Cultivar, site or harvest date: the gordian knot of wine terroir.

INTRODUCTION: The complex interactions of vine cultivars, and localised regional climate associated with specific vineyard sites are important attributes to the concept of terroir and significant contributors to grape maturity and wine sensory profiles. An improved understanding of the influence of each factor and their interactions is a challenging conundrum, and will enable more efficient production targeting specific wine styles. OBJECTIVES: To characterise the metabolic flux of grape berries and resulting wines to characterise the relative impact of site specific climate, cultivar, and grape maturity based upon berry sugar accumulation models that consistently target specific wine styles. METHODS: A spatial and temporal study of grape and wine composition was undertaken for two important cultivars in two distinct regions of New South Wales. Measures of composition and wine sensory ratings were simultaneously analysed using a multiblock algorithm taking advantage of the ANOVA framework to identify important contributions to wine style arising from grape maturity, vineyard site and cultivar. RESULTS: A consistent flux of grape and wine constituents is evident for wine made from sequentially harvested grapes from the same vineyard with increasing levels of grape maturity. Contributions of region and vineyard site to wine style could also be elucidated. Differences in metabolite flux in grapes and resulting wines between cultivars growing in similar conditions are evident. CONCLUSIONS: The combination of a metabolomics and multiblock data decomposition approach may be successfully used to profile and elucidate the contribution of abiotic factors to grape and wine composition and provide improved understanding of the terroir concept.

Profiling of steroid metabolites after transdermal and oral administration of testosterone by ultra-high pressure liquid chromatography coupled to quadrupole time-of-flight mass spectrometry.

The screening of testosterone (T) misuse for doping control is based on the urinary steroid profile, including T, its precursors and metabolites. Modifications of individual levels and ratio between those metabolites are indicators of T misuse. In the context of screening analysis, the most discriminant criterion known to date is based on the T glucuronide (TG) to epitestosterone glucuronide (EG) ratio (TG/EG). Following the World Anti-Doping Agency (WADA) recommendations, there is suspicion of T misuse when the ratio reaches 4 or beyond. While this marker remains very sensitive and specific, it suffers from large inter-individual variability, with important influence of enzyme polymorphisms. Moreover, use of low dose or topical administration forms makes the screening of endogenous steroids difficult while the detection window no longer suits the doping habit. As reference limits are estimated on the basis of population studies, which encompass inter-individual and inter-ethnic variability, new strategies including individual threshold monitoring and alternative biomarkers were proposed to detect T misuse. The purpose of this study was to evaluate the potential of ultra-high pressure liquid chromatography (UHPLC) coupled with a new generation high resolution quadrupole time-of-flight mass spectrometer (QTOF-MS) to investigate the steroid metabolism after transdermal and oral T administration. An approach was developed to quantify 12 targeted urinary steroids as direct glucuro- and sulfo-conjugated metabolites, allowing the conservation of the phase II metabolism information, reflecting genetic and environmental influences. The UHPLC-QTOF-MS(E) platform was applied to clinical study samples from 19 healthy male volunteers, having different genotypes for the UGT2B17 enzyme responsible for the glucuroconjugation of T. Based on reference population ranges, none of the traditional markers of T misuse could detect doping after topical administration of T, while the detection window was short after oral TU ingestion. The detection ability of the 12 targeted steroids was thus evaluated by using individual thresholds following both transdermal and oral administration. Other relevant biomarkers and minor metabolites were studied for complementary information to the steroid profile, including sulfoconjugated analytes and hydroxy forms of glucuroconjugated metabolites. While sulfoconjugated steroids may provide helpful screening information for individuals with homozygotous UGT2B17 deletion, hydroxy-glucuroconjugated analytes could enhance the detection window of oral T undecanoate (TU) doping.