TY - THES A1 - Drzymala, Sarah T1 - Instrumental analysis, metabolism and toxicity of cis- and trans-zearalenone and their biotransformation products N2 - Trans-Zearalenone(ZEN)is a non-steroidal estrogenic mycotoxin which frequently contaminates cereal grains worldwide. Ingestion of food and feed containing ZEN causes numerous mycotoxicoses in animals and possibly humans with pronounced estrogenic effects. Due to the trans-configurated double bond, ZEN isomerizes to the cis-configuration upon the influence of light. This work investigates the instrumental analysis, metabolism and toxicity of ZEN and cis- ZEN. The first part focused on the determination of ZEN in edible oils. Due to a maximum level of 400 µg/kg ZEN in the European Union (EU), reliable analytical methods are needed. A comprehensive method comparison proved dynamic covalent hydrazine chemistry (DCHC) to be the most suitable approach. Thus, an automated solid phase extraction (SPE) coupled online to high performance liquid chromatography (HPLC) was developed with the novelty of a covalent SPE step comprising the DCHC principle. The automated online system allows an accurate, selective and reliable quantification of ZEN in edible oils in compliance with EU performance criteria while significantly reducing workload and thereby personnel costs. In contrast to ZEN, reference standards and analytical methods are missing for cis- ZEN which causes a lack of data on the occurrence, fate and risks of cis-ZEN. Therefore, a native and an isotopically labeled cis-ZEN standard were synthesized and implemented in an existing stable isotope dilution analysis HPLC tandem mass spectrometry (HPLC-MS/MS) method. Using this method, a large extent of cis-ZEN formation was observed for ZEN contaminated maize germ oils when exposed to daylight which confirms that cis-ZEN can be a relevant food contaminant and should be considered in the analysis of food and feed. Furthermore, this work investigated the in vitro phase I metabolism of ZEN and cis-ZEN in rat and human liver microsomes by using HPLC-MS and -MS/MS analyses. The metabolic pathways of cis-ZEN were found to be essentially similar to ZEN including reduction and oxidation reactions generating α- and β-cis- zearalenol as well as 13- and 15-OH-cis-ZEN. A previously unidentified oxidative metabolic pathway for both isomers of ZEN results in the formation of cis-ZEN-11,12-oxide and ZEN-11,12-oxide in human liver microsomes. The estrogenicity of cis-ZEN and its reductive metabolites was assessed using the E-Screen assay. cis-ZEN proved to be slightly more estrogenic than ZEN. Biotransformation of cis-ZEN to β-cis-ZEL corresponds to a detoxification, whereas metabolism to α-cis-ZEL resembles a metabolic activation as its estrogenicity considerably exceeds that of cis-ZEN. The catecholic metabolites can be expected to show a decreased estrogenicity as demonstrated for 15-OH- ZEN. Independent of the estrogenic effects, the catecholic and epoxidic metabolites identified in this work can be expected to act genotoxic and carcinogenic. The epoxides in particular could fundamentally change the widely accepted view of ZEN causing adverse effects exclusively through endocrine disrupting actions. T3 - BAM Dissertationsreihe - 152 KW - Hydrazinchemie KW - Isotopenstandard KW - SPE-HPLC Automatisierung KW - Zearalenon KW - Isomerisierung PY - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:b43-386016 SN - 978-3-9817853-9-5 SN - 1613-4249 VL - 152 SP - 1 EP - 160 PB - Bundesanstalt für Materialforschung und -prüfung (BAM) CY - Berlin AN - OPUS4-38601 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Drzymala, Sarah A1 - Weiz, S. A1 - Heinze, J. A1 - Marten, S. A1 - Prinz, Carsten A1 - Zimathies, Annett A1 - Garbe, L.-A. A1 - Koch, Matthias T1 - Automated solid-phase extraction coupled online with HPLC-FLD for the quantification of zearalenone in edible oil N2 - Established maximum levels for the mycotoxin zearalenone (ZEN) in edible oil require monitoring by reliable analytical methods. Therefore, an automated SPE-HPLC online system based on dynamic covalent hydrazine chemistry has been developed. The SPE step comprises a reversible hydrazone formation by ZEN and a hydrazine moiety covalently attached to a solid phase. Seven hydrazine materials with different properties regarding the resin backbone, pore size, particle size, specific surface area, and loading have been evaluated. As a result, a hydrazine-functionalized silica gel was chosen. The final automated online method was validated and applied to the analysis of three maize germ oil samples including a provisionally certified reference material. Important performance criteria for the recovery (70–120 %) and precision (RSDr <25 %) as set by the Commission Regulation EC 401/2006 were fulfilled: The mean recovery was 78 % and RSDr did not exceed 8 %. The results of the SPE-HPLC online method were further compared to results obtained by liquid&–liquid extraction with stable isotope dilution analysis LC-MS/MS and found to be in good agreement. The developed SPE-HPLC online system with fluorescence detection allows a reliable, accurate, and sensitive quantification (limit of quantification, 30 µg/kg) of ZEN in edible oils while significantly reducing the workload. To our knowledge, this is the first report on an automated SPE-HPLC method based on a covalent SPE approach. KW - Dynamic covalent hydrazine chemistry (DCHC) KW - Maize germ oil KW - Food KW - Mycotoxin KW - ZEA KW - Isomerisation KW - Occurrence KW - Edible oil KW - SIDA KW - LC-MS/MS KW - Quantification PY - 2015 U6 - https://doi.org/10.1007/s00216-015-8541-5 SN - 1618-2642 SN - 1618-2650 VL - 407 IS - 12 SP - 3489 EP - 3497 PB - Springer CY - Berlin AN - OPUS4-33067 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Drzymala, Sarah A1 - Binder, J. A1 - Brodehl, Antje A1 - Penkert, M. A1 - Rosowski, M. A1 - Garbe, L.-A. A1 - Koch, Matthias T1 - Estrogenicity of novel phase I and phase II metabolites of zearalenone and cis-zearalenone N2 - Zearalenone and its cis-isomer, cis-zearalenone, are nonsteroidal mycotoxins that elicit an estrogenic response upon binding to the estrogen receptor. This study compares the estrogenicity of eleven congeners including novel metabolites as 15-OH-zearalenone, zearalenone-14-sulfate, α-cis-zearalenol and β-cis-zearalenol using the E-Screen assay. Overall, a change in the configuration from trans to cis retains significant estrogenic activity. In contrast, alterations of the aromatic moiety including hydroxylation and sulfation showed a markedly decreased estrogenicity when compared to zearalenone. KW - Zearalenone KW - Mycotoxin KW - MCF-7 KW - E-screen assay KW - Estrogenicity KW - Isomerization PY - 2015 U6 - https://doi.org/10.1016/j.toxicon.2015.08.027 SN - 0041-0101 SN - 1879-3150 VL - 105 SP - 10 EP - 12 PB - Elsevier CY - Oxford [u.a.] AN - OPUS4-34167 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Drzymala, Sarah A1 - Riedel, Juliane A1 - Köppen, Robert A1 - Garbe, L.-A. A1 - Koch, Matthias T1 - Preparation of 13C-labelled cis-zearalenone and its application as internal standard in stable isotope dilution analysis N2 - Pure U-[13C18]-labelled cis-zearalenone (cis-ZEA) has been prepared and characterised as internal standard (ISTD) for a reliable quantification of cis-ZEA in contaminated food and feed products. The cis-isomer of the naturally trans-configurated Fusarium mycotoxin zearalenone is often neglected. However, isomerisation easily occurs by exposure of ZEA to (UV-)light. Thus, the applicability of the new cis-ZEA ISTD was demonstrated in a long-term isomerisation study comparing naturally trans-ZEA-contaminated edible oil with spiked edible oil. To estimate the benefits of the newly prepared cis-ZEA ISTD, various approaches to quantify cis-ZEA by high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) were compared. As a result, a significant bias was revealed if no appropriate cis-ZEA standards are used. Furthermore, the new ISTD was applied to the analysis of 15 edible oils by stable isotope dilution analysis in combination with HPLC-electrospray ionisation-MS/MS. One of the maize germ oils showed the presence of cis-ZEA above LOD (>0.3 µg/kg), whereas two out of 15 maize germ oils were found to be contaminated with trans-ZEA (range 17.0-31.0 µg/kg). KW - ZEA KW - Zearalenone KW - Isomerisation KW - Occurrence KW - Edible oil KW - SIDA KW - HPLC-MS/MS KW - Quantification KW - LC-MS/MS PY - 2014 U6 - https://doi.org/10.3920/WMJ2013.1610 SN - 1875-0710 SN - 1875-0796 VL - 7 IS - 1 SP - 45 EP - 52 PB - Wageningen Academic Publishers CY - Wageningen AN - OPUS4-30126 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Weiz, S. A1 - Marten, S. A1 - Heinze, J. A1 - Drzymala, Sarah A1 - Koch, Matthias T1 - Zearalenon auf der Spur - Hochselektive Mykotoxin-Bestimmung in Speiseöl N2 - Das neue Verfahren ermöglicht die direkte Analyse des Mykotoxins Zearalenon aus Speiseölen in Kombination mit einer hochselektiven Probenvorbereitung. Dies wird in einem automatisierten Online-SPE-HPLC-Prozess abgebildet, sodass ene routinemäßige Kontrolle des EU-Grenzwertes möglich ist. KW - Mykotoxine KW - HPLC KW - Online-SPE KW - Hydrazine PY - 2014 SN - 0012-0413 SN - 1869-2214 IS - Januar SP - 24 EP - 27 PB - Wiss. Verl.-Ges. CY - Stuttgart AN - OPUS4-30080 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Drzymala, Sarah A1 - Herrmann, Antje A1 - Maul, Ronald A1 - Pfeifer, Dietmar A1 - Garbe, L.-A. A1 - Koch, Matthias T1 - In vitro phase I metabolism of cis-zearalenone N2 - The present study investigates the in vitro phase I metabolism of cis-zearalenone (cis-ZEN) in rat liver microsomes and human liver microsomes. cis-ZEN is an often ignored isomer of the trans-configured Fusarium mycotoxin zearalenone (trans-ZEN). Upon the influence of (UV-) light, trans-ZEN isomerizes to cis-ZEN. Therefore, cis-ZEN is also present in food and feed. The aim of our study was to evaluate the in vitro phase I metabolism of cis-ZEN in comparison to that of trans-ZEN. As a result, an extensive metabolization of cis-ZEN is observed for rat and human liver microsomes as analyzed by HPLC-MS/MS and high-resolution MS. Kinetic investigations based on the substrate depletion approach showed no significant difference in rate constants and half-lives for cis- and trans-ZEN in rat microsomes. In contrast, cis-ZEN was depleted about 1.4-fold faster than trans-ZEN in human microsomes. The metabolite pattern of cis-ZEN revealed a total of 10 phase I metabolites. Its reduction products, α- and β-cis-zearalenol (α- and β-cis-ZEL), were found as metabolites in both species, with α-cis-ZEL being a major metabolite in rat liver microsomes. Both compounds were identified by co-chromatography with synthesized authentic standards. A further major metabolite in rat microsomes was monohydroxylated cis-ZEN. In human microsomes, monohydroxylated cis-ZEN is the single dominant peak of the metabolite profile. Our study discloses three metabolic pathways for cis-ZEN: reduction of the keto-group, monohydroxylation, and a combination of both. Because these routes have been reported for trans-ZEN, we conclude that the phase I metabolism of cis-ZEN is essentially similar to that of its trans isomer. As trans-ZEN is prone to metabolic activation, leading to the formation of more estrogenic metabolites, the novel metabolites of cis-ZEN reported in this study, in particular α-cis-ZEL, might also show higher estrogenicity. KW - Cis-ZEN KW - Phase I metabolism KW - LC-MS/MS KW - HRMS KW - Depletion kinetics PY - 2014 U6 - https://doi.org/10.1021/tx500312g SN - 0893-228X SN - 1520-5010 VL - 27 IS - 11 SP - 1972 EP - 1978 PB - Soc. CY - Washington, DC, USA AN - OPUS4-32071 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Drzymala, Sarah A1 - Penkert, Martin A1 - Garbe, L.-A. A1 - Koch, Matthias T1 - In vitro Metabolism of Zearalenone T2 - 2. BAM/BfR-Kolloquium CY - Berlin DA - 2014-10-16 PY - 2014 AN - OPUS4-32052 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Drzymala, Sarah A1 - Herrmann, A. A1 - Binder, J. A1 - Riedel, Juliane A1 - Köppen, Robert A1 - Maul, Ronald A1 - Garbe, L.-A. A1 - Koch, Matthias T1 - Cis-Zearalenone: Instrumental Analysis, Metabolism and Toxicity T2 - Mycotoxin Workshop CY - Göttingen, Germany DA - 2014-06-16 PY - 2014 AN - OPUS4-30910 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Drzymala, Sarah A1 - Weiz, S. A1 - Heinze, J. A1 - Marten, S. A1 - Garbe, L.-A. A1 - Koch, Matthias T1 - Automated SPE-HPLC for Determination of Zearalenone in Edible Oil T2 - RAFA 2013 (Recent Advances in Food Analysis) CY - Prague, Czech Republic DA - 2013-11-04 PY - 2013 AN - OPUS4-29630 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Weiz, S. A1 - Marten, S. A1 - Heinze, J. A1 - Drzymala, Sarah A1 - Koch, Matthias T1 - Unpolarer Analyt, unpolare Matrix: das Mykotoxin Zearalenon in Öl KW - Mykotoxine KW - HPLC KW - Online-SPE KW - Hydrazine PY - 2013 SN - 1439-9598 SN - 1521-3854 VL - 61 IS - Dezember SP - 1241 EP - 1243 PB - Wiley-VCH Verl. CY - Weinheim AN - OPUS4-29811 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Drzymala, Sarah A1 - Marten, Silvia A1 - Risch, Stefan A1 - Koch, Matthias A1 - Panne, Ulrich T1 - SPE-HPLC Online Coupling for the Determination of Zearalenone in Edible Oil: First Results T2 - JCF Frühjahrssymposium CY - Berlin, Germany DA - 2013-03-06 PY - 2013 AN - OPUS4-27831 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Drzymala, Sarah A1 - Marten, Silvia A1 - Risch, Stefan A1 - Koch, Matthias A1 - Panne, Ulrich T1 - Development of a SPE-HPLC Online Coupling for the Determination of Zearalenone in Edible Oil T2 - ANAKON 2013 CY - Essen, Germany DA - 2013-03-04 PY - 2013 AN - OPUS4-27830 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Drzymala, Sarah A1 - Risch, St. A1 - Heinze, J. A1 - Marten, S. A1 - Garbe, L.-A. A1 - Koch, Matthias A1 - Panne, Ulrich T1 - Online Coupling of Covalent Solid Phase Extraction to Liquid Chromatography for Automated Analysis of Zearalenone T2 - Tag der Chemie CY - Berlin, Germany DA - 2013-06-27 PY - 2013 AN - OPUS4-28750 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Drzymala, Sarah A1 - Koch, Matthias A1 - Marten, Silvia A1 - Risch, Stefan T1 - Highly Efficient Extraction and Clean-up of Zearalenone from Edible Oil T2 - 34. Mykotoxin-Workshop der Gesellschaft für Mykotoxinforschung e. V. CY - Braunschweig, Germany DA - 2012-05-14 PY - 2012 AN - OPUS4-25990 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Drzymala, Sarah A1 - Marten, S. A1 - Risch, St. A1 - Koch, Matthias T1 - Optimized Extraction and Clean-up of Zearalenone from Edible Oils T2 - 4th Potsdam Colloquium on Bioanalysis CY - Potsdam, Germany DA - 2012-11-08 PY - 2012 AN - OPUS4-27406 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Drzymala, Sarah A1 - Kraus, Werner A1 - Emmerling, Franziska A1 - Koch, Matthias T1 - (3S,7R)-7,14,16-trihydroxy-3-methyl-3,4,5,6,7,8,9,10,11,12-decahydro-1H-2-benzoxacyclotetradecin-1-one N2 - The asymmetric unit of the title compound, C18H26O5, which is known as α-zearalanol, contains two molecules having the same conformation, with a r.m.s. deviation of less than 0.03 Å for all non-H atoms. In each independent molecule, an intramolecular O—H···O hydrogen bond stabilizes the molecular conformation. In the crystal, O—H···O hydrogen bonds link the molecules, forming infinite chains along [110] and [1¯10]. KW - Alpha-zearalanol KW - Zearalenon KW - Estrogen KW - Growth promoter KW - Ralgro PY - 2012 U6 - https://doi.org/10.1107/S1600536812041141 SN - 1600-5368 VL - 68 IS - 11 SP - o3071, sup-1 - sup-11 PB - Munksgaard CY - Copenhagen AN - OPUS4-26771 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Drzymala, Sarah A1 - Kraus, Werner A1 - Emmerling, Franziska A1 - Koch, Matthias T1 - (3S)-14,16-Dihydroxy-3-methyl-3,4,5,6,9,10,11,12-octa-hydro-1H-2-benzoxacyclotetradecine-1,7(8H)-dione (zearalanone) monohydrate N2 - The absolute configuration of the title compound, C18H24O5·H2O, was not been determined by anomalous-dispersion effects, but has been assigned by reference to an unchanging chiral centre in the synthetic procedure. Intramolecular O—H···O hydrogen bonds stabilize the molecular conformation. In the crystal, O—H···O hydrogen bonds link the main molecules and the water molecules, forming an infinite three-dimensional network. KW - Zearalanone KW - Crystal structure PY - 2012 U6 - https://doi.org/10.1107/S1600536812018168 SN - 1600-5368 VL - 68 IS - 5 SP - o1577, sup1 EP - sup7 PB - Munksgaard CY - Copenhagen AN - OPUS4-25832 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Köppen, Robert A1 - Riedel, Juliane A1 - Proske, Matthias A1 - Drzymala, Sarah A1 - Rasenko, Tatjana A1 - Durmaz, V. A1 - Weber, M. A1 - Koch, Matthias T1 - Photochemical trans-/cis-isomerization and quantitation of zearalenone in edible oils N2 - The emphasis of the present work was to investigate the photochemical conversion of trans- to cis-zearalenone in edible oils under real-life conditions. For quantitation purposes a cis-zearalenone standard was synthesized and characterized for its identity and purity (≥95%) by 1H NMR, X-ray crystallography, HPLC fluorescence and mass spectrometric detection. In a sample survey of 12 edible oils (9 corn oils, 3 hempseed oils) from local supermarkets all corn oils contained trans-zearalenone (median 194 µg/kg), but no cis-zearalenone was detected. For alteration studies trans-zearalenone contaminated corn oils were exposed to sunlight over 4 and 30 weeks, revealing an obvious shift toward cis-zearalenone up to a cis/trans ratio of 9:1 by storage in colorless glass bottles. Irradiation experiments of trans-zearalenone in different organic solvents confirmed the preferred formation of cis-zearalenone possibly caused by entropic effects rather than by enthalpic entities as investigated by quantum chemical and classical force field simulations. KW - Fusarium mycotoxins KW - Food KW - Analysis KW - Occurrence PY - 2012 U6 - https://doi.org/10.1021/jf3037775 SN - 0021-8561 SN - 1520-5118 VL - 60 IS - 47 SP - 11733 EP - 11740 PB - American Chemical Society CY - Columbus, Ohio AN - OPUS4-27375 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -