Error propagation in normalization of stable isotope data: a Monte Carlo analysis

A higher analytical precision of a stable isotope ratio mass spectrometer does not automatically guarantee accurate determination of the true isotope composition (δ‐value) of samples, since estimates of true δ‐values are obtained from the normalization of raw isotope data. We performed both Monte Carlo simulations and laboratory experiments to investigate aspects of error propagation during the normalization of carbon stable isotope data. We found that increasing both the number of different reference standards and the number of repetitions of each of these standards reduces the normalization error. A 50% reduction in the normalization error can be achieved over the two‐point normalization by either analyzing two standards four times each, or four standards two times each. If the true δ‐value of a sample is approximately known a priori, the normalization error may then be reduced through a targeted choice of locally optimal standards. However, the difference in improvement is minimal and, therefore, a more practical strategy is to use two or more standards covering the whole stable isotope scale. The selection of different sets of standards by different laboratories or for different batches of samples in the same laboratory may lead to significant differences in the normalized δ‐values of the same samples, leading to inconsistent results. Hence, the same set of standards should always be used for a particular element and a particular stable isotope analytical technique. Copyright © 2010 John Wiley & Sons, Ltd.     

Correlation between the synthetic origin of methamphetamine samples and their 15N and 13C stable isotope ratios

The active ingredient of ecstasy, N-methyl-3,4-methyldioxyphenylisopropylamine (MDMA) can be manufactured by a number of easy routes from simple precursors. We have synthesised 45 samples of MDMA following the five most common routes using N-precursors from 12 different origins and three different precursors for the aromatic moiety. The ¹³C and ¹⁵N contents of both the precursors and the MDMA samples derived therefrom were measured by isotope ratio mass spectrometry coupled to an elemental analyser (EA-IRMS). We show that within-pathway correlation between the ¹⁵N content of the precursor and that of the derived MDMA can be strong but that no general pattern of correlation can be defined. Rather, it is evident that the δ¹⁵N values of MDMA are strongly influenced by a combination of the δ¹⁵N values of the source of nitrogen used, the route by which the MDMA is synthesised, and the experimental conditions employed. Multivariate analysis (PCA) based on the δ¹⁵N values of the synthetic MDMA and of the δ¹⁵N and δ¹³C values of the N-precursors leads to good discrimination between the majority of the reaction conditions tested.     

Reconstructing bulk isotope ratios from compound‐specific isotope ratios

Carbon isotope analysis by bulk elemental analysis coupled with isotope ratio mass spectrometry has been the mainstay of δ¹³C analyses both at natural abundance and in tracer studies. More recently, compound‐specific isotope analysis (CSIA) has become established, whereby organic constituents are separated online by gas or liquid chromatography before oxidation and analysis of CO₂ for constituent δ¹³C. Theoretically, there should be concordance between bulk δ¹³C measurements and carbon‐weighted δ¹³C measurements of carbon‐containing constituents. To test the concordance between the bulk and CSIA, fish oil was chosen because the majority of carbon in fish oil is in the triacylglycerol form and ～95% of this carbon is amenable to CSIA in the form of fatty acids. Bulk isotope analysis was carried out on aliquots of oil extracted from 55 fish samples and δ¹³C values were obtained. Free fatty acids (FFAs) were produced from the oil samples by saponification and derivatised to fatty acid methyl esters (FAMEs) for CSIA by gas chromatography/combustion/isotope ratio mass spectrometry. A known amount of an internal standard (C15:0 FAME) was added to allow analyte quantitation. This internal standard was also isotopically calibrated in both its FFA (δ¹³C = ?34.30‰) and FAME (δ¹³C = ?34.94‰) form. This allowed reporting of FFA δ¹³C from measured FAME δ¹³C values. The bulk δ¹³C was reconstructed from CSIA data based on each FFA δ¹³C and the relative amount of CO₂ produced by each analyte. The measured bulk mean δ¹³C (SD) was ?23.75‰ (1.57‰) compared with the reconstructed bulk mean δ¹³C of ?23.76 (1.44‰) from CSIA and was not significantly different. Further analysis of the data by the Bland‐Altman method did not show particular bias in the data relative to the magnitude of the measurement. Good agreement between the methods was observed with the mean difference between methods (range) of 0.01‰ (?1.50 to 1.30). Copyright © 2010 John Wiley & Sons, Ltd.     

A simple, practical methodology for routine VSMOW/SLAP normalization of water samples analyzed by continuous flow methods

Normalization of stable isotope data is important for meaningful inter-laboratory comparisons of data, especially for waters where there may be large natural variations in isotope ratios of oxygen and hydrogen. As a result, large, systematic errors may arise in continuous flow applications without correction, whereas normalization to the VSMOW/SLAP scale can facilitate inter-laboratory comparison and can be accomplished by a simple procedure in which secondary laboratory standards, carefully calibrated, are analyzed along with unknown samples. Delta values for these standards, as analyzed, are plotted against the calibrated values and a linear regression is performed. The resulting equation is applied to unknown samples to achieve the normalization. The one-sigma [1sigma] standard deviation for replicate samples by this normalization method using a Finnigan Gasbenchll should be 相似文献   

δ 15N measurement of organic and inorganic substances by EA-IRMS: a speciation-dependent procedure

Little attention has been paid so far to the influence of the chemical nature of the substance when measuring δ ¹⁵N by elemental analysis (EA)–isotope ratio mass spectrometry (IRMS). Although the bulk nitrogen isotope analysis of organic material is not to be questioned, literature from different disciplines using IRMS provides hints that the quantitative conversion of nitrate into nitrogen presents difficulties. We observed abnormal series of δ ¹⁵N values of laboratory standards and nitrates. These unexpected results were shown to be related to the tailing of the nitrogen peak of nitrate-containing compounds. A series of experiments were set up to investigate the cause of this phenomenon, using ammonium nitrate (NH₄NO₃) and potassium nitrate (KNO₃) samples, two organic laboratory standards as well as the international secondary reference materials IAEA-N1, IAEA-N2—two ammonium sulphates [(NH₄)₂SO₄]—and IAEA-NO-3, a potassium nitrate. In experiment 1, we used graphite and vanadium pentoxide (V₂O₅) as additives to observe if they could enhance the decomposition (combustion) of nitrates. In experiment 2, we tested another elemental analyser configuration including an additional section of reduced copper in order to see whether or not the tailing could originate from an incomplete reduction process. Finally, we modified several parameters of the method and observed their influence on the peak shape, δ ¹⁵N value and nitrogen content in weight percent of nitrogen of the target substances. We found the best results using mere thermal decomposition in helium, under exclusion of any oxygen. We show that the analytical procedure used for organic samples should not be used for nitrates because of their different chemical nature. We present the best performance given one set of sample introduction parameters for the analysis of nitrates, as well as for the ammonium sulphate IAEA-N1 and IAEA-N2 reference materials. We discuss these results considering the thermochemistry of the substances and the analytical technique itself. The results emphasise the difference in chemical nature of inorganic and organic samples, which necessarily involves distinct thermochemistry when analysed by EA-IRMS. Therefore, they should not be processed using the same analytical procedure. This clearly impacts on the way international secondary reference materials should be used for the calibration of organic laboratory standards.

Concentration effects on laser-based δ18 O and δ2 H measurements and implications for the calibration of vapour measurements with liquid standards

Recently available isotope ratio infrared spectroscopy can directly measure the isotopic composition of atmospheric water vapour (δ¹⁸O, δ²H), overcoming one of the main limitations of isotope ratio mass spectrometry (IRMS) methods. Calibrating these gas‐phase instruments requires the vapourisation of liquid standards since primary standards in principle are liquids. Here we test the viability of calibrating a wavelength‐scanned cavity ring‐down spectroscopy (CRDS) instrument with vapourised liquid standards. We also quantify the dependency of the measured isotope values on the water concentration for a range of isotopic compositions. In both liquid and vapour samples, we found an increase in δ¹⁸O and δ²H with water vapour concentration. For δ¹⁸O, the slope of this increase was similar for liquid and vapour, with a slight positive relationship with sample δ‐value. For δ²H, we found diverging patterns for liquid and vapour samples, with no dependence on δ‐value for vapour, but a decreasing slope for liquid samples. We also quantified tubing memory effects to step changes in isotopic composition, avoiding concurrent changes in the water vapour concentration. Dekabon tubing exhibited much stronger, concentration‐dependent, memory effects for δ²H than stainless steel or perfluoroalkoxy (PFA) tubing. Direct vapour measurements with CRDS in a controlled experimental chamber agreed well with results obtained from vapour simultaneously collected in cold traps analysed by CRDS and IRMS. We conclude that vapour measurements can be calibrated reliably with liquid standards. We demonstrate how to take the concentration dependencies of the δ‐values into account. Copyright © 2010 John Wiley & Sons, Ltd.     

Simultaneous δ15N, δ13C and δ34S measurements of low‐biomass samples using a technically advanced high sensitivity elemental analyzer connected to an isotope ratio mass spectrometer

Conventional simultaneous CNS stable isotope abundance measurements of solid samples usually require high sample amounts, up to 1 mg carbon, to achieve exact analytical results. This rarely used application is often impaired by high C:S element ratios when organic samples are analyzed and problems such as incomplete conversion into sulphur dioxide occur during analysis. We introduce, as a technical innovation, a high sensitivity elemental analyzer coupled to a conventional isotope ratio mass spectrometer, with which CNS‐stable isotope ratios can be determined simultaneously in samples with low carbon content (<40 µg C corresponding to ～100 µg dry weight). The system includes downsized reactors, a temperature program‐controlled gas chromatography (GC) column and a cryogenic trap to collect small amounts of sulphur dioxide. This modified application allows for highly sensitive measurements in a fully automated operation with standard deviations better than ±0.47‰ for δ¹⁵N and δ³⁴S and ±0.12‰ for δ¹³C (n = 127). Samples collected from one sampling site in a Baltic fjord within a short time period were measured with the new system to get a first impression of triple stable isotope signatures. The results confirm the potential of using δ³⁴S as a stable isotope tracer in combination with δ¹⁵N and δ¹³C measurements to improve discrimination of food sources in aquatic food webs. Copyright © 2009 John Wiley & Sons, Ltd.     

An online method combining a Gasbench II with continuous flow isotope ratio mass spectrometry to determine the content and isotopic compositions of minor amounts of carbonate in silicate rocks

An online method using continuous flow isotope ratio mass spectrometry (CF‐IRMS) interfaced with a Gasbench II device was established to analyze carbon and oxygen isotopic compositions and to estimate the content of minor amounts of carbonate in silicate rocks. The mixtures of standard materials and high‐purity quartz are firstly used to calibrate different quantities of carbonate in silicates. The results suggest that the accuracy and precision of the online analysis are both better than those obtained using an offline method. There is a positive correlation between the carbonate weight and the Mass44 ion beam intensity (or peak area). When the weight of carbonate in the mixtures is greater than 70 µg (equal to ～1800 mV Mass44 ion beam intensity), the δ¹³C and δ¹⁸O values of samples usually have accuracy and precision of ±0.1‰ and ±0.2‰ (1σ), respectively. If the weight is less than 70 µg, some limitations (e.g., not perfectly linear) are encountered that significantly reduce the accuracy and precision. The measured δ¹⁸O values are systematically lower than the true values by ?0.3 to ?0.7‰; the lower the carbonate content, the lower the measured δ¹⁸O value. For samples with lower carbonate content, the required phosphoric acid doses are higher and more oxygen isotope exchanges with the water in the phosphoric acid. To guarantee accurate results with high precision, multiple analyses of in‐house standards and an artificial MERCK sample with δ¹³C values from ?35.58 to 1.61‰ and δ¹⁸O from 6.04 to 18.96‰ were analyzed simultaneously with the unknown sample. This enables correction of the measured raw data for the natural sample based on multiple‐point normalization. The results indicate that the method can be successfully applied to a range of natural rocks. Copyright © 2010 John Wiley & Sons, Ltd.     

A test of comparative equilibration for determining non‐exchangeable stable hydrogen isotope values in complex organic materials

Comparative equilibration has been proposed as a methodological approach for determining the hydrogen isotopic composition (δD) of non‐exchangeable hydrogen in complex organic materials, from feathers to blood and soils. This method depends on using homogenized standards that have been previously calibrated for their δD values of non‐exchangeable H, that are compositionally similar to unknown samples, and that span an appropriate isotopic range. Currently no certified organic reference materials with exchangeable H exist, and so isotope laboratories have been required to develop provisional internal calibration standards, such as the keratin standards currently used in animal migration studies. Unfortunately, the isotope ratios of some samples fall outside the range of keratin standards currently used for comparative equilibration. Here we tested a set of five homogenized keratin powders as well as feathers from Painted Buntings and Dark‐eyed Juncos to determine the effects of extrapolating comparative equilibration normalization equations outside the isotopic range of keratin standards. We found that (1) comparative equilibration gave precise results within the range of the calibration standards; (2) linear extrapolation of normalization equations produced accurate δD results to ～40‰ outside the range of the keratins standards used (?187 to ?108); and (3) for both homogenized keratin powders and heterogeneous unknown samples there was no difference in variance between samples within and outside the range of keratin standards. This suggested that comparative equilibration is a robust and practical method for determining the δD of complex organic matrices, although caution is required for samples that fall far outside the calibration range. Copyright © 2009 John Wiley & Sons, Ltd.     

Inter-laboratory calibration of new silver orthophosphate comparison materials for the stable oxygen isotope analysis of phosphates

Stable oxygen isotope compositions (δ¹⁸O values) of two commercial and one synthesized silver orthophosphate reagents have been determined on the VSMOW scale. The analyses were carried out in three different laboratories: lab (1) applying off‐line oxygen extraction in the form of CO₂ which was analyzed on a dual inlet and triple collector isotope ratio mass spectrometer, while labs (2) and (3) employed an isotope ratio mass spectrometer coupled to a high‐temperature conversion/elemental analyzer (TC/EA) where Ag₃PO₄ samples were analyzed as CO in continuous flow mode. The δ¹⁸O values for the proposed new comparison materials were linked to the generally accepted δ¹⁸O values for Vennemann's TU‐1 and TU‐2 standards as well as for Ag₃PO₄ extracted from NBS120c. The weighted average δ¹⁸O_VSMOW values for the new comparison materials UMCS‐1, UMCS‐2 and AGPO‐SCRI were determined to be + 32.60 (± 0.12), + 19.40 (± 0.12) and + 14.58 (± 0.13)‰, respectively. Copyright © 2011 John Wiley & Sons, Ltd.     

Coupled N,C and S stable isotope measurements using a dual‐column gas chromatography system

Many laboratories routinely analyze plant, animal and soil samples with elemental analyzers coupled to isotope ratio mass spectrometers, obtaining rapid results for nitrogen (%N, δ¹⁵N) and carbon (%C, δ¹³C) from the same sample. The coupled N and C measurements are possible because of a gas chromatography (GC) separation of N₂ and CO₂ gases produced in elemental analysis. Adding a second GC column allows additional measurement of sulfur (%S, δ³⁴S) from the same sample, so that combined N, C and S information is obtained routinely. Samples are 1–15 mg, and replicates generally differ by less than 0.1‰ for δ¹⁵N, δ¹³C or δ³⁴S. An example application shows that the N, C, and S measurement system allows a three‐dimensional view of element dynamics in estuarine systems that are undergoing pollution inputs from upstream watersheds. Extension of these GC principles should allow coupled H, C, N, and S isotope measurements in future work. Copyright © 2007 John Wiley & Sons, Ltd.     

Determination of δ18O in soils: measuring conditions and a potential application

The stable oxygen isotope signature (δ¹⁸O) of soil is expected to be the result of a mixture of components within the soil with varying δ¹⁸O signatures. Thus, the δ¹⁸O of soils should provide information about the soil's substrate, especially about the relative contribution of organic matter versus minerals. As there is no standard method available for measuring soil δ¹⁸O, the method for the measurement of single components using a high‐temperature conversion elemental analyser (TC/EA) was adapted. We measured δ¹⁸O in standard materials (IAEA 601, IAEA 602, Merck cellulose) and soils (organic and mineral soils) in order to determine a suitable pyrolysis temperature for soil analysis. We consider a pyrolysis temperature suitable when the yield of signal intensity (intensity of mass 28 per 100 µg) is at a maximum and the acquired raw δ¹⁸O signature is constant for the standard materials used and when the quartz signal from the soil is still negligible. After testing several substances within the temperature range of 1075 to 1375°C we decided to use a pyrolysis temperature of 1325°C for further measurements. For the Urseren Valley we have found a sequence of increasing δ¹⁸O signatures from phyllosilicates to upland soils, wetland soils and vegetation. Our measurements show that the δ¹⁸O values of upland soil samples differ significantly from wetland soil samples. The latter can be related to the changing mixing ratio of the mineral and organic constituents of the soil. For wetlands affected by soil erosion, we have found intermediate δ¹⁸O signatures which lie between typical signatures for upland and wetland sites and give evidence for the input of upland soil material through erosion. Copyright © 2008 John Wiley & Sons, Ltd.     

Nicotine,acetanilide and urea multi‐level 2H‐, 13C‐ and 15N‐abundance reference materials for continuous‐flow isotope ratio mass spectrometry

Accurate determinations of stable isotope ratios require a calibration using at least two reference materials with different isotopic compositions to anchor the isotopic scale and compensate for differences in machine slope. Ideally, the δ values of these reference materials should bracket the isotopic range of samples with unknown δ values. While the practice of analyzing two isotopically distinct reference materials is common for water (VSMOW‐SLAP) and carbonates (NBS 19 and L‐SVEC), the lack of widely available organic reference materials with distinct isotopic composition has hindered the practice when analyzing organic materials by elemental analysis/isotope ratio mass spectrometry (EA‐IRMS). At present only L‐glutamic acids USGS40 and USGS41 satisfy these requirements for δ¹³C and δ¹⁵N, with the limitation that L‐glutamic acid is not suitable for analysis by gas chromatography (GC). We describe the development and quality testing of (i) four nicotine laboratory reference materials for on‐line (i.e. continuous flow) hydrogen reductive gas chromatography‐isotope ratio mass‐spectrometry (GC‐IRMS), (ii) five nicotines for oxidative C, N gas chromatography‐combustion‐isotope ratio mass‐spectrometry (GC‐C‐IRMS, or GC‐IRMS), and (iii) also three acetanilide and three urea reference materials for on‐line oxidative EA‐IRMS for C and N. Isotopic off‐line calibration against international stable isotope measurement standards at Indiana University adhered to the ‘principle of identical treatment’. The new reference materials cover the following isotopic ranges: δ²H_nicotine ?162 to ?45‰, δ¹³C_nicotine ?30.05 to +7.72‰, δ¹⁵N_nicotine ?6.03 to +33.62‰; δ¹⁵N_acetanilide +1.18 to +40.57‰; δ¹³C_urea ?34.13 to +11.71‰, δ¹⁵N_urea +0.26 to +40.61‰ (recommended δ values refer to calibration with NBS 19, L‐SVEC, IAEA‐N‐1, and IAEA‐N‐2). Nicotines fill a gap as the first organic nitrogen stable isotope reference materials for GC‐IRMS that are available with different δ¹⁵N values. Comparative δ¹³C and δ¹⁵N on‐line EA‐IRMS data from 14 volunteering laboratories document the usefulness and reliability of acetanilides and ureas as EA‐IRMS reference materials. Published in 2009 by John Wiley & Sons, Ltd.     

δ13C, δ15N and δ2H isotope ratio mass spectrometry of ephedrine and pseudoephedrine: application to methylamphetamine profiling

Conventional chemical profiling of methylamphetamine has been used for many years to determine the synthetic route employed and where possible to identify the precursor chemicals used. In this study stable isotope ratio analysis was investigated as a means of determining the origin of the methylamphetamine precursors, ephedrine and pseudoephedrine. Ephedrine and pseudoephedrine may be prepared industrially by several routes. Results are presented for the stable isotope ratios of carbon (δ¹³C), nitrogen (δ¹⁵N) and hydrogen (δ²H) measured in methylamphetamine samples synthesized from ephedrine and pseudoephedrine of known provenance. It is clear from the results that measurement of the δ¹³C, δ¹⁵N and δ²H stable isotope ratios by elemental analyzer/thermal conversion isotope ratio mass spectrometry (EA/TC‐IRMS) in high‐purity methylamphetamine samples will allow determination of the synthetic source of the ephedrine or pseudoephedrine precursor as being either of a natural, semi‐synthetic, or fully synthetic origin. Copyright © 2009 Commonwealth of Australia. Published by John Wiley & Sons, Ltd.     

IRMS detection of testosterone manipulated with C labeled standards in human urine by removing the labeled C

Isotope ratio mass spectrometry (IRMS) is applied to confirm testosterone (T) abuse by determining the carbon isotope ratios (δ¹³C value). However, ¹³C labeled standards can be used to control the δ¹³C value and produce manipulated T which cannot be detected by the current method. A method was explored to remove the ¹³C labeled atom at C-3 from the molecule of androsterone (Andro), the metabolite of T in urine, to produce the resultant (A-nor-5α-androstane-2,17-dione, ANAD). The difference in δ¹³C values between Andro and ANAD (Δδ¹³C_Andro–ANAD, ‰) would change significantly in case manipulated T is abused. Twenty-one volunteers administered T manipulated with different ¹³C labeled standards. The collected urine samples were analyzed with the established method, and the maximum value of Δδ¹³C_Andro–ANAD post ingestion ranged from 3.0‰ to 8.8‰. Based on the population reference, the cut-off value of Δδ¹³C_Andro–ANAD for positive result was suggested as 1.2‰. The developed method could be used to detect T manipulated with 3-¹³C labeled standards.     

Ensuring the reliability of stable isotope ratio data—beyond the principle of identical treatment

The need for inter-laboratory comparability is crucial to facilitate the globalisation of scientific networks and the development of international databases to support scientific and criminal investigations. This article considers what lessons can be learned from a series of inter-laboratory comparison exercises organised by the Forensic Isotope Ratio Mass Spectrometry (FIRMS) network in terms of reference materials (RMs), the management of data quality, and technical limitations. The results showed that within-laboratory precision (repeatability) was generally good but between-laboratory accuracy (reproducibility) called for improvements. This review considers how stable isotope laboratories can establish a system of quality control (QC) and quality assurance (QA), emphasising issues of repeatability and reproducibility. For results to be comparable between laboratories, measurements must be traceable to the international δ-scales and, because isotope ratio measurements are reported relative to standards, a key aspect is the correct selection, calibration, and use of international and in-house RMs. The authors identify four principles which promote good laboratory practice. The principle of identical treatment by which samples and RMs are processed in an identical manner and which incorporates three further principles; the principle of identical correction (by which necessary corrections are identified and evenly applied), the principle of identical scaling (by which data are shifted and stretched to the international δ-scales), and the principle of error detection by which QC and QA results are monitored and acted upon. To achieve both good repeatability and good reproducibility it is essential to obtain RMs with internationally agreed δ-values. These RMs will act as the basis for QC and can be used to calibrate further in-house QC RMs tailored to the activities of specific laboratories. In-house QA standards must also be developed to ensure that QC-based calibrations and corrections lead to accurate results for samples. The δ-values assigned to RMs must be recorded and reported with all data. Reference materials must be used to determine what corrections are necessary for measured data. Each analytical sequence of samples must include both QC and QA materials which are subject to identical treatment during measurement and data processing. Results for these materials must be plotted, monitored, and acted upon. Periodically international RMs should be analysed as an in-house proficiency test to demonstrate results are accurate.     

Carbon and nitrogen isotope analysis of atrazine and desethylatrazine at sub-microgram per liter concentrations in groundwater

Environmental degradation of organic micropollutants is difficult to monitor due to their diffuse and ubiquitous input. Current approaches—concentration measurements over time, or daughter-to-parent compound ratios—may fall short, because they do not consider dilution, compound-specific sorption characteristics or alternative degradation pathways. Compound-specific isotope analysis (CSIA) offers an alternative approach based on evidence from isotope values. Until now, however, the relatively high limits for precise isotope analysis by gas chromatography—isotope ratio mass spectrometry (GC-IRMS) have impeded CSIA of sub-microgram-per-liter scale micropollutant concentrations in field samples. This study presents the first measurements of C and N isotope ratios of the herbicide atrazine and its metabolite desethylatrazine at concentrations of 100 to 1,000 ng/L in natural groundwater samples. Solid-phase extraction and preparative HPLC were tested and validated for preconcentration and cleanup of groundwater samples of up to 10 L without bias by isotope effects. Matrix interferences after solid-phase extraction could be greatly reduced by a preparative HPLC cleanup step prior to GC-IRMS analysis. Sensitivity was increased by a factor of 6 to 8 by changing the injection method from large-volume to cold-on-column injection on the GC-IRMS system. Carbon and nitrogen isotope values of field samples showed no obvious correlation with concentrations or desethylatrazine-to-atrazine ratios. Contrary to expectations, however, δ ¹³ C values of desethylatrazine were consistently less negative than those of atrazine from the same sites. Potentially, this line of evidence may contain information about further desethylatrazine degradation. In such a case, the common practice of using desethylatrazine-to-atrazine ratios would underestimate natural atrazine degradation.     

Normalization of measured stable isotopic compositions to isotope reference scales--a review

In stable isotope ratio mass spectrometry (IRMS), the stable isotopic composition of samples is measured relative to the isotopic composition of a working gas. This measured isotopic composition must be converted and reported on the respective international stable isotope reference scale for the accurate interlaboratory comparison of results. This data conversion procedure, commonly called normalization, is the first set of calculations done by the users. In this paper, we present a discussion and mathematical formulation of several existing routinely used normalization procedures. These conversion procedures include: single-point anchoring (versus working gas and certified reference standard), modified single-point normalization, linear shift between the measured and the true isotopic composition of two certified reference standards, two-point and multi-point linear normalization methods. Mathematically, the modified single-point, two-point, and multi-point normalization methods are essentially the same. By utilizing laboratory analytical data, the accuracy of the various normalization methods (given by the difference between the true and the normalized isotopic composition) has been compared. Our computations suggest that single-point anchoring produces normalization errors that exceed the maximum total uncertainties (e.g. 0.1 per thousand for delta(13)C) often reported in the literature, and, therefore, that it must not be used for routinely anchoring stable isotope measurement results to the appropriate international scales. However, any normalization method using two or more certified reference standards produces a smaller normalization error provided that the isotopic composition of the standards brackets the isotopic composition of unknown samples.     

Isotope shift measurements in zirconium by Doppler-free laser polarization spectroscopy

Isotope shifts in 17 transitions of ZrI have been measured by Doppler-free laser polarization spectroscopy in a hollow cathode discharge. The results were combined with other known isotope shift data of ZrI and tested for consistency in a multidimensional King plot. By including values ofδ〈r ²〉 deduced from optical isotope shifts mass and field shifts were separated, and improved values ofδ〈r ²〉 could be determined from the King plot. The results for the isotope pairs A, A′ are as follows; 90, 92: 0.244(43) fm²; 92, 94: 0.176(20) fm²; 94, 96: 0.126(23) fm².     

External calibration in gas chromatography-combustion-isotope ratio mass spectrometry measurements of endogenous androgenic anabolic steroids in sports doping control

An alternative calibration procedure for the Gas Chromatography–Combustion–Isotope Ratio Mass Spectrometry (GC–C–IRMS) measurements of the World Antidoping Agency (WADA) Accredited Laboratories is presented. To alleviate the need for externally calibrated CO₂ gas for GC–C–IRMS analysis of urinary steroid metabolites, calibration using an external standard mixture solution of steroids with certified isotopic composition was investigated. The reference steroids of the calibration mixture and routine samples underwent identical instrumental processes. The calibration standards bracketed the entire range of the relevant δ¹³C values for the endogenous and exogenous steroids as well as their chromatographic retention times. The certified δ¹³C values of the reference calibrators were plotted in relation to measured m/z¹³CO₂/¹²CO₂ (i.e. R(45/44)) mass spectrometric signals of each calibrator. δ¹³C values of the sample steroids were calculated from the least squares fit through the calibration curve. The effect of the external calibration on δ¹³C values, using the same calibration standards and set of urine samples but different brands of GC–C–IRMS instruments, was assessed by an interlaboratory study in the WADA Accredited Laboratories of Sydney, Australia and Athens, Greece. Relative correspondence between the laboratories for determination of androsterone, etiocholanolone, 5β-androstane-3α,17β-diacetate, and pregnanediacetate means were SD(δ¹³C) = 0.12‰, 0.58‰, −0.34‰, and −0.40‰, respectively. These data demonstrate that accurate intralaboratory external calibration with certified steroids provided by United States Antidoping Agency (USADA) and without external CO₂ calibration is feasible and directly applicable to the WADA Accredited Laboratories for the harmonization of the GC–C–IRMS measurements.