Optimization and Validation of a High Throughput UHPLC-MS/MS Method for Determination of the EU Regulated Lipophilic Marine Toxins and Occurrence in Fresh and Processed Shellfish.

Under the name of lipophilic marine toxins, there are included more than 1000 toxic secondary metabolites, produced by phytoplankton, with the common chemical property of lipophilicity. Due to toxicological effects and geographical distribution, in European legislation relevant compounds are regulated, and their determination is accomplished with the reference liquid chromatography-tandem mass spectrometry method. In this study a modified ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method has been developed for the identification and quantification of EU-regulated lipophilic toxins. The method optimization included a refinement of SPE-C18 clean-up, in order to reduce matrix interferences. Improved LC conditions and upgraded chromatographic ammonia-based gradient ensured the best separation of all analytes and, in particular, of the two structural isomers (OA and DTX2). Also, different MS parameters were tested, and confirmation criteria finally established. The validation studies confirmed that all parameters were satisfactory. The requirements for precision (RSD% < 11.8% for each compound), trueness (recoveries from 73 to 101%) and sensitivity (limits of quantification in the range 3−8 µg kg−1) were fulfilled. The matrix effect, ranging from −9 to 19%, allowed the use of a calibration curve in solvent (3−320 µg kg−1 in matrix) for quantification of real samples. Method relative uncertainty ranged from 12 to 20.3%. Additionally, a total of 1000 shellfish samples was analysed, providing a first preliminary surveillance study that may contribute to the knowledge of lipophilic marine toxins contamination. Increase in algae proliferation events and intoxication cases, EFSA suggestions for modification of maximum permitted levels and toxicity equivalency factors, and new studies of important toxic effects underline that implementation of reference methods still represents an important task for health and food safety laboratories.

Distribution of Aspergillus Fungi and Recent Aflatoxin Reports, Health Risks, and Advances in Developments of Biological Mitigation Strategies in China.

Aflatoxins (AFs) are secondary metabolites that represent serious threats to human and animal health. They are mainly produced by strains of the saprophytic fungus Aspergillus flavus, which are abundantly distributed across agricultural commodities. AF contamination is receiving increasing attention by researchers, food producers, and policy makers in China, and several interesting review papers have been published, that mainly focused on occurrences of AFs in agricultural commodities in China. The goal of this review is to provide a wider scale and up-to-date overview of AF occurrences in different agricultural products and of the distribution of A. flavus across different food and feed categories and in Chinese traditional herbal medicines in China, for the period 2000-2020. We also highlight the health impacts of chronic dietary AF exposure, the recent advances in biological AF mitigation strategies in China, and recent Chinese AF standards.

Sub-Acute Feeding Study of Saxitoxin to Mice Confirms the Effectiveness of Current Regulatory Limits for Paralytic Shellfish Toxins.

Regulatory limits for shellfish toxins are required to protect human health. Often these limits are set using only acute toxicity data, which is significant, as in some communities, shellfish makes up a large proportion of their daily diet and can be contaminated with paralytic shellfish toxins (PSTs) for several months. In the current study, feeding protocols were developed to mimic human feeding behaviour and diets containing three dose rates of saxitoxin dihydrochloride (STX.2HCl) were fed to mice for 21 days. This yielded STX.2HCl dose rates of up to 730 µg/kg bw/day with no effects on food consumption, growth, blood pressure, heart rate, motor coordination, grip strength, blood chemistry, haematology, organ weights or tissue histology. Using the 100-fold safety factor to extrapolate from animals to humans yields a dose rate of 7.3 µg/kg bw/day, which is well above the current acute reference dose (ARfD) of 0.5 µg STX.2HCl eq/kg bw proposed by the European Food Safety Authority. Furthermore, to reach the dose rate of 7.3 µg/kg bw, a 60 or 70 kg human would have to consume 540 or 630 g of shellfish contaminated with PSTs at the current regulatory limit (800 µg/kg shellfish flesh), respectively. The current regulatory limit for PSTs therefore seems appropriate.

Edible Oils Adulteration: A Review on Regulatory Compliance and Its Detection Technologies.

Various events of edible oils adulteration with inferior ingredients were reported regularly in recent years. This review is aimed to provide an overview of edible oils adulteration practices, regulatory compliance and detection technologies. Many detection technologies for edible oils adulteration were developed in the past such as methods that are based on chromatography or spectroscopy. Electrochemical sensors like electric nose and tongue are also gaining popularity in the detection of adulterated virgin olive oil and virgin coconut oil. It can be concluded that these detection technologies are essential in the combat with food adulterers and can be improved.

Food recalls associated with foodborne disease outbreaks, United States, 2006-2016.

About 800 foodborne disease outbreaks are reported in the United States annually. Few are associated with food recalls. We compared 226 outbreaks associated with food recalls with those not associated with recalls during 2006-2016. Recall-associated outbreaks had, on average, more illnesses per outbreak and higher proportions of hospitalisations and deaths than non-recall-associated outbreaks. The top confirmed aetiology for recall-associated outbreaks was Salmonella. Pasteurised and unpasteurised dairy products, beef and molluscs were the most frequently implicated foods. The most common pathogen-food pairs for outbreaks with recalls were Escherichia coli-beef and norovirus-molluscs; the top pairs for non-recall-associated outbreaks were scombrotoxin-fish and ciguatoxin-fish. For outbreaks with recalls, 48% of the recalls occurred after the outbreak, 27% during the outbreak, 3% before the outbreak, and 22% were inconclusive or had unknown recall timing. Fifty per cent of recall-associated outbreaks were multistate, compared with 2% of non-recall-associated outbreaks. The differences between recall-associated outbreaks and non-recall-associated outbreaks help define the types of outbreaks and food vehicles that are likely to have a recall. Improved outbreak vehicle identification and traceability of rarely recalled foods could lead to more recalls of these products, resulting in fewer illnesses and deaths.

Contaminação microscópica por matérias estranhas em noz-moscada, cúrcuma, gengibre, colorífico, pimenta-do-reino e páprica comercializados no estado de São Paulo, Brasil. (Parte I) / Microscopic contamination by foreign matter in nutmeg, turmeric, ginger, colorific, black pepper and paprika marketed in the State of Sao Paulo, Brazil. (Part I)

O objetivo deste trabalho foi avaliar a presença de matérias estranhas em noz-moscada, (Myristica fragrans Houttuyn), cúrcuma (Curcuma longa Linnaeus), gengibre (Zingiber officinale Roscoe), colorífico (mistura de urucum [Bixa orellana Linnaeus] com fubá), pimenta-do-reino (Piper nigrum Linnaeus) e páprica (Capsicum annuum Linnaeus) conforme a legislação sanitária. Foram analisadas 180 amostras empregando os métodos preconizados pela AOAC International, de maio de 2018 a maio de 2020. A presença de ao menos uma matéria estranha foi observada em 80% das amostras. Pelos animais foram observados em todos os produtos e fragmentos de insetos foram encontrados na maioria das amostras. Ácaro, inseto inteiro, larva de inseto, bárbula e exúvia também foram encontrados, além de fibras sintéticas e fragmentos de microplásticos. Quanto à legislação, 47,8% das amostras estavam acima dos limites de tolerância, destas, 90,7% por conterem matérias estranhas indicativas de falhas na aplicação das boas práticas e 9,3% por matérias estranhas indicativas de risco à saúde. Os resultados obtidos denotam ineficiência da aplicação das medidas de boas práticas na cadeia produtiva e alertam para a intensificação da fiscalização direcionada ao cumprimento das normas sanitárias, além de provocarem a reflexão sobre a necessidade de alterações na legislação referente às matérias estranhas. (AU)

The aim of this work was to evaluate the presence of foreign matter in nutmeg, (Myristica fragrans Houttuyn), turmeric (Curcuma longa Linnaeus), ginger (Zingiber officinale Roscoe), colorific (mixture of annatto [Bixa orellana Linnaeus] with cornmeal), black pepper (Piper nigrum Linnaeus) and paprika (Capsicum annuum Linnaeus) according to the health legislation. A total of 180 samples were analyzed using the methods recommended by AOAC International, from May 2018 to May 2020. The presence of at least one foreign matter was observed in 80% of the samples. Animal's hairs were observed in all the evaluated products and insect's fragments were found in the most of the samples. Mites, whole insects, insect larvae, barbules and exuvia were also found, as wellas synthetic fibers and microplastic fragments. About legislation, 47.8% of the samples were above the tolerance limits, of these, 90.7% for containing foreign matters indicating failure of good practices and 9.3% for foreign matters indicating health risk. The results obtained show inefficiency in the application of good practice measures in production chain and advertise for the intensification of inspection directed to fulfillment of sanitary norms, besides provoking the reflection on the necessity of alterations in the legislation about foreign matter in foods. (AU)

A Review on Mycotoxins and Microfungi in Spices in the Light of the Last Five Years.

Spices are imported worldwide mainly from developing countries with tropical and/or subtropical climate. Local conditions, such as high temperature, heavy rainfall, and humidity, promote fungal growth leading to increased occurrence of mycotoxins in spices. Moreover, the lack of good agricultural practice (GAP), good manufacturing practice (GMP), and good hygienic practice (GHP) in developing countries are of great concern. This review summarizes recent data from a total of 56 original papers dealing with mycotoxins and microfungi in various spices in the last five years. A total of 38 kinds of spices, 17 mycotoxins, and 14 microfungi are discussed in the review. Worldwide, spices are rather overlooked in terms of mycotoxin regulations, which usually only cover aflatoxins (AFs) and ochratoxin A (OTA). In this paper, an extensive attention is devoted to the limits on mycotoxins in spices in the context of the European Union (EU) as well as other countries. As proven in this review, the incidence of AFs and OTA, as well as other mycotoxins, is relatively high in many spices; thus, the preparation of new regulation limits is advisable.

Occurrence, sources, and pathways of chemical contaminants in infant formulas.

Infant formulas are manufactured products to meet specific nutritional requirements for infants. However, infant formulas can contain harmful substances, such as chemical contaminants and residues, normally due to possible contamination of the raw material or from the production chain. Some studies have demonstrated that veterinary drugs, pesticides, mycotoxins, heavy metals, packaging materials, within other chemicals are found in infant formulas from different sources of contamination. It is known that some of these substances can be hepatotoxic, carcinogenic, teratogenic, mutagenic, immunotoxic, contributing to antibiotic resistance, among other detrimental consequences for consumers' health. The purpose of this review is to assess the scientific evidence concerning the occurrence, sources, and pathways of contamination, as well as the detrimental impacts on infant health due to the possible presence of chemical contaminants and residues in infant formulas. Moreover, strategies to reduce the risk of contamination of infant formulas are presented to ensure the highest standards of quality of infant formulas. The entire infant formula manufacturing process should be monitored and controlled to minimize the risk of contamination during processing, storage, and distribution, besides ensuring the use of raw materials with as low as acceptable levels of harmful substances in order to assure that the final product shall comply with the maximum levels and maximum residue limits, when established, for residues and contaminants in the final product.

Repeated use food contact materials: A categorisation approach in support of risk assessment.

The current migration assessment requirements regarding safety of plastic food contact materials in Europe (e.g. kitchen utensils, kitchen appliances, packaging, etc.) widely rely on migration testing. According to Annexe V of Regulation (EU) No 10/2011 migration testing requirements consider the specificities of repeated use applications only to a limited extent. Repeated use food contact materials should be tested for three consecutive times at the worst-case contact time and temperature. If diffusion controlled, the migration decreases with increasing number of repeated uses. Compared to single use applications, repeated use food contact materials typically exhibit much shorter contact times, much lower ratios of surface in contact with a given amount of food, and in some cases higher temperatures. Compared to real use, in many cases highly overestimated migration testing result are observed. Overestimation by testing at the beginning of use may be coupled with underestimation at later times. National legislation of the Netherlands on food contact materials has established a classification for repeated use rubber materials based on the R-value indicating whether migration testing is required or not. The R-value considers in more detail specificities of repeated use applications. This publication investigates to which extent it is possible to apply the approach to plastics food contact materials in Europe. It is practically impossible to perform migration tests for various materials by putting them many times in contact with food over a long period of time, typically several years, at several temperatures and areas to food amount ratios migration. Modelling has therefore been used to predict migration from various food contact materials under different repeated use scenarios. Realistic diffusion properties of materials and migrant partitioning behaviour have been assumed to minimise the risk of underestimation.

Beauvericin and Enniatins: In Vitro Intestinal Effects.

Food and feed contamination by emerging mycotoxins beauvericin and enniatins is a worldwide health problem and a matter of great concern nowadays, and data on their toxicological behavior are still scarce. As ingestion is the major route of exposure to mycotoxins in food and feed, the gastrointestinal tract represents the first barrier encountered by these natural contaminants and the first structure that could be affected by their potential detrimental effects. In order to perform a complete and reliable toxicological evaluation, this fundamental site cannot be disregarded. Several in vitro intestinal models able to recreate the different traits of the intestinal environment have been applied to investigate the various aspects related to the intestinal toxicity of emerging mycotoxins. This review aims to depict an overall and comprehensive representation of the in vitro intestinal effects of beauvericin and enniatins in humans from a species-specific perspective. Moreover, information on the occurrence in food and feed and notions on the regulatory aspects will be provided.

[Microbiological safety of food: development of normative and methodical base].

The main results and prospects of fundamental and applied hygienic research of the laboratory of biosafety and nutrimicrobiome analysis of the Federal Research Centre of Nutrition, Biotechnology and Food Safety (hereinafter - the Institute of Nutrition) in the direction of developing a regulatory and methodological framework for assessing the microbiological safety of food are reviewed. The formation of microbiological regulation as a scientific analytical and administrative managerial process in the former USSR and the Russian Federation is considered in the context of historical data, including personal contribution of the scientists of the Institute of Nutrition and other specialists. The basic principles of regulation are emphasized: the scientific validity of the established criteria and requirements, the feasibility, technological attainability, differentiation according to the degree of danger to the health of consumers, preventive nature. The resource of the national normative and methodological base in the field of microbiological food safety at the turn of the century is characterized, the features of the introduction of the microbiological risk assessment (MRA) methodology in the substantiation of Russian norms and measures for the prevention of food infections are described. The information is given on the developed guidance documents on MRA and on the examples of norms adopted on its basis. The article covers the issues of reglamentation the requirements for food safety and reducing the spread of new pathogens Stx-Escherichia coli, Listeria monocytogenes, Enterobacter sakazakii, Campylobacter spp. in the food chain based on risk-oriented approaches. The necessity of taking specific measures for the prevention of cross-contamination in the poultry processing industry is substantiated, taking into account the evidence of the high adaptability of C. jejuni isolated from domestic raw poultry. In the sanitarian-mycological aspect, the monitoring perspective of mould fungi, taking into account their chemotypes, in cereals and non-grain plant products is shown to predict the risk of mycotoxin accumulation and take timely measures. The need to assess the impact on the population, taking into account the characteristics of consumption in the country, as well as the development of criteria for indirect risk of residues are argued for regulation of the antibiotics in food. In light of the challenges in the field of agro- and food technologies to public health at the present stage, contributing to the acceleration of microbial evolution and the emergence of new risks in food, the priority tasks of improving the regulatory and methodological base for assessing microbiological safety have been identified, with an emphasis on the introduction into the process of substantiating the norms of innovative OMICs-technologies based on the achievements of genomics, transcriptomics, proteomics, metabolomics, bioinformatics.

Significance of and Challenges in Regulating Endocrine Disruptors - How Regulators and Industry Can Conquer?

Endocrine disruptors (EDs) disrupt the standard operation of the endocrine systems, resulting in untoward effects. EDs have gained extensive consideration due to their severe adverse impacts on public and wildlife health. A variety of compounds from both natural and synthetic origin may cause endocrine disruptions. These may be found in industrial chemicals, persistent organic pollutants, and products of regular use including pharmaceuticals, medical equipment, implants, medical/surgical and dental devices, cosmetics, food products, other consumer goods, their packaging and processing materials. Apart from direct consumption or use, these chemicals may impact by entering our food chain or ecosystem. These chemicals act by mimicking the hormones or blocking their receptors or interfering in their normal production, absorption, distribution, metabolism and excretion. The implementation of a regulatory framework on the complex multidisciplinary field of EDs brings enormous challenges, which pose barriers to the regulatory process. This study aims to focus on the key public and ecological health concerns presented by EDs, challenges faced by regulators to achieve successful regulatory proposition and the importance of collaboration endeavours to potentially conquer such challenges. Endocrine-disrupting chemicals (EDCs) or EDs can impact at low exposure levels, bringing about a broad range of health issues including disorders related to reproductive, fetal development, neurological, immunological, metabolic and cancer, etc. They may cause health effects across generations. The regulatory frameworks available across major regulators are tackling the identification of EDs and their mechanisms to provide necessary guidance on the safety and disposal of such substances. However, the challenges faced outweigh the regulatory mechanisms in place. The major challenges are related to structural ranges at times leading to no representative structures, active metabolites, substantiate quantum, delayed effects, epigenetic changes, widespread existence, concentration correlation for different biological species, availability of appropriate methods, exposure to a mixture of chemicals, complex endocrinology principles, unknown sources, routes and mechanisms, impacts at early stages of life, geographical movement of EDs, hazard-based vs. risk-based approaches. Regulators of healthcare and environmentalists needs to collaborate amongst them and with wider stakeholders including industry sponsors to find ways of dealing with such challenges and capitalize on the research-based knowledge grid available across institutions. Existence of EDs, their impact on living beings and mechanism of influence are like a tangled web, which induces difficulties in regulating them with conventional mindset. Conquering these challenges necessitates that regulators should join forces amongst themselves, with other institutions operating for environment, with industry sponsors and researchers to achieve success in public health safety.

Mycotoxin Contamination Concerns of Herbs and Medicinal Plants.

Plants and medicinal herbs that are available on the market do not always meet quality and safety standards. One particular concern is the risk of contamination with mycotoxins. Aflatoxins and ochratoxin A are the most frequently described mycotoxins in herbal products and have repeatedly been reported to occur at concentrations which exceed regulatory levels set by the European Union (EU). Possible solutions include enforcing existing limits, and for the new materials, establishing tighter limits and mandate the growth of medicinal plants in EU member countries under more strict conditions.

Lead Concentrations in Mexican Candy: A Follow-Up Report.

Background: Lead is a neurotoxic metal potentially affecting the developing brain. Children are particularly susceptible since they can absorb between 50% and 100% of ingested lead. There is no safe level for lead, therefore preventing exposure is crucial. We previously reported a positive association between lead concentrations found in candy and concurrent blood lead levels in Mexican children. This first report garnered media and the general public's attention. Objective: To conduct a follow-up study to assess lead concentrations in candy brands that we previously reported with concentrations ≥0.1ppm the U.S. Food and Drug Administration's recommended maximum lead level in candy likely to be consumed frequently by small children. Methods: In 2018 we analyzed 50 additional candy samples. Lead concentrations were analyzed by an inductively coupled plasma mass spectrometer and lead content per candy unit was calculated. Findings: We found concentrations were typically low, with a marked decrease from prior levels (2008). Nevertheless two candy units had concentrations of 0.1 ppm of lead. Conclusions: Candy may have lead concentrations up to 0.1 ppm and 1.2 µg per unit. This is a concern because candies are exported and consumed in many countries worldwide potentially resulting in human exposure. Continued public health surveillance is needed to protect populations especially vulnerable to lead exposure, especially children.

Mycotoxins in maize harvested in Serbia in the period 2012-2015. Part 2: Non-regulated mycotoxins and other fungal metabolites.

The main objective of this study was to screen, for the first time, the natural occurrence of non-regulated fungal metabolites in 204 maize samples harvested in Serbia in maize growing seasons with extreme drought (2012), extreme precipitation and flood (2014) and moderate drought conditions (2013 and 2015). In total, 109 non-regulated fungal metabolites were detected in examined samples, whereby each sample was contaminated between 13 and 55 non-regulated fungal metabolites. Moniliformin and beauvericin occurred in all samples collected from each year. In samples from year 2012, oxaline, questiomycin A, cyclo (l-Pro-l-Val), cyclo (l-Pro-l-Tyr), bikaverin, kojic acid and 3-nitropropionic acid were the most predominant (98.0-100%). All samples from 2014 were contaminated with 7-hydroxypestalotin, 15-hydroxyculmorin, culmorin, butenolid and aurofusarin. Bikaverin and oxaline were quantified in 100% samples from 2013 and 2015, while 3-nitropropionic acid additionally occurred in 100% samples from 2015.

Acrylamide in human diet, its metabolism, toxicity, inactivation and the associated European Union legal regulations in food industry.

Nowadays acrylamide is known not only as synthetic material used in industry, but also as carcinogenic, cyto- and genotoxic compound which forms during heat-induced process (due to Maillard reaction) mostly in foodstuff such as potato, bakery, plant derivatives products and coffee. The International Agency for Research on Cancer in 1994 declared acrylamide as a probable carcinogenic agent in humans. After metabolic process, acrylamide is distributed to all organs and tissues in human body. Acrylamide is classified as human neurotoxin, because this effect was observed in humans occupationally exposed to this compound. Acrylamide was found to cause apoptosis by mitochondrial dysfunction. Methods of acrylamide inactivation by microorganisms and bioactive diet compounds have also been reviewed. Moreover, there is still deficit of the European Union legal regulation concerning acrylamide mitigation strategies in food. Regulation 2017/2158 from 20 November 2017 is a step in the right direction when it comes to ensuring food safety and maximum levels of acrylamide in foodstuffs, however when exceeding those, it should result in elimination of such food from the market.

Fumonisins at Doses below EU Regulatory Limits Induce Histological Alterations in Piglets.

Fumonisins (FBs) are mycotoxins produced by Fusarium species that can contaminate human food and animal feed. Due to the harmful effects of FBs on animals, the European Union (EU) defined a recommendation of a maximum of 5 mg FBs (B1 + B2)/kg for complete feed for swine and 1 µg FBs/kg body weight per day as the tolerable daily intake for humans. The aim of this study was to evaluate the toxicity of dietary exposure to low doses of FBs, including a dose below the EU regulatory limits. Four groups of 24 weaned castrated male piglets were exposed to feed containing 0, 3.7, 8.1, and 12.2 mg/kg of FBs for 28 days; the impact was measured by biochemical analysis and histopathological observations. Dietary exposure to FBs at a low dose (3.7 mg/kg of feed) significantly increased the plasma sphinganine-to-sphingosine ratio. FBs-contaminated diets led to histological modifications in the intestine, heart, lung, lymphoid organs, kidney, and liver. The histological alterations in the heart and the intestine appeared at the lowest dose of FBs-contaminated diet (3.7 mg/kg feed) and in the kidney at the intermediate dose (8.1 mg/kg feed). At the highest dose tested (12.2 mg/kg feed), all the organs displayed histological alterations. This dose also induced biochemical modifications indicative of kidney and liver alterations. In conclusion, our data indicate that FBs-contaminated diets at doses below the EU regulatory limit cause histological lesions in several organs. This study suggests that EU recommendations for the concentration of FBs in animal feed, especially for swine, are not sufficiently protective and that regulatory doses should be modified for better protection of animal health.