James F. Carter

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.

Stable isotope and trace metal compositions of Australian prawns as a guide to authenticity and wholesomeness.

This research has explored the potential of stable isotope and trace metal profiles to distinguish Australian prawns from prawns imported from neighbouring Asian countries. Australian prawns were collected mostly from the Brisbane area. Strong differences in Australian vs. imported prawns were evident from both the isotope and trace element data, with the differences most likely occurring because imported prawns are typically reared in aquaculture facilities and frozen prior to sale in Australia. The aquaculture origins are characterised by comparatively; low δHVSMOW, δ(13)CVPDB values, low concentrations of arsenic, zinc and potassium, and high water contents (>80%). Relatively high arsenic and cadmium contents were found within Australian prawns, but the concentrations did not exceed local human health guidelines.

Isotopic and Elemental Composition of Roasted Coffee as a Guide to Authenticity and Origin

This study presents the stable isotopic and elemental compositions of single-origin, roasted coffees available to retail consumers. The δ(13)C, δ(15)N, and δ(18)O compositions were in agreement with those previously reported for green coffee beans. The δ(15)N composition was seen to be related to organic cultivation, reflected in both δ(2)H and δ(18)O compositions. The δ(13)C composition of extracted caffeine differed little from that of the bulk coffee. Stepwise discriminant analysis with jackknife tests, using isotopic and elemental data, provided up to 77% correct classification of regions of production. Samples from Africa and India were readily classified. The wide range in both isotopic and elemental compositions of samples from other regions, specifically Central/South America, resulted in poor discrimination between or within these regions. Simpler X-Y and geo-spatial plots of the isotopic data provided effective visual means to distinguish between coffees from different regions.

Stable isotope and chemical compositions of European and Australasian ciders as a guide to authenticity.

This paper presents a data set derived from the analysis of bottled and canned ciders that may be used for comparison with suspected counterfeit or substitute products. Isotopic analysis of the solid residues from ciders (predominantly sugar) provided a means to determine the addition of C4 plant sugars. The added sugars were found to comprise cane sugar, high-fructose corn syrup, glucose, or combinations. The majority of ciders from Australia and New Zealand were found to contain significant amounts of added sugar, which provided a limited means to distinguish these ciders from European ciders. The hydrogen and oxygen isotopic compositions of the whole ciders (predominantly water) were shown to be controlled by two factors, the water available to the parent plant and evaporation. Analysis of data derived from both isotopic and chemical analysis of ciders provided a means to discriminate between regions and countries of manufacture.

The role of isotope ratio mass spectrometry as a tool for the comparison of physical evidence

This paper considers how likelihood ratios can be derived for a combination of physical, chemical and isotopic measurements. Likelihood ratios were formulated based on the characteristics of a small convenience sample of 20 duct tapes. The propositions considered were: The physical and isotopic characteristics of ten rolls of duct tape were shown to be consistent throughout each roll. The width and thickness of the tapes and the density of the scrim fibres provided equivalent information and the combined physical characteristics provided a basis upon which to discriminate between many of the samples. Scatter-plots and confidence ellipses provided a convenient method to group the isotopic composition of the tape backing material and provided a basis to discriminate between samples which were physically indistinguishable. Considering both the physical and isotopic characteristics it was possible, at best, to ascertain that the evidence provided moderately strong support for the proposition that two samples of tape were derived from the same batch (LR=400). Kernel density estimates were used to model the distribution of isotopic compositions of the backing material. Using this technique it was possible to estimate objectively the probability that a sample with given characteristics could be drawn, at random, from the background population and to calculate a likelihood ratio based on the propositions above. The strength of evidence which could be presented by either model was ultimately limited by the size of the background sample.

The evaluation of evidence relating to traces of cocaine on banknotes

Banknotes can be seized from crime scenes as evidence for suspected association with illicit drug dealing. Tandem mass spectrometry data are available from banknotes seized in criminal investigations, as well as from banknotes from general circulation. The aim of the research is to evaluate the support provided by the data gathered in a criminal investigation for the proposition that the banknotes from which the data were obtained are associated with a person who is associated with a criminal activity related to cocaine in contrast to the proposition that the banknotes are associated with a person who is not associated with a criminal activity involving cocaine. The data considered are the peak area for the ion count for cocaine product ion m/z 105. Previous methods for assessment of the relative support for these propositions were concerned with the percentage of banknotes contaminated or assume independence of measurements of quantities between adjacent banknotes. Methods which account for an association of the quantity of drug on a banknote with that on adjacent banknotes are described. The methods are based on an autoregressive model of order one and on two versions of a nonparametric approach. The results are compared with a standard model which assumes measurements on individual banknotes are independent; there is no autocorrelation. Performance is assessed using rates of misleading evidence and a recommendation made as to which method to use.

A global survey of the stable isotope and chemical compositions of bottled and canned beers as a guide to authenticity

This study presents a dataset, derived from the analysis of 162 bottled and canned beers from around the globe, which may be used for comparison with suspected counterfeit or substitute products. The data comprise δ2H and δ18O compositions of the whole beer and δ13C compositions of the dry residue (mostly sugar) together with the concentrations of five anions (F, Cl, NO3, SO4, PO4) and seven cations (Ca, K, Mg, SiO2, V, Mn, Sr). A strong correlation, consistent with natural waters but offset from the Global Meteoric Water Line, was observed between the δ2H/δ18O composition of the beers. The extent of the offset could be explained by the brewing process and the alcohol and sugars present in the beers. Correlations between inorganic analytes were consistent with the addition of salts in the brewing process. Beers were classified as follows: ale, lager, stout or wheat-beer and the chemical composition was found to be characteristic of the assigned type, with lagers being the most readily classified. A combination of chemical and isotopic data was found to be characteristic of the geographical origin (on a continental scale) and could most easily identify beers from Australasia or Europe. A global map of δ18O data revealed a geo-spatial distribution that mirrored existing maps of the isotopic composition of annual precipitation. This confirmed a commonsense view that local precipitation will be the primary source for the water used in brewing. Using this isoscape (or alcoscape) it may be possible to assess the geographical origins of samples for which genuine comparative samples cannot be obtained.

Isotopic profiling of seized benzylpiperazine and trifluoromethylphenylpiperazine tablets using δ13C and δ15N stable isotopes

This paper demonstrates the use of isotopic analysis of 23 benzylpiperazine (BZP) and trifluoromethylphenylpiperazine (TFMPP) containing tablets seized on two independent occasions by the Northern Territory (NT) Police, Australia. Isolation (High Performance Liquid Chromatography (HPLC)) of BZP and TFMPP followed by Isotope Ratio Mass Spectrometry (IRMS) (carbon and nitrogen stable isotopes) analysis was performed. Results are presented for δ13C and δ15N values of the respective piperazine analogues. The isotopic data and statistical analysis suggest a common source of manufacture for the BZP samples but suggest different sources for the TFMPP isolated from the corresponding BZP containing tablets investigated. The use of IRMS in this case study demonstrated the ability to obtain information regarding the BZP/TFMPP sources unattainable via conventional chemical analysis.

Prawn biomonitors of nutrient and trace metal pollution along Asia-Pacific coastlines

ABSTRACT To assess coastal ecosystem status and pollution baselines, prawns were collected from the commercial catches of eight Asia-Pacific countries (Australia, Bangladesh, Indonesia, Myanmar, Philippines, Pakistan, Sri Lanka and Thailand). Samples collected from 21 sites along regional coastlines were analysed for trace metal and stable isotopic compositions of H, C, N, O and S. A combination of simple averaging and multivariate analyses was used to evaluate the data. Sites could be assigned to easily recognise polluted and unpolluted groups based on the prawn results. Some filter-feeding clams were also collected and analysed together with the benthic-feeding prawns, and the prawns generally had lower trace metal burdens. Climate change effects were not strongly evident at this time, but altered ocean circulation and watershed run-off patterns accompanying future climate change are expected to change chemical patterns recorded by prawns along these and other coastlines. Stable isotopes, especially 15N, can help to distinguish between relatively polluted and unpolluted sites.

Do it yourself reference materials for δ13C determinations by isotope ratio mass spectrometry.

A method is described to prepare inexpensive organic reference materials for routine stable isotope work. The method blends isotope-enriched and isotope-depleted materials to match closely international measurement standards. Examples are presented showing how 13C-enriched and 13C-depleted glucose were blended to match the isotopic compositions of the international carbon isotope standards, NBS 19 and L-SVEC. Preparation of the blended standards was straightforward, and the prepared solutions proved easy to dispense into tin capsules and easy to combust in a conventional isotope analysis system. The blended materials allow a laboratory to perform a two-point calibration for isotopic composition on a daily basis, with the use of an international reference material recommended for routine quality assurance. The blending method can be applied widely to make isotope standards for other elements and compounds.