Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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National Reference Laboratories WFSR : annual report 2018
Noordam, M.Y. ; Silletti, E. ; Alewijn, A. ; Scholtens, I.M.J. ; Jong, J. de; Raamsdonk, L. aan; Lasaroms, J.J.P. ; Gerssen, A. ; Lee, M.K. van der; Mol, J.G.J. ; Leeuwen, S.P.J. van - \ 2019
Wageningen : Wageningen University & Research (WFSR-report 2018.011) - 51
Emerging marine biotoxins
Gerssen, Arjen ; Gago-Mart Í Nez, Ana - \ 2019
Toxins 11 (2019)6. - ISSN 2072-6651
Quantitative in vitro-to-in vivo extrapolation (QIVIVE) of estrogenic and anti-androgenic potencies of BPA and BADGE analogues
Punt, Ans ; Aartse, Aafke ; Bovee, Toine F.H. ; Gerssen, Arjen ; Leeuwen, Stefan P.J. van; Hoogenboom, Ron L.A.P. ; Peijnenburg, Ad A.C.M. - \ 2019
Archives of Toxicology 93 (2019)7. - ISSN 0340-5761 - p. 1941 - 1953.
Androgenic - Bisphenol A - BP analogues - Estrogenic - QIVIVE - Relative potencies

The goal of the present study was to obtain an in vivo relevant prioritization method for the endocrine potencies of different polycarbonate monomers, by combining in vitro bioassay data with physiologically based kinetic (PBK) modelling. PBK models were developed for a selection of monomers, including bisphenol A (BPA), two bisphenol F (BPF) isomers and four different bisphenol A diglycidyl ethers (BADGEs), using in vitro input data. With these models, the plasma concentrations of the compounds were simulated, providing means to estimate the dose levels at which the in vitro endocrine effect concentrations are reached. The results revealed that, whereas the in vitro relative potencies of different BADGEs (predominantly anti-androgenic effects) can be up to fourfold higher than BPA, the estimated in vivo potencies based on the oral equivalent doses are one to two orders of magnitude lower than BPA because of fast detoxification of the BADGEs. In contrast, the relative potencies of 2,2-BPF and 4,4-BPF increase when accounting for the in vivo availability. 4,4-BPF is estimated to be fivefold more potent than BPA in humans in vivo in inducing estrogenic effects and both 2,2-BPF and 4,4-BPF are estimated to be, respectively, 7 and 11-fold more potent in inducing anti-androgenic effects. These relative potencies were considered to be first-tier estimates, particularly given that the potential influence of intestinal metabolism on the in vivo availability was not accounted for. Overall, it can be concluded that both 2,2-BPF and 4,4-BPF are priority compounds.

Oesters controleren zonder proefdieren
Gerssen, Arjen - \ 2019
Testing shellfish without lab animals
Gerssen, Arjen - \ 2019
Mosselgif opsporen zonder proefdieren
Rietjens, Ivonne ; Gerssen, Arjen ; Bodero Baeza, Marcia - \ 2019
BPA, BADGE and analogues : A new multi-analyte LC-ESI-MS/MS method for their determination and their in vitro (anti)estrogenic and (anti)androgenic properties
Leeuwen, Stefan P.J. van; Bovee, Toine F.H. ; Awchi, Mohamad ; Klijnstra, Mirjam D. ; Hamers, Astrid R.M. ; Hoogenboom, Ron L.A.P. ; Portier, Liza ; Gerssen, Arjen - \ 2019
Chemosphere 221 (2019). - ISSN 0045-6535 - p. 246 - 253.
Androgenic - BADGE - BP-analogues - BPA - Estrogenic - In vitro - LC-MS/MS - Multimethod

Information on the occurrence and endocrine potencies of analogues of bisphenol A (BPA) and diglycidyl ester derivatives (BDGEs) of BPA and BPF is limited. Such information is, however, important as the current debate on BPA and the lowered BPA migration limit in Europe may provide an incentive for application of structural analogues. A new sensitive multi-analyte LC-ESI-MS/MS method was developed to measure 17 bisphenols (BPs) and 6 BDGEs in food, beverages and drinkware. Yeast based bioassays were used to determine the in vitro (anti)estrogenic and (anti)androgenic properties of these and 7 additional BPs and BDGEs. Drinkware of polycarbonate and other materials were analysed for BPs and BDGEs. Only BPA and BPS and both at trace levels were found in a few containers. A limited number of (canned) foods and beverages were also analysed. BPA was the most frequently detected BP (ranged from 0.03 ng mL−1 in a beverage sample to 68 ng g−1 in food). Other BPs detected were BPS, 2,2-BPF and 4,4-BPF. In addition BADGE, BADGE.HCl, BADGE.H2O and BADGE.2H2O were detected from 0.08 ng mL−1 in a beverage sample to 3.3 ng g−1 in food. In vitro testing showed that most BPs exhibited an equal or higher estrogenic potency than BPA and most of them also showed a higher anti-androgenic potency, i.e. BPB, BPCl, BPC, BPE, 4,4-BPF, BPP, BPAF, and BPTMC. Some BPs and BDGEs were not estrogenic, but showed an anti-estrogenic effect and were anti-androgenic too. BPS was only weakly estrogenic and BADGE.2H2O and BFDGE.2H2O showed no in vitro activity. The present data show that in addition to BPA, other BPs and BDGEs can be present in food and drinks, some displaying in vitro endocrine activities.

Food and feed safety : Cases and approaches to identify the responsible toxins and toxicants
Gerssen, Arjen ; Bovee, Toine H.F. ; Ginkel, Leendert A. van; Iersel, Marlou L.P.S. van; Hoogenboom, Ron L.A.P. - \ 2019
Food Control 98 (2019). - ISSN 0956-7135 - p. 9 - 18.
Effect directed analysis - Feed and food safety - Identification strategy - In-vitro bioassays - Intoxications

There are food and feed safety monitoring programs to protect consumers. These programs however, are strongly focused on known and regulated substances. New or unexpected substances that might be of risk for consumers will thus escape routine controls. These risks are therefore mainly discovered by human or animal intoxications. All kind of analytical chemical methods, in-vitro bioassays, tracking, and chain analysis are then used to reveal the substance(s) responsible for the intoxication. Only in a few occasions (new) risks were revealed in time by analytical chemical methods or cell based in-vitro bioassays. This paper describes some relevant food and feed safety cases and how the causative substances were identified. This overview strongly indicates that more intense monitoring, including the use of cell based effect bioassays, can reduce the number of intoxications. Moreover, registration and follow-up actions should be arranged in a better way, for example by sharing information within the scientific communities or by establishing a national contact point. In addition, a strategy based on broad screening and bioassay directed identification with liquid chromatography high resolution mass spectrometry is proposed to prevent intoxications and identify toxin and toxicants relevant for food and feed safety.

Sensitive mass spectrometric method for the detection of palytoxins in shellfish using lithium cationization
Gerssen, A. - \ 2018
Potentieel toxisch fytoplankton in productiegebieden van tweekleppige weekdieren : evaluatie van het Nederlandse monitoringsprogramma met voorstellen voor een alternatieve aanpak
Faassen, E. ; Bovee, T. ; Klijnstra, M. ; Alewijn, M. ; Gerssen, A. - \ 2018
Wageningen : RIKILT Wageningen University & Research (RIKILT-rapport 2018.002) - 55
National Reference Laboratories RIKILT : annual report 2017
Leeuwen, S.P.J. van; Mol, J.G.J. ; Lee, M.K. van der; Gerssen, A. ; Lasaroms, J.J.P. ; Sterk, S.S. ; Raamsdonk, L. aan; Jong, J. de; Scholtens, I.M.J. ; Alewijn, A. ; Silletti, E. ; Ginkel, L. van; Noordam, M.Y. ; Meijer, N. - \ 2018
Wageningen : RIKILT Wageningen University & Research (RIKILT-report 2018.009) - 51
Metabolism and absorption of marine biotoxins using in vitro systems and MS based analysis
Klijnstra, M.D. ; Portier-Onstenk, L. ; Bovee, T.F.H. ; Gerssen, A. - \ 2018
A sensitive LC-MS/MS method for palytoxin using lithium cationization
Klijnstra, Mirjam D. ; Gerssen, Arjen - \ 2018
Toxins 10 (2018)12. - ISSN 2072-6651
Cationization - Mass spectrometry - Ovatoxins - Palytoxin

Palytoxin (PlTX) and analogues are produced by certain dinoflagellates, sea anemones, corals and cyanobacteria. PlTX can accumulate in the food chain and when consumed it may cause intoxication with symptoms like myalgia, weakness, fever, nausea, and vomiting. The analysis of PlTXs is challenging, and because of the large molecular structure, it is difficult to develop a sensitive and selective liquid chromatography-mass spectrometry (LC-MS/MS) method. In this work, an LC-MS/MS method was developed to analyse PlTXs with use of lithium iodine and formic acid as additives in the mobile phase. For method development, initially, LC-hrMS was used to accurately determine the elemental composition of the precursor and product ions. The main adduct formed was [M + H + 2Li]3+. Fragments were identified with LC-hrMS and these were incorporated in the LC-MS/MS method. A method of 10 min was developed and a solid phase extraction clean-up procedure was optimised for shellfish matrix. The determined limits of detection were respectively 8 and 22 µg PlTX kg−1 for mussel and oyster matrix. Oysters gave a low recovery of approximately 50% for PlTX during extraction. The method was successfully in-house validated, repeatability had a relative standard deviation less than 20% (n = 5) at 30 µg PlTX kg−1 in mussel, cockle, and ensis, and at 60 µg PlTX kg−1 in oyster.

Ambient Ionization for direct food analysis by (trans)portable Mass Spectrometry
Gerssen, Arjen - \ 2018
A strategy to replace the mouse bioassay for detecting and identifying lipophilic marine biotoxins by combining the neuro-2a bioassay and LC-MS/MS analysis
Bodero, Marcia ; Gerssen, Arjen ; Portier, Liza ; Klijnstra, Mirjam D. ; Hoogenboom, Ron L.A.P. ; Guzmán, Leonardo ; Hendriksen, Peter J.M. ; Bovee, Toine F.H. - \ 2018
Marine Drugs 16 (2018)12. - ISSN 1660-3397
Lipophilic marine toxins - Mouse bioassay - Neuro-2a assay

Marine biotoxins in fish and shellfish can cause several symptoms in consumers, such as diarrhea, amnesia, or even death by paralysis. Monitoring programs are in place for testing shellfish on a regular basis. In some countries testing is performed using the so-called mouse bioassay, an assay that faces ethical concerns not only because of animal distress, but also because it lacks specificity and results in high amounts of false positives. In Europe, for lipophilic marine biotoxins (LMBs), a chemical analytical method using LC-MS/MS was developed as an alternative and is now the reference method. However, safety is often questioned when relying solely on such a method, and as a result, the mouse bioassay might still be used. In this study the use of a cell-based assay for screening, i.e., the neuro-2a assay, in combination with the official LC-MS/MS method was investigated as a new alternative strategy for the detection and quantification of LMBs. To this end, samples that had been tested previously with the mouse bioassay were analyzed in the neuro-2a bioassay and the LC-MS/MS method. The neuro-2a bioassay was able to detect all LMBs at the regulatory levels and all samples that tested positive in the mouse bioassay were also suspect in the neuro-2a bioassay. In most cases, these samples contained toxin levels (yessotoxins) that explain the outcome of the bioassay but did not exceed the established maximum permitted levels.

The mystery of Tetrodotoxinen shellfish in the Netherlands manage the non-understood
Poelman, M. ; Gerssen, A. ; Heuvel-Greve, M.J. van den; Blanco Garcia, A. ; Klijnstra, M.D. ; Murk, A.J. - \ 2018
First Report on the Occurrence of Tetrodotoxins in Bivalve Mollusks in The Netherlands
Gerssen, Arjen ; Bovee, Toine ; Klijnstra, Mirjam ; Poelman, Marnix ; Portier, Liza ; Hoogenboom, Ron - \ 2018
Toxins 10 (2018)11. - ISSN 2072-6651
tetrodotoxins - bivalve mollusks - marine biotoxins - LC-MS/MS - neuroblastoma bioassay
Tetrodotoxin (TTX) is traditionally associated with seafood from tropical regions, but recently TTX was detected in bivalve mollusks in more temperate European waters. In The Netherlands it was therefore decided to monitor TTX in shellfish harvested from Dutch production areas. All shellfish production areas were monitored in 2015, 2016 and 2017. Samples were analyzed using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). In total 1063 samples were investigated, and the highest concentrations were observed in 2016, i.e., 253 g TTX/kg in oysters and 101 g TTX/kg in mussels. No TTX analogues, with the exception of 4-epi-TTX in one single sample, were found and contaminated samples also showed positive results in the neuro-2a bioassay. The occurrence of TTX seems to be consistent over the last three years with the highest concentrations observed annually in late June. The causative organism and the reasons why specific Dutch production areas are affected while others are not, are still unclear. Initially in The Netherlands an action limit of 20 g TTX/kg was used to ensure the safety of consumers (2016), but recently
The European Food Safety Authority (EFSA) established an acute reference dose, and based on a high portion size of consuming 400 g mussels, this dose was translated into a safe concentration of 44 g TTX per kg for shellfish. This concentration is now used as an action limit and TTX is formally included in the Dutch shellfish monitoring program.
Sensitive MS method for the detection of palytoxins in shellfish using li+ cationization
Klijnstra, M.D. ; Gerssen, A. - \ 2018
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Screening for the presence of lipophilic marine biotoxins in shellfish samples using the neuro-2a bioassay
Bodero Baeza, Marcia ; Bovee, Toine F.H. ; Wang, S. ; Hoogenboom, Ron L.A.P. ; Klijnstra, Mirjam D. ; Portier, Liza ; Hendriksen, Peter J.M. ; Gerssen, Arjen - \ 2018
Food Additives & Contaminants. Pt. A, Chemistry, Analysis, Control, Exposure & Risk Assessment 35 (2018)2. - ISSN 1944-0049 - p. 351 - 365.
diarrhetic shellfish poisons - Lipophilic marine biotoxins - MTT-test - neuro-2a cells - screeningfood safety
The neuro-2a bioassay is considered as one of the most promising cell-based in vitro bioassays for the broad screening of seafood products for the presence of marine biotoxins. The neuro-2a assay has been shown to detect a wide array of toxins like paralytic shellfish poisons (PSPs), ciguatoxins, and also lipophilic marine biotoxins (LMBs). However, the neuro-2a assay is rarely used for routine testing of samples due to matrix effects that, for example, lead to false positives when testing for LMBs. As a result there are only limited data on validation and evaluation of its performance on real samples. In the present study, the standard extraction procedure for LMBs was adjusted by introducing an additional clean-up step with n-hexane. Recovery losses due to this extra step were less than 10%. This wash step was a crucial addition in order to eliminate false-positive outcomes due to matrix effects. Next, the applicability of this assay was assessed by testing a broad range of shellfish samples contaminated with various LMBs, including diarrhetic shellfish toxins/poisons (DSPs). For comparison, the samples were also analysed by LC-MS/MS. Standards of all regulated LMBs were tested, including analogues of some of these toxins. The neuro-2a cells showed good sensitivity towards all compounds. Extracts of 87 samples, both blank and contaminated with various toxins, were tested. The neuro-2a outcomes were in line with those of LC-MS/MS analysis and support the applicability of this assay for the screening of samples for LMBs. However, for use in a daily routine setting, the test might be further improved and we discuss several recommended modifications which should be considered before a full validation is carried out.
Development and validation of a maleimide-based enzyme-linked immunosorbent assay for the detection of tetrodotoxin in oysters and mussels
Reverté, Laia ; Rambla-Alegre, Maria ; Leonardo, Sandra ; Bellés, Carlos ; Campbell, Katrina ; Elliott, Christopher T. ; Gerssen, Arjen ; Klijnstra, Mirjam D. ; Diogène, Jorge ; Campàs, Mònica - \ 2018
Talanta 176 (2018). - ISSN 0039-9140 - p. 659 - 666.
Liquid chromatography-tandem mass spectrometry (LC-MS/MS) - Maleimide-based enzyme-linked immunosorbent assay (mELISA) - Mussel - Oyster - Solid-phase extraction (SPE) clean-up - Tetrodotoxin (TTX)

The recent detection of tetrodotoxins (TTXs) in puffer fish and shellfish in Europe highlights the necessity to monitor the levels of TTXs in seafood by rapid, specific, sensitive and reliable methods in order to protect human consumers. A previous immunoassay for TTX detection in puffer fish, based on the use of self-assembled monolayers (SAMs) for the immobilization of TTX on maleimide plates (mELISA), has been modified and adapted to the analysis of oyster and mussel samples. Changing dithiol for cysteamine-based SAMs enabled reductions in the assay time and cost, while maintaining the sensitivity of the assay. The mELISA showed high selectivity for TTX since the antibody did not cross-react with co-occurring paralytic shellfish poisoning (PSP) toxins and no interferences were observed from arginine (Arg). Moreover, TTX-coated maleimide plates stored for 3 months at −20 °C and 4 °C were stable, thus when pre-prepared, the time to perform the assay is reduced. When analyzing shellfish samples, matrix effects and toxin recovery values strongly depended on the shellfish type and the sample treatment. Blank oyster extracts could be directly analyzed without solid-phase extraction (SPE) clean-up, whereas blank mussel extracts showed strong matrix effects and SPE and subsequent solvent evaporation were required for removal. However, the SPE clean-up and evaporation resulted in toxin loss. Toxin recovery values were taken as correction factors (CFs) and were applied to the quantification of TTX contents in the analysis of naturally-contaminated shellfish samples by mELISA. The lowest effective limits of detection (eLODs) were about 20 and 50 µg/kg for oyster extracts without and with SPE clean-up, respectively, and about 30 µg/kg for mussel extracts with both protocols, all of them substantially below the eLOD attained in the previous mELISA for puffer fish (230 µg/kg). Analysis of naturally-contaminated samples by mELISA and comparison with LC-MS/MS quantifications demonstrated the viability of the approach. This mELISA is a selective and sensitive tool for the rapid detection of TTX in oyster and mussel samples showing promise to be implemented in routine monitoring programs to protect human health.

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