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Kinds of Cartridges Selected AbstractsSimultaneous determination of maraviroc and raltegravir in human plasma by HPLC-UVIUBMB LIFE, Issue 4 2009Stefania Notari Abstract Therapeutic drug monitoring is pivotal to improve the management of HIV infection. Here, a new HPLC,UV method to quantify simultaneously maraviroc and raltegravir levels in human plasma is reported. Remarkably, this is the first method for maraviroc determination in human plasma. The volume of the plasma sample was 600 ,L. This method involved automated solid-phase extraction with Oasis HLB Cartridge 1 cc (30 mg divinylbenzene and N -vinylpyrrolidone) and evaporation in a water bath under nitrogen stream. The extracted samples were reconstituted with 200 ,L 50/50 of mobile-phase solution (0.01 M KH2PO4 and acetonitrile). Twenty microliters of these samples were injected into a HPLC,UV system, the analytes were eluted on an analytical dC18 Atlantis column (150 mm × 4.6 mm I.D.) with a particle size of 5 ,m. The mobile phase (0.01 M KH2PO4 and acetonitrile) was delivered at 1.0 mL/min with isocratic elution. The total run time for a single analysis was 10 min; maraviroc and raltegravir were detected by UV at 197 and 300 nm. The calibration curves were linear up to 2,500 ng/mL. The absolute recovery ranged between 93 and 100%. The HPLC,UV method reported here has been validated and is currently applied to monitor plasma levels of maraviroc and raltegravir in HIV-infected patients. © 2009 IUBMB IUBMB Life, 61(4):470,475, 2009 [source] CEC-ESI ion trap MS of multiple drugs of abuseELECTROPHORESIS, Issue 7 2010Zeineb Aturki Abstract This article describes a method for the separation and determination of nine drugs of abuse in human urine, including amphetamines, cocaine, codeine, heroin and morphine. This method was based on SPE on a strong cation exchange cartridge followed by CEC-MS. The CEC experiments were performed in fused silica capillaries (100,,m×30,cm) packed with a 3,,m cyano derivatized silica stationary phase. A laboratory-made liquid junction interface was used for CEC-MS coupling. The outlet capillary column was connected with an emitter tip that was positioned in front of the MS orifice. A stable electrospray was produced at nanoliter per minute flow rates applying a hydrostatic pressure (few kPa) to the interface. The coupling of packed CEC columns with mass spectrometer as detector, using a liquid junction interface, provided several advantages such as better sensitivity, low dead volume and independent control of the conditions used for CEC separation and ESI analysis. For this purpose, preliminary experiments were carried out in CEC-UV to optimize the proper mobile phase for CEC analysis. Good separation efficiency was achieved for almost all compounds, using a mixture containing ACN and 25,mM ammonium formate buffer at pH 3 (30:70, v/v), as mobile phase and applying a voltage of 12,kV. ESI ion-trap MS detection was performed in the positive ionization mode. A spray liquid, composed by methanol,water (80:20, v/v) and 1% formic acid, was delivered at a nano-flow rate of ,200,nL/min. Under optimized CEC-ESI-MS conditions, separation of the investigated drugs was performed within 13,min. CEC-MS and CEC-MS2 spectra were obtained by providing the unambiguous confirmation of these drugs in urine samples. Method precision was determined with RSDs values ,3.3% for retention times and ,16.3% for peak areas in both intra-day and day-to-day experiments. LODs were established between 0.78 and 3.12,ng/mL for all compounds. Linearity was satisfactory in the concentration range of interest for all compounds (r2,0.995). The developed CEC-MS method was then applied to the analysis of drugs of abuse in spiked urine samples, obtaining recovery data in the range 80,95%. [source] Poly(dimethylsiloxane)-based microfluidic device with electrospray ionization-mass spectrometry interface for protein identificationELECTROPHORESIS, Issue 21 2003Wang-Chou Sung Abstract An easy method to fabricate poly(dimethylsiloxane) (PDMS)-based microfluidic chips for protein identification by tandem mass spectrometry is presented. This microchip has typical electrophoretic microchannels, a flow-through sampling inlet, and a sheathless nanoelectrospray ionization (ESI) interface. The surface of the microchannel was modified with 2-acrylamido-2-methyl-1-propanesulfonic acid (AMPS) and the generated electroosmotic flow under acidic buffer condition used for the separation was found to be more stable compared to that generated by the microchannel without modification. The feasibility of the device for flow-through sampling, separation, and ESI-MS/MS analysis was demonstrated by the analysis of a standard mixture composed of three tryptic peptides. Results show that four peaks corresponding to three peptide standards and acetylated products of the standard peptide were well resolved and the deduced sequences were consistent with those expected. Furthermore, the compatibility of this device with other miniaturized devices to integrate the whole process was also explored by connecting a miniaturized enzymatic digestion cartridge and a desalting cartridge in series to the sampling inlet of the microchip for the identification of a model protein, ,-casein. [source] DNA fragment analysis by an affordable multiple-channel capillary electrophoresis systemELECTROPHORESIS, Issue 1-2 2003Ming S. Liu Abstract We are demonstrating a cost-effective multichannel capillary electrophoresis system for a high-efficiency double-stranded DNA (dsDNA) fragments analysis. This bench-type high-performance DNA analysis (HDAÔ) system uses fluorescence-type detection with inexpensive solid-state light sources and nonmoving integrated emission collection micro-optics. DNA samples are analyzed simultaneously by using a multiple usage and disposable multicapillary cartridge, which contains integrated capillary channels, optical fibers and an integrated sieving gel reservoir. Using commercially available dsDNA size markers as indicators, the HDAÔ system provides high resolving power in 7 min separations. The system can hold a total of 192 samples in two 96-well polymerase chain reaction (PCR) plates, which can be automatically analyzed within 2.5 h. This affordable system can be used in laboratories to replace slab gel electrophoresis for routine and high-throughput dsDNA analysis. [source] Determination of acephate, methamidophos and monocrotophos in crude palm oilEUROPEAN JOURNAL OF LIPID SCIENCE AND TECHNOLOGY, Issue 11 2006Chee Beng Yeoh Abstract A method for the determination of acephate, methamidophos and monocrotophos in oil matrices is described. Pesticide residues in crude palm oil were extracted with acetonitrile, and a clean-up process was performed by cooling the entire extract below 10,°C, followed by a discolouring process using a Carbograph SPE cartridge. The extract was analysed using gas chromatography coupled with a pulsed flame photometry detector. The limit of detection for the method was calculated from regression data, and the recovery test results were in the range of 85,109%. [source] Highly Enantioselective Synthesis of No-Carrier-Added 6-[18F]Fluoro- L -dopa by Chiral Phase-Transfer AlkylationEUROPEAN JOURNAL OF ORGANIC CHEMISTRY, Issue 13 2004Christian Lemaire Abstract [18F]Fluoro- L -dopa, an important radiopharmaceutical for positron emission tomography (PET), has been synthesized using a phase-transfer alkylation reaction. A chiral quaternary ammonium salt derived from a Cinchona alkaloid served as phase-transfer catalyst for the enantioselective alkylation of a glycine derivative. The active methylene group of this Schiff-base substrate was deprotonated with cesium hydroxide and rapidly alkylated by the 2-[18F]fluoro-4,5-dimethoxybenzyl halide (X = Br, I). The reaction proceeded with high yield (> 90%) at 0 °C or room temperature in various solvents such as toluene or dichloromethane. Preparation of the [18F]alkylating agent on a solid support was developed. After labelling, the labeled [18F]fluoroveratraldehyde was trapped on a tC18 cartridge and then converted on the cartridge into the corresponding benzyl halide derivatives by addition of aqueous sodium borohydride and gaseous hydrobromic or -iodic acid. Hydrolysis and purification by preparative HPLC made 6-[18F]fluoro- L -dopa ready for human injection in a 25,30% decay-corrected radiochemical yield in a synthesis time of 100 min. The product was found to be chemically, radiochemically and enantiomerically pure (ee > 95%). (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004) [source] Identification of Major Alkaloids in Rat Urine by HPLC/DAD/ESI-MS/MS Method Following Oral Administration of Cortex Phellodendri DecoctionHELVETICA CHIMICA ACTA, Issue 2 2009Chun-Hui Ma Abstract A rapid, sensitive, and specific high-performance liquid chromatography (HPLC), diode-array detection, and mass-spectrometry techniques were developed for an identification of the constituents of Cortex Phellodendri and their metabolites in rat urine. The dose of 10,ml/kg of Cortex Phellodendri decoction was used for rats' oral administration. 0,24-h Urine was purified using a C18 solid-phase extraction cartridge, and then analyzed by an on-line MS detector. A total of 13,characteristic HPLC peaks were detected in the urine samples. Nine of them, including five alkaloids and four of their metabolites, were tentatively elucidated as magnoflorine (1), the glucuronide conjugate of demethyleneberberine (2), menisperine (3), jatrorrhizine 3- O -glucuronide (4), berberubine 9- O -glucuronide (5), jatrorrhizine (6), the monomethyl and monohydroxy catabolite of berberubine (7), palmatine (8), and berberine (9). Identification and structural elucidation of the metabolites were performed by comparing their MSn spectra data with those reported. [source] Prediction of hemodialysis sorbent cartridge urea nitrogen capacity and sodium release from in vitro testsHEMODIALYSIS INTERNATIONAL, Issue 2 2008Benjamin P. ROSENBAUM Abstract In sorbent-based hemodialysis, factors limiting a treatment session are urea conversion capacity and sodium release from the cartridge. In vitro experiments were performed to model typical treatment scenarios using various dialyzers and 4 types of SORBÔ sorbent cartridges. The experiments were continued to the point of column saturation with ammonium. The urea nitrogen removed and amount of sodium released in each trial were analyzed in a multi-variable regression against several variables: amount of zirconium phosphate (ZrP), dialysate flow rate (DFR), simulated blood flow rate (BFR), simulated patient whole-body fluid volume (V), initial simulated patient urea concentration (BUNi), dialyzer area permeability (KoA) product, initial dialysate sodium and bicarbonate (HCO3i) concentrations, initial simulated patient sodium (Nai), pH of ZrP, creatinine, breakthrough time, and average urea nitrogen concentration in dialysate. The urea nitrogen capacity (UNC) of various new SORBÔ columns is positively related to ZrP, BFR, V, BUNi, and ZrP pH and negatively to DFR with an R2adjusted=0.990. Two models are described for sodium release. The first model is related positively to DFR and V and negatively to ZrP, KoA product, and dialysate HCO3i with an R2adjusted=0.584. The second model incorporates knowledge of initial simulated patient sodium (negative relationship) and urea levels (negative relationship) in addition to the parameters in the first model with an R2adjusted=0.786. These mathematical models should allow for prediction of patient sodium profiles and the time of column urea saturation based on simple inputs relating to patient chemistries and the dialysis treatment. [source] Application of laser scanning cytometry followed by epifluorescent and differential interference contrast microscopy for the detection and enumeration of Cryptosporidium and Giardia in raw and potable watersJOURNAL OF APPLIED MICROBIOLOGY, Issue 4 2002M.-R. De Roubin Aims: The main goal of this study was to validate a new laser scanning cytometry method (ChemScanRDI) that couples immunofluorescence detection with differential interference contrast (DIC) confirmation, against manual microscopic enumeration of Giardia and Cryptosporidium (oo)cysts. This study also assessed the basic performance of the new Association Française de Normalisation (AFNOR) NF T 90-455 method for Giardia and Cryptosporidium (oo)cyst enumeration with respect to (oo)cyst yield, linearity, repeatability, influence of turbidity and detection limit in raw and potable waters. Methods and Results: The new standard method relies on cartridge (Envirocheck) filtration, immunomagnetic separation purification, immunofluorescence staining and detection followed by DIC confirmation. The recovery was 30,50% for both parasites at seeding levels from 30 to 230 (oo)cysts. The method is linear from 0 to around 400 seeded (oo)cysts and the yield does not significantly vary for turbidity levels from 10 to 40 Formazin Nephelometric Units (FNU). The results were obtained using manual microscopic enumeration of the (oo)cysts. The ChemScanRDI yielded counts that were at least equivalent to those obtained using manual microscopy for both parasites in raw and potable water concentrates, for seeding levels of 10,300 or 10,100, respectively. The purification and labelling method proposed by the supplier of theChemScanRDI (Chemunex) reached very similar recoveries to the AFNOR protocol (70,86% in both cases). Conclusions: Laser scanning cytometry can be used as a more standardized alternative to manual enumeration as part of the new AFNOR standard method. Significance and Impact of the Study: By using laser scanning cytometry instead of manual microscopy, laboratories could circumvent the limitations of manual microscopy, namely: low sample throughput, operator subjectivity and operator fatigue. The study further supports the drive to incorporate laser scanning cytometry in the standard methods for Giardia and Cryptosporidium enumeration. [source] ANALYSIS AND FORMATION OF ACRYLAMIDE IN FRENCH FRIES AND CHICKEN LEGS DURING FRYINGJOURNAL OF FOOD BIOCHEMISTRY, Issue 5 2006W.H. CHUANG ABSTRACT The analysis and formation of acrylamide in French fries and chicken legs during frying were studied. Results showed that the most appropriate extraction solvent was ethyl acetate, with C-18 cartridge for purification and 5-mL deionized water as elution solvent. Dibromination of acrylamide followed by dehydrobromination to 2-bromopropionamide in the presence of triethylamine was necessary for subsequent analysis by gas chromatography,mass spectrometry. The most appropriate temperature programming condition was as follows: 70C in the beginning, raised to 150C at a rate of 10C/min, maintained for 1 min and to 240C at a rate of 30C/min, maintained for 5 min. Detection was carried out using selected-ion monitoring mode, and N,N -dimethylacrylamide was used as internal standard for quantification. French fries and the outer flour portion of chicken legs fried at 180C generated a higher level of acrylamide than at 160C. Compared to soybean oil and palm oil, a lower amount of acrylamide was produced in French fries and the outer flour portion of chicken legs fried in lard. However, no acrylamide was detected in the inner meat portion of fried chicken legs. [source] Recent advances in the assessment of the ratios of cortisol to cortisone and of some of their metabolites in urine by LC-MS-MSJOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 4 2009Alessandro Saba Abstract A previously reported method for the assessment of the ratio of tetrahydrocortisol (THF) + allo-tetrahydrocortisol (A-THF) to tetrahydrocortisone (THE) by HPLC-MS-MS has been significantly improved, in order to increase either ruggedness and reliability. That was achieved by the introduction of an on-line sample cleanup stage, which made use of a perfusion column as a solid phase microextraction (SPE) cartridge. The set of analytes was expanded, by introducing cortisol and cortisone, whose ratio supply additional diagnostic information. The response factors of both THF and A-THF has been checked, resulting almost identical, as well as the influence of the matrix on the calibration curves which, although different for water and urine, had similar effect on the ratios of interest. As a consequence, the calibration solutions can be prepared in pure water. The influence of several different storage procedures has also been tested, resulting in no substantial effect on the final result. Finally, the improved method has been used to run real samples from healthy volunteers, with satisfactory results. Copyright © 2008 John Wiley & Sons, Ltd. [source] Determination of trace organophosphorus pesticides in water samples with TiO2 nanotubes cartridge prior to GC-flame photometric detectionJOURNAL OF SEPARATION SCIENCE, JSS, Issue 14 2010Yunrui Huang Abstract This article described a new method for the sensitive determination of organophosphorus pesticides in water samples using SPE in combination with GC-flame photometric detection. In the procedure of method development, TiO2 nanotubes were used as SPE adsorbents for the enrichment of organophosphorus pesticides from water samples. Several factors, such as eluent and its volume, sample pH, sample volume, sample flow rate, and concentration of humic acid, were optimized. Under the optimal conditions, the proposed method had good linear ranges as 0.1,40,,g/L for each of them, LOD of 0.11, 0.014, and 0.0025,,g/L, and LOQs of 0.37, 0.047, and 0.0083,,g/L for chlorpyrifos, phorate, and methyl parathion, respectively. The proposed method was validated with real environmental water samples and the spiked recoveries were over the range of 86.5,115.1%. All these results indicated that TiO2 nanotubes, as a new SPE adsorbent, would be used widespread for the preconcentraiton and determination of environmental pollutants in the future. [source] Determination of methylene blue residues in aquatic products by liquid chromatography-tandem mass spectrometryJOURNAL OF SEPARATION SCIENCE, JSS, Issue 23-24 2009Jin-Zhong Xu Abstract A method for the determination and confirmation of methylene blue (MB) in aquatic products was developed. Residues of MB were extracted from homogenized tissues with acetonitrile/sodium acetate buffer solution, and simply cleaned up with dichloromethane liquid/liquid extraction. After concentration and dissolution, the sample solutions were cleaned up by the neutral alumina and weak cation-exchange solid phase extraction (SPE) cartridge, prior to LC-MS/MS analysis. MB was determined at 1.0,20,,g/kg in eel, toasted eel and shrimp, with a limit of quantification of 0.5,,g/kg. Recovery for MB was between 73.0% and 108.3%. This method is fast, exact and sensitive. It can be applied to determine MB in aquatic products. [source] Evaluation of sulfobetaine-type polymer resin as an SPE adsorbent in the analysis of trace tetracycline antibiotics in honeyJOURNAL OF SEPARATION SCIENCE, JSS, Issue 21 2009Tomoyasu Tsukamoto Abstract A new sulfobetaine polymer resin SPE method combined with HPLC-MS/MS for the determination of tetracycline (TC) antibiotics residues from honey samples is presented. The sulfobetaine resin was synthesized and was packed into a syringe-type tube, which served as the SPE cartridge for selective adsorption of TCs. TCs were quantitatively adsorbed on the sulfobetaine cartridge, when the loading solvent was 95%,v/v acetonitrile solution, and TCs adsorbed were not eluted by aqueous acetonitrile washing solution. TFA aqueous solution was used for eluting the adsorbed TCs. The proposed SPE method has been applied to the determination of TCs in honey samples. The recoveries of TCs spiked in honey samples ranged from 70 to 80%. Reduction of the recoveries might be derived from low solubility of TCs in acetonitrile. Compared with other SPE resins, this resin was superior in terms of selectivity with simple pretreatment. [source] Molecular imprinting of AMP by an ionic-noncovalent dual approachJOURNAL OF SEPARATION SCIENCE, JSS, Issue 19 2009Florent Breton Abstract In order to mimic recognition properties of adenylate kinase, molecularly imprinted polymers (MIPs) were prepared for adenosine 5,-monophosphate (AMP), a substrate of the enzyme. Different functional monomers interacting with the phosphate moiety were tested, and the MIP giving the best specific binding of AMP was composed with one equivalent of 2-(dimethylamino)ethyl methacrylate and ten equivalents of acrylamide compared to AMP. Packed into solid phase cartridge, this polymer showed similar characteristics than the enzyme, since it was specific for AMP toward other nucleotides. [source] Determination of fluoroquinolone antibiotics in surface waters from Mondego River by high performance liquid chromatography using a monolithic columnJOURNAL OF SEPARATION SCIENCE, JSS, Issue 17 2007Angelina Pena Abstract A novel LC,fluorescence detection method based on the use of a monolithic column for the determination of norfloxacin, ciprofloxacin, and enrofloxacin antibiotic residues in environmental waters was developed. Fluoroquinolones (FQs) were isocratically eluted using a mobile phase consisting of 0.025 M phosphoric acid solution at pH 3.0 with tetrabutylammonium and methanol (960:40, v/v) through a Chromolith Performance RP-18e column (100×4.6 mm) at a flow rate of 2.5 mL/min and detected at excitation and emission wavelengths of 278 and 450 nm, respectively. After acidification and addition of EDTA, water samples were extracted using an Oasis HLB cartridge. Linearity was evaluated in the range of 0.05 to 1 ,g/mL and correlation coefficients of 0.9945 for norfloxacin, 0.9974 for ciprofloxacin, and 0.9982 for enrofloxacin were found. The limit of quantification was 25 ng/L for the three FQs. The recovery of FQs spiked into river water samples at 25, 50, and 100 ng/L fortification levels ranged from 76.5 to 91.0% for norfloxacin, 78.5 to 97.2% for ciprofloxacin, and 79.4 to 93.6% for enrofloxacin. This method was successfully applied to the analysis of water samples from the Mondego River, and ciprofloxacin and enrofloxacin residues were detected in eight water samples. [source] Determination of cholesterol oxides content in milk products by solid phase extraction and gas chromatography-mass spectrometryJOURNAL OF SEPARATION SCIENCE, JSS, Issue 9-10 2003María V. Calvo Abstract A method for quick and reliable analysis of cholesterol oxidation products (COPs) in dairy products has been developed. After lipid extraction, fat was transesterified under mild conditions to avoid degradation of the target compounds. Isolation of the COPs studied from other components in the lipid fraction was carried out on an aminopropyl SPE cartridge. Finally, analytes were analysed by GC-MS without derivatisation. The method developed provides high specificity and good sensitivity, allowing the direct and unambiguous determination of the underivatized COPs investigated. Application of the method to dairy food analysis revealed the presence of COPs in powder milks and milk-based infant formulas commercially available in Spain, showing that attention should be focused on reduction of cholesterol oxidation levels during both the processing and the storage of these types of foodstuffs. [source] HPLC,SPE,NMR hyphenation in natural products research: optimization of analysis of Croton membranaceus extract,MAGNETIC RESONANCE IN CHEMISTRY, Issue 9 2005Maja Lambert Abstract The HPLC,SPE,NMR technique was used for the analysis of a root-bark extract of Croton membranaceus. The components of the extract were separated on an analytical-size reversed-phase HPLC column, the chromatographic peaks were trapped on SPE (solid-phase extraction) cartridges after post-column dilution of the eluate with water and the compounds were eluted from the cartridges with acetonitrile- d3 into a 30 µl 600 MHz NMR probe in a fully automated procedure. The trapping efficiency of scopoletin (1), the major extract constituent, was much higher on a GP (general phase, a polystyrene-type polymer) SPE phase than on a C18 phase. Thus, under the conditions used, up to 100 µg of scopoletin per cartridge could be accumulated linearly after repeated trappings. The maximum achievable NMR signal-to-noise ratio using the GP cartridges was at least four times higher than that achievable with the C18 cartridges. It was shown that excessively long T1 relaxation times may compromise experiments in which acetonitrile- d3 is used as the cartridge eluent. Nevertheless, the sensitivity gain provided by the HPLC,SPE,NMR technique through repeated peak trappings allowed the acquisition of good-quality proton-detected 2D NMR spectra without the need for solvent suppression. Copyright © 2005 John Wiley & Sons, Ltd. [source] Inkjet printed, conductive, 25 ,m wide silver tracks on unstructured polyimidePHYSICA STATUS SOLIDI (A) APPLICATIONS AND MATERIALS SCIENCE, Issue 7 2009Henning Meier Abstract 25 ,m wide, conductive silver tracks on untreated and unstructured polyimide have been produced by inkjet printing using standard 10 pL printheads. This was achieved by reducing the droplet size to ,2 pL by tailoring the waveform and by heating the print cartridge and the substrate to 55 °C. A simple formula was used to predict the track width. (© 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source] Investigation of the Decomposition Mechanism and Thermal Stability of Nitrocellulose/Nitroglycerine Based Propellants by Electron Spin ResonancePROPELLANTS, EXPLOSIVES, PYROTECHNICS, Issue 2 2007Anton Chin Abstract Nitrocellulose based (NC) and nitroglycerin based (NG) propellants often have a fixed acid and water content during the manufacturing time. After manufacture, the quantity and ratio of acid/water will continue to vary depending upon the conditions of storage and operation. The level of variation depends on many factors such as loading density, temperature, volume of ullage and sealing condition of the containing cartridge, just to name a few. As described in this paper and other literature, the degradation mechanisms and aging processes of NC/NG based propellants are extremely complicated. This paper describes the details of the application of Electron Spin Resonance (ESR) to study if the free-radical mechanism is involved in the decomposition of nitrocellulose and nitroglycerin. Due to the high free-radical intensity possessed by the propellant composition, we believe that a , complex intermediate may be formed between DPA and NG and/or NC. The formation of a , complex intermediate is not preferred because it may enhance the rate of decomposition of nitrate esters. [source] A mass spectrometric strategy for profiling glycoproteinoses, Pompe disease, and sialic acid storage diseasesPROTEOMICS - CLINICAL APPLICATIONS, Issue 4 2008Valegh Faid Abstract Glycoproteinoses, Pompe disease, and sialic acid storage diseases are characterized by a massive accumulation of unprocessed oligosaccharides and/or glycoconjugates in urine. The identification of these glycocompounds is essential for a proper diagnosis. In this study, we investigated the potential of MALDI-TOF-MS to identify glycocompounds present in urine from patients with different inborn errors of glycan metabolism. Urinary glycocompounds were permethylated, and analyzed using GC-MS and MALDI-TOF-MS. In order to confirm tentative assignments, a second aliquot of urine was purified on a C18 Sep-Pak cartridge and glycocompounds were desalted on a column of nonporous graphitized carbon. The glycocompounds were then sequentially on-plate digested using an array of exoglycosidases. A range of disease-specific oligosaccharides as well as glycopeptides was identified for all oligosacchariduria models. In addition, free sialic acid accumulated in urine from a patient suffering from French-type sialuria, has been detected by a GC-MS approach, which could be applied to other sialic acid storage diseases. This procedure is simple, and can be performed in few simple steps in less than 24,h. This current method can be applied for newborn screening for other inherited metabolic diseases as well as for assessing treatments in clinical trials. [source] Liquid chromatography/electrospray ionization tandem mass spectrometry validated method for the simultaneous quantification of sibutramine and its primary and secondary amine metabolites in human plasma and its application to a bioequivalence studyRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 23 2006Deepak S. Jain A high-throughput and sensitive bioanalytical method using liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) has been developed for the estimation of sibutramine and its two metabolites (M1 and M2). The extraction of sibutramine, its metabolites and imipramine (internal standard (IS)) from the plasma involved treatment with phosphoric acid followed by solid-phase extraction (SPE) using a hydrophilic-lipophilic balanced HLB cartridge. The SPE eluate without drying and reconstitution was analyzed by LC/MS/MS, equipped with a with turbo ion spray (TIS) source, operating in the positive and multiple reaction monitoring (MRM) acquisition mode. Sample preparation by this method yielded extremely clean extracts with quantitative and consistent mean recoveries; 95.12% for sibutramine, 92.74% for M1, 95.97% for M2 and 96.60% for the IS. The total chromatographic run time was 3.0,min with retention times of 2.51, 2.13, 2.09,min for sibutramine, M1, M2 and imipramine, respectively. The developed method was validated in human plasma matrix, with a sensitivity of 0.1,ng/mL (coefficient of variance (CV), 2.07%) for sibutramine, 0.1,ng/mL (CV, 3.59%) for M1 and 0.2,ng/mL (CV, 4.93%) for M2. Validation of the method for its accuracy, precision, recovery, matrix effect and stability was carried out especially with regard to real subject sample analysis. The response was linear over the dynamic range 0.1 to 8.0,ng/mL for sibutramine and M1, and 0.2 to 16.0,ng/mL for M2 with correlation coefficients of r,,,0.9959 (sibutramine), 0.9935 (M1) and 0.9943 (M2). The method was successfully applied for bioequivalence studies in 40 human subjects with 15,mg capsule formulations. Copyright © 2006 John Wiley & Sons, Ltd. [source] Determination of levetiracetam in human plasma by liquid chromatography/electrospray tandem mass spectrometry and its application to bioequivalence studiesRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 17 2006Deepak S. Jain The first liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the determination of levetiracetam, an antiepileptic drug, in human plasma is described. The plasma filtrate obtained after solid-phase extraction (SPE), using a polymer-based, hydrophilic-lipophilic balanced (HLB) cartridge, was submitted directly to a short column LC/MS/MS assay. There was no significant matrix effect on the analysis. For validation of the method, the recovery of the free analytes was compared to that from an optimized extraction method, and the analyte stability was examined under conditions mimicking sample storage, handling, and analytical procedures. The extraction procedure yielded extremely clean extracts with a recovery of 79.95% and 89.02% for levetiracetam and the internal standard (IS), respectively. The intra-assay and inter-assay precision for the samples at the lower limit of quantitation (LLOQ) were 6.33 and 6.82%, respectively. The calibration curves were linear for the dynamic range of 0.5 to 50,µg/mL with a correlation coefficient r,,,0.9971. The intra-assay accuracy at LLOQ, LQC, MQC, and HQC levels ranged from 81.60 to 95.40, 93.00 to 103.47, 95.97 to 104.09, and 91.15 to 95.18%, respectively, while the inter-assay accuracy at LLOQ, LQC, MQC and HQC levels varied from 80.20 to 95.40, 88.53 to 107.53, 95.97 to 108.45, and 91.15 to 112.70%, respectively. The method is rugged and fast with a total instrumental run time of 2,min. The method was successfully applied for bioequivalence studies in human subject samples after oral administration of 1000,mg immediate release (IR) formulations. Copyright © 2006 John Wiley & Sons, Ltd. [source] Development of a multiresidue method for analysis of major Fusarium mycotoxins in corn meal using liquid chromatography/tandem mass spectrometryRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 14 2005Chiara Cavaliere A sensitive and reliable liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) method has been developed to determine, in a single run, eight trichothecenes, three fumonisins, zearalenone and , -zearalenol, in corn meal samples. LC and MS conditions were varied to find the best compromise in terms of sensitivity and separation. An acceptable compromise was obtained using a C18 column thermostatted at 45°C and a mobile phase gradient of methanol/water with 10,mmol/L formate buffer (pH 3.8). A multiple reaction monitoring program, in which fumonisins and trichothecenes (except nivalenol and deoxynivalenol) are acquired in positive ESI as [M+H]+ or [M+NH4]+, and all other compounds in negative ESI, was developed to match appropriate retention time windows. Sample preparation used a simple homogenization of the corn meal sample with acetonitrile/water (75:25, v/v) followed by extraction on a C18 cartridge and clean-up on a cartridge containing graphitized carbon black. Method detection limits were in the range 2,14,ng/g, with the exception of nivalenol (27,ng/g), deoxynivalenol (40,ng/g) and 15-acetyldeoxynivalenol (30,ng/g). Good accuracy (recoveries 81,104%) and precision (RSD 4,11%) were obtained by performing calibration using a spiked analyte-free extract. Copyright © 2005 John Wiley & Sons, Ltd. [source] Determination of atrazine, deethylatrazine and simazine in water at parts-per-trillion levels using solid-phase extraction and gas chromatography/ion trap mass spectrometryRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 24 2003W. T. Ma Methods for trace analysis of atrazine and simazine in water have been developed by using stable-isotope dilution with detection by gas chromatography/mass spectrometry. D5 -Atrazine was used as the internal standard for the determination of atrazine and deethylatrazine, while 13C3 -simazine was used for simazine analysis. Water samples were fortified with known amounts of the internal standards and submitted to solid-phase extraction with a C18 bonded-silica cartridge. A gas chromatograph coupled with an ion-trap mass spectrometer was used to analyze the water sample extracts. Method detection limits were 38 parts-per-trillion (ppt) for atrazine and deethylatrazine and 75 ppt for simazine. The accuracy of the method, represented by relative analytical errors, was less than 15%, and the method precision was less than 5% (relative standard deviation, n,=,9). The method was successfully applied to analyze surface water samples collected from a reservoir and a river at ppt levels. Copyright © 2003 John Wiley & Sons, Ltd. [source] Determination of non-steroidal estrogens in breast milk, plasma, urine and hair by gas chromatography/mass spectrometryRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 24 2002Man Ho Choi It is suspected that all the natural estrogens occurring in the human body, as well as dietary and synthetic estrogens, diversely affect the endocrine system depending on their exposure patterns. More rapid, reliable and accurate measurements of these compounds in various biological matrices are thus becoming an important task. After solid-phase extraction using an Oasis HLB extraction cartridge, the estrogen concentrates were derivatized with a mixture of N -methyl- N -trifluorotrimethylsilylacetamide/ammonium iodide/dithioerythritol (1000:4:5, v/w/w) for analysis by gas chromatography/mass spectrometry in the selected ion-monitoring (SIM) mode. The qualitative identification of estrogens detected in SIM mode was further confirmed by tandem mass spectrometry using low-energy collision-induced dissociation (CID) mode. The method for the assay of the 20 estrogens was linear over the ranges of 1,1000,µg/L for biological fluids and 1,200,µg/kg for hair with high correlation coefficient (>0.99). The limits of quantitation (LOQ) ranged from 1.0,10,µg/L (or,µg/kg) and the limit of detection ranged from 0.2,3,µg/L (or,µg/kg). The average precision (% CV) and accuracy (% bias) of the method determined at the LOQ, low, and medium concentrations were in the ranges 2.6,9.2 and ,4.1,7.7, respectively. The average extraction recovery of the estrogens from plasma and hair at the three concentration levels varied in the ranges 77,103% (1.9,14.3% CV) and 73,104% (3.1,14%), respectively. The distribution patterns of the estrogens were characteristic of each biosample. Five estrogens in the range 1.5,44.9,µg/L were measured in breast milk, 8 estrogens in the range 3.5,322,µg/L in plasma, 12 estrogens at 1.2,442,µg/L in urine, and biochanin-A at 13.2,39.1,µg/kg in hair. Because of its high sensitivity, good precision and specificity, the present method was found suitable for the trace analysis of dietary and synthetic estrogens in complex biosamples such as breast milk, plasma, urine and hair. Copyright © 2002 John Wiley & Sons, Ltd. [source] An original approach to determining traces of tetracycline antibiotics in milk and eggs by solid-phase extraction and liquid chromatography/mass spectrometryRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 14 2002Federica Bruno An original and highly specific method able to identify and quantify traces of five tetracycline antibiotics (TCAs) in milk and eggs is presented. This method uses a single solid-phase extraction (SPE) cartridge for simultaneous extraction and purification of TCAs in the above matrices. After diluting 5,mL of intact whole milk or 2,g egg samples with Na2EDTA-containing water, samples are passed through a 0.5-g Carbograph 4 extraction cartridge. After analyte elution from the SPE cartridge, an aliquot of the final extract is injected into a liquid chromatography/mass spectrometry (LC/MS) instrument equipped with an electrospray ion source and a single quadrupole. MS data acquisition is performed in the positive-ion mode and by a time-scheduled multiple-ion selected ion-monitoring program. With methanol as organic modifier, the in-source collision-induced dissociation (CID) process generated fragment ions able to pick up one methanol molecule. In several cases, these methanol-adduct fragment ions have m/z values higher than those of the protonated molecules. This event is rarely encountered in MS, thus making the analysis of TCAs by this method extremely specific. Compared with a conventional published method, the present protocol extracted larger amounts of TCAs from both milk and egg and decreased the analysis time by a factor of 3. Recovery of TCAs in milk at the 25-ppb level ranged between 81 and 96% with relative standard deviation (RSD) no larger than 9%. Recovery of TCAs in egg at the 50-ppb level ranged between 72 and 92% with RSD no larger than 7%. Estimated limits of quantification(S/N,=,10) of the method were 2,9 ppb TCAs in whole milk and 2,19 ppb TCAs in eggs. Copyright© 2002 John Wiley & Sons, Ltd. [source] High-performance liquid chromatography/tandem mass spectrometry for the quantitative analysis of a novel taxane derivative (BAY59-8862) in biological samples and characterisation of its metabolic profile in rat bile samplesRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 19 2001Cristina Sottani A sensitive, specific, accurate and reproducible high-performance liquid chromatography (HPLC) analytical method was developed and validated for the quantification of the novel oral taxane analogue BAY59-8862 in mouse plasma and tissue samples. A fully automated solid-phase extraction procedure was applied to the plasma after internal standard (IS) addition, with only 0.2,mL volume of the sample loaded on a CN-Sep-pak cartridge. In the case of the tissues a very simple acetonitrile extraction was used to recover BAY59-8862 and its internal standard from liver. The procedure for the quantification of BAY59-8862 and its IS (IDN5127) is based on high-performance liquid chromatography/ion spray-tandem mass spectrometry, operating in selected ion monitoring mode. The retention times of BAY and IS were 7.21 and 10.36,min, respectively. In both plasma and tissue specimens the assay was linear in the range 50,5000,ng/mL (ng/g). The overall precision and accuracy were assessed on three different days. The results for plasma were within 6.1% (precision) and between 99 and 112% (accuracy), and for the liver samples within 7.3% and between 104 and 118%, respectively. The LOD was 5,ng/mL and 20,ng/g in the plasma and liver, respectively. In addition, the biliary excretion of the compound in rats was studied. The study showed that an oxidative chemical reaction was the preferred metabolic pathway for biliary excretion, and two sets of mono- and dihydroxylated metabolites were detected by LC/ISP-MS/MS experiments. With this method, BAY59-8862 pharmacokinetic was determined in mice. The combined results demonstrate that the methodology can be considered a valid approach to conduct pharmacokinetic and metabolic studies during preclinical and clinical investigations. Copyright © 2001 John7 Wiley & Sons, Ltd. [source] Testing extraction and storage parameters for a fecal hormone methodAMERICAN JOURNAL OF PRIMATOLOGY, Issue 11 2010David J. Pappano Abstract Four experiments were conducted to test different aspects of a "field-friendly" fecal hormone extraction method that utilizes methanol extraction in the field followed by storage on C18 solid-phase extraction cartridges. Fecal samples were collected from geladas (Theropithecus gelada) housed at the Bronx Zoo, and the experiments were conducted in a laboratory setting to ensure maximum control. The experiments were designed to either simulate the conditions to which fecal samples are subjected during fieldwork or improve on an existing protocol. The experiments tested the relationship between fecal hormone metabolite preservation/recovery and: (1) the amount of time a sample is stored at ambient temperature; (2) the number of freeze/thaw cycles a sample undergoes; (3) the effectiveness of different extraction solutions; and (4) the effectiveness of different cartridge washes. For each experiment, samples were assayed by radioimmunoassay for fecal glucocorticoid (GC) and testosterone (T) metabolites. Results for each of the experiments were as follows. First, storage at ambient temperature did not affect hormone levels until 4 weeks of storage, with significant increases for both GC and T metabolites at 4 weeks. Second, hormone levels significantly decreased in samples after two freeze/thaw cycles for GCs and six freeze/thaws cycles for T. Third, for both GCs and T, hormone extraction using various methanol solutions was significantly higher than using 100% ethanol. Finally, using a 20% methanol solution to wash cartridges significantly increased GC levels but had no effect on T levels. These results suggest that, when utilizing C18 cartridges for fecal steroid storage, researchers should consider several methodological options to optimize hormone preservation and recovery from fecal samples. Am. J. Primatol. 72:934,941, 2010. © 2010 Wiley-Liss, Inc. [source] High-performance liquid chromatography with solid-phase extraction for the quantitative determination of nilotinib in human plasmaBIOMEDICAL CHROMATOGRAPHY, Issue 7 2010Masatomo Miura Abstract A simple, rapid and sensitive high-performance liquid chromatography (HPLC)-based method with ultraviolet detection was developed for the quantitation of nilotinib, a tyrosine kinase inhibitor, in human plasma. Nilotinib and the internal standard dasatinib were separated using a mobile phase of 0.5% KH2PO4 (pH2.5),acetonitrile,methanol (55:25:20, v/v/v) on a Capcell Pak MG II column (250 × 4.6,mm) at a flow rate of 0.5,mL/min and optical measurement at 250,nm. Analysis required only 100,,L of plasma and involved a rapid and simple solid-phase extraction with an Oasis HLB cartridge, which gave recoveries from 72 to 78% for nilotinib and from 74 to 76% for dasatinib. The lower limit of quantification for nilotinib was 10,ng/mL. The linear range of this assay was between 10 and 5000,ng/mL (r2 > 0.9992 for the regression line). Intra- and inter-day coefficients of variation were less than 10.0% and accuracies were within 10.4% over the linear range. Our results indicate that this method is applicable to the monitoring of plasma levels of nilotinib in a clinical setting. Copyright © 2009 John Wiley & Sons, Ltd. [source] | ||||||||||||||||||||||||||||||||||