Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Analytic Sample Preparation Methods”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 379 records · Page 21Linked to original sources

Potentiality of gas chromatography-triple quadrupole mass spectrometry in vanguard and rearguard methods of pesticide residues in vegetables.

A new analytical strategy for the screening and confirmation/quantification of multiclass pesticide residues in vegetables has been established and validated. No complicated sample preparation was needed, but only a simple and rapid extraction using ethyl acetate and sodium sulfate, which required no cleanup. The approach is based on the use of the triple quadrupole (QqQ) mass spectrometry (MS) as detection system in gas chromatography (GC). In a first step, a GC-QqQ-MS screening method, which monitors only one MS/MS transition by compound, allows the identification of approximately 130 pesticides in 11.6 min. In this way, the differentiation between negative and potentially nonnegative samples is carried out. In the second step, the nonnegative samples are reanalyzed by the GC-QqQ-MS confirmation/quantification method, which monitors two or three MS/MS transitions by compound. Confirmation of pesticides was based on the comparison of intensity ratios for the main ions in samples with those obtained on the same day from the standard in a matrix containing the pesticides at a preestablished concentration level. Quantification of the identified and confirmed pesticides was based on the addition standard method, which avoids matrix effect. The proposed analytical strategy allowed a reliable identification and confirmation of the target pesticides at trace levels, reducing analysis time and increasing sample throughput in routine analytical laboratories.

Food Contamination↗

Development of high-performance liquid chromatographic determination of salicylaldehyde isonicotinoyl hydrazone in rabbit plasma and application of this method to an in vivo study.

An analytical methodology appropriate for the determination of the novel drug candidate salicylaldehyde isonicotinoyl hydrazone (SIH) in rabbit plasma has been developed and validated. Desirable chromatographic separation was achieved on a C18 column employing a mixture of phosphate buffer (0.01 M NaH2PO4 x 2 H2O with 2 mM EDTA, pH 6.0) and methanol (53:47; v/v) as the mobile phase. In order to develop a suitable sample preparation procedure, different methods have been tested (solid-phase extraction, liquid-liquid extraction, and protein precipitation). Protein precipitation using 0.1 M HClO4 and acetonitrile allowed the highest recoveries of the analyte to be reproducibly attained. The analytical methodology developed in this study was validated with respect to linearity (0.26-30.0 microg/mL), accuracy, precision, selectivity, recovery, and stability. A concentration of 0.26 microg/mL was determined as the LLOQ. The chromatographic method was applied to a preliminary plasma pharmacokinetic study. This study has provided the first information about the concentrations of SIH in plasma of a living subject. These results could have a significant impact on further progress in the development of this promising compound.

Aldehydes↗

Alternative high-performance liquid chromatographic assay for p-aminohippuric acid (PAH): effect of aging on PAH excretion in the isolated perfused rat kidney.

Para-aminohippuric acid (PAH), an indicator of renal plasma flow, is a commonly used marker of organic anion transport by the kidney. An analytical method for PAH using HPLC was developed. The method is simple, fast and requires a minimum amount of organic solvent. Sample preparation involved protein precipitation with zinc sulfate. Para-amino benzoic acid was utilized as an internal standard (IS). Chromatography was performed using a reversed-phase phenyl column with UV detection at a wavelength of 254 nm. Mobile phase consisted of 0.1 M acetic acid and acetonitrile (99:1) at a flow rate of 1 ml/min. The assay was validated over a standard concentration range from 1 to 25 microg/ml. Accuracy, precision, reproducibility and specificity of the method was established with coefficients of variation <10%. The method was sensitive and showed linear response in peak height ratio (analyte:IS) over the concentration range studied (r(2)>0.99). The assay was used to study the effect of aging on PAH excretion in the isolated perfused rat kidney model. Experiments were conducted in kidneys from young (2-3 months, n=6), adult (6-9 months, n=5) and aged (12-16 months, n=3) male Sprague-Dawley rats at an initial drug concentration of 20 microg/ml. Significant differences in kidney function (e.g. glomerular filtration rate and glucose reabsorption) were observed in aged kidneys. Despite a 5-fold reduction in glomerular filtration rate, PAH renal clearance (kidney weight-corrected) decreased by only 2-fold in aged (2.2+/-0.42 ml/min per gram) compared to young (4.6+/-0.70 ml/min per gram, P<0.05) rats. Furthermore, renal excretion ratio was significantly higher in aged rats (27+/-8.0 vs. 15+/-5.0, P<0.05). These preliminary findings challenge the 'Whole Nephron Hypothesis' that assumes parallel reductions in renal filtration and secretory capacity secondary to disease or aging.

Aging↗

Simultaneous determination of Z-3-[(2,4-dimethylpyrrol-5-yl)methylidenyl]-2-indolinone (SU5416) and its interconvertible geometric isomer (SU5886) in rat plasma by LC/MS/MS.

Z-3-[(2,4-Dimethylpyrrol-5-yl)methylidenyl]-2-indolinone (SU5416) is a cytostatic substance in development as an anti-angiogenic agent. SU5416 exists as the thermodynamically stable cis or Z-isomer as a solid. Studies have shown that in light exposed solutions of SU5416, the unstable trans or E-isomer, namely SU5886, is formed. The E-isomer converts back to the Z-isomer when protected from light. The E-isomer is unstable for synthesis and isolation; therefore, the analytical standard of the E-isomer is not available. In this study, a simple, fast and reliable LC/MS/MS method has been developed to determinate both isomers simultaneously in rat plasma samples to support the study of disposition kinetics of SU5416. This method is sensitive (limit of quantitation (LOQ=0.5 ng/mL)), reproducible and has a wide linear range (0.5-2500 ng/mL). There was no conversion between E- and Z-isomer during sample preparation procedure and sample determination with LC/MS/MS. Experimental results proved that SU5416 and SU5886 have identical detection response. Therefore, SU5416 (Z-isomer) was used successfully as analytical standard for SU5886 (E-isomer). This method has been applied to rat plasma samples obtained from a pharmacokinetic study. This study underscores the use of LC/MS/MS technique for bioanalytical methods where analytical standards are not available and analytes are interconvertible.

Angiogenesis Inhibitors↗

Determination of nicotine in mainstream smoke on the single puff level by liquid-phase microextraction coupled to matrix-assisted laser desorption/ionization Fourier transform mass spectrometry.

A method coupling liquid-phase microextraction with matrix-assisted laser desorption/ionization Fourier transform mass spectrometry (LPME/MALDI-FTMS) was developed to measure the content of nicotine in mainstream smoke at the single puff level. Glycerol was utilized as a matrix additive in the sample preparation to improve the homogeneity of analyte distribution in a sample spot. Good repeatability of the MALDI-MS signal (RSD <9%) was achieved by the method. Selective LPME facilitated the separation and purification of basic components from cigarette smoke. The LPME device was coupled to a smoking machine, and each puff of one cigarette could be gathered by this modified machine. The amount of nicotine in the mainstream smoke was measured at the single puff level by LPME/MALDI-FTMS. The method was simple and selective and was sufficiently sensitive to detect nicotine in each puff of one cigarette. The method thus offered an alternative approach to the study of the formation mechanism of cigarette smoke constituents.

Journal Article↗

Determination of pilocarpine, isopilocarpine, pilocarpic acid and isopilocarpic acid in human plasma and urine by high-performance liquid chromatography with tandem mass spectrometric detection.

A method is described for the determination of pilocarpine and its degradation products isopilocarpine, pilocarpic acid and isopilocarpic acid in human plasma and urine. The method is based on a simple sample preparation step -- ultrafiltration for plasma and dilution for urine samples -- followed by a reversed-phase liquid chromatographic separation of the analytes and detection by means of tandem mass spectrometry. Parameters affecting the performance of these steps are discussed. The high sensitivity and selectivity of the method allow low ng/ml concentrations to be determined for all compounds in plasma and undiluted urine, which enables the investigation of the metabolic fate and elimination of pilocarpine after oral administration to humans.

Chromatography, High Pressure Liquid↗

A new on-line sample preparation system for the liquid chromatography/time-of-flight mass spectrometry simultaneous analysis of pesticides in river water.

Current on-line solid-phase extraction methods combined with HPLC for shortening the clean-up operation are not suitable for simultaneously detecting compounds that have a wide variety of hydrophobicities. To solve these problems, we designed a new on-line sample preparation system. The system consists of an eluting pump, a mixing TEE connector, a 10-port 2-position valve and a solid-phase extraction precolumn. The eluate from the precolumn is diluted with a weak solvent from the HPLC at the TEE connection to load low hydrophobic compounds onto the analytical column. The proposed on-line sample preparation system was successfully applied to the simultaneous analysis of 21 pesticides in river water using LC/TOF-MS. In this method, the recoveries from river water samples were 67 to 126% (mean 83%), the reproducibility (CV%) was in the range from 1.1 to 11% (mean 5.6%), the calibration curve was linear in the range from 1 ppb to 500 ppb (r > 0.999) and the detection limits (S/N = 3) were in the range from 0.0034 ppb (daimuron) to 3.3 ppb (oxine-copper).

Automation↗

Capillary columns in liquid chromatography: between conventional columns and microchips.

Liquid chromatography on columns with small internal diameters has been reviewed as the intermediate technique between conventional liquid chromatography and microchip separations. The development of micro column separations in the early years has been described, starting with the papers of Horváth and co-workers and Ishii and co-workers, continuing into the first part of the eighties, then making a leap in time to recent innovations with small-bore columns. Based on internal diameters a classification of the different analytical HPLC columns has been suggested. The advantages of small-bore columns have been discussed, with particular emphasis on the advantage of coupling to concentration sensitive detectors when the sample amount is limited. Open tubular columns are treated as a part of the historic background. The recent developments include a brief look into the current status of monolithic columns, the use of packed nano columns and micro columns with electrospray mass spectrometry, and the potential of two-dimensional comprehensive liquid chromatography. Finally, the coupling of sample preparation to analytical columns and the future applications of the novel technological improvements to the microchip separation methods have been discussed.

Journal Article↗

Determination of isotope ratios in human tissues enriched with zinc stable isotope tracers using inductively coupled plasma-mass spectrometry (ICP-MS).

Enriched fecal and urine samples were prepared using ion-exchange column chromatography for analysis by Inductively Coupled and Fast Atom Bombardment Mass spectrometry (ICP-MS, FAB-MS) to compare precision between methods. Unenriched samples of human milk, feces, and whole blood were prepared similarly to monitor instrumental precision and analytical error. A least squares fit of the ICP-MS results vs the FAB-MS for 70Zn/64Zn gave a slope of 0.98, with a relative standard deviation (RSD) of only 0.7%. The results for 68Zn/64Zn gave a slope of 0.82, with a RSD of 14%. For unenriched tissues, all potential interferences were removed by the preparation procedure with no significant differences between preparation for isotope ratios of 70Zn/64Zn, 68Zn/64Zn, 67Zn/64Zn, and 66Zn/64Zn. Poisson counting statistics are a major contribution to the total analytical error indicating the usefulness of this procedure for enrichment studies.

Animals↗

Development of an ombrotrophic peat bog (low ash) reference material for the determination of elemental concentrations.

Given the increasing interest in using peat bogs as archives of atmospheric metal deposition, the lack of validated sample preparation methods and suitable certified reference materials has hindered not only the quality assurance of the generated analytical data but also the interpretation and comparison of peat core metal profiles from different laboratories in the international community. Reference materials play an important role in the evaluation of the accuracy of analytical results and are essential parts of good laboratory practice. An ombrotrophic peat bog reference material has been developed by 14 laboratories from nine countries in an inter-laboratory comparison between February and October 2002. The material has been characterised for both acid-extractable and total concentrations of a range of elements, including Al, As, Ca, Cd, Cr, Cu, Fe, Hg, Mg, Mn, Na, Ni, P, Pb, Ti, V and Zn. The steps involved in the production of the reference material (i.e. collection and preparation, homogeneity and stability studies, and certification) are described in detail.

Environmental Monitoring↗

Forensic application of an automated drug-profiling system.

The rapid emergency drug identification (REMEDi) system is an automated drug-profiling system that employs high-performance liquid chromatography with a multicolumn design. It has previously proven successful in emergency toxicology situations and in the clinical analysis of urine and serum. Its capabilities include the broad spectrum identification of more than 500 basic, neutral, and slightly acidic drugs and metabolites. Forensic applications, including analysis of whole blood and tissue, were investigated, and comparisons with more traditional laboratory methods are reported. The whole blood and tissue samples require offline sample extraction prior to system analysis. Approximately 50 drugs were used as standards to test the preparation method, analytical system, and limit of detection. More than 50 cases from the medical examiner's office were analyzed with the combination extraction and automated drug-profiling system; these cases were compared with previously reported findings. Results showed that the REMEDi system is a useful complimentary tool for screening forensic cases; the current range of detectable drugs was expanded by using the system.

Bile↗

The design of analytical methods for use in topical epidermal growth factor product development.

Data from a reverse phase gradient elution HPLC assay for human epidermal growth factor (EGF) was compared and correlated with data obtained from a competitive heterogeneous radioimmunoassay (RIA). The RIA was established to measure very low concentrations of EGF in formulation drug release and compatibility studies. The HPLC assay, capable of resolving parent and possible modified or fragmented forms of EGF isolated from human urine, was studied as a potential development tool for stability and final product evaluation. As independent analytical methods, the HPLC and the RIA procedures produced correlated results when quantifying freshly prepared and certain degraded samples of EGF. The capacity of the HPLC method to serve as a stability indicating assay was examined. Degradation of EGF was induced by storage in 0.05 M phosphate buffer pH 7.4 at 25, 37 or 50 degrees C. The same three degradation products were detected at each temperature by the HPLC method, one of which was identified as L-isoaspartyl EGF.

Amino Acids↗

[Studies for the assessment of the herbicide prometryne in the air].

An analytical method for the quantitative determination of the triazine herbicide prometryne in air is described. In the sample preparation procedure developed, the ambient air is drawn through the adsorber tube packed with the prepurified crosslinked polystyrene resin Wofatit EP 61 (0.2-0.5 mm) for collecting the pesticide with a high efficiency. A special modified cellulose filter is of use to sample particulate matter. The trapped vapours and aerosol are nearly quantitatively eluted from the sorbent by diethyl ether and then analyzed by gas chromatography. A greenhouse experiment yielded prometryne concentrations in air up to 146 ng.m-3 after spraying on soil. The described procedure permits the measurement of airborne prometryne at trace levels up to 1 ng.m-3.

Air Pollutants↗

Application of inductively coupled plasma sector field mass spectrometry for elemental analysis of urine.

An analytical method using double focusing sector field inductively-coupled plasma mass spectrometry (sector field ICP-MS) for rapid simultaneous determination of 42 elements in urine is described. Sample preparation consisted of 20-fold dilution with 0.14 mol/l nitric acid in ultrapure water. The importance of controlling possible contamination sources at different sample preparation and analysis stages in order to achieve adequate method detection limits (DL) is emphasized. Correction for matrix effects was made using indium and lutetium as internal standards. Different approaches for accuracy assessment in urine analysis are evaluated. Additional information on trace element concentrations in a urine reference material is given. Between-batch precision was assessed from the analysis of separately prepared aliquots of the reference material and was better than 10% RSD for 32 of the elements. The robustness of the procedure was tested by analysis of about 250 samples in one analytical run lasting more than 50 hours. A statistical summary of results for 19 urine samples from non-exposed subjects is presented. For a majority of the elements tested concentrations were higher than the detection limit of the method.

Calibration↗

Validation of a high-performance liquid chromatographic assay for lysylpyridinoline in urine: a potential biomarker of bone resorption.

We developed and validated a high-performance liquid chromatographic (HPLC) method for quantifying the bone-specific collagen crosslink, lysylpyridinoline (LP), in urine, LP was purified from cortical bone and characterized by spectrophotometry, HPLC, and 1H NMR spectroscopy. Our HPLC detected urinary LP independently of the sample volume and the range of quantification was between 63 nM and 1 microM. Average recovery of added LP standard to human urine samples was 106 +/- 21%. Mean inter- and intraassay CVs, respectively, for urines containing low, medium, and high concentrations of the crosslink LP were 7.4 and 6.3%. This analytical method is more efficient than previously published HPLC assays for LP because of the significant 24-h reduction in urinary sample preparation time. There was agreement between urinary LP concentrations measured with this method and the Metra Pyrilinks-D enzyme immunoassay (r2 = 0.714). These results emphasize the importance of using a thoroughly standardized HPLC assay as the "gold standard" for comparison of results with newly developed immunoassays.

Aged↗

Fluoride determination in plasma by ion selective electrodes: a simplified method for the clinical laboratory.

A potentiometric method for the determination of fluoride (F-) in serum and plasma is proposed; it is based on a combination of the single-known-addition method and the electrode slope-by-dilution method. This procedure provides reliable results in extremely low measuring ranges down to 2.5 mug/l, where the electrode slope deviates markedly from Nernstian behaviour. In this method no electrode calibration is required and only one standard is necessary. 1 ml of plasma is sufficient for one analysis. Apart from a 5% enrichment of all samples with a concentrated total ionic strength adjustment buffer, no further preparation of the sample is required. The simplicity of the various pipetting and analytical steps, and also of the evaluation of the readings, may render this method highly suitable for the clinical laboratory. Investigations into the accuracy and precision of the method produced satisfactory results. The recovery in plasma amounted to 99.7%, even in the low measuring ranges. The discrimination capacity of the method amounts to 0.1 mug/l. With the apparatus and experimental procedures described, 18 plasma analyses per day can be performed even at low F- concentrations with which longer electrode stabilization periods are required. Storage of the plasma samples frozen at --20 degrees C for up to 8 weeks exerts no effect on the F- concentration. Problems of sample contamination and other disturbances are discussed. Determinations of normal values in 20 test subjects resulted in a mean value of 10.4 plus or minus 4.01 mug/l (Mean plus or minus S.D.). The modal value amounted to 9.5 mug/l, and the range was between 5.9 and 18.8 mug/l. The F- content of the drinking water supplied to this group of persons amounted to 180 mug/l. The importance of the method has been illustrated using a clinico-nephrological study as an example.

Adult↗

Liquid chromatographic determination of ivermectin in feed.

An analytical method for determining ivermectin in feed at 0.50-3 ppm is presented. The method is based on liquid chromatographic measurement after sample preparation by adsorption chromatography on alumina and solid-phase extraction. Two complete, final, finished medicated feeds and the corresponding control feeds used in their preparation were analyzed. Recoveries from feeds fortified at 50-150% of the 2 ppm ivermectin use concentration also were determined. Mean recoveries from replicate analyses ranged from 90 to 100%, and coefficients of variation (CVs) were less than 4.5%. No significant interferences were found in control feeds. The pooled distribution of individual analytical results (n = 100) gave a mean recovery of 100%, a recovery range of 90-111%, and an overall CV of 5.5%. Resolution of the total variance into its 2 components gave a within-laboratory CV of 4.1% and a between-laboratory CV of 3.4%. There was no significant difference in recoveries among laboratories, days, concentrations, and feed base or between fortified and medicated feeds (P > 0.2).

Animal Feed↗

Evaluation of ursodeoxycholic acid bioavailability from immediate- and sustained-release preparations using gas chromatography-mass spectrometry and high-performance liquid chromatography.

An improved procedure is presented for the determination of ursodeoxycholic acid (CAS 128-13-2, UDCA) in human plasma and bile after oral administration of UDCA-containing dosage forms. The plasma samples after solid-phase extraction with silica-based C18- and strong anion exchange cartridges were assayed by gas chromatography-mass spectrometry (GC-MS) using selected-ion monitoring. The hexafluoroisopropyl trifluoroacetate ester derivative of UDCA was selected for GC analysis since it is easily and rapidly prepared by a one-step reaction. Biliary UDCA levels were determined by a rapid and simple high-performance liquid chromatographic (HPLC) method with on-line sample purification. This analytical protocol was used to investigate the pharmacokinetic of a new sustained-release capsule of UDCA in comparison with a reference immediate-release preparation after single oral administration. Statistical evaluation of the area under the plasma concentration-time curves indicated that two formulations are equivalent with regard to the amount of drug absorbed. However, pharmacokinetic data showed that with the sustained-release preparation a significantly delayed mean peak plasma level was reached compared with the reference preparation. Moreover, the immediate- and extended-release capsules were found to achieve a comparable degree of biliary enrichment with UDCA.

Adult↗