Comparison of volatile organic compounds from lung cancer patients and healthy controls-challenges and limitations of an observational study.

This paper outlines the design and performance of an observational study on the profiles of volatile organic compounds (VOCs) in the breath of 37 lung cancer patients and 23 healthy controls of similar age. The need to quantify each VOC considered as a potential disease marker on the basis of individual calibration is elaborated, and the quality control measures required to maintain reproducibility in breath sampling and subsequent instrumental trace VOC analysis using solid phase microextraction-gas chromatography-mass spectrometry over a study period of 14 months are described. Twenty-four VOCs were quantified on the basis of their previously suggested potential as cancer markers. The concentration of aromatic compounds in the breath was increased, as expected, in smokers, while lung cancer patients displayed significantly increased levels of oxygenated VOCs such as aldehydes, 2-butanone and 1-butanol. Although sets of selected oxygenated VOCs displayed sensitivities and specificities between 80% and 90% using linear discriminant analysis (LDA) with leave-one-out cross validation, the effective selectivity of the breath VOC approach with regard to cancer detection is clearly limited. Results are discussed against the background of the literature on volatile cancer marker investigations and the prospects of linking increased VOC levels in patients' breath with approaches that employ sniffer dogs. Experience from this study and the literature suggests that the currently available methodology is not able to use breath VOCs to reliably discriminate between cancer patients and healthy controls. Observational studies often tend to note significant differences in levels of certain oxygenated VOCs, but without the resolution required for practical application. Any step towards the exploitation of differences in VOC profiles for illness detection would have to solve current restrictions set by the low and variable VOC concentrations. Further challenges are the technical complexity of studies involving breath sampling and possibly the limited capability of current analytical procedures to detect unstable marker candidates.

Narrow-Band Emitting Solid Fluorescence Reference Standard with Certified Intensity Pattern.

The development of a lanthanum-phosphate glass doped with several rare-earth-ions for use as solid fluorescence standard is described. The cuvette-shaped reference material which shows a characteristic emission intensity pattern upon excitation at 365 nm consisting of a multitude of relatively narrow emission bands in the wavelength region between 450 and 700 nm is intended for the day-to-day performance validation of fluorescence measuring devices. Evaluation of the fluorescent glass includes the determination of all properties which can affect its relative emission intensity profile or contribute to the uncertainty of the certified values like absorption spectra, fluorescence anisotropy, excitation wavelength, and temperature dependence of the spectroscopic features, homogeneity of fluorophore distribution, and photo- and long-term stability. Moreover, a certification procedure was developed including the normalization of the intensity profile consisting of several narrow emission bands and the calculation of wavelength-dependent uncertainties. Criteria for the design, characterization, and working principle of the new reference material BAM-F012 are presented, and possible applications of this ready-to-use fluorescence standard are discussed.

T-2 and HT-2 toxins in oat flakes: development of a certified reference material.

Trichothecene mycotoxins, with T-2 and HT-2 toxins being the main representatives of the type A subgroup, are naturally and worldwide occurring contaminants frequently found in grain-based food and feed. Due to the high consumption of these products and the potential health risk associated herewith, concerns about the safety and quality of food and feed have increased over the last decades at both governmental and consumer levels. Since it is not possible to avoid their occurrence, tremendous efforts have been performed to identify and monitor mycotoxins in food and feed to make their consumption safe. However, suitable certified reference materials (CRMs) intended for quality assurance and quality control purposes are still lacking for many mycotoxin-matrix combinations. Therefore, in the framework of a European Reference Material (ERM®) project, the first CRM for T-2 and HT-2 toxin in ground oat flakes (ERM®-BC720) was developed according to the requirements of ISO Guide 35. The whole process of ERM®-BC720 development, including sample preparation, homogeneity and stability studies and value assignment, is presented. The assignment of the certified mass fractions was based upon an in-house study using high-performance liquid chromatography isotope-dilution tandem mass spectrometry. Simultaneously, an interlaboratory comparison study involving 24 expert laboratories was conducted in order to support the in-house certification study. The certified values and their corresponding expanded uncertainties (k = 2) for both T-2 and HT-2 toxin in ERM®-BC720, traceable to the international system of units, are (82 ± 4) µg kg(-1) and (81 ± 4) µg kg(-1), respectively.

Certification of a reference material for determination of total cyanide in soil to support implementation of the International Standard ISO 11262:2011.

Cyanides are among the most important inorganic pollutants to be tested and monitored in environmental compartments. They can be distinguished and determined as free cyanide, weak acid dissociable cyanide or as total cyanide. However, in any case obtained, measurement results are operationally defined referring to the applied analytical method. In 2011, the International Standard ISO 11262 has been published which specifies a normative analytical method for the determination of total cyanide in soil. The objective of the project described in this paper was to provide the first soil reference material (CRM) certified for its mass fraction of total cyanide on the basis of this standard. A total of 12 German laboratories with proven experience in the determination of cyanides in environmental samples participated in the certification study. Measurement results were evaluated in full compliance with the requirements of ISO Guide 35. Taking into account the results of the inter-laboratory comparison as well as the outcome of the homogeneity and stability studies, a certified mass fraction of total cyanide of 21.1 mg/kg and an expanded uncertainty (k = 2) of 1.3 mg/kg were assigned to the material. The reference material has been issued as CRM BAM-U114.

Development and certification of a reference material for Fusarium mycotoxins in wheat flour.

Deoxynivalenol (DON), nivalenol (NIV) and zearalenone (ZEN) are toxic secondary metabolites produced by several species of Fusarium fungi. These mycotoxins are often found together in a large variety of cereal-based foods, which are regulated by maximum content levels of DON and ZEN. To date, suitable certified reference materials (CRM) intended for quality control purposes are lacking for these Fusarium mycotoxins. In order to overcome this lack, the first CRM for the determination of DON, NIV and ZEN in naturally contaminated wheat flour (ERM®-BC600) was developed in the framework of a European Reference Materials (ERM®) project. This article describes and discusses the whole process of ERM®-BC600 development, including material preparation, homogeneity and stability studies, and an interlaboratory comparison study for certification. A total of 21 selected expert laboratories from different European countries with documented expertise in the field of mycotoxin analysis took part in the certification study using various gas and liquid chromatographic methods. The certified values and their corresponding expanded uncertainties (k = 2) were assigned in full compliance with the requirements of ISO Guide 35 and are as follows: 102 ± 11 µg kg(-1) for DON, 1000 ± 130 µg kg(-1) for NIV and 90 ± 8 µg kg(-1) for ZEN.

State-of-the art comparability of corrected emission spectra. 1. Spectral correction with physical transfer standards and spectral fluorescence standards by expert laboratories.

The development of fluorescence applications in the life and material sciences has proceeded largely without sufficient concern for the measurement uncertainties related to the characterization of fluorescence instruments. In this first part of a two-part series on the state-of-the-art comparability of corrected emission spectra, four National Metrology Institutes active in high-precision steady-state fluorometry performed a first comparison of fluorescence measurement capabilities by evaluating physical transfer standard (PTS)-based and reference material (RM)-based calibration methods. To identify achievable comparability and sources of error in instrument calibration, the emission spectra of three test dyes in the wavelength region from 300 to 770 nm were corrected and compared using both calibration methods. The results, obtained for typical spectrofluorometric (0°/90° transmitting) and colorimetric (45°/0° front-face) measurement geometries, demonstrated a comparability of corrected emission spectra within a relative standard uncertainty of 4.2% for PTS- and 2.4% for RM-based spectral correction when measurements and calibrations were performed under identical conditions. Moreover, the emission spectra of RMs F001 to F005, certified by BAM, Federal Institute for Materials Research and Testing, were confirmed. These RMs were subsequently used for the assessment of the comparability of RM-based corrected emission spectra of field laboratories using common commercial spectrofluorometers and routine measurement conditions in part 2 of this series (subsequent paper in this issue).

State-of-the art comparability of corrected emission spectra. 2. Field laboratory assessment of calibration performance using spectral fluorescence standards.

In the second part of this two-part series on the state-of-the-art comparability of corrected emission spectra, we have extended this assessment to the broader community of fluorescence spectroscopists by involving 12 field laboratories that were randomly selected on the basis of their fluorescence measuring equipment. These laboratories performed a reference material (RM)-based fluorometer calibration with commercially available spectral fluorescence standards following a standard operating procedure that involved routine measurement conditions and the data evaluation software LINKCORR developed and provided by the Federal Institute for Materials Research and Testing (BAM). This instrument-specific emission correction curve was subsequently used for the determination of the corrected emission spectra of three test dyes, X, QS, and Y, revealing an average accuracy of 6.8% for the corrected emission spectra. This compares well with the relative standard uncertainties of 4.2% for physical standard-based spectral corrections demonstrated in the first part of this study (previous paper in this issue) involving an international group of four expert laboratories. The excellent comparability of the measurements of the field laboratories also demonstrates the effectiveness of RM-based correction procedures.

How to improve reliability in groundwater analysis: over a decade of experience with external quality control in field campaigns on volatile halogenated compounds.

The reliability in measurement results obtained during environmental monitoring is crucial for the assessment and further planning of remediation efforts on the respective contaminated sites by the responsible authorities. A case study concerned with groundwater contaminated with perchloroethylene, trichloroethylene and 1,1,2-trichlorotrifluoroethane including their degradation products which involves private contract laboratories and an independent provider of quality assurance (QA) is presented. The experience gained with biannual monitoring campaigns over 14 years indicates that the selection of contractors on basis of accreditation status and successful performance in interlaboratory comparisons are not sufficient. Rather the auditing of the contractors by the QA provider prior to each campaign and the crosschecking of selected monitoring samples by the QA provider led to a lasting improvement of reliability in the contractors' measurement results. A mean deviation of 20% from the reference value determined by the QA provider for the crosschecked samples was reached.

Temporal trend (1988-2008) of hexabromocyclododecane enantiomers in herring gull eggs from the German coastal region.

Levels of α-, ß-, and γ-hexabromocyclododecane (HBCD) were determined in pooled eggs from herring gulls (Larus argentatus) sampled on three bird sanctuaries near the German North Sea coast between 1988 and 2008 (Mellum and Trischen) and the German Baltic Sea coast between 1998 and 2008 (Heuwiese) and archived by the German Environmental Specimen Bank. Pressurized fluid extraction, gel permeation chromatography, and LC-MS/MS using (13)C(12)-labelled isotope standards and a chiral column were applied. α-HBCD was the dominating diastereomer and ranged between 3.7 and 107 ng g(-1)lw while ß- and γ-HBCD were throughout close to LOQ. The highest α-HBCD concentration was found in eggs from Mellum sampled in the year 2000. Interestingly, HBCD in eggs from the three islands displayed similar time courses with levels increasing to a peak contamination around 2000 and decreasing levels ever since. Chiral signatures of α-HBCD in eggs differed among the islands but indicated a preferential enrichment of the first eluting enantiomer (-)-α-HBCD.

HBCD stereoisomer pattern in mirror carps following dietary exposure to pure gamma-HBCD enantiomers.

1,2,5,6,9,10-hexabromocyclododecane (HBCD) is a brominated flame retardant consisting of a mixture of diastereomeric pairs of enantiomers that is a known omnipresent, environmental contaminant. The present study investigated the possibility of bioisomerization of HBCD stereoisomers. Therefore, mirror carps (Cyprinus carpio morpha noblis) were exposed to pure (+)- and (-)-gamma-HBCD, randomly sampled biweekly over a period of three and a half months and the fillets were subjected to enantiomer-specific determination of HBCD. Considering the background contamination of the fish at the beginning of the feeding period, significant enrichment of the respectively fed gamma-enantiomer was already detectable after two weeks of exposure. However, no significant enrichment of the respectively expected alpha-enantiomer was observed within this period. Thus, no evidence for the isomerization of HBCD stereoisomers was found in mirror carp under the applied conditions.

Degradation kinetics of the Alternaria mycotoxin tenuazonic acid in aqueous solutions.

The degradation kinetics of the Alternaria mycotoxin tenuazonic acid (l-TA) in aqueous buffer were studied over a period of 4 months at different pH levels (3.5 and 7.0) and temperatures (4, 25 and 40 degrees C). l-TA and its degradation products were quantified by newly developed high-performance liquid chromatography methods with UV or electrospray multistage mass spectrometry detection. At pH 3.5, significant degradation occurred at 25 and 40 degrees C, the respective l-TA half-lives being 73.8 +/- 0.4 and 14.0 +/- 0.1 days. Two degradation processes, epimerization and hydrolysis, were evaluated kinetically. The hydrolytically formed iso-deacetyl TA (iso-DTA, epimeric mixture) was found to be the stable end product of l-TA degradation under the conditions of this study. This indicates that iso-DTA as well as the l-TA epimer u-TA are formed in aqueous beverage matrices.

Development of two certified reference materials for acrylamide determination in foods.

Certified reference materials (CRMs) are a versatile tool for quality assurance in the chemical laboratory. In the case of acrylamide analysis, the availability of appropriate materials was rather limited. This lack of acrylamide matrix CRMs has now been overcome by the development of two European reference materials (ERM) for the determination of acrylamide in food (crispbread, ERM-BD272, and rusk, ERM-BD274). This article describes the preparation of the materials, provides the results of the homogeneity and stability studies, and presents and discusses the outcome of the certification studies. Expert laboratories from different European countries took part in the certification studies using various analytical methods. The acrylamide mass fractions were certified to 980 microg kg(-1) for crispbread and 74 microg kg(-1) for rusk.

Mineral oil content in sediments and soils: comparability, traceability and a certified reference material for quality assurance.

The performance of twelve laboratories with previously established proficiency in the determination of the mineral oil content in a fresh water sediment is described. The summation parameter total petrol hydrocarbon (TPH) is defined according to ISO 16703:2004 with regard to the sample preparation to be applied, the flame ionisation detection (FID) and the boiling range of C10-C40 to be integrated. Comprehensive tests of homogeneity and stability have been carried out on the candidate material using appropriate models. The outcome of the study served as the basis for the certification of the candidate reference material as ERM-CC015a. The certified mass fraction is 1,820+/-130 mg kg(-1) and traceability was established by using an appropriate calibration standard certified for the mass fraction of C10-C40. The interlaboratory scatter of measurement results in this exercise can largely be explained by the variability of the individual calibrations based on this common calibration standard.

Analysis of natural gas by gas chromatography reduction of correlated uncertainties by normalisation.

The results of gas chromatographic analysis of natural gas mixtures reveal strong correlations (Pearson correlation coefficient of >0.96) between the uncertainty of each component and variations in the ambient pressure. Although correction for ambient pressure variations can reduce this variability, normalisation of the results of each analysis using the assumption that the sum of all component amount fractions is unity provides significantly greater reductions in the uncertainty of each measured component. We show that the uncertainty in normalised components can be estimated approximately using the correlation coefficient as a measure of the correlation present in the measurements, or exactly using a full calculation of the variance/covariance (V/C) structure of the data.