2 0 1 5 Development and validation of analytical methods for the analysis of 3-MCPD (both in free and ester form) and glycidyl esters in various food matrices and performance of an ad-hoc survey on specific food groups in support to a scientific opinion on comprehensive risk assessment on the presence of 3-MCPD and glycidyl esters in food Reproduction is authorised provided the source is acknowledged.Abstract 3-Monochloropropane-1,2-diol (3-MCPD), and 2-monochloropropane-1,3-diol (2-MCPD), are substances that might be generated in the processing of food. EU legislation specifies maximum levels for 3-MCPD in hydrolysed vegetable proteins and soya sauce. However, besides the free forms of 2-and 3-MCPD high levels of esterified MCPD forms were found in fats and oils. Another group of substances identified in fats and oils are glycidyl esters (GE). In order to provide reliable occurrence data on the levels of both bound and free forms of those substances, the European Food Safety Authority (EFSA) requested the Joint Research Centre (JRC) to develop suitable analysis methods and test the analysis methods on different kinds of food. Consequently two analytical methods were developed. One of the developed methods allows the determination of ester-bound analytes, whereas the other analysis method is suitable to determine free 2-MCPD and free 3-MCPD. Reliability of analysis results and robustness of the analysis methods were the main focus during method development and optimisation. The analytes were extracted with organic solvents under mild conditions. GEs are converted to monobromopropanediol esters (MBPD esters) prior to transesterification. MCPD esters and MBPD esters were transesterified followed by derivatisation of the analytes with phenyl boronic acid (PBA) in organic solvent. The PBA derivatives were measured by gas chromatography mass spectrometry (GC-MS) applying stable isotope labelled analogues of the analytes for quantification. The performance of both analysis methods was compliant with criteria specified by EFSA. The analytical methods were applied for the analysis of breads and bread rolls, fine bakery wares, smoked fish and meat products, fried and roasted meat, potato-based snacks and fried potato products, cereal-based snacks, and margarines. Analysis results were compiled and reported to EFSA in standard sample description (SSD) format.
ABSTRACT3-Monochloropropane-1,2-diol (3-MCPD), and 2-monochloropropane-1,3-diol (2-MCPD), are substances that might be generated in the processing of food. EU legislation specifies maximum levels for 3-MCPD in hydrolysed vegetable proteins and soya sauce. However, besides the free forms of 2-and 3-MCPD high levels of esterified MCPD forms were found in fats and oils. Another group of substances identified in fats and oils are glycidyl esters (GE). In order to provide reliable occurrence data on the levels of both bound and free forms of those substances, the European Food Safety Authority (EFSA) requested the Joint Research Centre (JRC) to develop suitable analysis...
Fatty acid esters of 3-monochloro-1,2-propanediol (3-MCPDEs), of 2-monochloro-1,3-propanediol (2-MCPDEs) and of 2,3-epoxy-1-propanol or glycidol (GEs), which are considered to be deleterious to human health, may occur in a broad variety of food samples. A proper risk assessment of those substances requires the availability of robust occurrence data; in this respect concerns have been raised regarding the reliability of results obtained with the currently available methods to determine those substances in processed food. This article presents an indirect analytical procedure for the simultaneous determination of 3-MCPDEs, 2-MCPDEs and GEs in a wide variety of food products after extraction by pressurised liquid extraction (PLE) and determination by gas chromatography mass-spectrometry (GC-MS). For the differentiation of MCPDEs and GEs, the latter were first converted to monobromopropanediol esters (MBPDEs) in acid aqueous solution of sodium bromide. MCPDEs and MBPDEs were then hydrolysed under acidic conditions followed by derivatisation of the released free (non-esterified) form in ethyl acetate with phenyl boronic acid (PBA). Quantification of the analytes was carried out using the isotopic labelled analogues of both MCPDEs and GEs. Limits of detection (LODs) and limits of quantitation (LOQs) were in the range of 7-17mgkg(-1) and 13-31mgkg(-1) respectively, while the working range of the method was between LOQ and 1850mgkg(-1) expressed on fat basis. The developed method was successfully applied for the analysis of the target compounds in more than 650 different food samples covering the following commodities: bread and rolls, fine bakery wares, smoked fish products, fried and roasted meat, potato based snacks and fried potato products, cereal-based snacks and margarines.
Multiple lineages of SARS-CoV-2 have been identified featuring distinct sets of genetic changes that confer to the virus higher transmissibility and ability to evade existing immunity. The continuous evolution of SARS-CoV-2 may pose challenges for current treatment options and diagnostic tools. In this study, we have first evaluated the performance of the 14 WHO-recommended real-time reverse transcription (RT)-PCR assays currently in use for the detection of SARS-CoV-2 and found that only one assay has reduced performance against Omicron. We then developed a new duplex real-time RT-PCR assay based on the amplification of two ultra-conserved elements present within the SARS-CoV-2 genome. The new duplex assay successfully detects all of the tested SARS-CoV-2 variants of concern (including Omicron sub-lineages BA.4 and BA.5) from both clinical and wastewater samples with high sensitivity and specificity. The assay also functions as a one-step droplet digital RT-PCR assay. This new assay, in addition to clinical testing, could be adopted in surveillance programs for the routine monitoring of SARS-CoV-2’s presence in a population in wastewater samples. Positive results with our assay in conjunction with negative results from an Omicron-specific assay may provide timely indication of the emergence of a novel SARS-CoV-2 variant in a certain community and thereby aid public health interventions.
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