European Interlaboratory Comparison Research Articles

Reproducibility of the wet part of the soil water retention curve: a European interlaboratory comparison

Abstract. The soil water retention curve (SWRC) is a key soil property required for predicting basic hydrological processes. The SWRC is often obtained in the laboratory with non-harmonized methods. Moreover, procedures associated with each method are not standardized. This can induce a lack of reproducibility between laboratories using different methods and procedures or using the same methods with different procedures. The goal of this study was to estimate the inter- and intralaboratory variability of the measurement of the wet part (from 10 to 300 hPa) of the SWRC. An interlaboratory comparison was carried out between 14 laboratories, using artificially constructed, porous reference samples that were transferred between laboratories according to a statistical design. The retention measurements were modelled by a series of linear mixed models using a Bayesian approach. This allowed the detection of sample-to-sample variability, interlaboratory variability, intralaboratory variability and the effects of sample changes between measurements. The greatest portion of the differences in the measurement of SWRCs was due to interlaboratory variability. The intralaboratory variability was highly variable depending on the laboratory. Some laboratories successfully reproduced the same SWRC on the same sample, while others did not. The mean intralaboratory variability over all laboratories was smaller than the mean interlaboratory variability. A possible explanation for these results is that all laboratories used slightly different methods and procedures. We believe that this result may be of great importance regarding the quality of SWRC databases built by pooling SWRCs obtained in different laboratories. The quality of pedotransfer functions or maps that might be derived is probably hampered by this inter- and intralaboratory variability. The way forward is that measurement procedures of the SWRC need to be harmonized and standardized.

European interlaboratory comparison investigations (ICI) and external quality assurance schemes (EQUAS) for the analysis of bisphenol A, S and F in human urine: Results from the HBM4EU project

The Human Biomonitoring for Europe initiative (HBM4EU) aims to study the exposure of citizens to chemicals and potentially associated health effects. One objective of this project has been to build a network of laboratories able to answer to the requirements of European human biomonitoring studies. Within the HBM4EU quality assurance and quality control scheme (QA/QC), a number of interlaboratory comparison investigations (ICIs) and external quality assurance schemes (EQUASs) were organized to ensure data consistency, comparability and reliability. Bisphenols are among the prioritized substance groups in HBM4EU, including bisphenol A (BPA), bisphenol S (BPS) and bisphenol F (BPF) in human urine. In four rounds of ICI/EQUAS, two target concentration levels were considered, related to around P25 and P95 of the typical exposure distribution observed in the European general population. Special attention was paid to the conjugated phase II metabolites known to be most dominant in samples of environmentally exposed individuals, through the analysis of both native samples and samples fortified with glucuronide forms. For the low level, the average percentage of satisfactory results across the four rounds was 83% for BPA, 71% for BPS and 62% for BPF. For the high level, the percentages of satisfactory results increased to 93% for BPA, 89% for BPS and 86% for BPF. 24 out of 32 participating laboratories (75%) were approved for the analyses of BPA in the HBM4EU project according to the defined criterion of Z-scores for both low and high concentration levels in at least two ICI/EQUAS rounds. For BPS and BPF, the number of qualified laboratories was 18 out of 27 (67%) and 13 out of 28 (46%), respectively. These results demonstrate a strong analytical capability for BPA and BPS in Europe, while improvements may be needed for BPF.

Reliability of Morphological and PCR Methods for the Official Diagnosis of Aethina tumida (Coleoptera: Nitidulidae): A European Inter-Laboratory Comparison.

Simple SummaryAethina tumida, also called the Small Hive Beetle, is an insect that multiplies primarily in honeybee hives, causing honey losses and weakening colonies. It is native to sub-Saharan Africa and was introduced into different countries and continents over the last 20 years, posing a threat to beekeeping internationally. In case of introduction into a new area, officially approved laboratories (certified by government services) carry out analyses to confirm the outbreak. The reliability of the results is essential in the implementation of management measures. Therefore, a study was organised at the European level to compare the results between official laboratories for two types of methods, used routinely for the identification of A. tumida: morphological examination (form and structure) and DNA testing (genetics). The 22 participants analysed in a blinded way a panel of 12 samples (positive and negative samples). The results were very satisfactory, with the exception of one participant who encountered several anomalies for negative samples and especially for DNA tests, probably related to his inexperience with the method. This study proved the ability of laboratories and analytical methods to identify A. tumida, which is a key element in monitoring and managing this risk.The Small Hive Beetle (Aethina tumida Murray, 1867) is an invasive scavenger of honeybees. Originally endemic in sub-Saharan Africa, it is regulated internationally in order to preserve the areas still free from this species. To ensure the reliability of official diagnoses in case of introduction, an inter-laboratory comparison was organised on the identification of A. tumida by morphology and real-time PCR. Twenty-two National Reference Laboratories in Europe participated in the study and analysed 12 samples with adult coleopterans and insect larvae. The performance of the laboratories was evaluated in terms of sensitivity and specificity. Sensitivity was satisfactory for all the participants and both types of methods, thus fully meeting the diagnostic challenge of confirming all truly positive cases as positive. Two participants encountered specificity problems. For one, the anomaly was minor whereas, for the other, the issues concerned a larger number of results, especially real-time PCR, which probably were related to inexperience with this technique. The comparison demonstrated the reliability of official diagnosis, including the entire analytical process of A. tumida identification: from the first step of the analysis to the expression of opinions. The performed diagnostic tools, in parallel with field surveillance, are essential to managing A. tumida introduction.

Volatile organic compounds from building products—Results from six round robin tests with emission test chambers conducted between 2008 and 2018

Emission testing of volatile organic compounds (VOC) from materials and products is commonly based on emission test chamber measurements. To ensure the comparability of results from different testing laboratories, their measurement performance must be verified. For this purpose, Bundesanstalt für Materialforschung und -prüfung (BAM) organizes an international proficiency test (round robin test, RRT) every two years using well-characterized test materials (one sealant, one furniture board, and four times a lacquer) with defined VOC emissions. The materials fulfilled the requirements of homogeneity, reproducibility, and stability. Altogether, 36 VOCs were included of which 33 gave test chamber air concentrations between 13 and 83µg/m3 . This is the typical concentration range to be expected and to be quantified when performing chamber tests. Three compounds had higher concentrations between 326 and 1105µg/m3 . In this paper, the relative standard deviations (RSD) of BAM round robin tests since 2008 are compared and the improvement of the comparability of the emission chamber testing is shown by the decrease of the mean RSD down to 28% in 2018. In contrast, the first large European interlaboratory comparison in 1999 showed a mean RSD of 51%.

An international inter-laboratory study on Nosema spp. spore detection and quantification through microscopic examination of crushed honey bee abdomens

Nosemosis is a microsporidian disease causing mortality and weakening of honey bee colonies, especially in the event of co-exposure to other sources of stress. As a result, the disease is regulated in some countries. Reliable and harmonised diagnosis is crucial to ensure the quality of surveillance and research results. For this reason, the first European Interlaboratory Comparison (ILC) was organised in 2017 in order to assess both the methods and the results obtained by National Reference Laboratories (NRLs) in counting Nosema spp. spores by microscopy. Implementing their own routine conditions of analysis, the 23 participants were asked to perform an assay on a panel of ten positive and negative samples of crushed honey bee abdomens. They were asked to report results from a qualitative and quantitative standpoint. The assessment covered specificity, sensitivity, trueness and precision. Quantitative results were analysed in compliance with international standards NF ISO 13528 (2015) and NF ISO 5725-2 (1994). Three results showed a lack of precision and five a lack of trueness. However, overall results indicated a global specificity of 98% and a global sensitivity of 100%, thus demonstrating the advanced performance of the microscopic methods applied to Nosema spores by the NRLs. Therefore, the study concluded that using microscopy to detect and quantify spores of Nosema spp. was reliable and valid.

Optimization of the liquid scintillation counting technique for the European interlaboratory comparison on gross α/β activity concentrations in water

A liquid scintillation counting (LSC) measurement method optimization for the gross α/β activity analysis in drinking waters with different chemical and radionuclide composition was performed. The optimized method was suitable to provide gross radioactivity results in drinking waters with the levels of the accuracy and precision similar to those obtained using other radioactivity screening techniques. Robust LSC results contributed to the calculation of gross α/β activity reference values of EC-JRC interlaboratory comparison water samples in 2012. Some of the most common errors in the determination of the gross radioactivity using the LSC are presented.

60Co in cast steel matrix: A European interlaboratory comparison for the characterisation of new activity standards for calibration of gamma-ray spectrometers in metallurgy

Two series of activity standards of 60Co in cast steel matrix, developed for the calibration of gamma-ray spectrometry systems in the metallurgical sector, were characterised using a European interlaboratory comparison among twelve National Metrology Institutes and one international organisation. The first standard, consisting of 14 disc shaped samples, was cast from steel contaminated during production (“originally”), and the second, consisting of 15 similar discs, from artificially-contaminated (“spiked”) steel. The reference activity concentrations of 60Co in the cast steel standards were (1.077±0.019) Bqg−1 on 1 January 2013 12h00 UT and (1.483±0.022) Bqg−1 on 1 June 2013 12h00 UT, respectively.

Results of a European interlaboratory comparison on gross alpha/beta activity determination in drinking water

An interlaboratory comparison was organised by JRC-IRMM among environmental radioactivity monitoring laboratories for the determination of gross alpha/beta activity concentration in drinking water. Independent standard methods were used for the reference value determination. The performance of participating laboratories was evaluated with respect to the reference values using relative deviations. Sample preparation and measurement methods used by the participating laboratories are detailed, in particular in the view of method-dependency of the results. Many of the participants’ results deviate by more than two orders of magnitude from the reference values regardless of the techniques used. This suggests that gross methods need revision.

European interlaboratory comparison exercise for the analysis of PAHs on PM10quartz filters

Eighteen European reference laboratories of air quality had been involved in a comparison of polycyclic aromatic hydrocarbon analysis based on particulate matter (PM) field samples collected on quartz filter. Four PM10 samples and two blanks were selected for the comparison with the aim of covering a representative range of concentrations typical of ambient air. Laboratory results were treated according to International Organization for Standardization 5725 from which repeatability and reproducibility values for the comparison were calculated. In general, intercompound median repeatability and reproducibility values ranged between the samples from 2.1 to 7.4% and 23.6 to 61%, respectively. The median relative overall expanded uncertainty (OEU) was 30% for the highest concentration filter sample reaching 50% for the lowest concentration filter samples. The OEU was higher for the lighter compounds (phenanthrene and anthracene), characterized by a higher blank level, and for compounds (chrysene or dibenzo[a,h]anthracene), which showed difficult analytical resolution. The median OEU and the analytical relative expanded uncertainties estimated by the participants were in line with the data quality objectives of the European Directive 2004/107/EC. No significant differences were observed when comparing the different analytical techniques from the exercise's results. However, other factors such as the use of certified reference materials and the laboratory blank were important causes of biases. Moreover, the chromatographic separation of several isomers was noted suggesting the importance of the use of specific columns more appropriate to the separation of unresolved isomers.

Critical remarks on gross alpha/beta activity analysis in drinking waters: Conclusions from a European interlaboratory comparison

The most common gross alpha/beta standard methods used for drinking water analysis are discussed, and sources of interferences are reviewed from a metrological point of view. Our study reveals serious drawbacks of gross methods on the basis of an interlaboratory comparison analyzing commercial mineral water samples with the participation of 71 laboratories. A proposal is made to obtain comparable measurement results using true standardized methods.

Radiochemical characterization of mineral waters for a European interlaboratory comparison

The objective of this work was to select waters as realistic test items for an interlaboratory comparison on gross alpha/beta measurement in drinking waters. For this reason, a preliminary radioanalytical survey studying the naturally occurring alpha emitting radionuclides was carried out in 11 popular and regularly consumed mineral waters from the European market. The activity concentrations of the main naturally occurring alpha-emitting radionuclides (226Ra, 210Po, 234U, 235U, 238U and 228Th) were determined by using alpha spectrometry after separation from the matrix elements. Additionally, the committed effective dose was also estimated from the water consumption. Average activity concentrations of radionuclides in the mineral waters were in the following order: 226Ra>234U>238U>210Po>228Th>235U. The estimated committed effective doses for the analyzed mineral water samples are in the range 2.5–134.2μSv a−1 with an average 41.5μSv a−1, which is well below 100μSv a−1, the total indicative dose recommended by EC/WHO for drinking water (excluding mineral waters). Only one mineral water sample did not meet this requirement, its constant consumption would result in a dose higher than 100μSv a−1, mainly due to the contribution of 226Ra and 210Po. Apart from the main radiological characteristics of the analyzed waters, this study provides additional information for consumers and authorities about the potential internal exposure risk from mineral water intake. Based on the radionuclide composition and the total dissolved solids of the mineral waters two candidates were selected as potential test items for an interlaboratory comparison exercise.

FIRST EUROPEAN INTERLABORATORY COMPARISON OF TETRACYCLINE AND AGE DETERMINATION WITH RED FOX TEETH FOLLOWING ORAL RABIES VACCINATION PROGRAMS

The first European interlaboratory comparison of tetracycline and age determination with red fox (Vulpes vulpes) tooth samples was organized by the European Union Reference Laboratory for rabies. Performance and procedures implemented by member states were compared. These techniques are widely used to monitor bait uptake in European oral rabies vaccination campaigns. A panel of five red fox half-mandibles comprising one weak positive juvenile sample, two positive adult samples, one negative juvenile sample, and one negative adult sample were sent, along with a technical questionnaire, to 12 laboratories participating on a voluntary basis. The results of only three laboratories (25%) were 100% correct. False-negative results were more frequently seen in weak positive juvenile samples (58%) but were infrequent in positive adult samples (4%), probably due to differences in the ease of reading the two groups of teeth. Four laboratories (44%) had correct results for age determination on all samples. Ages were incorrectly identified in both adult and juvenile samples, with 11 and 17% of discordant results, respectively. Analysis of the technical questionnaires in parallel with test results suggested that all laboratories cutting mandible sections between the canine and first premolar obtained false results. All the laboratories using longitudinal rather than transverse sections and those not using a mounting medium also produced false results. Section thickness appeared to affect the results; no mistakes were found in laboratories using sections <150 μm thick. Factors having a potential impact on the success of laboratories were discussed, and recommendations proposed. Such interlaboratory trials underline the importance of using standardized procedures for biomarker detection in oral rabies vaccination campaigns. Several changes can be made to improve analysis quality and increase the comparability of bait uptake frequencies among member states.

Results of an European inter-laboratory comparison study on the determination of the 15 + 1 EU priority polycyclic aromatic hydrocarbons (PAHs) in liquid smoke condensates

A collaborative study on the determination of the 15 + 1 EU priority PAHs in Primary Smoke Condensate (PSC) investigating the performance profile of EU Member States’ laboratories and supporting the work carried out by the European Food Safety Authority on smoke flavourings was organised. Two spiked liquid smoke condensate materials were employed in this study. The results of 25 laboratories from across the European Union, using either high performance liquid chromatography with fluorescence detection or gas chromatography with mass selective detection, were evaluated by application of robust statistics. The assessment of the data indicated broadened Gaussian distributions for all analytes. For benzo[ a]pyrene (BaP), benzo[ a]anthracene (BaA), chrysene and 5-methylchrysene more than 80% of the respective reported values gave rise to a satisfactory score of | z| ⩽ 2. For benzo[ b]fluoranthene, benzo[ k]fluoranthene, and dibenzo[ a, l]pyrene 70–80%, for the remaining analytes less than 70%, and for dibenzo[ a, i]pyrene 52% of the scores were satisfactory. No systematic differences could be detected between values reported by laboratories using methods based on HPLC and the values related to methods based on GC for most of the analytes, except for benzo[ k]fluoranthene and benzo[ c]fluorene. In both cases laboratories using GC based methods reported about 50% higher values than laboratories using HPLC based methods. The overview on all z-scores sorted by laboratory revealed broad distributions and/or laboratory biases for several laboratories. An assessment of the reported method performance parameters revealed that for the two regulated compounds, BaP and BaA, only two thirds of the reported data were in compliance with Regulation (EC) 627/2006. Overall the methods used in the participating laboratories were – with ample room for improvement – well on the way to comply with European legislation.

European inter-laboratory comparison of high pressure CO 2 sorption isotherms. I: Activated carbon

In order to assess and improve the quality of high-pressure sorption isotherms of carbon dioxide (CO 2) on coals, an inter-laboratory study (“Round Robin”) has been conducted among four European research laboratories. In a first round of measurements, excess sorption isotherms were determined on Filtrasorb 400 (F400) activated carbon at 318 K using the manometric (TU Delft and RWTH Aachen University) and the gravimetric (FP Mons and INERIS) method up to 16 MPa. The study shows that CO 2 sorption in the supercritical range can be determined accurately with both gravimetric and manometric equipment but requires thorough optimization of instrumentation and measuring as well as proper sample preparation procedures. For the characterization of the activated carbon F400, which we used as benchmark, we have determined a surface area of 1063 m 2 g −1, and Dubinin-Radushkevich (DR) micropore volume of 0.51 cm 3 g −1. Additionally, we analysed the elementary near-surface composition by energy dispersive X-ray spectroscopy (EDX). To characterise the bulk composition of the F400 activated carbon, a proximate and ultimate analysis was performed. The observed excess sorption maxima around 5 MPa have values around 8.0 mol kg −1, which are consistently higher (by upto 0.8 mol kg −1) than literature data.

A multiplex RT-PCR assay for the rapid and differential diagnosis of classical swine fever and other pestivirus infections

Classical swine fever is a highly contagious viral disease causing severe economic losses in pig production almost worldwide. All pestivirus species can infect pigs, therefore accurate and rapid pestivirus detection and differentiation is of great importance to assure control measures in swine farming. Here we describe the development and evaluation of a novel multiplex, highly sensitive and specific RT-PCR for the simultaneous detection and rapid differentiation between CSFV and other pestivirus infections in swine. The universal and differential detection was based on primers designed to amplify a fragment of the 5′ non-coding genome region for the detection of pestiviruses and a fragment of the NS5B gene for the detection of classical swine fever virus. The assay proved to be specific when different pestivirus strains from swine and ruminants were evaluated. The analytical sensitivity was estimated to be as little as 0.89TCID50. The assay analysis of 30 tissue homogenate samples from naturally infected and non-CSF infected animals and 40 standard serum samples evaluated as part of two European Inter-laboratory Comparison Tests conducted by the European Community Reference Laboratory, Hanover, Germany proved that the multiplex RT-PCR method provides a rapid, highly sensitive, and cost-effective laboratory diagnosis for classical swine fever and other pestivirus infections in swine.

Results of a European inter-laboratory comparison study on the determination of EU priority polycyclic aromatic hydrocarbons (PAHs) in edible vegetable oils

A collaborative study on the analysis for 15 + 1 EU priority PAHs in edible oils was organised to investigate the state-of-the-art of respective analytical methods. Three spiked vegetable oils, one contaminated native sunflower oil, and one standard solution were investigated in this study. The results of 52 laboratories using either high-performance liquid chromatography with fluorescence detection or gas chromatography with mass-selective detectors were evaluated by application of robust statistics. About 95% of the laboratories were able to quantify benzo[a]pyrene together with five other PAHs included in the commonly known list of 16 US-EPA PAHs. About 80% of the participants also quantified seven additional PAHs in most samples, two of which were benzo[b]fluoranthene and benzo[k]fluoranthene, which were also known from the EPA list. Only about 50% of the participants quantified cyclopenta[cd]pyrene, benzo[j]fluoranthene, and benzo[c]fluorene. The robust relative standard deviations of the submitted results without discrimination between the methods applied ranged between 100% for 5-methylchrysene in spiked olive oil and 11% for the same analyte in spiked sunflower oil. The results clearly showed that for these analytes the methods of analysis are not yet well established in European laboratories, and more collaborative trials are needed to promote further development and to improve the performances of the respective methods.

First European interlaboratory study of the analysis of benzoxazinone derivatives in plants by liquid chromatography

Six laboratories from four different countries participated in the first European interlaboratory comparison exercise within the framework of the “Fate and toxicity of allelochemicals (natural plant toxins) in relation to environment and consumer” (FATEALLCHEM) European Union Project. The study, organized between November 2002 and March 2003, involved the analyses of seven benzoxazinone derivatives in two standard solutions and one purified extract of root material. Results are reported from the first phase of the study that examined the variability associated with different detection methods and different laboratories. The analytical strategies were based on liquid chromatography (LC) with diode array detection, LC coupled to mass spectrometry (MS) and LC coupled to tandem MS. When data from all laboratories were pooled, the relative standard deviation values ranged from 2 to 14% for the determination of target compounds in standard solutions, and between 19 and 47% for the analysis in root material. Comparison of the three detection techniques leads to the conclusion that MS approaches are the most accurate and precise techniques for the determination of benzoxazinone derivatives at ng/μL level in plant material.

Determination of uranium in urine - measurement of isotope ratios and quantification by use of inductively coupled plasma mass spectrometry.

For analysis of uranium in urine determination of the isotope ratio and quantification were investigated by high-resolution inductively coupled plasma mass spectrometry (HR ICP-MS). The instrument used (ThermoFinniganMAT ELEMENT2) is a single-collector MS and, therefore, a stable sample-introduction system was chosen. The methodical set-up was optimized to achieve the best precision for both the isotope ratio and the total uranium concentration in the urine matrix. Three spiked urine samples from an European interlaboratory comparison were analyzed to determine the (235)U/(238)U isotope ratio. The ratio was found to be in the range 0.002116 to 0.007222, the latter being the natural uranium isotope ratio. The first ratio indicates the abundance of depleted uranium. The effect of storage conditions and the stability for the matrix urine were investigated by using "real-life" urine samples from unexposed persons in the Netherlands. For samples stored under refrigeration and acidified the results (range 0.8 to 5.3 ng L(-1) U) were in the normal fluctuation range whereas a decrease in uranium concentration was observed for samples stored at room temperature without acidification.

State-of-the-art in the measurement of volatile organic compounds emitted from building products: results of European interlaboratory comparison.

Eighteen laboratories from 10 European countries participated in a comparison organized as part of the VOCEM project, a 2.5-year research collaboration among 4 research institutes and 4 industrial companies. The scope of the project was to improve the procedure used to measure volatile organic compounds (VOC) emitted from building materials and products in small test chambers. The interlaboratory comparison included the GC-MS determination of 5 target compounds from carpet, 8 from polyvinyl chloride (PVC) cushion vinyl and 2 from paint; for the first time, chamber recovery (sinks), homogeneity of solid materials and possible contamination during transport were tested. The results show that the intralaboratory variance (random errors) is much smaller than the interlaboratory variance (systematic errors). Causes of the largest interlaboratory discrepancies were: (i) analytical errors; (ii) losses of the heaviest compounds due to sorption on the chamber walls; and (iii) non homogeneity of the materials. The output of this work concerns both the objective of labelling materials with regard to their VOC emissions and the pre-standard drafted by the European Commitee for Standardization (CEN) for this type of determination.

European Interlaboratory Comparison of Lyme Borreliosis Serology

Serological testing for Lyme borreliosis was compared in 5 European reference laboratories with a total of 79 sera in order to determine variations in laboratory performance. A considerable range of methods were used and several laboratories employed 2 or 3 genomospecies of Borrelia burgdorferi sensu lato. No laboratory relied routinely on a single test and each weighted the significance of the findings of the various tests differently. A difference in strategy between laboratories in high and low prevalence areas was apparent in that laboratories in low prevalence areas emphasised specificity more than sensitivity and therefore produced fewer false positives, but also missed some cases. Overall agreement between the laboratories was poor and it was concluded that there is a need for a quality assurance scheme within Europe.