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 415 records · Page 23Linked to original sources

Laboratory and analytical method performance of lead measurements in paint chips, soils, and dusts.

The National Lead Laboratory Accreditation Program (NLLAP) recognizes laboratories capable of analyzing lead in paints, soils, and dusts. NLLAP requires successful participation in the Environmental Lead Proficiency Analytical Testing (ELPAT) program. For paint chip analyses, laboratory-to-laboratory variability is about 10% relative standard deviation (RSD) for lead levels near 0.5%, the HUD definition of lead-based paint. For soil analyses, RSDs are about 9 to 10% near relevant federal soil standards and 16% near the lowest state bare soil standard that currently exists. For dust wipe analyses, RSDs range from 10 to 16% for lead levels near relevant HUD standards. Of participating laboratories, 92 to 93% consistently meet ELPAT performance limits. A variety of analytical methods gives similar results. No conclusive significant differences were found among most frequently used hotplate and microwave sample preparation techniques. In addition, several participating laboratories have successfully used ultrasonic extraction methods, a method suitable for use at abatement sites. The three most frequently used instrumental techniques, flame atomic absorption (FAA), inductively coupled plasma-atomic emission spectroscopy (ICP-AES), and graphite furnace atomic absorption show no statistically significant differences in ability to meet ELPAT performance limits. However, small statistically significant biases between these methods sometimes occur. The magnitude of biases is less than 5% of the corresponding laboratory mean near relevant federal standards except for lead levels near the lowest HUD lead wipe standard, where biases can be as high as 8%. Other instrumental methods that have been used successfully include ICP-mass spectroscopy, direct current plasma-atomic emission spectroscopy, dithizone spectrophotometry, and anodic stripping voltametry.

Accreditation↗

Online coupling of solid-phase microextraction and capillary electrophoresis.

Solid-phase microextraction (SPME) and capillary electrophoresis (CE) are two of the main inventions that shaped 20th Century analytical chemistry. SPME is an effective microscale sampling and sample preparation technique, and CE is a high-efficiency microanalytical method. Online coupling of SPME with CE can be a powerful combination because of the significant advantages of the two techniques. The progress in the development of online SPME-CE coupling is surveyed in this review. Problems encountered and solutions reported are highlighted.

Journal Article↗

Inductively coupled plasma mass spectrometric analysis of calcium isotopes in human serum: a low-sample-volume acid-equilibration method.

BACKGROUND: Analytical methods for measuring the calcium isotope distribution in enriched human serum samples that use low blood volumes, simple preparation methods, and rapid analysis are important in clinical studies of calcium kinetics. Previously, sample preparation by oxalate precipitation typically required 500 micro L of serum. This method was time-consuming, and the blood volume required was limiting in circumstances when only a small amount of serum could be obtained. METHODS: Serum was collected from humans who were administered (42)Ca, and 20 micro L of serum was mixed with 2 mL of 0.22-0.67 mol/L HNO(3) at room temperature for between 1 min and 16 h. The (42)Ca/(43)Ca ratio in the supernatant was measured by a magnetic sector inductively coupled plasma mass spectrometer (ICP-MS). Calcium isotope ratios from these equilibration solutions were compared with data from oxalate-precipitated serum samples to determine the optimum equilibrium time and the effect of acid concentration on equilibrium. RESULTS: Various amounts of aggregated particles developed in different acid-serum mixtures. These affected the time required for isotope equilibration in the mixture. The shortest equilibrium time needed for the calcium isotopes varied from 1 to 6 h for samples acidified with 0.22-0.45 mol/L HNO(3). Data obtained from these solutions were consistent with data from oxalate-precipitated calcium. The precision of (42)Ca/(43)Ca ratio measurements was better than 0.5%. CONCLUSIONS: We have developed a simple, rapid sample preparation technique for ICP-MS analysis in which 20 micro L of serum can be used for accurate measurement of the calcium isotope distribution in a sample with good precision and a rapid analysis time.

Calcium↗

Bile acids and conjugates identified in metabolic disorders by fast atom bombardment and tandem mass spectrometry.

From a study of the collision-activated fragmentation of bile acids, a qualitative analytical method based on negative ion fast atom bombardment tandem mass spectrometry has been developed. The times for sample preparation and analyses are short. Both free and conjugated bile acids are detected as they occur in biological fluids, without derivatization. For identifying bile acids and conjugates, the method offers better specificity and sensitivity than does the fast atom bombardment mass spectrometric technique alone. Specific scan modes have been developed for the selective detection of taurine conjugates, delta 4-unsaturated taurine conjugates, delta 4-3-keto free acids and their glycine conjugates, free acids and glycine conjugates bearing a hydroxyl group at the C-12 position, sulfates of glycine and taurine conjugates, and a C29 dicarboxylic bile acid, specific for generalized peroxisomal disorders. Applications of this technique demonstrate its potential usefulness, principally in the diagnosis of several peroxisomal disorders.

Adrenoleukodystrophy↗

The determination of histamine in challenged human leukocyte preparations by high-performance liquid chromatography.

A highly sensitive and rapid method was developed for the determination of histamine in challenged human leukocyte preparations by high-performance liquid chromatography. The assay is based on the Shore's OPT-reaction of the unpurified sample and on a specific separation of the derivate with analytical reversed phase phenyl column combined with spectrofluorometric detection. The detection limit of histamine by this method was 0.07 pmol (signal to noise ratio 2:1) and the within-day variation for peak height was 3.6% and for retention time 0.8%. A good linear standard curve ranging from 12.5 pg to 500 pg (0.07 pmol-2.7 pmol) was obtained with correlation coefficient of 0.998. The histamine release from human basophils in mixed leukocyte preparation was induced by the calcium ionophore A 23187. A concentration of 0.4 micrograms/ml ionophore was required for 50% histamine release with a Ca2(+)-concentration of 1.8 mmol/l. The measured total histamine content was 1.5 pg/basophil.

Basophils↗

The use of low-resolution FT-IR spectrometry for the analysis of alcohols in breath.

Fast and reliable diagnostic methods are needed for detection or exclusion of industrial solvents as a cause of intoxication. Analyzing human breath reveals the presence of any volatile substance. A portable Fourier transform infrared (FT-IR) multicomponent point-of-care analyzer was developed for exhaled breath. The analyzer proved to be accurate and precise in laboratory tests for simultaneous measurement of methanol and ethanol in water. Ethanol, in addition to normal contents of breath, was simultaneously analyzed in human experiments, and the results correlated well with blood samples. FT-IR method has a traceable calibration to physical properties of the analyte. The measured spectra can also be saved and analyzed later. Breath analysis with FT-IR is fast and easy, and no preparation of the sample is needed.

Adult↗

Immunoassay and antibody microarray analysis of the HUPO Plasma Proteome Project reference specimens: systematic variation between sample types and calibration of mass spectrometry data.

Four different immunoassay and antibody microarray methods performed at four different sites were used to measure the levels of a broad range of proteins (N = 323 assays; 39, 88, 168, and 28 assays at the respective sites; 237 unique analytes) in the human serum and plasma reference specimens distributed by the Plasma Proteome Project (PPP) of the HUPO. The methods provided a means to (1) assess the level of systematic variation in protein abundances associated with blood preparation methods (serum, citrate-anticoagulated-plasma, EDTA-anticoagulated-plasma, or heparin-anticoagulated-plasma) and (2) evaluate the dependence on concentration of MS-based protein identifications from data sets using the HUPO specimens. Some proteins, particularly cytokines, had highly variable concentrations between the different sample preparations, suggesting specific effects of certain anticoagulants on the stability or availability of these proteins. The linkage of antibody-based measurements from 66 different analytes with the combined MS/MS data from 18 different laboratories showed that protein detection and the quality of MS data increased with analyte concentration. The conclusions from these initial analyses are that the optimal blood preparation method is variable between analytes and that the discovery of blood proteins by MS can be extended to concentrations below the ng/mL range under certain circumstances. Continued developments in antibody-based methods will further advance the scientific goals of the PPP.

Blood Proteins↗

Direct determination of methylmercury and inorganic mercury in biological materials by solid sampling-electrothermal vaporization-inductively coupled plasma-isotope dilution-mass spectrometry.

This paper reports on the use of solid sampling-electrothermal vaporization-inductively coupled plasma mass spectrometry (SS-EIV-ICPMS) for the direct and simultaneous determination of methylmercury and inorganic mercury in biological materials. The main advantage of this fast and sensitive method is that no sample preparation is required. In this way, the sample throughput can be considerably increased, problems of contamination and analyte losses are kept to a minimum and, even more important, the original chemical form of the different analyte species in the solid samples is preserved. To achieve this goal, a solid sample is inserted into a graphite furnace of the boat-in-tube type and is subsequently submitted to an appropriate temperature program, leading to the separate vaporization of methylmercury and inorganic mercury, which are transported into the ICP by means of an argon carrier gas. The separation was accomplished within 75 s. For the quantification of the two peaks, species-unspecific isotope dilution was used. For this purpose, a stable flow of argon loaded with gaseous Hg isotopically enriched in 200Hg was generated using a permeation tube that was constructed in-house. Its emission rate was determined by collecting the mercury released during a given time interval on a gold-coated silica absorber, after which the amount collected was released by heating of the absorber and determined by cold vapor atomic absorption spectrometry (CVAAS) and cold vapor atomic fluorescence spectrometry (CVAFS). A reference material from the Canadian National Research Council (NRC) (TORT-2) was used to assess the accuracy of the method. For the application of the method to samples with diverse mercury contents, the spike/sample ratio can be optimized by varying the emission rate of the permeation tube simply by adapting its temperature. To prove the feasibility of this approach, two reference materials (BCR 463 and DORM-2) with a methylmercury content more than 10 times higher than that of TORT-2 were also analyzed. The detection limits obtained for 1 mg of sample (2 ng g(-1) and 6 ng g(-1) for methylmercury and inorganic mercury, respectively) were found to be sufficiently low for this kind of application and are competitive when compared to other techniques.

Animals↗

Analytical characteristics of the determination of benzene, toluene, ethylbenzene and xylenes in water by headspace solvent microextraction.

Headspace solvent microextraction (HSM) is a novel method of sample preparation for chromatographic analysis. It involves exposing a microdrop of high-boiling point organic solvent extruded from the needle tip of a gas chromatographic syringe to the headspace above a sample. Volatile organic compounds are extracted and concentrated in the microdrop. Next, the microdrop is retracted into the microsyringe and injected directly into the chromatograph. HSM has a number of advantages, including renewable drop (no sample carryover), low cost, simplicity and ease of use, short time of analysis, high sensitivity and low detection limits, good precision, minimal solvent use, and no need for instrument modification. This paper presents analytical characteristics of HSM as applied to the determination of benzene, toluene, ethylbenzene and xylenes in water.

Benzene↗

High performance liquid chromatographic determination of Picumast and two active metabolites in plasma using on-line sample preparation.

A method for determining Picumast, an antiallergic drug, in plasma by HPLC and column switching has been developed. The system consisted of two precolumns, an analytical column, three pumps, an autosampler and a fluorescence detector. The precolumns (17 x 4.6 mm i.d.) were packed with LiChroprep RPR (a moderately polar reversed phase) and the analytical column with Nucleosil ODS (RP 18, 5 microns). The columns were connected according to the alternating precolumn technique. The mobile phase consisted of 30% CH3CN/70% 0.05 M KH2PO4, pH 2.5, with a flow gradient. Detection wavelengths were 333 nm for excitation and 383 nm for emission. The retention times of Picumast, M1 and M2 were 12, 3.6 and 4.0 min, respectively. Total run time was 15 min. The limit of detection was 3 ng/mL for M1 and 1 ng/mL for M2 and Picumast using an injection volume of 150 microL. The recoveries vary between 89% and 97% with standard deviations between 2.4 and 3.3%.

Chromatography, High Pressure Liquid↗

Simultaneous determination of seventeen glucocorticoids residues in milk and eggs by ultra-performance liquid chromatography/electrospray tandem mass spectrometry.

A comprehensive analytical method has been developed and validated for the simultaneous determination of seventeen glucocorticoid residues in eggs and milk. The mass spectrometer parameters, the composition of the mobile phase and the sample preparation method were firstly optimized to obtain maximum sensitivity. The samples were deconjugated with beta-glucuronidase/arylsulfatase enzyme and concentrated using an Oasis HLB solid-phase extraction cartridge, followed by cleanup with a dual Sep-pak silica and aminopropyl cartridge. The analytes were quantified by ultra-performance liquid chromatography (using a C18 column)/electrospray ionization tandem mass spectrometry (UPLC/ESI-MS/MS) operating in the negative ion mode. The assay for the 17 glucocorticoids was linear over the range of 1-200 microg/L for milk and egg samples with a high correlation coefficient (>0.99). The limits of quantification (LOQs) for the target analytes were 0.04-1.27 microg/kg for the egg samples and 0.03-0.73 microg/kg for the milk samples. The average extraction recoveries of the glucocorticoids from eggs and milk at two concentration levels (spiked at 0.40 and 2.00 microg/kg) were 65.6-118.7% and 61.5-119.6%, respectively, with relative standard deviations between 1.8-17.0% and 2.4-18.4%, respectively. Because of its high sensitivity, good precision and specificity, the method was found to be suitable for trace analysis of synthetic and natural glucocorticoids in complex biosamples such as eggs and milk.

Animals↗

Recognizing heterogeneous distribution of platinum group elements (PGE) in geological materials by means of the Re-Os isotope system.

The identification of uncertainties caused by sample inhomogeneity, as distinct from those caused by sample preparation and measurement, is a challenging task. Use of chemometric methods to separate and estimate these contributions to the combined standard uncertainty of a measurement (uc) of an analytical result requires complex experiments. The difficulty of platinum group element measurement makes this task even more complex. But unless it can be demonstrated that sample inhomogeneity is the major contributor to the high variability of an analytical result one should be careful not to mistakenly attribute this to a nugget effect. In this contribution we are able to demonstrate in two special cases that irreproducible results (up to 90% RSD) for analysis of Os and Re in the pg g(-1) to ng g(-1) range are truly caused by a nugget effect and not by inadequacies of the analytical method.

Journal Article↗

Electrowetting-based microfluidics for analysis of peptides and proteins by matrix-assisted laser desorption/ionization mass spectrometry.

A new technique for preparing samples for matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is reported. The technique relies on electrowetting-on-dielectric (EWOD) to move droplets containing proteins or peptides and matrix to specific locations on an array of electrodes for analysis. Standard MALDI-MS reagents, analytes, concentrations, and recipes are demonstrated to be compatible with the technique. Mass spectra are comparable to those collected by conventional methods. Nonspecific adsorption of analytes to device surfaces is demonstrated to be negligible. The results suggest that EWOD may be a useful tool for automating sample preparation for high-throughput proteomics and other applications of MALDI-MS.

Equipment Design↗

Sample preparation for biomedical analysis.

A review of sample preparation techniques for biomedical analysis is presented. Firstly, sample preparation techniques can be divided into unit operations which can be classified into four groups: release of the analyte from the matrix, removal of endogenous material, liquid handling procedures and the enhancement of selectivity and sensitivity. The concept of unit operations gives an analyst a tool with which to evaluate critically any method for preparing a sample for analysis. Secondly, the major techniques of sample preparation (protein precipitation, liquid-liquid extraction, liquid-solid extraction and high-performance liquid chromatography, HPLC) are discussed and their advantages and disadvantages presented. Thirdly, the rationale for the automation of sample preparation is reviewed; in general liquid-solid extraction and HPLC are the best techniques for automation. The means by which this can be effected (either flexible or dedicated automation) is discussed. Finally, techniques such as supercritical fluid extraction, micellar liquid chromatography, microwave energy and immunoextraction which may be applied to biomedical sample preparation are evaluated briefly.

Chemistry Techniques, Analytical↗

Single-step extraction of fluconazole from plasma by ultra-filtration for the measurement of its free concentration by high performance liquid chromatography.

High performance liquid chromatography (HPLC) is the reference method for measuring concentrations of antimicrobials in blood. This technique requires careful sample preparation. Protocols using organic solvents and/or solid extraction phases are time consuming and entail several manipulations, which can lead to partial loss of the determined compound and increased analytical variability. Moreover, to obtain sufficient material for analysis, at least 1 ml of plasma is required. This constraint makes it difficult to determine drug levels when blood sample volumes are limited. However, drugs with low plasma-protein binding can be reliably extracted from plasma by ultra-filtration with a minimal loss due to the protein-bound fraction. This study validated a single-step ultra-filtration method for extracting fluconazole (FLC), a first-line antifungal agent with a weak plasma-protein binding, from plasma to determine its concentration by HPLC. Spiked FLC standards and unknowns were prepared in human and rat plasma. Samples (240 microl) were transferred into disposable microtube filtration units containing cellulose or polysulfone filters with a 5 kDa cut-off. After centrifugation for 60 min at 15000g, FLC concentrations were measured by direct injection of the filtrate into the HPLC. Using cellulose filters, low molecular weight proteins were eluted early in the chromatogram and well separated from FLC that eluted at 8.40 min as a sharp single peak. In contrast, with polysulfone filters several additional peaks interfering with the FLC peak were observed. Moreover, the FLC recovery using cellulose filters compared to polysulfone filters was higher and had a better reproducibility. Cellulose filters were therefore used for the subsequent validation procedure. The quantification limit was 0.195 mgl(-1). Standard curves with a quadratic regression coefficient > or = 0.9999 were obtained in the concentration range of 0.195-100 mgl(-1). The inter and intra-run accuracies and precisions over the clinically relevant concentration range, 1.875-60 mgl(-1), fell well within the +/-15% variation recommended by the current guidelines for the validation of analytical methods. Furthermore, no analytical interference was observed with commonly used antibiotics, antifungals, antivirals and immunosuppressive agents. Ultra-filtration of plasma with cellulose filters permits the extraction of FLC from small volumes (240 microl). The determination of FLC concentrations by HPLC after this single-step procedure is selective, precise and accurate.

Animals↗

Direct extract derivatization for determination of amino acids in human urine by gas chromatography and mass spectrometry.

The purpose of this study was to develop a simple and accurate analytical method to determine amino acids in urine samples. The developed method involves the employment of an extract derivatization technique together with gas chromatography-mass spectrometry (GC-MS). Urine samples (300 microl) and an internal standard (10 microl) were placed in a screw tube. Ethylchloroformate (50 microl), methanol-pyridine (500 microl, 4:1, v/v) and chloroform (1 ml) were added to the tube. The organic layer (1 microl) was injected to a GC-MS system. In this proposed method, the amino acids in urine were derivatized during an extraction, and the analytes were then injected to GC-MS without an evaporation of the organic solvent extracted. Sample preparation was only required for ca. 5 min. The 15 amino acids (alanine, aspartic acid, cysteine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, tyrosine, tryptophan, valine) quantitatively determined in this proposed method. However, threonine, serine, asparagine, glutamine, arginine were not derivatized using any tested derivatizing reagent. The calibration curves showed linearity in the range of 1.0-300 microg/ml for each amino acid in urine. The correlation coefficients of the calibration curves of the tested amino acids were from 0.966 to 0.998. The limit of detection in urine was 0.5 microg/ml except for aspartic acid. This proposed method demonstrated substantial accuracy for detection of normal levels. This proposed method was limited for the determination of 15 amino acids in urine. However, the sample preparation was simple and rapid, and this method is suitable for a routine analysis of amino acids in urine.

Amino Acids↗

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↗

Laboratory database to manage electrophoresis and chromatography separations and the associated samples.

A database was developed to store, organize, and retrieve the data associated with electrophoresis and chromatography separations. It allows laboratories to store extensive data on separation techniques (analytical and preparative). The data for gel electrophoresis includes gel composition, staining methods, electric fields, analysis, and samples loaded. The database stores data on chromatography conditions, the samples used, and the fractions collected. The data structure of this database was designed to maintain the link between samples (including fractions) from chromatography separations and their analysis by gel electrophoresis. The database will allow laboratories to organize and maintain a large amount of separation and sample data in a uniform data environment. It will facilitate the retrieval of the separation history of important samples and the separation conditions used.

Chromatography↗