Effect of changes in the deuterium content of drinking water on the hydrogen isotope ratio of urinary steroids in the context of sports drug testing.

The hydrogen isotope ratio (HIR) of body water and, therefore, of all endogenously synthesized compounds in humans, is mainly affected by the HIR of ingested drinking water. As a consequence, the entire organism and all of its synthesized substrates will reflect alterations in the isotope ratio of drinking water, which depends on the duration of exposure. To investigate the effect of this change on endogenous urinary steroids relevant to doping-control analysis the hydrogen isotope composition of potable water was suddenly enriched from -50 to 200 0/00 and maintained at this level for two weeks for two individuals. The steroids under investigation were 5β-pregnane-3α,20α-diol, 5α-androst-16-en-3α-ol, 3α-hydroxy-5α-androstan-17-one (ANDRO), 3α-hydroxy-5β-androstan-17-one (ETIO), 5α-androstane-3α,17β-diol, and 5β-androstane-3α,17β-diol (excreted as glucuronides) and ETIO, ANDRO and 3β-hydroxyandrost-5-en-17-one (excreted as sulfates). The HIR of body water was estimated by determination of the HIR of total native urine, to trace the induced changes. The hydrogen in steroids is partly derived from the total amount of body water and cholesterol-enrichment could be calculated by use of these data. Although the sum of changes in the isotopic composition of body water was 150 0/00, shifts of approximately 30 0/00 were observed for urinary steroids. Parallel enrichment in their HIR was observed for most of the steroids, and none of the differences between the HIR of individual steroids was elevated beyond recently established thresholds. This finding is important to sports drug testing because it supports the intended use of this novel and complementary methodology even in cases where athletes have drunk water of different HIR, a plausible and, presumably, inevitable scenario while traveling

A novel high-temperature combustion based system for stable isotope analysis of dissolved organic carbon in aqueous samples. : II optimization and assessment of analytical performance

RATIONALE: Dissolved organic carbon (DOC) plays an important role in carbon cycling, making precise and routine measurement of delta C-13 values and DOC concentration highly desirable. A new promising system has been developed for this purpose. However, broad-scale application of this new technique requires an in-depth assessment of analytical performance, and this is described here. METHODS: A high-temperature combustion Total Organic Carbon analyzer was interfaced with continuous flow isotope ratio mass spectrometry (TOC/IRMS) for the simultaneous analysis of the bulk DOC concentration and delta C-13 signature. The analytical performance (precision, memory effects, linearity, volume/concentration effects, accuracy) was thoroughly evaluated, including realistic and challenging conditions such as low DOC concentrations and natural DOC. RESULTS: High precision (standard deviation, SD predominantly ≤0.15 ‰) and accuracy (R2 = 0.9997) were achieved for the δ13C analysis of a broad diversity of DOC solutions. Simultaneously, good results were obtained for the measurement of DOC concentration. Assessment of natural abundance and slightly 13C-enriched DOC, a wide range of concentrations (~0.2–150 mgC/L) and injection volumes (0.05–3 mL), demonstrated minor/negligible memory effects, good linearity and flexible usage. Finally, TOC/IRMS was successfully applied to determine low DOC concentrations (<2 mgC/L) and DOC from diverse terrestrial, freshwater and marine environments (SD ≤0.23 ‰). CONCLUSIONS: TOC/IRMS enables fast and reliable measurement of DOC concentrations and delta C-13 values in aqueous samples, without pre-concentration and freeze-drying. Further investigations should focus on complex, saline matrices and very low DOC concentrations, to achieve a potential lower limit of 0.2 mgC/L. Thus, TOC/IRMS will give DOC research in terrestrial and aquatic environments a huge impulse with high-resolution, routine delta C-13 analysis. Copyright (C) 2014 John Wiley & Sons, Ltd

A novel high-temperature combustion based system for stable isotope analysis of dissolved organic carbon in aqueous samples.: I development and validation

RATIONALE: Traditionally, dissolved organic carbon (DOC) stable isotope analysis (SIA) is performed using either offline sample preparation followed by elemental analyzer/isotope ratiomass spectrometry (EA/IRMS) or a wet chemical oxidation (WCO)-based device coupled to an isotope ratio mass spectrometer. The first method is time-consuming and laborious. The second involves the risks of underestimation of DOC concentration and isotopic fractionation due to incomplete oxidation. The development of an analytical method for accurate and sensitive DOC SIA is described in this study. METHODS: A high-temperature combustion (HTC) system improves upon traditional methods. A novel total organic carbon (TOC) system, specially designed for SIA, was coupled to an isotope ratio mass spectrometer. An integrated purge and trap technique (peak focusing), flexible injection volume (0.05-3 mL), favorable carrier gas flow, modified ash crucible, new design of combustion tube and optimized drying system were used to achieve the necessary performance. RESULTS: The system can reliably measure concentrations up to 1000 mgC/L. Compounds resistant to oxidation, such as barbituric acid, melamine and humic acid, were analyzed with recovery rates of 100 +/- 1% proving complete oxidation. In this initial testing, the delta C-13 values of these compounds were determined with precision and trueness of CONCLUSIONS: The novel HTC system coupled to an isotope ratio mass spectrometer resulted in significantly improved sensitivity. The system is suitable for salt-containing liquids and compounds that are resistant to oxidation, and it offers a large concentration range. A second paper (which follows this one in this issue) will present a more comprehensive assessment of the analytical performance with a broad set of solutions and real samples. This highly efficient TOC stable isotopic analyzer will probably open up new possibilities in biogeochemical carbon cycle research. Copyright (C) 2014 John Wiley & Sons, Ltd