Simultaneous quantification of ergot and tropane alkaloids in bread in the Netherlands by LC-MS/MS.

Atropine and scopolamine are tropane alkaloids (TAs), which are regulated for cereal-based foods for children in the EU. For ergot alkaloids (EAs) in cereals and cereal-based food harmonised legislation is not yet established. A fast and straightforward method, which employs extraction by acidified water/methanol followed by ultra-filtration prior to analysis by LC-MS/MS, was validated in bread for 20 EAs and six TAs. LOQs for individual alkaloids ranged from 0.3 to 1.2 µg kg-1, while recoveries ranged from 65% to 94% and repeatability from 3.4% to 17%. A survey was conducted in the Netherlands on 40 retail samples of bread (wheat, rye, wheat-rye, multi-grain) collected in 2014 and 2018. TAs, including atropine and scopolamine, were not detected. Eighteen different EAs were detected and total levels varied between

Straightforward analytical method to determine opium alkaloids in poppy seeds and bakery products.

A straightforward method to determine the content of six opium alkaloids (morphine, codeine, thebaine, noscapine, papaverine and narceine) in poppy seeds and bakery products was developed and validated down to a limit of quantification (LOQ) of 0.1mg/kg. The method was based on extraction with acetonitrile/water/formic acid, ten-fold dilution and analysis by LC-MS/MS using a pH 10 carbonate buffer. The method was applied for the analysis of 41 samples collected in 2015 in the Netherlands and Germany. All samples contained morphine ranging from 0.2 to 240mg/kg. The levels of codeine and thebaine ranged from below LOQ to 348mg/kg and from below LOQ to 106mg/kg, respectively. Sixty percent of the samples exceeded the guidance reference value of 4mg/kg of morphine set by BfR in Germany, whereas 25% of the samples did not comply with the limits set for morphine, codeine, thebaine and noscapine by Hungarian legislation.

Survey of moniliformin in wheat- and corn-based products using a straightforward analytical method.

A straightforward analytical method was developed and validated to determine the mycotoxin moniliformin in cereal-based foods. Moniliformin is extracted with water and quantified with liquid chromatography tandem mass spectrometry, and its presence confirmed with liquid chromatography-Orbitrap-high-resolution mass spectrometry. The method was validated for flour, bread, pasta and maize samples in terms of linearity, matrix effect, recovery, repeatability and limit of quantification. Quantification was conducted by matrix-matched calibration. Positive samples were confirmed by standard addition. Recovery ranged from 77 to 114% and repeatability from 1 to 14%. The limit of quantification, defined as the lowest concentration tested at which the validation criteria of recovery and repeatability were fulfilled, was 10 µg/kg. The method was applied to 102 cereal-based food samples collected in the Netherlands and Germany. Moniliformin was not detected in bread samples. One of 22 flour samples contained moniliformin at 10.6 µg/kg. Moniliformin occurred in seven out of 25 pasta samples at levels around 10 µg/kg. Moniliformin (MON) was present in eight out of 23 maize products at levels ranging from 12 to 207 µg/kg.

Fate of enniatins and deoxynivalenol during pasta cooking.

The fate of deoxynivalenol and enniatins was studied during cooking of commercially available dry pasta in the Netherlands in 2014. Five samples containing relatively high levels of deoxynivalenol and/or enniatins were selected for the cooking experiment. Cooking was performed in duplicate on different days, under standardised conditions, simulating house-hold preparation. Samples were extracted with a mixture of acetonitrile/water followed by salt-induced partitioning. The extracts were analysed by LC-MS/MS. The method limits of detection were 8µg/kg for deoxynivalenol, 10µg/kg for enniatin A1 and 5µg/kg for enniatins A, B and B1. During the cooking of the five dry pasta samples, 60% of the deoxynivalenol and 83-100% of the enniatins were retained in the cooked pasta. It is recommended to study food processing fate of mycotoxins through naturally contaminated materials (incurred materials).

Tropane and ergot alkaloids in grain-based products for infants and young children in the Netherlands in 2011-2014.

An LC-MS/MS multi-method was developed to simultaneously quantify ergot alkaloids (EAs) and tropane alkaloids (TAs) in 113 cereal-based food for infants and young children. To assess yearly variation, samples were collected in 2011, 2012 and 2014. EAs were detected in 54% and TAs in 22% of the samples. Mean EA levels in the three sampling years were 10.6, 6.2 and 8.6 µg kg(-1), respectively (maximum: 115.4 µg kg(-1)), indicating that exposure to EAs would not have exceeded the health-based guidance values set by EFSA in 2012. Mean TA levels were 3.9, 2.4 and 0.4 µg kg(-1), respectively (maximum: 80.8 µg kg(-1)). The acute reference dose for TAs, derived by EFSA in 2013, would have been exceeded by young children when consuming some of the products sampled in 2011-2012. TA levels had decreased drastically in 2014, possibly due to measures taken by producers as response to the EFSA Opinion.

Safe addition of vitamins and minerals to foods: setting maximum levels for fortification in the Netherlands.

BACKGROUND: In 2004, the European Court of Justice decided that the prohibition of fortification with vitamin A, vitamin D, folic acid, selenium, copper, and zinc in the Netherlands conflicts with the principle of free movement of goods in the European Union. This decision led to a change in the Dutch policy, resulting in a more flexible handling of requests for exemption from this prohibition to fortify. Therefore, an investigation was proposed in which it would be determined whether a general exemption could be granted for food fortification with a certain maximum safe amount per micronutrient. AIM OF THE STUDY: To develop a risk assessment model to estimate maximum safe fortification levels (MSFLs) of vitamins and minerals to foods on the Dutch market, and to evaluate these levels to derive allowed fortification levels (AFLs), which can be used for a general exemption. METHODS: We developed a risk assessment model to estimate MSFLs of vitamins and minerals to foods on the basis of existing models. We used European tolerable upper intake levels in combination with national food consumption data to estimate MSFLs for fortification of foods for several age groups. Upon extensive stakeholder dialogue, the risk manager considered these estimated MSFLs and the final AFLs for a general exemption were set. RESULTS: For folic acid, vitamin A, and vitamin D, the MSFLs were calculated in the risk-assessment model. Children up to 6-years old were the group most sensitive to folic acid fortification, and they had an MSFL of 0 microg/100 kcal, but following a risk management evaluation, this was upgraded to an AFL of 100 microg/100 kcal. The MSFL for vitamin D was 3.0 microg/100 kcal (children 4-10 years old), and the risk manager increased this to an AFL of 4.5 microg/100 kcal. Children up to 10 years old, men, and postmenopausal women were the groups most sensitive to vitamin A fortification (MSFL = 0 microg/100 kcal). Because these groups represent a large part of the population and because of the seriously harmful effects of excessive vitamin A, the risk manager did not allow a general exemption. CONCLUSIONS: The combination of a risk assessment model and risk manager evaluation led to the setting of AFLs for general exemption of fortification with folic acid and vitamin D. This model is also applicable for other micronutrients, for which an UL is derived, and in other countries.