DETERMINATION OF TOTAL VICINAL DIKETONES IN BEER BY HEADSPACE CAPILLARY GAS CHROMATOGRAPHY

The determination of total vicinal diketones in beer by the headspace capillary gas chromatography method of the Institute of Brewing Analysis Committee has been collaboratively tested by the members of the Analysis Committee of the European Brewery Convention. Two collaborative trials on three beer samples were performed in 14 laboratories. Repeatability (r₉₅) and reproducibility (Ras) values were obtained for 9 laboratories over the range of 20 to 90 μg. It was judged that reproducibility coefficients ranging after the second test from 32 to 94 were poor but it was decided that the method should nevertheless be included in the Analytica EBC as a guideline method with notice on its poor precision .     

DETERMINATION OF THE MOISTURE AND NITROGEN CONTENTS OF BARLEY AND MALT BY NEAR INFRARED SPECTROSCOPY (NIRS)

The determination of the moisture and nitrogen contents of barley and malt by near infrared spectroscopy (NIRS) has been tested by the Analysis Committee of the European Brewery Convention. In the collaborative trial four samples of barley and malt were analysed by 17 laboratories. Repeatability (r₉₅) and reproducibility (R₉₅) values of 0.3 and 1.5% m/m respectively were obtained for barley moisture over the range 12.7 to 15.8% m/m. For malt moisture these values were 0.2 and 1.3% m/m over the range 4.0 to 4.3% m/m, for barley nitrogen 0.1 and 0.3% m/m on dry matter over the range 1.57 to 2.14% m/m, and for malt nitrogen 0.1 and 0.2% m/m on dry matter over the range 1.58 to 1.82% m/m, respectively.     

DETERMINATION OF THE VOLATILE COMPONENTS OF BEER

A method relying on headspace sampling and gas chromatography for the determination of the lower boiling point volatile compounds (fusel oils) in beer, has been approved by the Analysis Committee of the Institute of Brewing (IOB), for inclusion in Recommended Methods of Analysis. In view of the large values obtained for precision in terms of repeatability (r₉₅) and reproducibility (R₉₅) and the differences in gas chromatographic equipment used by individual brewing laboratories, the method is considered not to be ideal, but is given as a guideline only. The IOB Analysis Committee has recommended a method relying on gas chromatography and flame photometric detection for the determination of dimethyl sulphide in beer. Repeatability (r₉₅) and reproducibility (R₉₅) values of 3.3 μg/litre and 3.66+0.168 m μg/litre (where m is concentration), respectively, were obtained over the range 20–50 μg/litre. Methods currently used in brewing laboratories, for the measurement of vicinal diketones, are being surveyed with a view to obtaining a suitable method for collaborative testing by the Analysis Committee.     

Determination of Boiled Wort Colour

A method for the determination of boiled wort colour has been collaboratively tested by the Analysis Committee of the European Brewery Convention. In this trial the 14th EBC Standard Malt and two commercial malts were tested by 21 laboratories. Repeatability (r₉₅) and reproducibility (R₉₅) values of 0.33 ? 0.42 and 1.12 ? 1.44 EBC units were obtained respectively at a mean level of 4.7 ? 5.2 EBC units. At a mean level of 8.9 EBC units these respective values were 0.44 and 2.25 EBC units. The data sets could be split into two sub?sets for laboratories using either the included heating plate boiling procedure or the glycerol/oil bath boiling procedure. Statistical analysis of both sub?sets revealed that reproducibility is unambiguously better for the glycerol/oil bath procedure at all levels. For repeatability the situation is less clear. The evaluated method is recommended for inclusion in Analytical?EBC.     

DETERMINATION OF REPEATABILITY AND REPRODUCIBILITY OF EBC ACCEPTED METHODS IV—COLOURED MALTS

Methods for the determination of caramel and roasted malt moisture, extract and colour published in Analytica EBC have been collaboratively tested by the European Brewery Convention Analysis Committee, according to the ISO standard 5725. Repeatability (r₉₅) and reproducibility (R₉₅) values are presented.     

Determination of the Total Soluble Nitrogen Content of Malt and Beer by the Dumas Combustion Method: Collaborative Trial

The determination of the total soluble nitrogen content of malt and beer, by the Dumas procedure, has jointly been collaboratively tested by the Analysis Committee of the Institute of Brewing (IOB) and the European Brewery Convention (EBC). Five samples of beer (range 362 to 1159 mg/l) and five samples of malt (range EBC 0.598 to 0.798 %m/m (dry basis) and IOB 0.534 to 0.706 %m/m (dry basis)) were distributed to eighteen participating laboratories for analysis. Precision values were judged to be independent of the mean soluble nitrogen content for malt by both IOB and EBC methodologies. Values for r₉₅ and R₉₅ were 0.047 and 0.136%m/m for EBC laboratory wort and 0.039 and 0.144 %m/m for IOB laboratory wort respectively. Precision values for beer were judged to be dependent upon the mean nitrogen content (m) in the case of r₉₅ and independent of the mean nitrogen content in the case of R₉₅. Values for r₉₅ and R₉₅ were 0.074m and 120 mg/l respectively.     

Determination of DMS and other Volatile Compounds in Beer by Headspace Capillary Gas Chromatography and Flame Ionisation Detection

The determination of beer volatiles by the headspace capillary gas chromatography with flame ionisation detection method of the Institute of Brewing Analysis Committee has been collaboratively tested by the members of the Analysis Committee of the European Brewery Convention and some members of the American Society of Brewing Chemists. Determined analytes were acetaldehyde, dimethylsulphide, ethylacetate, n-propanol, isobutanol, isoamylalcohols and isoamylacetates. The internal standards used were ethylmethyl sulphide and n-butanol. After three preliminary collaborative trials, a last fourth one was performed on three beer samples in 15 laboratories. Repeatability (r₉₅) and reproducibility (R₉₅) values were obtained for 12 laboratories. Assuming that too many members did not follow exactly the proposed method and judging that reproducitibility values were poor, it was decided to not include the method in the Analytica EBC as an official method, but to include it as a guideline method with the aforementioned notice.     

THE DETERMINATION OF DIMETHYL SULPHIDE IN BEER BY HEADSPACE GAS CHROMATOGRAPHY — A COLLABORATIVE INVESTIGATION OF PRECISION

A method for the determination of dimethyl sulphide in beer by headspace gas chromatography has been collaboratively tested within ten laboratories of one brewing group at 3 levels from 19.8 to 52.4 μg/litre. The repeatability values (r₃₅ were found to be independent of concentration, but a strong linear relationship was found between the mean values (m) and the reproducibility value (R₃₅). The precision data can be summarised as r₃₅ = 3.3 μg/litre, R₃₅ = 3.66 + 0.168 μg/litre .     

Detection of Salmonella in food,feed and veterinary samples by EU laboratories

The European Union Reference Laboratory (EURL) for Salmonella organises interlaboratory comparison studies on bacteriological detection of Salmonella in different matrices (veterinary, food and animal feed) amongst National Reference Laboratories for Salmonella of the 27 European Member States (EU-MS). The EURL also gives non-EU Member States (EU candidate countries, member countries of the European Free Trade Association (EFTA) and third countries from outside Europe) the opportunity to participate in these studies. The objective of the studies is to test the ability of the participating laboratories to detect Salmonella at different contamination levels in the various matrices. In addition a comparison is made between the prescribed method of the food and animal feed studies (ISO 6579: RVS and MKTTn) and the prescribed method of the veterinary studies (annex D of ISO 6579: MSRV).This article describes the performance of the individual laboratories in the interlaboratory comparison studies as organised by the EURL for Salmonella in the period 2004-2010.The performance of the majority of the NRLs improved during the course of the studies. Overall the NRL laboratories performed well and were able to detect Salmonella in the different tested matrices. After improvement of the samples, it was possible to define criteria for good performance in the studies organised since 2006. In the eight interlaboratory comparison studies organised in the period 2006–2010, 15 laboratories found in total 19 deviations of which eight concerned a false positive blank result. The other deviations concerned sensitivity problems (false negative results). After checking and improving several technical aspects, the underperforming participating laboratories were able to improve their performance in a follow-up study.     

PRODUCTION OF PALM OIL REFERENCE MATERIALS FOR THE DETERMINATION OF SOLID FAT CONTENT

ABSTRACT

Three palm oil reference materials were produced for solid fat content analysis employing the Malaysian Palm Oil Board (MPOB) Test Methods. Thirteen laboratories participated in the characterization study, where the solid fat content in palm oil, palm olein and palm stearin were identified at temperature range from 0 to 45C. The consensus values were calculated based on the acceptability of statistical results from collaborating laboratories. Both of the consensus values and their uncertainties (%) for each reference material are as follows: 71.12 ± 1.82% (0C), 54.07 ± 1.05% (10C), 37.44 ± 0.88% (15C), 24.18 ± 1.09% (20C), 13.32 ± 0.86% (25C), 7.64 ± 0.28% (30C), 4.05 ± 0.14% (35C) and 1.25 ± 0.49% (40C) for SFC of palm oil, 66.32 ± 1.93% (0C), 43.53 ± 1.32% (10C), 22.50 ± 0.78% (15C), 6.68 ± 1.14% (20C) and 1.40 ± 0.35% (25C) for SFC of palm olein, and 78.09 ± 1.23% (0C), 67.66 ± 0.71% (10C), 56.29 ± 1.18% (15C), 44.37 ± 1.13% (20C), 30.22 ± 1.25% (25C), 20.08 ± 0.58% (30C), 13.72 ± 0.32% (35C), 8.97 ± 0.72% (40C) and 5.06 ± 0.41% (45C) for solid fat content of palm stearin.

PRACTICAL APPLICATIONS

The implementation of the ISO 17025 accreditation should be encouraged for testing laboratories to assure the traceability of analysis performed. This accreditation requires the use of certified reference materials to ensure that the analysis performed is parallel to the ISO 17025 standards. However, the unavailability of palm oil reference materials will affect the implementation of such standards for laboratories that are performing palm oil analyses. Hence, this study is aimed to characterize and certify the palm oil reference materials for determination of solid fat content. The establishment of this study is hoped to increase the credibility of laboratories, which are involved in the palm oil analysis. This will also promote the Malaysia Palm Oil Board as a center for the certification of palm oil reference materials.     

Inter-laboratory comparison of an analytical method for the determination of the feed additive semduramicin in poultry feed at authorised level by liquid chromatography single quadrupole mass spectrometry

A collaborative study was carried out according to internationally recognised guidelines in order to establish the performance characteristics of an LC/MS method for the determination of the feed additive semduramicin (SEM) in poultry feed at the level (20–25?mg?kg^?1) authorised within the European Union. Fifteen laboratories participated in the validation study, and all reported results. The content of SEM in the tested materials, provided as blind duplicates, ranged from 11.5?mg?kg^?1, which corresponds to half the mean authorised level, to 45.0?mg?kg^?1, which corresponds to twice the mean authorised level. All the materials were analysed by the participating laboratories using two different quantification approaches: standard addition and external standard calibration. The relative standard deviation of reproducibility (RSD_R) for both quantification approaches varied from 8% to 18%, corresponding to HORRAT values ranging from 0.8 to 1.5, which were therefore in all cases below the critical value of 2.0. Consequently, the proposed analytical method and both quantification approaches can be considered to be fully validated and transferable to the control laboratories and applied for the determination of SEM in poultry compound feed at authorised level within the frame of official control. Further steps in the administrative procedure aiming to adopt the method as part of an ISO/CEN standard are currently ongoing.     

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.     

DETERMINATION OF THE SOLUBLE IRON CONTENT OF FILTER AIDS: A COLLABORATIVE TRIAL

The international method for the determination of the soluble iron content of filter aids has been collaboratively tested by members of the Analysis Committee of the European Brewery Convention to obtain repeatability (r₉₈) and reproducibility (R₉₈) values. It was Judged that precision values were dependent on the quantity of the soluble iron content of the filter aids over the range 32 to 220 mg/kg. Repeatability (r₉₈) value of 0.050 m and reproducibility value (R₉₈) of 0.255 m were obtained over this range.     

ESTIMATION OF ENDO BETA-GLUCANASE ACTIVITY IN MALT USING A VISCOMETRIC METHOD

A viscometric method for estimation of endo beta-glucanase activity in malt has been collaboratively tested by the Institute of Brewing Analysis Committee. Ten participating laboratories analysed five pairs of malt samples in the range 176 to 1238 IRV units. The repeatability (r₉₅) values ranged from 22 to 122 IRV units and the reproducibility (R₉₅) values from 93 to 650 IRV units. The results indicated that R₉₅, but not r₉₅, was dependent on endo beta-glucanase activity. Overall precision values were 76.8 for r₉₅ and 0.506Mean + 4.786 for R₉₅. It was decided that the method would be included in IOB Methods of Analysis but that attention would be drawn to the poor precision values obtained in this and previous collaborative trials.     

Applicability of thermoluminescence techniques to identify irradiated seafoods using different methods of mineral separation: An interlaboratory blind trial

Thermoluminescence (TL) technique for identifying γ-irradiated (0–10 kGy) anchovies (dried), kelp (dried), and mackerel (fresh) was validated in an interlaboratory blind trial. Different irradiation detection laboratories were involved by using 2 methods of mineral separation (density separation and acid hydrolysis) for the analysis. Key TL parameters, including the TL glow-curve shape, intensity, and the TL ratio (TL₁/TL₂) were used to characterize the irradiation status. All irradiated samples exhibited an intense TL peak at approximately 200°C, which was absent in non-irradiated samples. TL glow curve interpretations were also confirmed by determining the TL ratio. Different participating laboratories reported 89–100% correct results. Both methods of mineral separation were equally effective; however, some variation was observed in the results from different laboratories for irradiated mackerel, which might be due to a lack of isolated minerals, differences in personnel expertise, and different TL instruments.     

AN INTER-LABORATORY STUDY OF STARCH DEGRADING ENZYME ASSAYS

The reliability of specific colorimetric assays for the main starch degrading enzymes, α-amylase, β-amylase and limit dextrinase was investigated by conducting an inter-laboratory collaborative study. Three malt samples were assayed in duplicate in 14 laboratories. The repeatability (r) within laboratories was good for α-amylase and β-amylase assays. Between-laboratory reproducibility (R) was acceptable (CV_R 9.2–14%). Errors for limit dextrinase were higher. Repeatability was good at high enzyme concentrations (CV_r = 2%) but increased at low enzyme concentrations. Reproducibility was relatively poor (CV_R-27–33%).     

SCREENING AND QUANTIFICATION OF AFLATOXINS AND OCHRATOXIN A IN DIFFERENT CEREALS CULTIVATED IN ROMANIA USING THIN-LAYER CHROMATOGRAPHY-DENSITOMETRY

ABSTRACT

The main focus of our study was to implement a rapid, inexpensive and reliable method that could be utilized to check the cereals for safety (i.e., screening for total aflatoxins, as well as individual B₁, B₂, G₁, G₂ aflatoxins and ochratoxin A). We developed a protocol by which we were able to isolate mycotoxins from cereals collected from different regions of Romania. After extraction in chloroform, the mycotoxins were separated by thin‐layer chromatography (TLC) and quantified using densitometry. Forty‐three samples of different cereals (wheat, maize, rye and Triticale) were analyzed. Twenty‐five of the 43 samples (58.14% of the total number) were found to be contaminated with different mycotoxins in various concentrations: aflatoxin B₁ (1.6–5.7 µg/kg), aflatoxin B₂ (0.89–4 µg/kg), aflatoxin G₁ (1.2–5.76 µg/kg), aflatoxin G₂ (0.96–3.4 µg/kg) and/or 4.3–30 µg/kg ochratoxin A. The concentration of total aflatoxin contamination ranged from 11.2 to 10.8 µg/kg. Among the different cereals, the highest number of contaminated samples was found to be in the wheat samples (62.5%). The method outlined in this study has been adopted already by our laboratory for current analyses of mycotoxins. The results obtained using this method is in compliance with the strict regulatory guidelines developed both in Romania, as well as in the European Union.

PRACTICAL APPLICATIONS

Thin‐layer chromatography (TLC) is a rapid, inexpensive and convenient method that can be used routinely to screen for mycotoxins. This article describes the detailed procedures for the extraction, purification and quantification of aflatoxins and ochratoxin A, using TLC. Using this method one can identify concomitantly five different mycotoxins and by coupling it with densitometry a quantitative determination is also possible. Therefore, this could become a routine technique in regional laboratories responsible for checking agrifood safety.     

DETERMINATION OF DIMETHYL SUPLHIDE IN BEER BY GAS CHROMATOGRAPHY — COLLABORATIVE TRIAL

The Analysis Committee has collaboratively tested both routine headspace gas chromatographic methods and a standardised method for the determination of dimethyl sulphide in beer using the flame photometric and flame ionisation detectors available in the participating laboratories. The mean repeatability value (r₉₅) was 7.8 μg/litre for the concentration range 12–65 μg/litre, and the reproducibility value (R₉₅) showed the relationship R₉₅ = 18.1 + 0.452m μg/litre to the concentration (m). No differences in precision were attributed either to the methods or the types of detectors used.     

DETERMINATION OF ALUMINIUM IN BEER BY GF-AAS

A method for the determination of aluminum in beer, has been collaboratively tested. The method tested relies on atomic absorption spectrometry with atomization in a graphite furnace. Five pairs of beers, with concentrations ranging from 96 to 1000 μg/litre, were analysed by eleven laboratories. The repeatability (r₉₅) values ranged from 10 to 69 μg/l, and the reproducibility (R₉₅) values ranged from 70 to 465 μg/l,. A second collaborative trial with a slightly different method protocol gave no improvement. Due to the high reproducibility values the method was not adopted for inclusion in Analytica-EBC.     

Relationships between milk coagulation property traits analyzed with different methodologies

Milk coagulation properties (MCP) analysis is performed using a wide range of methodologies in different countries and laboratories, using different instruments, coagulant activity in the milk, and type of coagulant. This makes it difficult to compare results and data from different research. The aims of this study were to propose a method for the transformation of values of rennet coagulation time (RCT) and curd firmness (a₃₀) and to predict the noncoagulation (NC) probability of milk samples analyzed using different methodologies. Individual milk samples were collected during the morning milking in October 2010 from each of 165 Holstein-Friesian dairy cows in 2 freestall barns in Italy, and sent to 3 laboratories for MCP analysis. For each laboratory, MCP analysis was performed using a different methodology: A, with a computerized renneting meter instrument using 0.051 international milk clotting units (IMCU)/mL of coagulant activity; B, with a Lattodinamografo (Foss-Italia, Padova, Italy) using 0.051 IMCU/mL of coagulant activity; and C, with an Optigraph (Ysebaert, Frépillon, France) using 0.120 IMCU/mL of coagulant activity. The relationships between MCP traits were analyzed with correlation and regression analyses for each pair of methodologies. For each MCP trait, 2 regression models were applied: model 1 was a single regression model, where the dependent and independent variables were the same MCP trait determined by 2 different methodologies; in model 2, both a₃₀ and RCT were included as independent variables. The NC probabilities for laboratories with the highest number of NC samples were predicted based on the RCT and a₃₀ values measured in the laboratories with lower number of NC samples using logistic regression and receiver operating characteristic analysis. The percentages of NC samples were 4.2, 11.5, and 0.6% for A, B, and C, respectively. The transformation of MCP traits was more precise with model 1 for RCT (R²: 0.77-0.82) than for a₃₀ (R²: 0.28-0.63). The application of model 2 was needed when the C measurements were transformed into the other scales. The analyses of NC probabilities of milk samples showed that NC samples from one methodology were well distinguishable (with an accuracy of 0.972-0.996) based on the rennet coagulation time measured with the other methodology. A standard definition for MCP traits analysis is needed to enable reliable comparisons between MCP traits recorded in different laboratories and in different animal populations and breeds.