Unknown Sample (unknown + sample)

Distribution by Scientific Domains

Selected Abstracts

Comparative metabolite profiling of carboxylic acids in rat urine by CE-ESI MS/MS through positively pre-charged and 2H-coded derivatization

Wen-Chu Yang
Abstract A new approach to the selective comparative metabolite profiling of carboxylic acids in rat urine was established using CE-MS and a method for positively pre-charged and 2H-coded derivatization. Novel derivatizing reagents, N -alkyl-4-aminomethyl-pyridinum iodide (alkyl=butyl, butyl-d9 or hexyl), containing quaternary amine and stable-isotope atoms (deuterium), were introduced for the derivatization of carboxylic acids. CE separation in positive polarity showed high reproducibility (0.99,1.32% RSD of migration time) and eliminated problems with capillary coating known in CE-MS anion analyses. Essentially complete ionization and increased hydrophobicity after the derivatization also enhanced MS detection sensitivity (e.g. formic acid was detected at 0.5,pg). Simultaneous derivatization of one sample using two structurally similar reagents, N -butyl-4-aminomethyl-pyridinum iodide (BAMP) and N -hexyl-4-aminomethyl-pyridinum iodide, provided additional information for recognizing a carboxylic acid in an unknown sample. Moreover, characteristic fragmentation acquired by online CE-MS/MS allowed for identification and categorization of carboxylic acids. Applying this method on rat urine, we found 59 ions matching the characteristic patterns of carboxylic acids. From these 59, 32 ions were positively identified and confirmed with standards. For comparative analysis, 24 standard carboxylic acids were derivatized by chemically identical but isotopically distinct BAMP and N -butyl-d9-4-aminomethyl-pyridinium iodide, and their derivatization limits and linearity ranges were determined. Comparative analysis was also performed on two individual urine samples derivatized with BAMP and N -butyl-d9-4-aminomethyl-pyridinium iodide. The metabolite profiling variation between these two samples was clearly visualized. [source]

Linearization of second-order calibration curves in stable isotope dilution,mass spectrometry

Laurent B. Fay
Abstract The quantification of compounds using isotope dilution mass spectrometry requires the establishment of calibration curves prior to determination of any unknown sample. When calibration over a wide concentration range is required and/or when an overlap exists between internal standard and analyte ions (if mono- or di-isotopically-labelled internal standards are used), second-order calibration curves are obtained. In this paper we have compared several calculation methods to linearize such calibration curves. We found that the method published by Bush and Trager6 gives a satisfactory linear relationship between the corrected amount ratio y = Ql(Qu+tQl) (the value Qu being the amount of unlabelled analyte, Ql the amount of labelled internal standard and t, the fixed fraction of the internal standard, which is identical to the unlabelled analyte) and the ratio of unlabelled to labelled ion intensities. All the other calculation methods that have been published so far have failed to linearize the second-order calibration curve build-up over a wide concentration range. Copyright © 2001 John Wiley & Sons, Ltd. [source]

Structural determination of ethylene-propylene-diene rubber (EPDM) containing high degree of controlled long-chain branching

Susanta Mitra
Abstract This work highlights an attempt to characterize the degree and nature of long-chain branching (LCB) in an unknown sample of ethylene-propylene-diene rubber (EPDM). Two EPDM rubbers selected for this study were comparable in comonomer compositions but significantly different with respect to molar mass and the presence of LCB. Both rubbers contained 5-ethylidene-2-norbornene (ENB) as diene. Solution cast films of pure EPDM samples were used for different characterization techniques. 1H-NMR, and 13C-NMR were used for assessing the comonomer ratios and LCB. Size exclusion chromatography (SEC) equipped with triple detector system was used to determine the molar mass (both absolute and relative) and polydispersity index (PDI). Presence of branching was also detected using sec-viscometry. Rheological analysis has also been used for characterizing LCB. Finally, on the basis of the experimental findings and the available theories, an attempt was made to identify the chemical nature and degree of LCB. This study reveals the possibility of detailed characterization of molecular architecture of EPDM containing LCB by comparing with an essentially linear EPDM in light of an existing theory. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2009 [source]

Characterization by mass spectrometry of an unknown polysiloxane sample used under uncontrolled medical conditions for cosmetic surgery

Cédric Schneider
For a complete understanding of the raw material used for cosmetic surgery under uncontrolled medical conditions, an unknown sample of polydimethylsiloxanes has been investigated utilizing a combination of analytical techniques: pyrolysis/gas chromatography/mass spectrometry (Py/GC/MS), electrospray ionization (ESI)-MS, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF)MS, and liquid chromatography (LC)/MS. Among these techniques, the LC/APCI-MS coupling allowed the fastest and more effective analysis. In addition, the complexity of the mass spectra deduced from these LC/MS experiments was simplified compared to the mass spectra obtained by MALDI-TOF. In this work, we have demonstrated how the LC/APCI-MS coupling applied to polydimethylsiloxane samples permits the full characterization of samples where end groups of different nature can be present in very small quantities. Copyright © 2008 John Wiley & Sons, Ltd. [source]

Bayesian Analysis of Serial Dilution Assays

BIOMETRICS, Issue 2 2004
Andrew Gelman
Summary. In a serial dilution assay, the concentration of a compound is estimated by combining measurements of several different dilutions of an unknown sample. The relation between concentration and measurement is nonlinear and heteroscedastic, and so it is not appropriate to weight these measurements equally. In the standard existing approach for analysis of these data, a large proportion of the measurements are discarded as being above or below detection limits. We present a Bayesian method for jointly estimating the calibration curve and the unknown concentrations using all the data. Compared to the existing method, our estimates have much lower standard errors and give estimates even when all the measurements are outside the "detection limits." We evaluate our method empirically using laboratory data on cockroach allergens measured in house dust samples. Our estimates are much more accurate than those obtained using the usual approach. In addition, we develop a method for determining the "effective weight" attached to each measurement, based on a local linearization of the estimated model. The effective weight can give insight into the information conveyed by each data point and suggests potential improvements in design of serial dilution experiments. [source]

Identification of CpG methylation of the SNRPN gene by methylation-specific multiplex PCR

Chia-Cheng Hung
Abstract In this article, we show that methylation-specific multiplex PCR (MS-multiplex PCR) is a sensitive and specific single assay for detecting CpG methylation status as well as copy number aberrations. We used MS-multiplex PCR to simultaneously amplify three sequences: the 3, ends of the SNRPN gene (for unmethylated sequences), the KRITI gene (as internal control), and the promoter of the SNRPN gene containing CpG islands (for methylated sequences) after digestion with a methylation-sensitive restriction enzyme (HhaI). We established this duplex assay for the analysis of 38 individuals with Prader,Willi syndrome, 2 individuals with Angelman syndrome, and 28 unaffected individuals. By comparing the copy number of the three regions, the methylation status and the copy number changes can be easily distinguished by MS-multiplex PCR without the need of bisulfite treatment of the DNA. The data showed that MS-multiplex PCR allows for the estimation of the methylation level by comparing the copy number aberrations of unknown samples to the standards with a known methylated status. The in-house-designed MS-multiplex PCR protocol is a relatively simple, cost-effective, and highly reproducible approach as a significant strategy in clinical applications for epigenetics in a routine laboratory. [source]

Predicting pasture root density from soil spectral reflectance: field measurement

This paper reports the development and evaluation of a field technique for in situ measurement of root density using a portable spectroradiometer. The technique was evaluated at two sites in permanent pasture on contrasting soils (an Allophanic and a Fluvial Recent soil) in the Manawatu region, New Zealand. Using a modified soil probe, reflectance spectra (350,2500 nm) were acquired from horizontal surfaces at three depths (15, 30 and 60 mm) of an 80-mm diameter soil core, totalling 108 samples for both soils. After scanning, 3-mm soil slices were taken at each depth for root density measurement and soil carbon (C) and nitrogen (N) analysis. The two soils exhibited a wide range of root densities from 1.53 to 37.03 mg dry root g,1 soil. The average root density in the Fluvial soil (13.21 mg g,1) was twice that in the Allophanic soil (6.88 mg g,1). Calibration models, developed using partial least squares regression (PLSR) of the first derivative spectra and reference data, were able to predict root density on unknown samples using a leave-one-out cross-validation procedure. The root density predictions were more accurate when the samples from the two soil types were separated (rather than grouped) to give sub-populations (n = 54) of spectral data with more similar attributes. A better prediction of root density was achieved in the Allophanic soil (r2 = 0.83, ratio prediction to deviation (RPD ) = 2.44, root mean square error of cross-validation (RMSECV ) = 1.96 mg g ,1) than in the Fluvial soil (r2 = 0.75, RPD = 1.98, RMSECV = 5.11 mg g ,1). It is concluded that pasture root density can be predicted from soil reflectance spectra acquired from field soil cores. Improved PLSR models for predicting field root density can be produced by selecting calibration data from field data sources with similar spectral attributes to the validation set. Root density and soil C content can be predicted independently, which could be particularly useful in studies examining potential rates of soil organic matter change. [source]

FULLPAT: a full-pattern quantitative analysis program for X-ray powder diffraction using measured and calculated patterns

Steve J. Chipera
FULLPAT is a quantitative X-ray diffraction methodology that merges the advantages of existing full-pattern fitting methods with the traditional reference intensity ratio (RIR) method. Like the Rietveld quantitative analysis method, it uses complete diffraction patterns, including the background. However, FULLPAT can explicitly analyze all phases in a sample, including partially ordered or amorphous phases such as glasses, clay minerals, or polymers. Addition of an internal standard to both library standards and unknown samples eliminates instrumental and matrix effects and allows unconstrained analyses to be conducted by direct fitting of library standard patterns to each phase in the sample. Standard patterns may include data for any solid material including glasses, and calculated patterns may also be used. A combination of standard patterns is fitted to observed patterns using least-squares minimization, thereby reducing user intervention and bias. FULLPAT has been coded into Microsoft EXCEL using standard spreadsheet functions. [source]

Electronic Nose Technology in Quality Assessment: Predicting Volatile Composition of Danish Blue Cheese During Ripening

Jeorgos Trihaas
ABSTRACT This work describes for the 1st time the use of an electronic nose (e-nose) for the determination of changes of blue cheeses flavor during maturation. Headspace analysis of Danish blue cheeses was made for 2 dairy units of the same producer. An e-nose registered changes in cheeses flavor 5, 8, 12, and 20 wk after brining. Volatiles were collected from the headspace and analyzed by gas chromatography-mass spectrometry (GC-MS). Features from the chemical sensors of the e-nose were used to model the volatile changes by multivariate methods. Differences registered during ripening of the cheeses as well as between producing units are described and discussed for both methods. Cheeses from different units showed significant differences in their e-nose flavor profiles at early ripening stages but with ripening became more and more alike. Prediction of the concentration of 25 identified aroma compounds by e-nose features was possible by partial least square regression (PLS-R). It was not possible to create a reliable predictive model for both units because cheeses from 1 unit were contaminated by Geotrichum candidum, leading to unstable ripening patterns. Correction of the e-nose features by multiple scatter correction (MSC) and mean normalization (MN) of the integrated GC areas made correlation of the volatile concentration to the e-nose signal features possible. Prediction models were created, evaluated, and used to reconstruct the headspace of unknown cheese samples by e-nose measurements. Classification of predicted volatile compositions of unknown samples by their ripening stage was successful at a 78% and 54% overall correct classification for dairy units 1 and 2, respectively. Compared with GC-MS, the application of the rapid and less demanding e-nose seems an attractive alternative for this type of investigation. [source]

Use of evaporative light scattering detector in the detection and quantification of enantiomeric mixtures by HPLC

Tong Zhang
Abstract Routinely used in our laboratories at analytical scale, an evaporative light scattering detector (ELSD) has proved to be versatile in the detection of enantiomeric resolution using chiral stationary phases by HPLC. Though this kind of detector has been widely used in various domains, its application in enantiomeric resolution has not been discussed in the literature and is found to have very specific features especially in the quantitative perspective. In contrast with the UV detection, the peak area from ELSD for both enantiomers of a racemic mixture may not be the same. This complicates the assessment of the enantiomeric purity of unknown samples. This current work deals with some practical aspects in the detection of enantiomers and in accurate quantitative determination of enantiomeric purity by ELSD. Effects of analyte nature (more precisely molecular weight and volatility), peak shape and peak shape difference between enantiomers on the quantitative integration by ELSD are discussed in connection with the UV-detection results. The calibration for quantitative enantiomeric analysis and its effectiveness are demonstrated. [source]