Development of a reference material for residues of chlorfluazuron and fluazuron in beef fat ACSL CRM 3

The certification by interlaboratory testing of a candidate reference material (ACSL CRM 3) for the concentration (mass fraction) of the pesticides chlorfluazuron (CFZ) and fluazuron (FZN) is described. The certification process was based on a two-stage nested design described in ISO Guide 35. Analysis of results reported by collaborating laboratories provided an assessment of the homogeneity of the RM. The certified values were chlorfluazuron -1.01 mg/kg and fluazuron -1.02 mg/kg. The upper and lower 95% confidence limits for CFZ and FZN were calculated to be: CFZ upper=1.05 mg/kg, lower=0.97 mg/kg; FZN upper=1.08 mg/kg, lower=0.95 mg/kg. A certificate was prepared according to guidelines set out in ISO Guide 31.     

Development of the NIST Rocky Flats Soil Standard

The National Institute of Standards and Technology (NIST) Rocky Flats Soil-II Standard reference material (SRM) is being certified through a statistical analysis of results from an interlaboratory comparison of fourteen laboratories from four countries. Mean values were calculated as the most robust and reliable certified values for each of the radionuclides. Twenty two radionuclides and four isotopes ratios were evaluated based on reported data. This article describes the approach for establishing the certified values and uncertainties for the radionuclides in the Rocky Flats Soil II SRM.     

Virginia Tobacco Leaves (CTA-VTL-2) – new Polish CRM for inorganic trace analysis including microanalysis

Preparation and certification of a new Polish reference material (CTA-VTL-2) for inorganic trace analysis including microanalysis is described. Virginia Tobacco Leaves of Bulgarian origin were hand picked, dried, comminuted and sieved at conditions preventing contamination of the material with metals. Ca. 13 kg of the material with particle size φ≤ 80 μm was prepared. Certification was based on world-wide interlaboratory comparison in which 60 laboratories from 18 countries participated. Data evaluation was done using an approach which includes rejection of outliers by concurrent use of four different outlier tests followed by calculation of overall means, confidence intervals etc. Several criteria were employed to qualify or disqualify the obtained overall mean as a certified or information value, respectively. 33 elements were certified and information values were established for 10 others. Particle size distribution of CTA-VTL-2 was investigated microscopically. The modal particle diameter was in the range of 15–35 μm. Extensive homogeneity checking by neutron activation analysis revealed that the material is sufficiently homogeneous for mg sample sizes.     

Creatinine determination in urine from the point of view of reference values

In this paper, the creation of a certified reference material for urinary creatinine is described. We used the Jaffe method and HPLC method for establishment of the certified value. Homogeneity tests are also described. We obtained material with sufficient homogeneity, stability, and with certified value (expanded uncertainty, k=2 for CI 95%) (7.77±0.27) mmol·L⁻¹. This material was consequently used for the interlaboratory comparison (EQA Czech Republic for clinical chemistry). Twenty-nine percent of the participants obtained measurement results within the interval of the certified value ± expanded uncertainty, while 85% of the participants obtained values inside the interval of the certified value ± target measurement uncertainty. Direct use of the certified reference materials for method evaluation in EQA programs means a significant advance for monitoring and documentation traceability of results in routine measurements.     

Development of clenbuterol reference materials: lyophilized bovine eye samples free of clenbuterol (CRM 673) and containing clenbuterol (CRM 674)

The certification by inter-laboratory testing of two candidate reference materials (RMs) for the mass concentration of the anabolic agent clenbuterol in bovine eye material is described: RM 674 with ca 10 microg clenbuterol per kg of eye matrix and RM 673 clenbuterol-free eye matrix as the negative control (<0.50 microg kg(-1)). Both candidate RMs were certified by eleven EU laboratories, and sixty-six accepted replicate measurements were included in the "Certification Study". The precision of the measurement process was assessed by calculation of the standard variation determined within each laboratory during the certification step. The study was performed according to the "Guidelines for the production and certification of BCR reference materials" and to "ISO guide 31, 33, and 35". The certified clenbuterol mass concentration for clenbuterol-free eye material CRM 673 (calculated on the basis of clenbuterol as the free base) was <0.50 microg kg(-1). The corresponding concentration for clenbuterol-containing eye material CRM 674 was 9.42 +/- 0.88 microg kg(-1). These certified values are very close to the desired target concentration of <0.5 microg kg(-1) and ca 10 microg kg(-1). This study has demonstrated that successful certification of clenbuterol-containing and clenbuterol-free bovine eye materials is possible.     

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.     

Certification of reference materials for Cd, Cr, Hg and Pb in polypropylene

Two reference materials, at relatively low and high concentrations (GBW08404 and GBW08405), for analysis of the mass fractions of Cd, Cr, Hg and Pb in polypropylene were developed. The reference materials were prepared by doping blank polypropylene base material with Cd, Cr, Hg and Pb in the form of oxides, salts or pigments. Homogeneity and stability studies were performed by inductively coupled plasma mass spectrometry. The certification of the four analytes was carried out by isotope-dilution mass spectrometry (IDMS) with microwave-assisted digestion. Combined uncertainties were calculated from the IDMS uncertainty evaluation budget and the uncertainty of the homogeneity. The mass fractions of Cd, Cr, Hg and Pb of the two certified reference materials (CRMs) were from 8 to 1,000 mg kg⁻¹. The two samples were also used in an interlaboratory comparison scheme in which National Institute of Metrology, China, National Metrological Institute of Japan and Korea Research Institute of Standards and Science participated. The agreement of the comparison results proved that the certification procedure of the CRMs is valid and that the certified values of Cd, Cr, Hg and Pb are accurate and reliable.     

Preparation and preliminary certification of two new Polish CRMs for inorganic trace analysis

Preparation and characterization of two new reference materials of biological origin, namely: Tea Leaves (INCT-TL-1) and Mixed Polish Herbs (INCT-MPH-2) is described. The raw materials were ground in an agate ball mill, sieved through a nylon sieve, collecting fraction of particle size: Æ£67 ·m, and carefully homogenized. Preliminary homogeneity testing by XRF method and final checking of homogeneity by NAA after distribution of the materials into containers revealed that they are sufficiently homogeneous at least for sample size ≥100 mg. Both materials were prepared in amounts exceeding 40 kg and certified on the basis of a worldwide interlaboratory comparison, in which 109 laboratories from 19 countries participated. The method of data evaluation leading to assignment of certified values was essentially the same as that used previously in this Laboratory, but supplemented by additional data from the analysis of a CRM which was sent to the participants and analyzed by them along with the candidate reference materials. In addition the results for a few elements by very accurate developed methods in this Laboratory were obtained and used to support the certification process. Analytical uncertainties and stability uncertainties were quantified to arrive at combined uncertainties of the certified values. So far 18 elements in INCT-TL-1 and 21 in INCT-MPH-2 could be certified.     

Preliminary study to prepare a reference material of styrene metabolites – mandelic acid and phenylglyoxylic acid – in human urine

 A preliminary batch of the reference material was prepared by freeze-drying pooled urine samples obtained from healthy persons occupationally exposed to styrene. Tests for homogeneity and stability were performed by determining urine concentrations of mandelic (MA) and phenylglyoxylic acids (PGA). The urinary MA and PGA concentrations were followed over an 8-month period using high performance liquid chromatography (HPLC). No changes of the concentration values were found. Pure PA and PGA from Merck and Fluka, respectively, were used for traceability purposes, because certified or standard reference materials for MA and PGA do not exist. Control material ClinChek-Urine Control (Recipe) was analysed simultaneously. The mean values of MA and PGA compared well with the means of control samples and fell within the control range. The certified values and their uncertainties were evaluated from the results of interlaboratory comparisons, homogeneity (277.0 ± 7.4 mg L⁻¹ for MA and 148.0 ± 4.7 mg L⁻¹ for FGA) and stability tests. The values are unweighted arithmetical averages of accepted results and their uncertainties are combined uncertainties enlarged by coefficient k=1, evaluated from the standard uncertainties of the interlaboratory comparison, homogeneity and stability tests. Received: 17 September 2002 Accepted: 1 November 2002 Acknowledgement This work was supported by the Internal Grant Agency of Ministry of Health of the Czech Republic (Grant NJ/6784–3). Presented at CERMM-3, Central European Reference Materials and Measurements Conference: The function of reference materials in the measurement process, May 30–June 1, 2002, Rogaška Slatina, Slovenia Correspondence to I. Šperlingová     

Improvements in efficiency of production and traceability for certification of reference materials

Issues of current interest to certified reference material producers are addressed. Alternative strategies for certification of matrix reference materials are discussed and the benefits of adopting a flexible, cost-effective approach are described. The difficulty of undertaking homogeneity testing where certification is to be carried out with definitive techniques capable of providing very small measurement uncertainty is discussed. Methodology is described which combines conventional screening of the candidate material for homogeneity with an additional, precise assessment of homogeneity based on isotope dilution mass spectrometry measurements. A systematic procedure for evaluating the commutability (horizontal traceability or scope) of matrix reference materials has been evaluated and shows that in some circumstances matrix effects may be less pervasive than is generally believed. This offers the possibility, especially for trace analysis applications, of more efficient use of existing reference materials without compromising measurement reliability. Vertical traceability of matrix reference material data is of growing interest but is difficult to achieve with present interlaboratory certification exercises. A modification is described which attempts to address this issue. It also offers the possibility of improved identification of outliers and reduced variation of data between the participating laboratories.Presented at BERM-9—9th International Symposium on Biological and Environmental Reference Materials, 15–19June 2003, Berlin, Germany     

International Measurement Evaluation Programme: IMEP-9, trace elements in water

The International Measurement Evaluation Programme (IMEP) is an interlaboratory comparison scheme, founded, owned and coordinated by the Institute for Reference Materials and Measurements (IRMM) since 1988. IMEP-9 is the third round of trace elements in water evaluation following IMEP-3 and IMEP-6. Reference values for 15 elements stating total concentrations and combined uncertainties (according to GUM) were established. The reference values were established mainly by isotope dilution mass spectrometry (IDMS) as a primary method of measurement, and values traceable to the SI were obtained. The four elements that could not be certified by IDMS were assigned values by means of other measurement techniques. Results from 201 laboratories from 35 countries and four continents were evaluated against the reference values and the comparability between the laboratories is presented graphically.     

Development and certification of reference materials for residues of organochlorine and organophosphorus pesticides in beef fat ACSL CRM 1 and 2

Beef fat samples were prepared and tested as candidate reference materials for organochlorine and organophosphorus pesticides. The CRMs consisted of beef fat spiked with pesticide solutions. One sample (ACSL CRM 1) was prepared containing close to 0.2 mg/kg of each of the organochlorine pesticides dieldrin and heptachlor epoxide. A second sample (ACSL CRM 2) was prepared containing close to 0.8 mg/kg of each of the organophosphorus pesticides diazinon, chlorpyrifos and ethion. The spiking levels and homogeneity of the materials were verified. The coefficients of variation of 5 analyses carried out to test between-jar homogeneity for each reference material were dieldrin, 3.5%; heptachlor epoxide, 1.1%; diazinon, 2.1%; chlorpyrifos, 1.2% and ethion, 3.1%. No instability in any of these compounds was detected over a twelve month period. The candidate reference materials were found to be suitable for certification by interlaboratory testing. The certification process was based on a two-stage nested design described in ISO Guide 35. Analysis of results reported by collaborating laboratories provided an assessment of the homogeneity of the reference materials. The certified values together with their upper and lower 95% confidence limits are: ACSL CRM 1*Dieldrin*0.199 mg/kg **(0.188, **0.210 mg/kg) *Heptachlor epoxide*0.194 mg/kg **(0.176, **0.212 mg/kg) ACSL CRM 2*Diazinon*0.805 mg/kg **(0.755, **0.855 mg/kg) *Chlorpyrifos*0.790 mg/kg **(0.728, **0.852 mg/kg) *Ethion*0.813 mg/kg **(0.746, **0.879 mg/kg). A certificate for each material was prepared according to the guidelines set out in ISO Guide 31. Received: 25 April 1997 / Revised: 19 August 1997 / Accepted: 28 August 1997     

Development and certification of reference materials for residues of organochlorine and organophosphorus pesticides in beef fat ACSL CRM 1 and 2

Beef fat samples were prepared and tested as candidate reference materials for organochlorine and organophosphorus pesticides. The CRMs consisted of beef fat spiked with pesticide solutions. One sample (ACSL CRM 1) was prepared containing close to 0.2 mg/kg of each of the organochlorine pesticides dieldrin and heptachlor epoxide. A second sample (ACSL CRM 2) was prepared containing close to 0.8 mg/kg of each of the organophosphorus pesticides diazinon, chlorpyrifos and ethion. The spiking levels and homogeneity of the materials were verified. The coefficients of variation of 5 analyses carried out to test between-jar homogeneity for each reference material were dieldrin, 3.5%; heptachlor epoxide, 1.1%; diazinon, 2.1%; chlorpyrifos, 1.2% and ethion, 3.1%. No instability in any of these compounds was detected over a twelve month period. The candidate reference materials were found to be suitable for certification by interlaboratory testing. The certification process was based on a two-stage nested design described in ISO Guide 35. Analysis of results reported by collaborating laboratories provided an assessment of the homogeneity of the reference materials. The certified values together with their upper and lower 95% confidence limits are: ACSL CRM 1*Dieldrin*0.199 mg/kg **(0.188, **0.210 mg/kg) *Heptachlor epoxide*0.194 mg/kg **(0.176, **0.212 mg/kg) ACSL CRM 2*Diazinon*0.805 mg/kg **(0.755, **0.855 mg/kg) *Chlorpyrifos*0.790 mg/kg **(0.728, **0.852 mg/kg) *Ethion*0.813 mg/kg **(0.746, **0.879 mg/kg). A certificate for each material was prepared according to the guidelines set out in ISO Guide 31.     

A DANREF certified reference plastic for measurement of overall migration into the food simulant olive oil by single sided testing

A reference material for the determination of overall migration from a plastic coextrudate into the fatty food simulant olive oil was produced and certified in an interlaboratory study. The analyses were carried out according to the ENV 1186 standard from the European Committee for Standardization (CEN) [ 1, 2, 3] with exposure of the coextrudate to olive oil for 10 days at 40 degrees C. After an initial preliminary interlaboratory study eight laboratories participated in the certification round, and two different methods were used to obtain single sided exposure of the plastic to the oil. The certified value was determined as the mean of laboratory mean values. No outliers were found. A reference value of 8.6 mg/dm2 +/- 1.4 mg/dm2 (+/- half width of the 95% confidence interval) was obtained which is within the range relevant for the regulatory limit (10 mg/ dm2), making this reference material suitable for laboratories measuring according to the EU overall migration limit [4]. The material has been found stable over 45 months.     

Use of uncertainty information for estimation of certified values. Comparison of four approaches using data from a recent certification exercise

Four approaches for estimation of reference values and their respective uncertainties of characterisation were compared using data from the recently finalised certified reference materials ERM-EC680k and ERM-EC681k, elements in plastics. Reference values and uncertainties of characterisation were estimated as mean of laboratory means and their respective standard deviations, using equal weights and the weighting procedure of Mandel–Paule. In addition, two approaches taking into consideration uncertainty information reported by the participants, namely the consistency check and simulation procedure proposed by Cox for CCQM Key comparisons and an approach suggested by Pauwels et al. (Accred Qual Assur 3:180–184, 2000) were used. No difference between the equally-weighted and Mandel–Paule consensus means was observed and the reference value from the Cox approach was in all cases within ±2 u _char of each consensus mean. Uncertainties varied between the three approaches. Uncertainties derived from equally-weighted mean of means approach are on average 14% above uncertainties using the Mandel–Paule consensus mean, 36% above the uncertainties estimated by Pauwels et al., and 54% above the uncertainties from the Cox approach. Robustness of the uncertainty estimation against incorrect estimation of uncertainties was assessed. Assumption of a 50% uncertainty of the individual uncertainties resulted in an uncertainty of 30% of the uncertainty of characterisation. Differences between the four approaches are negligible for this dataset when combined with the uncertainty contribution from heterogeneity and stability as prescribed in ISO Guide 35. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

k0-NAA, a valuable tool for reference-material producers.

The main concern of producers of certified reference materials (CRM) is the preparation of high-quality products with demonstrated homogeneity and stability, combined with a well established set of certified characteristics. CRM producers must, furthermore, comply with other constraints imposed by the ISO Guide 34: production processes, production control, and certification analyses should be performed by expert laboratories, using validated protocols documented in their respective quality assurance manuals; laboratory mean values and the corresponding "expanded" uncertainties, must be used for the determination of the certified values, as recommended by the ISO Guide to the Expression of Uncertainties in Measurements (GUM); and when possible, traceability of the certified value to the SI units, using appropriately validated and/or primary methods, must be ensured. k0-NAA, i.e. neutron activation analysis with k0 standardization, is one of the analytical techniques implemented at the Reference Material Unit of IRMM; it meets the first two requirements.     

Lifetime of the traceability chain in chemical measurement

Since the uncertainty of each link in the traceability chain (measuring analytical instrument, reference material or other measurement standard) changes over the course of time, the chain lifetime is limited. The lifetime in chemical analysis is dependent on the calibration intervals of the measuring equipment and the shelf-life of the certified reference materials (CRMs) used for the calibration of the equipment. It is shown that the ordinary least squares technique, used for treatment of the calibration data, is correct only when uncertainties in the certified values of the measurement standards or CRMs are negligible. If these uncertainties increase (for example, close to the end of the calibration interval or shelf-life), they are able to influence significantly the calibration and measurement results. In such cases regression analysis of the calibration data should take into account that not only the response values are subjects to errors, but also the certified values. As an end-point criterion of the traceability chain destruction, the requirement that the uncertainty of a measurement standard should be a source of less then one-third of the uncertainty in the measurement result is applicable. An example from analytical practice based on the data of interlaboratory comparisons of ethanol determination in beer is discussed. Received: 5 October 2000 Accepted: 3 December 2000     

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±130mgkg⁻¹and 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.     

Determination of measurement uncertainty for the determination of triazines in groundwater from validation data

Laboratories are increasingly urged to submit full uncertainties of their analytical results rather than only standard deviations. The determination of measurement uncertainties in compliance with the Guide to the Expression of Uncertainty in Measurement (GUM) is demonstrated using the validation approach explicitly endorsed by the recent edition of the EURACHEM guide for the determination of measurement uncertainty. Measurement uncertainty was split into uncertainty of the sample mass, uncertainty of the concentration of the stock standard solution, uncertainty of the calibration and uncertainty connected to within- and between-series precision. Uncertainties of sample mass and of the concentration of the stock standard solution were 0.26 and 1.14% for all analytes, which is negligible compared with the contributions of precision and calibration. Uncertainty of calibration was estimated from the calibration graph. Relative uncertainty of calibration was found to be strongly concentration dependent and to be the main uncertainty contribution below 0.2 microgram L-1. Precision was split into within-series and between-series standard deviation, which dominate the combined standard uncertainty at higher concentrations. The results obtained from these calculations are compared with results for a certified reference material and with the performance in an interlaboratory comparison. It was found that all results agreed within their uncertainty with the target values, showing that the estimated uncertainties are realistic.     

Certification of trace and major elements and methylmercury concentrations in a macroalgae (Fucus sp.) reference material, IAEA-140

A marine reference material, IAEA-140, prepared with a macroalgae (Fucus sp.) was recently produced by the International Atomic Energy Agency and certified for trace and major elements and for methylmercury (MeHg). Certification of this material was achieved as an outcome of an international analytical intercomparison study which resulted in 116 independent sets of results reported by participants from 54 countries. The statistical evaluation of the collected data and the criteria used for assignment of the mean and uncertainty values are described. The analysis of data allowed to certify concentration values for 24 elements and MeHg, and to provide information values for another 10 elements. Regarding the elements which could be given certified values, between two and eight different instrumental methods were used to measure metal concentrations, and four independent analytical procedures were used to measure MeHg concentrations. In order to assess the results of the certification procedure, a comparison was made between the certified values obtained from the world-wide intercomparison results and the values obtained from a small subgroup of well-qualified laboratories. The means and 95% confidence intervals for reference values obtained by the two methods are similar for most elements showing that the usual method of certification used by the IAEA, based on large groups of participants, is indeed pertinent.