An innovative approach to the analysis of VOCs exhaled by mice under regular housing conditions at the German Mouse Clinic using hs-PTR-MS.

The noninvasive method of breath gas analysis is widely used in human studies to explore the diagnostic potential of endogenous VOCs [1]. Proton transfer reaction mass spectrometry (PTR-MS) measures a wide spectrum of VOCs online, fast and nearly simultaneously from ppm (part-per-million) down to ppt (part-per-trillion) levels [2]. In the German Mouse Clinic (GMC) mutant mouse models for human diseases are comprehensively phenotyped to better understand the underlying molecular mechanisms and to develop new therapeutic strategies. Despite the advances in PTR-MS instrumentation the exploration of exhaled VOCs of small animals is still challenging and, hence, rather uncommon and furthermore restricted to specific VOCs as acetone or ethanol [3, 4]. Therefore, the aim of our study was to develop a PTR-MS setup suitable for unrestrained screening of exhaled VOCs in mouse models of human disease

Discrimination of cancerous and non-cancerous cell lines by headspace-analysis with PTR-MS.

Proton transfer reaction mass spectrometry (PTR-MS) has been used to analyze the volatile organic compounds (VOCs) emitted by in-vitro cultured human cells. For this purpose, two pairs of cancerous and non-cancerous human cell lines were selected:1. lung epithelium cells A-549 and retinal pigment epithelium cells hTERT-RPE1, cultured in different growth media; and 2. squamous lung carcinoma cells EPLC and immortalized human bronchial epithelial cells BEAS2B, cultured in identical growth medium. The VOCs in the headspace of the cell cultures were sampled: 1. online by drawing off the gas directly from the culture flask; and 2. by accumulation of the VOCs in PTFE bags connected to the flask for at least 12 h. The pure media were analyzed in the same way as the corresponding cells in order to provide a reference. Direct comparison of headspace VOCs from flasks with cells plus medium and from flasks with pure medium enabled the characterization of cell-line-specific production or consumption of VOCs. Among all identified VOCs in this respect, the most outstanding compound was m/z = 45 (acetaldehyde) revealing significant consumption by the cancerous cell lines but not by the non-cancerous cells. By applying multivariate statistical analysis using 42 selected marker VOCs, it was possible to clearly separate the cancerous and non-cancerous cell lines from each other

Assessment of Intestinal Absorption of Trace Metals in Humans by Means of Stable Isotopes.

This study is aimed to demonstrate the feasibility of stable isotopes for the assessment of reliable data on fractional intestinal absorption of trace metals in healthy humans. Among the various methods available, the double isotope technique, i.e. one isotope given orally together with the test substance to be investigated and another isotope injected intravenously to correct for retention and endogenous excretion of the particular trace metal, provides quantitative figures of intestinal absorption at reasonable expenses with regard to costs for materials and number of samples to be evaluated. The trace metals exemplarily included in this study, i.e. iron, cobalt and molybdenum show diverging relations between absorbed fractions and amounts administered which are indicative for different regulatory mechanisms of their body content. Food ligands influence the fractional absorption significantly so that the uptake from a composite meal cannot be derived from results on uptake from particular foodstuffs. Therefore, validated data on the behaviour of intestinal absorption will significantly contribute to a better understanding of human trace metal metabolism

Measurements of daily urinary uranium excretion in German peacekeeping personnel and residents of the Kosovo region to assess potential intakes of depleted uranium (DU)

A study designed to determine the extent of military and civilian population exposure to munitions-derived depleted uranium in Kosovo and southern Serbia. Used advanced multi-collector ICPMS to measure DU excretion and uranium isotope ratios. No current DU exposure indicated cf. previous publication where excretion found early after conflict

Investigations on the activity concentrations of U-238, Ra-226, Ra-228, Pb-210 and K-40 in Jordan phosphogypsum and fertilizers.

The activity concentrations of naturally occurring radionuclides (U-238, Ra-226, Ra-228, Pb-210 and K-40) in Jordanian phosphate ore, fertilizer material and phosphogypsum piles were investigated. The results show the partitioning of radionuclides in fertilizer products and phosphogypsum piles. The outcome of this study will enrich the Jordanian radiological map database, and will be useful for an estimation of the radiological impact of this industrial complex on the immediate environment. The activity concentration of Pb-210 was found to vary from 95 +/- 8 to 129 +/- 8 Bq kg(-1) with a mean value of 111 +/- 14 Bq kg(-1) in fertilizer samples, and from 364 +/- 8 to 428 +/- 10 Bq kg(-1) with a mean value of 391 +/- 30 Bq kg(-1) in phosphogypsum samples; while in phosphate wet rock samples, it was found to vary between 621 +/- 9 and 637 +/- 10 Bq kg(-1), with a mean value of 628 +/- 7 Bq kg(-1). The activity concentration of Ra-226 in fertilizer samples (between 31 +/- 4 and 42 +/- 5 Bq kg(-1) with a mean value of 37 +/- 6 Bq kg(-1)) was found to be much smaller than the activity concentration of Ra-226 in phosphogypsum samples (between 302 +/- 8 and 442 +/- 8 Bq kg(-1) with a mean value of 376 +/- 62 Bq kg(-1)). In contrast, the activity concentration of U-238 in fertilizer samples (between 1011 +/- 13 and 1061 +/- 14 Bq kg(-1) with a mean value of 1033 +/- 22 Bq kg(-1)) was found to be much higher than the activity concentration of U-238 in phosphogypsum samples (between 14 +/- 5 and 37 +/- 7 Bq kg(-1) with a mean value of 22 +/- 11 Bq kg(-1)). This indicates that Pb-210 and Ra-226 show similar behaviour, and are concentrated in phosphogypsum piles. In addition, both isotopes enhanced the activity concentration in phosphogypsum piles, while U-238 enhanced the activity concentration in the fertilizer. Due to the radioactivity released from the phosphate rock processing plants into the environment, the highest collective dose commitment for the lungs was found to be 1.02 person nGy t(-1). Lung tissue also shows the highest effect due the presence of Ra-226 in the radioactive cloud (0.087 person nGy t(-1))

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

Improvements in routine internal monitoring— an overview of the IDEA project.

The IDEA project aimed to improve the assessment of incorporated radionuclides through developments of advanced in vivo and bioassay monitoring techniques and making use of such enhancements for improvements in routine monitoring. Many of these findings are not new in the sense that they are being already employed in advanced laboratories or for specialised applications. The primary goal was to categorise those new developments regarding their potential and eligibility for the routine monitoring community. Attention has been given to in vivo monitoring techniques with respect to detector characteristics and measurement geometry to improve measurement efficiency with special attention to low energy gamma emitters. Calibration—specifically supported by or through methods of numerical simulation—have been carefully analysed to reduce overall measurement uncertainties and explore ways to accommodate the individual variability based on characteristic features of a given person. For bioassay measurements at low detection limits, inductively coupled plasma mass spectroscopy offers significant advantages both in accuracy, speed, and sample preparation. Specifically, the determination of U and Th in urine and the associated models have been investigated. Finally, the scientific achievements have been analysed regarding their potential to offer benefits for routine monitoring. These findings will be presented in greater detail in other papers at this conference, whereas this paper intends to give an overview and put both the scientific achievements as well as the derived benefits into perspective

Implementation of bioassay methods to improve assessment of incorporated radionuclides.

The present work which was carried out in the framework of an EU project (IDEA: Internal Dosimetry—Enhancements in Application; Contract Number: FIKR CT2001 00164) shall provide commonly acceptable guidelines for optimum performance of ICP-MS measurements with focus on urinary measurements of uranium, thorium and actinides. From the results of this work it is recommended that, whenever feasible, 24 h urine sampling should be conducted to avoid large uncertainties in the quantitation of daily urinary excretion values. For storage, urine samples should be acidified and kept frozen before analysis. Measurement of total uranium in urine by ICP-MS at physiological levels (<10 ng·l–1) requires no sample preparation besides UV photolysis and/or dilution. For the measurement of thorium in urine by ICP-MS, it can be concluded, that salt removal from the urine samples is not recommended. For the measurement of actinides in urine it is shown that ICP-MS is well-suited and a good alternative to alpha-spectrometry for isotopes with T1/2>5x104 years. In general, ICP-MS measurements are an easy, fast and cost-saving methodology. New improved measuring techniques (HR-SF-ICP-MS) with detection limits in urine of 150 pg·l–1 (1.9 µBq·l–1) for 238U, 30 pg·l–1 (2.4 µBq·l–1) for 235U and 100 pg·l–1 (0.4 µBq·l–1) for 232Th, respectively, meet all necessary requirements. This method should therefore become the routine technique for incorporation monitoring of workers and of members of the general public, in particular for uranium contamination

Practical implications of procedures developed in IDEA project—comparison with traditional methods.

The idea of the IDEA project aimed to improve assessment of incorporated radionuclides through developments of more reliable and possibly faster in vivo and bioassay monitoring techniques and making use of such enhancements for improvements in routine monitoring. In direct in vivo monitoring technique the optimum choice of the detectors to be applied for different monitoring tasks has been investigated in terms of material, size and background in order to improve conditions namely to increase counting efficiency and reduce background. Detailed studies have been performed to investigate the manifold advantageous applications and capabilities of numerical simulation method for the calibration and optimisation of in vivo counting systems. This calibration method can be advantageously applied especially in the measurement of low-energy photon emitting radionuclides, where individual variability is a significant source of uncertainty. In bioassay measurements the use of inductively coupled plasma mass spectrometry (ICP-MS) can improve considerably both the measurement speed and the lower limit of detection currently achievable with alpha spectrometry for long-lived radionuclides. The work carried out in this project provided detailed guidelines for optimum performance of the technique of ICP-MS applied mainly for the determination of uranium and thorium nuclides in the urine including sampling procedure, operational parameters of the instruments and interpretation of the measured data. The paper demonstrates the main advantages of investigated techniques in comparison with the performances of methods commonly applied in routine monitoring practice