid-liquid extraction with affinity interaction has been developed to selectively and efficiently purify and separate glycoproteins. The basis for the separation is the selective extraction of glycoproteins from an aqueous solution into a reverse micellar organic phase by using concanavalin A (a sugar-binding lectin) as a facilitative carrier. Specifically, horseradish peroxidase (a common glycoprotein) can be bound to concanavalin A in an aqueous phase and then extracted into an AOT-isooctane organic phase with negligible loss in enzyme activity. Virtually no extraction of peroxidase occurs in the absence of concanavalin A. Electron spin resonance studies have shown that the large lectin-glycoprotein complex (96,000 daltons) resides in a nonaqueous environment within the reverse micelle, perhaps at the surfactant, water-pool interface; hence, extraction of the large complex is feasible. The facilitative extraction has been extended to selective transport of peroxidase from a mixture of peroxidase and alkaline phosphatase (a nonglycosylated protein). This results in an efficient separation strategy with a separation factor of 16.

Chemistry Techniques, Analytical↗
PubMed1977

Selective inhibition of in vitro DNA synthesis dependent on phiX174 compared with fd DNA. I. Protein requirements for selective inhibition.

Crude extracts of Escherichia coli selectively convert fd viral DNA and not phiX174 DNA to duplex DNA via a complex series of reactions one of which involves RNA polymerase. Reactions leading to formation of fd duplex-replicative (RFII) structures have been reconstituted with purified proteins from E. coli. Maximal synthesis requires the combined action of E. coli binding protein, DNA elongation factor I, DNA elongation factor II preparations (which are a mixture of dna Z and DNA elongation factor III), DNA polymerase III, DNA-dependent RNA polymerase, Mg2+, dATP, dGTP, dCTP, dTTP, and ATP, GTP, CTP, and UTP. In contrast to crude extracts of E. coli, purified protein fractions do not distinguish between fd DNA and phiX174 DNA in duplex DNA formation. The addition of crude fractions of E. coli to the purified components listed above selectively permits fd RFII formation and prevents phiX RFII formation. This selective inhibition was used as an assay to isolate proteins essential for this phenomenon; they include RNase H, discriminatory factor alpha, and discriminatory factor beta.

Adenosine Triphosphatases↗