Associations Between Sugars Intakes and Urinary Sugars Excretion and Carbon Stable Isotope Ratios in Red Blood Cells as Biomarkers of Sugars Intake in a Predominantly Maori Population.

Determining the extent to which added sugars intake contribute to non-communicable disease in various populations is challenging because it is difficult to accurately measure intakes. Biomarkers may provide a reliable and easily measured method of assessing intakes. In a predominantly Maori population we compared various sugars intake estimates derived from a 36 item sugar-specific food frequency questionnaire (FFQ) with biomarkers of sugars intake; urinary sugars excretion in random spot collections (n = 153) and carbon stable isotope ratios (n = 36) in red blood cells (RBCs, δ13CRBC) and in the alanine fraction of the RBCs (δ13Calanine). Estimated 24 h urinary sucrose+fructose excretion was statistically significantly correlated with intakes of total sugars (r = 0.23), sucrose (r = 0.26) and added sugars from sugar-sweetened beverages (SSBs; r = 0.26). δ13Calanine was correlated with added sugars (r = 0.40). In log linear multiple regression models adjusted with HbA1C and eGFR δ13Calanine predicted added sugars intakes (r 2 = 0.29) and estimated 24 h urinary sucrose+fructose excretion predicted intakes of total sugars (r 2 = 0.14), sucrose (r 2 = 0.17), added sugars (r 2 = 0.17) and sugars from SSBs (r 2 = 0.14). These biomarkers have potential for improving assessment of sugars intake in New Zealand populations enabling monitoring of the effectiveness of sugar reduction strategies designed to reduce risk of NCDs. However, further validation is required to confirm these preliminary findings.

Feasibility of Casein to Record Stable Isotopic Variation of Cow Milk in New Zealand.

Dairy products occupy a special place among foods in contributing to a major part of our nutritional requirements, while also being prone to fraud. Hence, the verification of the authenticity of dairy products is of prime importance. Multiple stable isotopic studies have been undertaken that demonstrate the efficacy of this approach for the authentication of foodstuffs. However, the authentication of dairy products for geographic origin has been a challenge due to the complex interactions of geological and climatic drivers. This study applies stable isotope measurements of d2H, d18O, d13C and d15N values from casein to investigate the inherent geo-climatic variation across dairy farms from the South and North Islands of New Zealand. The stable isotopic ratios were measured for casein samples which had been separated from freeze-dried whole milk samples. As uniform feeding and fertilizer practices were applied throughout the sampling period, the subtropical (North Island) and temperate (South Island) climates were reflected in the variation of d13C and d15N. However, highly correlated d2H and d18O (r = 0.62, p = 6.64 × 10-10, a = 0.05) values did not differentiate climatic variation between Islands, but rather topographical locations. The highlight was the strong influence of d15N towards explaining climatic variability, which could be important for further discussion.

Natal origin of the invasive biosecurity pest, brown marmorated stink bug (Halyomorpha halys: Penatomidae), determined by dual-element stable isotope-ratio mass spectrometry.

BACKGROUND: Post-border detection of a single brown marmorated stink bug (BMSB) in New Zealand warranted a biosecurity response, the nature of which would be influenced by its status as part of an established population or as a new arrival. Stable isotope analysis has the potential to determine natal origins, but is difficult to achieve for samples as small as a single insect. Here an analytical modification to measure small samples was successfully trialled as a means to supply evidence as to the local or exotic natal origin of the intercepted BMSB specimen. RESULTS: Sufficient analytical sensitivity was achieved using a modified isotope ratio mass spectrometry method, involving thermolysis and carbon monoxide cryofocusing, to enable the simultaneous analysis of δ2 H and δ18 O from wings of the post-border BMSB sample. The values were much lower than those of the New Zealand green vegetable bug, used as a local reference. However, they fell within the range of those for BMSB of Northern Hemisphere origin intercepted at the New Zealand border over the same time period, specifically overlapping with the USA and Italy, but not China. CONCLUSION: The isotope signature of the post-border detected BMSB suggested a significantly cooler climate than the North Island of New Zealand, indicating that it was a new arrival and did not represent an established population. © 2019 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Extending the Limit of Measurement for Dual H and O Isotope Ratios Using Thermolysis.

Hydrogen and oxygen isotope ratios are of use to determine the origin of matter. Thermolysis is used to convert matter to H2 and CO gases, which are the respective substrates for measurement of these two isotope ratios, using isotope ratio mass spectrometry (IRMS). This work was done in response to the need for analysis of small invasive insects, requiring a decrease in the limit of measurement for isotope ratiometry of hydrogen and oxygen, while determining both isotope ratios on the same sample. Miniaturization of a thermolysis reactor using commercially available components is presented that results in improvement in the limit of measurement for both hydrogen and oxygen isotope ratios. δ2H was determined on 0.4 µg of H and δ18O determined on 5 µg of O with precisions of 3 mUr and 0.7 mUr, respectively. To extend the usable sample size range or increase the resolution of sampling gives obvious advantages in forensic and environmental sciences. The technique has been applied to determining the natural origin of Tephritidae fruit flies for which only the wing is suitable for analysis and provides just 60 µg of material for analysis.

Stable isotope measurements to differentiate sources of monofluoroacetate in a blackmail case.

RATIONALE: Stable isotope ratios can provide a 'fingerprint' to enable differentiation of sources of monofluoroacetate (MFA), hence providing a means to eliminate potential sources of MFA in a blackmail case involving the contamination of milk. METHODS: The stable isotopic compositions (δ2 H, δ13 C and δ18 O values) of a library of 43 samples of MFA were determined and multivariate models constructed to differentiate samples of different composition. The data from the MFA library were compared with those obtained from MFA extracted from contaminated milk powder (the case samples). The isotopic composition of the extracted samples was measured on dichloroaniline derivatives. RESULTS: A wide range of values was found for δ2 H, δ13 C and δ18 O of the MFA samples, much greater than the analytical repeatability between subsamples. Stable isotope data, therefore, provide a means of distinguishing samples of MFA. Of the 43 MFA samples tested, all but 6 could be eliminated as potential sources of the contamination, i.e. they had a distinctly different isotopic composition such that they must have had different histories and/or origins. CONCLUSIONS: Stable isotope measurements of bulk and derivatized MFA provide an effective means of discriminating MFA samples. Three of the library samples that could not be differentiated from the case samples were directly connected to the suspect, and this evidence contributed to the suspect's admission of guilt.

Precipitation isoscapes for New Zealand: enhanced temporal detail using precipitation-weighted daily climatology.

Predictive understanding of precipitation δ(2)H and δ(18)O in New Zealand faces unique challenges, including high spatial variability in precipitation amounts, alternation between subtropical and sub-Antarctic precipitation sources, and a compressed latitudinal range of 34 to 47 °S. To map the precipitation isotope ratios across New Zealand, three years of integrated monthly precipitation samples were acquired from >50 stations. Conventional mean-annual precipitation δ(2)H and δ(18)O maps were produced by regressions using geographic and annual climate variables. Incomplete data and short-term variation in climate and precipitation sources limited the utility of this approach. We overcome these difficulties by calculating precipitation-weighted monthly climate parameters using national 5-km-gridded daily climate data. This data plus geographic variables were regressed to predict δ(2)H, δ(18)O, and d-excess at all sites. The procedure yields statistically-valid predictions of the isotope composition of precipitation (long-term average root mean square error (RMSE) for δ(18)O = 0.6 ‰; δ(2)H = 5.5 ‰); and monthly RMSE δ(18)O = 1.9 ‰, δ(2)H = 16 ‰. This approach has substantial benefits for studies that require the isotope composition of precipitation during specific time intervals, and may be further improved by comparison to daily and event-based precipitation samples as well as the use of back-trajectory calculations.

The Geographic Origin of an Intercepted Biosecurity Pest Beetle Assigned Using Hydrogen Stable Isotopes.

Arhopalus ferus (Mulsant) (Coleoptera: Cerambycidae) is a forest pest that does not occur in Australia. In February 2010, the container ship Tatiana Schulte, en route from New Zealand, was refused permission to enter Australia following the discovery of numerous A. ferus aboard. The place where the infestation occurred was unknown, representing an uncontrolled biosecurity-risk pathway. Hydrogen isotope analysis of the beetles' wings showed that the infestation most likely originated from Auckland, New Zealand.

Assessing carbon and hydrogen isotopic fractionation of diesel fuel n-alkanes during progressive evaporation.

Compound-specific isotope analysis offers potential for fingerprinting of diesel fuels, however, possible confounding effects of isotopic fractionation due to evaporation need to be assessed. This study measured the fractionation of the stable carbon and hydrogen isotopes in n-alkane compounds in neat diesel fuel during evaporation. Isotope ratios were measured using a continuous flow gas chromatograph/isotope ratio mass spectrometer. Diesel samples were progressively evaporated at 24 ± 2°C for 21 days. Increasing depletion of deuterium in nC12-nC17 alkanes in the remaining liquid with increasing carbon chain length was observed. Negligible carbon isotope fractionation was observed. Preferential vaporization was measured for the shorter chain n-alkanes and the trend decreased with increasing chain length. The decrease in δ(2) H values indicates the preferential vaporization of the isotopically heavier species consistent with available quantitative data for hydrocarbons. These results are most important in the application of stable isotope technology to forensic analysis of diesel.

Isotopes and trace elements as natal origin markers of Helicoverpa armigera--an experimental model for biosecurity pests.

Protecting a nation's primary production sector and natural estate is heavily dependent on the ability to determine the risk presented by incursions of exotic insect species. Identifying the geographic origin of such biosecurity breaches can be crucial in determining this risk and directing the appropriate operational responses and eradication campaigns, as well as ascertaining incursion pathways. Reading natural abundance biogeochemical markers using mass spectrometry is a powerful tool for tracing ecological pathways as well as provenance determination of commercial products and items of forensic interest. However, application of these methods to trace insects has been underutilised to date and our understanding in this field is still in a phase of basic development. In addition, biogeochemical markers have never been considered in the atypical situation of a biosecurity incursion, where sample sizes are often small, and of unknown geographic origin and plant host. These constraints effectively confound the interpretation of the one or two isotope geo-location markers systems that are currently used, which are therefore unlikely to achieve the level of provenance resolution required in biosecurity interceptions. Here, a novel approach is taken to evaluate the potential for provenance resolution of insect samples through multiple biogeochemical markers. The international pest, Helicoverpa armigera, has been used as a model species to assess the validity of using naturally occurring δ2H, 87Sr/86Sr, 207Pb/206Pb and 208Pb/206Pb isotope ratios and trace element concentration signatures from single moth specimens for regional assignment to natal origin. None of the biogeochemical markers selected were individually able to separate moths from the different experimental regions (150-3000 km apart). Conversely, using multivariate analysis, the region of origin was correctly identified for approximately 75% of individual H. armigera samples. The geographic resolution demonstrated with this approach has considerable potential for biosecurity as well as other disciplines including forensics, ecology and pest management.

Modified sugar adulteration test applied to New Zealand honey.

The carbon isotope method (AOAC 998.12) compares the bulk honey carbon isotope value with that of the extracted protein; a difference greater than 1‰ suggesting that the protein and the bulk carbohydrate have different origins. New Zealand Manuka honey is a high value product and often fails this test. It has been suggested such failures are due to the pollen in the Manuka honey and an adaptation of the method to remove pollen prior to testing has been proposed. Here we test 64 authentic honey samples collected directly from the hives and find that a large proportion (37%) of Manuka honeys fail the test. Of these 60% still fail the adapted method. These honey samples were collected and processed under stringent conditions and have not been adulterated post-harvest. More work is required to ascertain the cause of these test failures.

Correlation of geographical location with stable isotope values of hydrogen and carbon of fatty acids from New Zealand milk and bulk milk powder.

The aim of this study was to investigate the correlation of δ²H and δ¹³C of bulk milk powder and milk powder fatty acids to their production region. A total of 46 milk powder samples from across New Zealand were collected and analyzed. Principal component analysis (PCA) showed that the δ²H and δ¹³C of four fatty acids (C4:0, C14:0, C16:0, C18:1) and bulk milk powder were found to be correlated with regional production area. Linear discriminant analysis (LDA) models were prepared using different combinations of bulk and fatty acid δ²H and δ¹³C. All models were effective in discriminating samples from the North and South Islands. The LDA model using just fatty acid δ²H and δ¹³C provided the best separation. Therefore, the isotopic composition of the aforementioned fatty acids can be utilized as a good biomarker in milk powder that conveys reliable isotopic information to track milk powders to their regional origin.