Differences in the stable isotope signatures of seabird egg membrane and albumen – implications for non‐invasive studies

In many bird species, egg membranes can be obtained non‐invasively after the chicks have hatched, and stable isotope analysis of egg membranes can be used to study the diet and foraging distribution of these birds during egg formation. It has been suggested that the enrichment factors of albumen and egg membranes differ for ¹³C, but are similar for ¹⁵N. In this study, we compared carbon and nitrogen stable isotopes of the membranes and albumen of individual eggs of three wild seabird species, the Southern Rockhopper penguin Eudyptes chrysocome, the Imperial shag Phalacrocorax atriceps albiventer, and the Thin‐billed prion Pachyptila belcheri. We also included chicken eggs for comparison. Egg membranes were generally enriched in ¹³C, compared with albumen. The difference varied between species, with 2.1‰ in Rockhopper penguins, 1.6‰ in Imperial shags, but only 0.5‰ in Thin‐billed prions and 0.4‰ in chicken eggs. Egg membranes were slightly enriched in ¹⁵N in Imperial shags (0.9‰) and chickens (0.5‰), compared with albumen, while there was no difference for Thin‐billed prions and Rockhopper penguins. The isotopic values of carbon and nitrogen were correlated between albumen and egg membranes of individual eggs, suggesting that egg membranes can be used reliably to investigate trophic differences between individuals, seasons or colonies. Species‐specific mathematical corrections could be used to compare results across studies that use different egg components. Copyright © 2009 John Wiley & Sons, Ltd.     

A fast and accurate isotope dilution GC‐IT‐MS/MS method for determination of eugenol in different tissues of fish: Application to a depletion study in mandarin fish

An accurate, rapid and effective method was established for determination of eugenol in plasma, muscle, skin, liver, kidney and gill of fish using gas chromatography–ion trap tandem mass spectrometry. Samples of muscle, skin, liver, kidney and gill were prepared using the modified QuEChERS (quick, easy, cheap, effective, rugged and safe) procedure, and a plasma sample was prepared by a liquid–liquid extraction procedure. Eugenol was monitored in <7 min using an electron‐ionization source in MS/MS mode and quantified by an internal standard of eugenol‐d₃. The limit of detection was 5.0 μg/kg, and the limit of quantification was 10.0 μg/kg. The calibration curve was linear in the range of 5–1000 μg/L (R² = 0.9996). Intra‐ and inter‐day precisions of eugenol expressed as relative standard deviation were within 9.74%, and the accuracy exhibited a relative error ranging from −2.20 to 8.89%. The developed method was successfully used to study the elimination regularity of eugenol in mandarin fish.     

Impact of non‐ideal analyte behavior on the separation of protein aggregates by asymmetric flow field‐flow fractionation

Asymmetric flow field‐flow fractionation is a valuable tool for the characterization of protein aggregates in biotechnology owing to its broad size range and unique separation principle. However, in practice asymmetric flow field‐flow fractionation is non‐trivial to use due to the major deviations from theory and the influence on separation by various factors that are not fully understood. Here, we report methods to assess the non‐ideal effects that influence asymmetric flow field‐flow fractionation separation and for the first time identify experimentally the main factors that impact it. Furthermore, we propose new approaches to minimize such non‐ideal behavior, showing that by adjusting the mobile phase composition (pH and ionic strength) the resolution of asymmetric flow field‐flow fractionation separation can be drastically improved. Additionally, we propose a best practice method for new proteins.     

Stable isotope ratio analysis to differentiate temporal diets of a free‐ranging herbivore

Stable isotope ratio analysis (SIRA) of carbon (δ¹³C) and nitrogen (δ¹⁵N) in tissue samples of herbivores can identify photosynthetic pathways (C₃ vs. C₄) of plants consumed. We present results from free‐ranging Rocky Mountain elk (Cervus elaphus) that highlight the ability to differentiate diets using tissue δ¹³C and δ¹⁵N. The signatures of δ¹³C and δ¹⁵N differed in tissues of varying metabolic activity: muscle, a short‐term dietary indicator (i.e., 1–2 months) and hoof, a long‐term dietary indicator (i.e., 3–12 months). We also documented that δ¹³C and δ¹⁵N values along elk hooves (proximal, middle, distal sections) elucidated temporal shifts in dietary selection. The carbon isotopes of the composite hoof were similar to those of the middle section, but the composite hoof differed in δ¹³C from the distal and proximal sections. The δ¹³C and δ¹⁵N signatures also differed among elk populations, indicating temporal dietary shifts of individuals occupying disparate native range and human‐derived agricultural landscapes. Analyses of stable isotopes in various tissues highlighted carbon and nitrogen assimilation through time and differences in the foraging ecology of a rangeland herbivore. Published in 2009 by John Wiley & Sons, Ltd.     

Stable isotope ratios of carbon and hydrogen to distinguish olive oil from shark squalene‐squalane

Squalene and its hydrogenated derivate squalane are widely used in the pharmaceutical and cosmetic fields. The two compounds are mainly produced from the liver oil of deep sea sharks and from olive oil distillates. Squalene and squalane from shark cost less than the same compounds derived from olive oil, and the use of these shark‐derived compounds is unethical in cosmetic formulations. In this work we investigate whether ¹³C/¹²C and ²H/¹H ratios can distinguish olive oil from shark squalene/squalane and can detect the presence of shark derivates in olive oil based products. The ¹³C/¹²C ratios (expressed as δ¹³C values) of bulk samples and of pure compounds measured using isotope ratio mass spectrometry (IRMS) were significantly lower in authentic olive oil squalene/squalane (N: 13; ?28.4 ± 0.5‰; ?28.3 ± 0.8‰) than in shark squalene/squalane samples (N: 15; ?20.5 ± 0.7‰; ?20.4 ± 0.6‰). By defining δ¹³C threshold values of ?27.4‰ and ?26.6‰ for olive oil bulk and pure squalene/squalane, respectively, illegal addition of shark products can be identified starting from a minimum of 10%. ²H/¹H analysis is not useful for distinguishing the two different origins. δ¹³C analysis is proposed as a suitable tool for detecting the authenticity of commercial olive oil squalene and squalane samples, using IRMS interfaced to an elemental analyser if the purity is higher than 80% and IRMS interfaced to a gas chromatography/combustion system for samples with lower purity, including solutions of squalane extracted from cosmetic products. Copyright © 2010 John Wiley & Sons, Ltd.     

A system for high‐quality CO2 isotope analyses of air samples collected by the CARIBIC Airbus A340‐600

In 2007, JRC‐IRMM began a series of atmospheric CO₂ isotope measurements, with the focus on understanding instrumental effects, corrections as well as metrological aspects. The calibration approach at JRC‐IRMM is based on use of a plain CO₂ sample (working reference CO₂) as a calibration carrier and CO₂‐air mixtures (in high‐pressure cylinders) to determine the method‐related correction under actual analytical conditions (another calibration carrier, in the same form as the samples). Although this approach differs from that in other laboratories, it does give a direct link to the primary reference NBS‐19‐CO₂. It also helps to investigate the magnitude and nature for each of the instrumental corrections and allows for the quantification of the uncertainty introduced. Critical tests were focused on the instrumental corrections. It was confirmed that the use of non‐symmetrical capillary crimping (an approach used here to deal with small samples) systematically modifies δ¹³C(CO₂) and δ¹⁸O(CO₂), with a clear dependence on the amount of extracted CO₂. However, the calibration of CO₂‐air mixtures required the use of the symmetrical dual‐inlet mode. As a proof of our approach, we found that δ¹³C(CO₂) on extracts from mixtures agreed (within 0.010‰) with values obtained from the ‘mother’ CO₂ used for the mixtures. It was further found that very low levels of hydrocarbons in the pumping systems and the isotope ratio mass spectrometry (IRMS) instrument itself were critical. The m/z 46 values (consequently the calculated δ¹⁸O(CO₂) values) are affected by several other effects with traces of air co‐trapped with frozen CO₂ being the most critical. A careful cryo‐distillation of the extracted CO₂ is recommended. After extensive testing, optimisation, and routine automated use, the system was found to give precise data on air samples that can be traced with confidence to the primary standards. The typical total combined uncertainty in δ¹³C(CO₂) and δ¹⁸O(CO₂) on the VPDB‐CO₂ scale, estimated on runs of CO₂‐air mixtures, is ±0.040‰ and 0.060‰ (2‐σ values). Inter‐comparison with MPI‐BGC resulted in a scale discrepancy of a similar magnitude. Although the reason(s) for this discrepancy still need to be understood, this basically confirms the approach of using specifically prepared CO₂‐air mixtures as a calibration carrier, in order to achieve scale unification among laboratories. As important practical application and as a critical test, JRC‐IRMM took part in the passenger aircraft‐based global monitoring project CARIBIC ( http://www.caribic‐atmospheric.com ). In this way, reliable CO₂ isotope data for the tropopause region and the free troposphere were obtained. From June 2007 to January 2009, approximately 500 CARIBIC air samples have been analysed. Some flights demonstrated a compact correlation of both δ¹³C(CO₂) and δ¹⁸O(CO₂) with respect to CO₂ concentration, demonstrating mixing of tropospheric and stratospheric air masses. These excellent correlations provide an independent, realistic data quality check. Copyright © 2009 John Wiley & Sons, Ltd.     

Simple evaluation of Franck–Condon factors and non‐Condon effects in the Morse potential

The calculation of Franck–Condon factors between different 1‐D Morse potential eigenstates using a formula derived from the Wigner function is discussed. Our numerical calculations using a simple program written in Mathematica are compared with other calculations. We show that our results have a similar accuracy as those calculations performed with more sophisticated methods. We discuss the extension of our method to include non‐Condon effects in the calculation. © 2002 Wiley Periodicals, Inc. Int J Quantum Chem 88: 280–295, 2002     

Stereochemical heterogeneity of biodegradable poly(L‐lactide) homopolymer as revealed by temperature rising elution fractionation and successive self‐nucleation/annealing thermal fractionation

Commercial poly(L ‐lactide) is typically heterogeneous in chain structure due to the existence of a small amount of D ‐lactyl units that are produced by the racemization reactions during the synthesis. In this article, the stereochemical heterogeneity of two commercial poly(L ‐lactide) was investigated with temperature rising elution fractionation (TREF) and successive self‐nucleation/annealing (SSA) thermal fractionation. For both samples, three fractions were collected and characterized with rotatory power analysis and DSC. The fractions show distinct optical purity and DSC results, which reflect the structure differences among them directly. After SSA treatment, the observation of multiple endotherms for each physically separated fraction confirms the fractionated sample contains a heterogeneous intermolecular and intramolecular distribution of defects. © 2012 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys, 2012