Vera Höllriegl

Investigations on the variability of breath gas sampling using PTR-MS

Breath gas analysis is a promising technology in the frame of medical diagnostics. By identifying disease-specific biomarkers in the breath of patients, a non-invasive and easy method for early diagnosis or therapy monitoring might be developed. However, to verify this potential and develop diagnostic tools based on breath gas analysis one essential prerequisite is a low variability in measurement of exhaled volatile organic compounds. Therefore, a study has been undertaken in order to identify possible artefacts within the application of a breath gas test in practice, for which the breath gas is analysed by proton transfer reaction-mass spectrometry (PTR-MS). After validating the low instrumental variability by repeatedly measuring standard gas, the variability of breath gas sampling has been evaluated. The latter has been carried out by measuring single breath gas samples (mixed expiratory breath) collected over different periods of time such as 1 min (10 volunteers, 4 breath gas samples each), 1 h (10 volunteers, 11 breath gas samples each) and several days (11 volunteers, 10 breath gas samples each). The breath gas samples were collected in Teflon bags and consecutively measured with PTR-MS. It was found that those samples collected within 1 min and 1 h show a low variability. This was, however, not the case for samples being collected over longer periods of time (15-70 days). Under these circumstances, many volatile organic compounds (VOCs) showed significant day-to-day variation in concentration, although the breath collection had been performed under the same conditions (similar sampling time, sampling technique, sample storage time, measurement conditions, etc). This large variation might be assigned to the influence of room air VOCs, which have been investigated in this work, or with other parameters which will be discussed. It was also found that the variability in the measurement of exhaled concentrations of methanol, acetone and isoprene within different individuals (inter individual variability) is much higher than differences in the same volunteer (intra individual variability) measured over a longer time interval.

Measurement of cerium in human breast milk and blood samples.

The aim of this study was to evaluate the relationship between cerium content in human breast milk and blood plasma or serum. Blood samples and breast milk at various stages of lactation, from 5 days to 51 weeks post partum, were donated by 42 healthy breast-feeding mothers from Munich, Germany and by 26 lactating Spanish mothers from Madrid at 4 weeks post partum. Inductively coupled plasma mass spectrometry was applied for the determination of cerium in the biological samples. Cerium concentration in the digested milk samples from Munich showed low values and the arithmetic mean values ranged between the quantification limit of 5 ng/L up to 65 ng/L. The median value amounted to 13 ng/L. The cerium concentrations in the Spanish breast milk samples amounted to similar low values. The data were about a factor of eight lower than values found in a former study of samples from an eastern German province. All cerium concentrations in the German plasma samples, except for two, were at the quantification limit of 10 ng/L. Interestingly, the serum samples of the Spanish mothers showed cerium values ranging between 21.6 and 70.3 ng/L; these higher data could be explained by an enhanced intake of cerium by humans in Madrid. This could be caused by increased cerium concentrations in particulate matter due to a higher traffic volume in Madrid compared to Munich. The results obtained in this study contribute to setting reference baseline values of cerium in human breast milk and blood plasma/serum and indicate a varying cerium amount depending on the cerium environmental pollution. Possibly, the cerium content in plasma/serum could be an indicator for environmental cerium, which is not valid for breast milk.

Effects of diet-matrix on volatile organic compounds in breath in diet-induced obese mice

Breath gas analysis in humans proved successful in identifying disease states and assessing metabolic functions in a non-invasive way. While many studies report diagnostic capability using volatile organic compounds (VOC) in breath, the inter-individual variability even in healthy human cohorts is rather large and not completely understood in its biochemical origin. Laboratory mice are the predominant animal model system for human disorders and are analysed under highly standardized and controlled conditions. We established a novel setup to monitor VOCs as biomarkers for disease in the breath gas of non-anesthetized, non-restrained mice using a proton transfer reaction mass spectrometer with time of flight detection. In this study, we implemented breath gas analysis in a dietary intervention study in C57BL/6J mice with the aim to assess the variability in VOC signatures due to a change in the diet matrix. Mice were fed a standard laboratory chow and then exposed to four semi-purified low- or high-fat diets for four weeks. Random forest (RF++) was used to identify VOCs that specifically respond to the diet matrix change. Interestingly, we found that the change from a chow diet to semi-purified diets resulted in a considerable drop of several VOC levels. Our results suggest that the diet matrix impacts VOC signatures and the underlying metabolic functions and may be one source of variability in exhaled volatiles.

Solubility of uranium and thorium from a healing earth in synthetic gut fluids: A case study for use in dose assessments

The aim of this case study was to estimate the bioaccessibility of uranium ((238)U) and thorium ((232)Th) from a healing earth by analysing the solubility of these radionuclides in synthetic gastric and intestinal fluids. An easy applicable in vitro test system was used to investigate the fractional mobilization of the soil contaminants being potentially available for absorption under human in vivo conditions. These findings provided the basis for a prospective dose assessment. The solubility experiments were performed using two different in vitro digestion methods. The concentrations of (238)U and (232)Th in the solutions extracted from the soil were measured by inductively coupled plasma mass spectrometry (ICP-MS). The dissolved fractions in the synthetic gastrointestinal fluid ranged in average from 10.3% to 13.8% for (238)U and from 0.3% to 1.6% for (232)Th, respectively, depending on the digestion method. Subsequently, the committed effective doses from intake of (238)U and (232)Th after ingestion of the healing earth during 1 year were evaluated for adult persons. Thereby ingestion dose coefficients calculated as a function of bioaccessibility were used. The dose assessments ranged between 4.3 × 10(-7)-1.9 × 10(-6) Sv y(-1) for (238)U and 5.6 × 10(-7)-3.3 × 10(-6) Sv y(-1) for (232)Th, respectively. On the basis of the assumptions and estimations made, the present work indicates a relatively low radiation risk due to (238)U and (232)Th after internal exposure of the healing earth.

Online breath gas analysis in unrestrained mice by hs-PTR-MS.

The phenotyping of genetic mouse models for human disorders may greatly benefit from breath gas analysis as a noninvasive tool to identify metabolic alterations in mice. Phenotyping screens such as the German Mouse Clinic demand investigations in unrestrained mice. Therefore, we adapted a breath screen in which exhaled volatile organic compounds (VOCs) were online monitored by proton transfer reaction mass spectrometry (hs-PTR-MS). The source strength of VOCs was derived from the dynamics in the accumulation profile of exhaled VOCs of a single mouse in a respirometry chamber. A careful survey of the accumulation revealed alterations in the source strength due to confounders, e.g., urine and feces. Moreover changes in the source strength of humidity were triggered by changes in locomotor behavior as mice showed a typical behavioral pattern from activity to settling down in the course of subsequent accumulation profiles. We demonstrated that metabolic changes caused by a dietary intervention, e.g., after feeding a high-fat diet (HFD) a sample of 14 male mice, still resulted in a statistically significant shift in the source strength of exhaled VOCs. Applying a normalization which was derived from the distribution of the source strength of humidity and accounted for varying locomotor behaviors improved the shift. Hence, breath gas analysis may provide a noninvasive, fast access to monitor the metabolic adaptation of a mouse to alterations in energy balance due to overfeeding or fasting and dietary macronutrient composition as well as a high potential for systemic phenotyping of mouse mutants, intervention studies, and drug testing in mice.

Estimating the absorption of soil-derived uranium in humans.

The aim of the present study was to improve the estimation of soil-derived uranium absorption in humans. For this purpose, an in vitro solubility assay was combined with a human study by using a specific edible soil low in uranium. The mean bioaccessibility of the soil-derived uranium, determined by the solubility assay in artificial gastrointestinal fluid, was found to be 7.7% with a standard deviation of 0.2%. The corresponding bioavailability of the soil-derived uranium in humans was assumed to be log-normal distributed with a geometric mean of 0.04% and a 95% confidence interval ranging from 0.0049% to 0.34%. Both results were used to calculate a factor, denoted as fA(sol), which describes the relation between the bioaccessibility and the bioavailability of soil-derived uranium. The geometric mean of fA(sol) was determined to be 0.53% with a 95% confidence interval ranging from 0.06% to 4.43%. Based on fA(sol), it is possible to estimate more realistic values on the bioavailability of uranium for highly uranium-contaminated soils in humans by just performing the applied solubility assay. The results of this study can be further used to obtain more reliable results on the internal dose assessment of ingested highly uranium-contaminated soils.

Human biokinetic data and a new compartmental model of zirconium — A tracer study with enriched stable isotopes

Biokinetic models describing the uptake, distribution and excretion of trace elements are an essential tool in nutrition, toxicology, or internal dosimetry of radionuclides. Zirconium, especially its radioisotope (95)Zr, is relevant to radiation protection due to its production in uranium fission and neutron activation of nuclear fuel cladding material. We present a comprehensive set of human data from a tracer study with stable isotopes of zirconium. The data are used to refine a biokinetic model of zirconium. Six female and seven male healthy adult volunteers participated in the study. It includes 16 complete double tracer investigations with oral ingestion and intravenous injection, and seven supplemental investigations. Tracer concentrations were measured in blood plasma and urine collected up to 100 d after tracer administration. The four data sets (two chemical tracer forms in plasma and urine) each encompass 105-240 measured concentration values above detection limits. Total fractional absorption of ingested zirconium was found to be 0.001 for zirconium in citrate-buffered drinking solution and 0.007 for zirconium oxalate solution. Biokinetic models were developed based on the linear first-order kinetic compartmental model approach used by the International Commission on Radiological Protection (ICRP). The main differences of the optimized systemic model of zirconium to the current ICRP model are (1) recycling into the transfer compartment made necessary by the observed tracer clearance from plasma, (2) different parameters related to fractional absorption for each form of the ingested tracer, and (3) a physiologically based excretion pathway to urine. The study considerably expands the knowledge on the biokinetics of zirconium, which was until now dominated by data from animal studies. The proposed systemic model improves the existing ICRP model, yet is based on the same principles and fits well into the ICRP radiation protection approach.

Radiation dose assessment of exposure to depleted uranium

Depleted uranium (DU) is claimed to contribute to human health problems, known as the Gulf War Syndrome and the Balkan Syndrome. Quantitative radiation dose is required to estimate the health risk of DU materials. The influences of the solubility parameters in the human alimentary tract and the respiratory tract systems and the aerosol particles size on the radiation dose of DU materials were evaluated. The dose conversion factor of daily urinary excretion of DU is provided. The retention and excretion of DU in the human body after a contamination at a wound site were predicted. Dose coefficients of DU after ingestion and inhalation were calculated using the solubility parameters of the DU corrosion products in simulated gastric and simulated lung fluid, which were determined in the Helmholtz Zentrum München. 238U is the main radiation dose contributor per 1 Bq of DU materials. The dose coefficients of DU materials were estimated to be 3.5 × 10−8 and 2.1 × 10−6 Sv Bq−1 after ingestion and inhalation for members of the public. The ingestion dose coefficient of DU materials is about 75% of the natural uranium value. The inhalation dose coefficient of DU material is in between those for Type M and Type S according to the category for inhaled materials defined by the International Commission on Radiological Protection. Radiation dose possibly received from DU materials can directly be estimated by using the dose conversion factor provided in this study, if daily urinary excretion of DU is measured.

Preparation of Ca-alginate biopolymer beads and investigation of their decorporation characteristics for 85Sr, 238U and 234Th by in vitro experiments

The aim of this work was to investigate whether Ca-alginate biopolymer beads (CaABBs) can be used to reduce the bioavailability of radionuclides in the gastrointestinal tract of humans. The uptake of strontium, uranium and thorium from a simulated gastrointestinal system was studied by in vitro techniques using CaABBs. This agent was prepared from Na-alginate through cross-linking with divalent calcium ions according to the egg-box model. The effects of process variables such as pH of the gastrointestinal juice, incubation time and solid-to-solution ratio for the removal of radionuclides from the gastrointestinal juice were investigated. The results suggest that CaABBs are a potent material for reducing the bioavailability of radionuclides with a high uptake efficiency in the gastrointestinal tract.

Daily urinary excretion of uranium in members of the public of Southwest Nigeria

The main aim of this study was to determine and evaluate urinary excretion values of uranium in members of the public of Southwest Nigeria living in areas of low environmental uranium. As several uranium mines are running in Nigeria and the operations could be a risk of contamination for the workers as well as for the members of the public, biomonitoring of urine could provide information about the exposure to uranium for the subjects. Therefore, baseline values of uranium in urine are needed from subjects living in areas without mining activities. Volunteers of both genders (age range 3 to 78 years) were asked to collect 24h-urine samples. The concentration measurements of uranium in urine were performed by Inductively Coupled Plasma Mass Spectrometry (ICP-MS). In addition, urinary creatinine values were determined for normalization of the renal uranium relative to the creatinine concentrations. The urinary uranium concentrations and their creatinine normalized values ranged from <10.4 to 150 ng L(-1) (median 13.8 ng L(-1)) and from 2.52 to 252.7 ng g(-1) creatinine (median 33.4 ng g(-1) creatinine), respectively, for adult subjects above 15 years of both genders. An increased uranium excretion value of 61.6 ng L(-1) (median), and of 76.0 ng g(-1) creatinine, respectively, were found in young subjects below 15 years. The median of daily excreted uranium was estimated to be 14.2 ng d(-1) for adults and of 45.1 ng d(-1) for children, respectively. The uranium excretion from males and females living in Nigeria in a non-mining area was comparable to reference values reported from other countries with low level of environmental uranium. The data can be considered as baseline values of urinary uranium in unexposed subjects in Nigeria.