TY - CONF A1 - Schneider, Rudolf T1 - Anthropogenic markers quantified by rapid immunochemical methods – what can their occurrence in wastewater, surface water, and drinking water tell us? N2 - In urban waters, a multitude of organic micropollutants, often termed emerging pollutants, has been found over the last decades. Analytical methods suitable for trace analysis are needed that are desirably also fast, inexpensive and, if possible, robust and portable. Immunoanalytical, i.e., antibody-based, methods which are available in a broad range of formats, can be profitably used here to screen for the distribution and to monitor the trends of concentration levels of contaminants of emerging concern in the environment. Some of these formats are single-analyte but high-throughput methods. To use them wisely, indicator substances, sometimes called anthropogenic markers, should be selected and used in screening approaches, i.e., as indicators for contamination and the pre-selection of samples at which to have a closer look by multiplex methods like LC-MS/MS. Other methods are suitable to be performed on portable instrumentation in the field (on-site) or in facilities such as wastewater treatment plants for on-line monitoring of the treatment and elimination process. Furthermore, array technologies have been established that allow for parallel (multiplex) analysis of several analytes of interest. The microtiter-plate based ELISA (Enzyme-linked Immunosorbent Assay) is the method of choice for the analysis of a large number of samples [1]. ELISAs are available to monitor for anthropogenic markers such as the antiepileptic carbamazepine, the analgesic diclofenac, the antihistaminic cetirizine, the steroid hormone estrone, the antimicrobial sulfamethoxazole, psychoactive caffeine and cocaine, the priority pollutant bisphenol A, and the bile acid isolithocholic acid. For on-site screening and monitoring, simpler formats, like mix-and-read assays, e.g., the Fluorescence Polarization Immunoassay (FPIA) [2] or Lateral-flow Immunoassays (LFIA) [3] are more suitable tools, the latter based on dipsticks or little cassettes, with which users have become very familiar during the COVID-19 pandemic via rapid antigen tests. The suitability of multi-analyte formats such as immunomicroarrays depends on the choice of a signal-producing system that provides small uncertainties and good reproducibility of the measurements. Bead-based (“suspension”) arrays, read out in flow cytometers, are a powerful platform for multiplex assays [4]. Electrochemical formats, run on portable devices, provide additional advantages as no light source is required. They are most promising for stand-alone analysers and biosensors [5]. The speed, low cost and on-site capabilities of these methods allow to gather a lot more data on anthropogenic compounds which enables to quantify inputs, differences in degradation power of elimination processes, dilution phenomena and a more precise image of individual water cycles which is demonstrated by several examples. T2 - EuChemS Chemistry Congress ECC8 CY - Lisbon, Portugal DA - 28.08.2022 KW - Biosensor KW - Immunoassay KW - Antibody PY - 2022 AN - OPUS4-56783 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schneider, Rudolf T1 - Environmental Analysis at BAM N2 - The talk is to introduce BAM, the Department of Analytical Chemistry; Reference Materials and the Division of Environmental Analysis and its works to the audience. T2 - Seminar des Centre for Environmental and Marine Studies (CESAM) der Universität Aveiro CY - Aveiro, Portugal DA - 12.09.2022 KW - Immunoassay KW - Biosensor KW - Antibody PY - 2022 AN - OPUS4-56784 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schneider, Rudolf T1 - Field Analysis by Antibody-based Analytical Methods: The Importance of Advanced Materials N2 - Analytical methods based on the selectivity of antibodies, often called immunoassays, are a back-bone of clinical laboratory diagnostics. To bring them to the field, i.e., to make immunoanalytical methods portable, hopefully even faster, more sensitive, and robust, advanced materials are re-quired. Materials can be novel labels, e.g., chemical or particle labels, such as fluorophores or na-noparticles. Carrier particles, such as magnetic or polymer beads, make it possible to adopt the as-says to meso- or microfluidic set-ups and encoding them opens the path to multiplex analysis. Spe-cialty electrodes can enable for higher sensitivity in electrochemical detection. Without research into better materials, efforts to bring analysis to the point-of-need will not bear fruit. T2 - Kolloquium des Aveiro Institut of Materials (CICECO) der Universität Aveiro CY - Aveiro, Portugal DA - 20.09.2022 KW - Immunoassay KW - Biosensor KW - Antibody PY - 2022 AN - OPUS4-56785 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Frenzel, Olivia T1 - Identification of Metabolites and Transformation Products of Bisphenols N2 - Bisphenol A (BPA) is used in Polycarbonate, Polyacrylic resins, Polysulfones, Epoxy resins, and Polyetherimides. It is also used in recycled Polyvinyl chloride [1–3]. BPA has been classified as a substance of very high concern (SVHC) under REACH [4] due to its endocrine disrupting properties. The German competent authorities want to reduce the content of BPA in the environment. Possible substitutes for BPA are Bisphenol B (BPB), Bisphenol E (BPE), Bisphenol F (BPF), or Bisphenol S (BPS), as they are similar in structure. BPA-based materials have a wide area of application especially outside. These outdoor applications are exposed to different external influences, including physical, biological, mechanical and chemical influences. This results in damage and aging of the material with leaching or migration into the environment. There, the substance is transformed by various transformation processes. The emerging metabolites and transformation products (TPs) can have different properties than the parent substance. Understanding the fate and behavior of the emerging pollutants is very important. Therefore, different transformation products of selected bisphenols will be generated and analyzed: To investigate the fate of different bisphenols in water treatment plants technical transformation products [5] will be generated by chlorination, ozonization, the Fenton reaction, and UV-radiation. For the investigation of the fate of Bisphenols in surface water global radiation will be simulated with UVA-lamps Simulation of phase-I-metabolism with an electrochemical cell coupled to mass spectrometry (EC-MS). T2 - Anakon 2023 CY - Vienna, Austria DA - 11.04.2023 KW - Transformation products KW - Bisphenols KW - Metabolites KW - Wastewater KW - Environment PY - 2022 AN - OPUS4-57374 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Bilsel, M. A1 - Gökçen, T. A1 - Binici, B. A1 - Isleyen, A. A1 - Piechotta, Christian A1 - Kar-wai, A. A1 - Krylov, A. A1 - Miheeva, A. A1 - Beliakov, M. A1 - Palagina, M. A1 - chenko, Irina Tka A1 - Perkola, N. A1 - Lewin, M. A1 - Hua, T. T1 - High polarity analyte(s) in aqueous media: determination of L-PFOA and L-PFOS in ground water N2 - The CCQM-K156 comparison was coordinated by TUBITAK UME on behalf of the Organic Analysis Working Group (OAWG) of the Comité Consultatif pour la Quantité de Matière (CCQM) for National Measurement Institutes (NMIs) and Designated Institutes (DIs) which provide measurement services in organic analysis under the 'Comité International des Poids et Mesures' Mutual Recognition Arrangement (CIPM MRA). Perfluoro alkyl substances (PFAS) such as PFOS and PFOA have been used in numerous industrial applications and products. Because of their high stability and resistance to biodegradation, atmospheric photooxidation, direct photolysis and hydrolysis, they are extremely persistent in the environment. The European Union (EU) Water Framework Directive lists PFOS as a priority hazardous substance that poses a significant risk to the aquatic environment. The use of PFOS-containing Aqueous Film-Forming Foams (AFFFs) has been banned since June 2011 in the EU. As relatively water-soluble, effectively non-degradable compounds, PFOS and PFOA migrate to ground water. They are not removed in the conventional drinking water treatment, and therefore cause health risks in polluted areas. The EU Drinking Water Directive and the European Commission has proposed a limit value of 100 ng/L for the sum of 20 PFAS, including PFOS and PFOA. This study provides the means for assessing measurement capabilities for determination of high polarity measurands in a procedure that requires extraction, clean-up, analytical separation and detection. Successful participation in CCQM-K156 demonstrates measurement capabilities in determining mass fraction of organic compounds, with a molecular mass of 200 g/mol to 700 g/mol, having high polarity pKow -2, in a mass fraction range from 0.5 ng/kg to 500 ng/kg in aqueous media. Nine NMIs and DIs participated in the CCQM-K156 key comparison. Seven institutes reported their results. SPE was applied in the sample pre-treatment and LC-MS was applied for detection. All participating laboratories applied isotope dilution mass spectrometry (IDMS) techniques for quantification. Participants established the metrological traceability of their results using certified reference materials (CRMs) from NMIs with stated traceability; where commercially available high purity materials were used the purity was determined in-house. The CCQM-K156 results for L-PFOA and L-PFOS range from 2.75 ng/kg to 5.50 ng/kg with a % RSD of 19.5 % for L-PFOA and from 2.04 ng/kg to 4.45 ng/kg with a % RSD of 21.3 % for L-PFOS. The KCRV was assigned using a Hierarchical Bayesian Random Effects Model (HB REM) estimator from the values reported by six of the participants. One participant result of L-PFOS and one result of L-PFOA were excluded from the KCRV for technical reasons. The KCRV was 4.9 ng/kg ± 0.4 ng/kg for L-PFOA and 3.8 ng/kg ± 0.4 ng/kg for L-PFOS. The six institutes that were included in the assignment of consensus KCRV all agreed within their standard uncertainties. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database https://www.bipm.org/kcdb/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA). KW - PFAS KW - Surface water KW - ILC KW - CCQM PY - 2022 DO - https://doi.org/10.1088/0026-1394/59/1A/08016 VL - 59 IS - 1A SP - 1 EP - 3 PB - IOP Publishing AN - OPUS4-58941 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Piechotta, Christian T1 - EDC-WFD: A project to deliver reliable measurements for better monitoring survey and risks assessments. N2 - Monitoring programs should generate high-quality data on the concentrations of substances and other pollutants in the aquatic environment to enable reliable risk assessment. Furthermore, the need for comparability over space and time is critical for analysis of trends and evaluation of restoration of natural environment. Additionally, research work and exercises at the European level have highlighted that reliable measurements of estrogenic substances at the PNEC level are still challenging to achieve. The project EDC-WFD Metrology for monitoring endocrine disrupting compounds under the EU Water Framework Directive aims to develop traceable analytical methods for determining endocrine disrupting compounds and their effects, with a specific focus on three estrogens of the first watch list (17-beta-estradiol (17βE2), 17-alpha-ethinylestradiol (EE2), and estrone (E1)). Estrogens 17-alpha-estradiol (17E2) and estriol (E3) will be included to demonstrate the reliability of the developed methods and to support the requirements of Directive 2013/39/EC, Directive 2009/90/EC and Commission Implementation Decision (EU) 2018/840, hence improving the comparability and compatibility of measurement results within Europe. During the EDC-WFD project four selected effect-based methods (EBM) will be deeply investigated in order to improve their rationale use and their support in water quality assessment. In particular, the EBM sensitivity, specificity and accuracy on reference materials with single or mixture solutions of the five substances at a concentration of EQS values will be explored. This contribution will present the objectives and methods applied within the EDC-WFD project. T2 - EuChemS CY - Lisbon, Portugal DA - 29.08.2022 KW - WFD KW - Estrogens KW - Whole water samples KW - EQS PY - 2022 AN - OPUS4-57096 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Piechotta, Christian T1 - A method for monitoring estrogens in whole surface waters by GC-MS/MS N2 - Natural and synthetic estrogens are key endocrine-disrupting chemicals. Despite occurring at ultra-trace levels (below ng L-1), it is believed that they are contributing to an increase in feminized fish and other endocrine disruptive effects, and hence, their inclusion in the Watch list was not unexpected. One of the main sources ofestrogens to surface waters is wastewater effluent. Once in surface waters, they can partition into different compartments, i.e., water and suspended particulate matter. For this reason, there is an urgent need for a methodology to monitor estrogen levels below the environmental quality standards (EQS) set by the Water Framework Directive requirements. In this study, a precise and accurate gas chromatography-mass spectrometry method (GC-MS/MS) for the analysis of estrone (E1), 17β-estradiol (17β-E2), 17α-estradiol (17α- E2), 17-alpha-ethinylestradiol (EE2), and estriol (E3) in whole water samples with ng L-1 limit of quantification (LOQ) was developed and validated in accordance with CEN/TS 16800:2020 guidelines. T2 - IMSC 2022 CY - Maastricht, Netherlands DA - 27.08.2022 KW - EDC KW - WFD KW - GC-MS PY - 2022 AN - OPUS4-57097 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schneider, Rudolf T1 - Bestimmung von anthropogenen Markerkonzentrationen für die SARS-CoV-2 Quantifizierung mittels eines Hochdurchsatzverfahrens (ELISA) N2 - Vortrag zum Kick-off des Projektes. Ziel des Projektes ist es, die Eignung der Bestimmung eines oder mehrerer der genannten anthropogenen Marker im Abwasser zur Volumenkorrektur („Kalibration“) im SARS-CoV-2-Monitoring von Abwässern auf der Basis eines von der BAM zu erhebenden Messdatensatzes zu evaluieren. T2 - Kick-off Meeting zum Projekt MARKERIA (VH1802) CY - Berlin, Germanyy DA - 01.04.2022 KW - Biosensoren KW - Immunoassay KW - SARS-CoV-2 PY - 2022 AN - OPUS4-57109 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schneider, Rudolf T1 - Bestimmung von anthropogenen Markerkonzentrationen für die SARS-CoV-2 Quantifizierung mittels eines Hochdurchsatzverfahrens (ELISA) N2 - ESI-CorA ist ein nationales Verbundprojekt mit dem Ziel des Nachweises von SARS-CoV-2 im Abwasser. Das UBA hat als Teilprojekt das Projekt MARKERIA (VH1802) an die BAM vergeben. Ziel des Projektes ist es, die Eignung der Bestimmung eines oder mehrerer der genannten anthropogenen Marker im Abwasser zur Volumenkorrektur („Kalibration“) im SARS-CoV-2-Monitoring von Abwässern auf der Basis eines von der BAM zu erhebenden Messdatensatzes zu evaluieren. T2 - 12. Sitzung des Begleitkreises des nationalen Verbundprojektes ESI-CorA CY - Online meeting DA - 13.06.2022 KW - Biosensoren KW - Immunoassay KW - SARS-CoV-2 PY - 2022 AN - OPUS4-57110 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schneider, Rudolf T1 - Bestimmung von anthropogenen Markerkonzentrationen für die SARS-CoV-2 Quantifizierung mittels eines Hochdurchsatzverfahrens (ELISA) N2 - 1. Projektreffen mit dem Umweltbundesamt für das Projekt MARKERIA (VH1802). Ziel des Projektes ist es, die Eignung der Bestimmung eines oder mehrerer der genannten anthropogenen Marker im Abwasser zur Volumenkorrektur („Kalibration“) im SARS-CoV-2-Monitoring von Abwässern auf der Basis eines von der BAM zu erhebenden Messdatensatzes zu evaluieren. T2 - 1. Projekttreffen mit dem UBA zum Projekt MARERIA (VH1802) CY - Berlin, Germany DA - 15.09.2022 KW - Biosensoren KW - Immunoassay KW - SARS-CoV-2 PY - 2022 AN - OPUS4-57111 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schneider, Rudolf T1 - Bestimmung von anthropogenen Markerkonzentrationen für die SARS-CoV-2 Quantifizierung mittels eines Hochdurchsatzverfahrens (ELISA) N2 - 2. Projekttreffen mit dem Umweltbundesamt zum Projekt MARKERIA (VH1802). Ziel des Projektes ist es, die Eignung der Bestimmung eines oder mehrerer der genannten anthropogenen Marker im Abwasser zur Volumenkorrektur („Kalibration“) im SARS-CoV-2-Monitoring von Abwässern auf der Basis eines von der BAM zu erhebenden Messdatensatzes zu evaluieren. T2 - 2. Projekttreffen mit dem UBA zum Projekt MARKERIA (VH1802) CY - Berlin, Germany DA - 25.11.2022 KW - Biosensoren KW - Immunoassay KW - SARS-CoV-2 PY - 2022 AN - OPUS4-57112 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Philipp, Rosemarie A1 - Lalere, B. A1 - Gantois, F. A1 - Sánchez, C. A1 - Sáez, A. A1 - Bebić, J. A1 - Banjanac, K. A1 - Alexopoulos, Ch. A1 - Kakoulides, E. A1 - Claramunt, A. V. A1 - Janko, P. A1 - Jotanovic, A. A1 - Hafner-Vuk, K. A1 - Buzoianu, M. A1 - Mihail, R. A1 - Fernández, M. M. A1 - Etcheverry, J. A1 - Mbithi Muendo, B. A1 - Muriira Karau, G. A1 - Silva, A. A1 - Almirón, F. A1 - Marajh, D. A1 - Makgatho, P. A1 - Visser, R. A1 - Alaskar, A. R. A1 - Alosaimi, A. A1 - Alrashed, M. A1 - Yılmaz, H. A1 - Ün, İ. A1 - Gündüz, S. A1 - Topal, K. A1 - Bilsel, M. A1 - Karasinski, J. A1 - Torres, J. T1 - Supplementary comparison study - measurement capabilities for the quantification of ethanol in water N2 - The accurate quantification of ethanol in water is essential for forensic applications such as blood and breath alcohol testing and for commercial applications such as the assessment of alcoholic beverages. The intercomparison EURAMET.QM-S14 is part of a capacity building project named ALCOREF “Certified forensic alcohol reference materials” that is running within the European Metrology Programme for Innovation and Research (EMPIR). The intercomparison should allow project partners and other interested National Metrology Institutes (NMIs) and Designated Institutes (DIs) to benchmark their analytical methods for the quantification of ethanol in water. The study plan was agreed by the European Association of National Metrology Institutes (EURAMET) Subcommittee Bio- and Organic Analysis (SCBOA) and the Organic Analysis Working Group (OAWG) of the Comité Consultatif pour la Quantité de Matière (CCQM) in February and April 2019, respectively. The intercomparison was coordinated by BAM. Two concentration levels relevant for the calibration and verification of evidential breath alcohol analysers were distributed to study participants. Fifteen institutes from 15 countries registered for the intercomparison and returned results. Participants mostly applied gas chromatography with flame ionisation detection (GC-FID) or mass spectroscopy (GC-MS), one participant used titrimetry and one participant employed a test bench for breath analyser calibration (“bubble train”). Participants did either in-house purity assessment of their commercial ethanol calibrants by Karl-Fischer titration, chromatographic methods, quantitative nuclear magnetic resonance spectroscopy (qNMR) and/or density measurements; or they used ethanol/water Certified Reference Materials (CRMs) from NMIs/DIs for calibration. CCQM OAWG agreed to use a consensus value from participants results that utilizes the reported uncertainties as Key Comparison Reference Value (KCRV). The Gaussian Random effects model with Hierarchical Bayesian solution (HB-REM) is a reasonable approach in this case. The KCRVs and Degrees of Equivalence (DoEs) were calculated with the NIST consensus builder version 1.2 Hierarchical Bayes procedure. Successful participation in the interlaboratory comparison has demonstrated the capabilities in determining the mass fraction of ethanol in aqueous matrices in the range 0.1 mg/g to 8 mg/g. Fourteen out of 15 participants have successfully quantified both samples, one participant successfully quantified only the lower-level (0.6 mg/g) sample. KW - Certified reference material KW - EURAMET KW - EMPIR KW - ALCOREF KW - Ethanol in water KW - Supplementary comparison PY - 2022 DO - https://doi.org/10.1088/0026-1394/59/1A/08015 VL - 59 IS - 1A SP - 08015 PB - IOP Publishing AN - OPUS4-55889 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Steinhäuser, Lorin A1 - Piechotta, Christian A1 - Westphalen, Tanja A1 - Kaminski, Katja T1 - Evaluation, comparison and combination of molecularly imprinted polymer solid phase extraction and classical solid phase extraction for the preconcentration of endocrine disrupting chemicals from representative whole water samples N2 - Estrogens are endocrine disrupting chemicals and of high concerns due to demonstrated harmful effects on the environment and low effect levels. For monitoring and risk assessment, several estrogens were included in the "watch list" of the EU Water Framework Directive which sets very low environmental quality standard (EQS) levels for Estrone (E1) and 17β-Estradiol (E2) of 0.4 ng L−1 and for 17α-Ethinylestradiol (EE2) of 0.035 ng L−1 requiring sensitive detection methods, as well as extensive sample preparation. A sensitive, derivatization-free, isotope dilution calibration HPLC-MS/MS method for a panel of 5 selected estrogens (including the 3 estrogens of the EU WFD watchlist), and a procedure for the reproducible preparation of a representative whole water matrix including mineral water, humic acids and solid particulate matter are presented. These are used in a diligent comparison of classical solid phase extraction (SPE) on hydrophilic-lipophilic balanced (HLB) phase to SPE on an estrogen-specific molecularly imprinted polymer phase (MISPE) for ultra-trace levels of the analytes (1–10 ng L−1). Additionally, a two-step procedure combining HLB SPE disks followed by MISPE is evaluated. The tow-step procedure provides superior enrichment, matrix removal and sample throughput while maintaining comparable recovery rates to simple cartridge SPE. Estimated method quantification limits (MQLs) range from 0.109–0.184 ng L−1 and thus meet EQS-levels for E1 and E2, but not EE2. The representative whole water matrix provides a reproducible comparison of sample preparation methods and lays the foundation for a certified reference material for estrogen analysis. The presented method will serve as the basis for an extended validation study to assess its use for estrogen monitoring in the environment. KW - Estrogens KW - Whole water samples KW - Molecular imprinted polymers KW - EU-WFD PY - 2022 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-563547 DO - https://doi.org/10.1016/j.talo.2022.100163 SN - 2666-8319 VL - 6 SP - 1 EP - 5 PB - Elsevier B.V. CY - Amsterdam, Niederlande AN - OPUS4-56354 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Riedel, Soraya T1 - Tracing toxic fumonisins - Fumonisin sensing by Aspergillus niger Fumonisin Amine Oxidase (AnFAO) and amperometric hydrogen peroxide detection N2 - Fumonisins are a class of toxic secondary metabolites produced by various Fusarium species. The two most important producers of fumonisins are F. verticillioides and F. proliferatum but also Aspergillus niger is known to produce fumonisins. Most frequently they occur on maize, but also other grains can be contaminated with this group of mycotoxins. Exposure to fumonisins by dietary intake can have serious health effects on farm animals such as equine leukoencephalomalacia and porcine pulmonary oedema and is associated with neural tube defects and esophageal cancer in humans. Thus, the European Commission sets legal limits for fumonisins in foodstuffs. The detection of fumonisins is frequently performed in laboratories by chromatographic methods, which are costly and require trained personnel. Simplifying the analysis is therefore a major goal using portable detection systems. Electrochemical enzymatic biosensors offer great promise to meet this demand. Here we report for the first time an enzymatic fumonisin sensing approach with amperometric detection. For this purpose, an Aspergillus niger fumonisin amine oxidase (AnFAO) catalyzing the oxidative deamination of fumonisins, producing hydrogen peroxide, was recombinantly produced in E. coli. For the first time, the specific enzyme activity of AnFAO was determined using a horseradish peroxidase-based fluorescence assay. It was found that the specific activity of AnFAO using 20 μM Fumonisin B1 as substrate is higher than for 20 μM Fumonisin B2 with 0.122 U mg-1 and 0.058 U mg-1, respectively. It was possible to show a dependence of enzyme activity with enzyme – and substrate-concentration. For fumonisin B1 detection, the enzyme was coupled covalently to magnetic particles and the enzymatically produced H2O2 was detected amperometrically in a flow injection system using Prussian blue carbon electrodes. The developed method allows to quantify fumonisin B1 concentrations down to 1.5 µM and demonstrates that the recombinantly produced AnFAO was able to deaminate different concentrations of fumonisin even in immobilized form. Thus, this enzyme is well suited to develop an enzyme based electrochemical biosensor for fumonisin contaminated food and feed. T2 - Posterschau Adlershofer Forschungsforum CY - Berlin, Germany DA - 11.11.2022 KW - Biosensor KW - Amperometry KW - Mycotoxins KW - Electrochemical sensor KW - Recombinant protein expression PY - 2022 N1 - Geburtsname von Riedel, Soraya: Höfs, S. - Birth name of Riedel, Soraya: Höfs, S. AN - OPUS4-56248 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Döring, Sarah T1 - Cost-effective method for full-length sequencing of monoclonal antibodies N2 - With the development of hybridoma technology, monoclonal antibodies are increasingly important in therapeutic and analytical applications. For recombinant expression in mammalian cells, knowledge of the variable regions of both, heavy (VH) and light (VL) chain, is a necessary prerequisite for generating expression plasmids. Furthermore, cloning antibody sequences including constant regions CH1 to CH3 and CL reduces impact of hybridoma cell loss and associated full natural antibody sequence leakage. Here, we show a cost-effective workflow for amplification of IgG antibody variable regions in combination with advanced methods for full-length cloning of monoclonal antibodies. T2 - Adlershofer Forschungsforum CY - Berlin, Germany DA - 11.11.2022 KW - Monoclonal Antibodies KW - DNA-Sequencing KW - PCR PY - 2022 AN - OPUS4-56275 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Frenzel, Olivia T1 - Identification of Metabolites and Transformation Products of Bisphenols N2 - Bisphenol A (BPA) is widely used in Polycarbonate (PC), Polyacrylic resins (PAR), Polysulfones (PSU), Epoxy resins (ER), and Polyetherimides (PEI). It is also used in recycled Polyvinyl chloride (PVC) [1–3]. These BPA-containing materials have a wide area of application, especially outside. All outdoor applications are exposed to a variety of environmental impacts, like temperature, solar radiation (physical influences), chemical influences (rain or ozone), biological influences (microorganisms), and mechanical influences (hail, sand). These impacts are damaging and aging the material which can be followed by leaching or migration of pollutants like Bisphenol A into the environment. Understanding the fate and behavior of the released pollutants is very important. Therefore, different transformation products of selected Bisphenols will be generated and analyzed. T2 - 33rd International Symposium on Chromatography – ISC 2022 CY - Budapest, Hungary DA - 18.09.2022 KW - Environment KW - Transformation products KW - Bisphenols KW - Metabolites KW - Wastewater PY - 2022 AN - OPUS4-56202 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Frenzel, Olivia T1 - Identification of transformation products and metabolites of selected Bisphenols N2 - Bisphenol A (BPA) is widely used in many Polymers. Many outdoor applications are made from BPA-based materials which are exposed to a variety of environmental impacts, like temperature, solar radiation, rain, and others. This could lead to damaging and aging of the material which might cause leaching or migration of pollutants into the environment. To understand the fate and behavior different transformation products of selected Bisphenols will be generated and analyzed. T2 - Umwelt2022 CY - Emden, Germany DA - 05.09.2022 KW - Environment KW - Bisphenols KW - Transformation products KW - Metabolites KW - Wastewater PY - 2022 AN - OPUS4-56203 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Ecke, Alexander T1 - An Immunosensor for Pharmaceutical Contaminants in Water Supply N2 - Contamination of drinking water with pharmaceuticals is an emerging problem and a potential health threat. Immunochemical methods based on the binding of the analyte to specific antibodies enable fast & cost-effective on-site analyses. Magnetic bead-based immunoassays (MBBAs) allow for implementation into an immunosensor for online testing. Particles are prepared by either coupling the analyte molecule (diclofenac) or the respective antibody (amoxicillin) to the surface. For miniaturization, detection is performed electrochemically (chronoamperometry) on a microfluidic chip. The developed immunosensor will enable detection of pharmaceuticals directly in water supply pipes. T2 - Analytica Conference CY - München, Germany DA - 21.06.2022 KW - Immunosensor KW - Magnetic Particles KW - Microfluidics KW - Amperometry KW - Diclofenac KW - Amoxicillin PY - 2022 AN - OPUS4-55381 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Langenhan, Jennifer A1 - Jaeger, Carsten A1 - Baum, K. A1 - Simon, M. A1 - Lisec, Jan T1 - A Flexible Tool to Correct Superimposed Mass Isotopologue Distributions in GC‐APCI‐MS Flux Experiments N2 - The investigation of metabolic fluxes and metabolite distributions within cells by means of tracer molecules is a valuable tool to unravel the complexity of biological systems. Technological advances in mass spectrometry (MS) technology such as atmospheric pressure chemical ionization (APCI) coupled with high resolution (HR), not only allows for highly sensitive analyses but also broadens the usefulness of tracer‐based experiments, as interesting signals can be annotated de novo when not yet present in a compound library. However, several effects in the APCI ion source, i.e., fragmentation and rearrangement, lead to superimposed mass isotopologue distributions (MID) within the mass spectra, which need to be corrected during data evaluation as they will impair enrichment calculation otherwise. Here, we present and evaluate a novel software tool to automatically perform such corrections. We discuss the different effects, explain the implemented algorithm, and show its application on several experimental datasets. This adjustable tool is available as an R package from CRAN. KW - Mass Spectrometry KW - Isotopologue Distribution KW - Metabolic Flux KW - R package PY - 2022 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-547318 DO - https://doi.org/10.3390/metabo12050408 VL - 12 IS - 5 SP - 1 EP - 10 PB - MDPI AN - OPUS4-54731 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Jaeger, Carsten A1 - Lisec, Jan T1 - Towards Unbiased Evaluation of Ionization Performance in LC-HRMS Metabolomics Method Development N2 - As metabolomics increasingly finds its way from basic science into applied and regulatory environments, analytical demands on nontargeted mass spectrometric detection methods continue to rise. In addition to improved chemical comprehensiveness, current developments aim at enhanced robustness and repeatability to allow long-term, inter-study, and meta-analyses. Comprehensive metabolomics relies on electrospray ionization (ESI) as the most versatile ionization technique, and recent liquid chromatography-high resolution mass spectrometry (LC-HRMS) instrumentation continues to overcome technical limitations that have hindered the adoption of ESI for applications in the past. Still, developing and standardizing nontargeted ESI methods and instrumental setups remains costly in terms of time and required chemicals, as large panels of metabolite standards are needed to reflect biochemical diversity. In this paper, we investigated in how far a nontargeted pilot experiment, consisting only of a few measurements of a test sample dilution series and comprehensive statistical analysis, can replace conventional targeted evaluation procedures. To examine this potential, two instrumental ESI ion source setups were compared, reflecting a common scenario in practical method development. Two types of feature evaluations were performed, (a) summary statistics solely involving feature intensity values, and (b) analyses additionally including chemical interpretation. Results were compared in detail to a targeted evaluation of a large metabolite standard panel. We reflect on the advantages and shortcomings of both strategies in the context of current harmonization initiatives in the metabolomics field. KW - Mass Spectrometry KW - Non-targeted analysis KW - Method development PY - 2022 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-548065 DO - https://doi.org/10.3390/metabo12050426 VL - 12 IS - 5 SP - 1 EP - 13 PB - MDPI AN - OPUS4-54806 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - GEN A1 - Lisec, Jan T1 - CorMID - Correct Mass Isotopologue Distribution Vectors N2 - The investigation of metabolic fluxes and metabolite distributions within cells by means of tracer molecules is a valuable tool to unravel the complexity of biological systems. Technological advances in mass spectrometry (MS) technology such as atmospheric pressure chemical ionization (APCI) coupled with high resolution (HR), not only allows for highly sensitive analyses but also broadens the usefulness of tracer-based experiments, as interesting signals can be annotated de novo when not yet present in a compound library. However, several effects in the APCI ion source, i.e., fragmentation and rearrangement, lead to superimposed mass isotopologue distributions (MID) within the mass spectra, which need to be corrected during data evaluation as they will impair enrichment calculation otherwise. Here, we present and evaluate a novel software tool to automatically perform such corrections. We discuss the different effects, explain the implemented algorithm, and show its application on several experimental datasets. This adjustable tool is available as an R package from CRAN. KW - Mass Spectrometry KW - Software KW - R package KW - Mass Isotopomer Distribution KW - Flux Analysis PY - 2022 UR - https://github.com/cran/CorMID/ PB - GitHub CY - San Francisco, CA, USA AN - OPUS4-56442 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Döring, Sarah T1 - Nontarget mass spectrometry and recombinant antibody panels for optimized protein metrology N2 - The metrological analysis uses an unbroken chain of comparative measurements to trace results back to the national or international standard. This enables comparable, absolute quantification between laboratories. In 2020/2021, a pilot study for the quantification of SARS-CoV-2 antibodies was initiated with the involvement of BAM. Despite the consistent use of ID-MS, a large discrepancy in the results of the participating laboratories was found. This was the motivation for a project to systematically investigate and optimize traceable methods of protein quantification using mass spectrometric nontarget analysis (NTA) and recombinant antibody panels. T2 - St Nicholas Mass Spectrometry Symposium CY - Berlin, Germany DA - 08.12.2022 KW - Antibody quantification KW - Metrology KW - Traceability PY - 2022 AN - OPUS4-56515 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Riedel, Soraya T1 - Application of 3,3’,5,5’-Tetramethylbenzidine (TMB) in Amperometric Immunoassays for Mycotoxin Detection N2 - Electrochemical methods make great promise to meet the demand for user-friendly on-site devices for monitoring important parameters. Food industry often runs own lab procedures, e.g., for mycotoxin analysis, but it is a major goal to simplify analysis, linking analytical methods with miniaturized technologies. Enzyme-linked immunosorbent assays, with photometric detection of the horseradish peroxidase (HRP) substrate 3,3’,5,5’-tetramethylbenzidine (TMB), form a good basis for sensitive detection. To provide a straight-forward approach for the miniaturization of the detection step, we have studied the pitfalls of the electrochemical TMB detection. By cyclic voltammetry, it could be shown that TMB electrochemistry is strongly dependent on the pH and the electrode material. It was found that screen-printed gold electrodes and a very low pH value (pH 1) are well-suited to perform the electrochemical detection of TMB, due to the reversible character of the redox reaction under these conditions. Under these conditions, a good signal stability over several measuring cycles is achieved, providing the basis for analyzing multiple samples. In contrast to this, for carbon screen-printed electrodes, it was found that the signal response has changed after the electrochemical reaction with TMB at pH 1. At moderately acidic conditions (pH 4), neither with carbon nor with gold electrodes a reproducible electrochemical detection of TMB could be achieved. Based on these findings, we created a smartphone-based, electrochemical, immunomagnetic assay for the detection of ochratoxin A (OTA) and ergometrine in food samples. A competitive assay is performed on magnetic beads using HRP and TMB/H2O2 to generate the signal. Enzymatically oxidized TMB is quantified after addition of H2SO4 by amperometry with screen-printed gold electrodes in a custom-made wall-jet flow cell. The results are in good correlation with the established photometric detection method, providing a solid basis for sensing of further analytes in HRP-based assays using the newly developed miniaturized smartphone-based, electrochemical, immunomagnetic assay. T2 - 73rd Annual Meeting of the ISE CY - Online meeting DA - 12.09.2022 KW - Cylic Voltammetry KW - Immunoassay KW - Mycotoxins KW - Amperometry KW - Electrochemistry PY - 2022 N1 - Geburtsname von Riedel, Soraya: Höfs, S. - Birth name of Riedel, Soraya: Höfs, S. AN - OPUS4-55793 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Hülagü, Deniz A1 - Tobias, Charlie A1 - Climent Terol, Estela A1 - Gojani, Ardian A1 - Rurack, Knut A1 - Hodoroaba, Vasile-Dan T1 - Generalized Analysis Approach of the Profile Roughness by Electron Microscopy with the Example of Hierarchically Grown Polystyrene–Iron Oxide–Silica Core–Shell–Shell Particles N2 - The roughness as a property of core–shell (CS) microparticles plays a key role in their functionality. Quantitative evaluation of the roughness of CS microparticles is, however, a challenging task with approaches using electron microscopy images being scarce and showing pronounced differences in terms of methodology and results. This work presents a generalized method for the reliable roughness determination of nonplanar specimens such as CS particles from electron microscopic images, the method being robust and reproducible with a high accuracy. It involves a self-written software package (Python) that analyzes the recorded images, extracts corresponding data, and calculates the roughness based on the deviation of the identified contour. Images of single particles are taken by a dual mode scanning electron microscopy (SEM) setup which permits imaging of the same field-of-view of the sample with high resolution and surface sensitive in SE InLens mode as well as in transmission mode (TSEM). Herein, a new type of polystyrene core–iron oxide shell–silica shell particles is developed to serve as a set of lower micrometer-sized study objects with different surface roughness; the analysis of their images by the semiautomatic workflow is demonstrating that the particles’ profile roughness can be quantitatively obtained. KW - Core–shell particles KW - Image analysis KW - Nanoparticles KW - Roughness KW - SEM KW - transmission mode PY - 2022 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-542576 DO - https://doi.org/10.1002/adem.202101344 SP - 1 EP - 9 PB - Wiley-VCH AN - OPUS4-54257 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Hülagü, Deniz A1 - Tobias, Charlie A1 - Gojani, Ardian B. A1 - Rurack, Knut A1 - Hodoroaba, Vasile-Dan T1 - From 2D and Single Particle to 3D and Batch Analysis as a Routine Quality Check Procedure for the Morphological Characterization of Core-Shell Microparticles N2 - CS particles show unique properties by merging individual characteristics of the core and the shell materials. An alteration particularly in their surface roughness affects the final performance of the particles in the targeted application. Quantitative evaluation of the roughness of CS microparticles is, however, a challenging task employing microscopic techniques being scarce and showing large differences in terms of methodology and results. In our previous work, we have reported a systematic study with a reliable analysis tool, which evaluates profile roughness quantitatively, for individual core-shell microparticles using electron microscopy (EM) images of both types, Scanning Electron Microscopy (SEM) and transmission mode SEM (or TSEM). The SEM images contain two-dimensional (2D) information, therefore, provide profile roughness data only from the projection in the horizontal plane (in other words, from the “belly”) of a spherical particle. The present study offers a practical procedure to give access to more information by tilting the sample holder and hence allowing images of a single particle to be recorded at different orientations under the same view angle. From the analysis of these images, extended information on surface roughness of the particle can be extracted. Thus, instead of obtaining 2D information from a single SEM image, three-dimensional (3D) information is obtained from 2D projections recorded at different particle orientations. T2 - Microscopy and Microanalysis 2022 CY - Oregon, Portland, USA DA - 31.07.2022 KW - Core-shell particles KW - Image processing KW - Roughness KW - Scanning electron microscopy KW - Tilting PY - 2022 DO - https://doi.org/10.1017/S1431927622002094 SN - 1431-9276 VL - 28 IS - S1 SP - 332 EP - 334 PB - Cambridge University Press AN - OPUS4-55373 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Riedel, Soraya T1 - Steps Forward in the Application of 3,3’,5,5’- Tetramethylbenzidine (TMB) in Amperometric Assays N2 - Electrochemical methods make great promise to meet the demand for user-friendly on-site devices for monitoring important parameters. Food industry often runs own lab procedures, e.g. for mycotoxin analysis, but it is a major goal to simplify analysis, linking analytical methods with miniaturized technologies. Enzyme-linked immunosorbent assays, with photometric detection of the horseradish peroxidase (HRP) substrate, 3,3’,5,5’-tetramethylbenzidine (TMB), form a good basis for sensitive detection. To provide a straight-forward approach for the miniaturization of the detection step, we have studied the pitfalls of the electrochemical TMB detection. By cyclic voltammetry it was found that the TMB electrochemistry is strongly dependent on the pH and the electrode material. It was found that screen-printed gold electrodes and a highly acidic pH value (pH 1) are well-suited to perform the electrochemical detection of TMB, due to the reversible character of the redox reaction under these conditions. This set-up provides a good signal stability over several measuring cycles, providing the basis for analysing multiple samples. In contrast to this, for carbon screen-printed electrodes, it was found that the signal response has changed after the electrochemical reaction with TMB at pH 1. At a weakly acidic pH value (pH 4), neither with carbon nor with gold electrodes a reproducible electrochemical detection of TMB could be achieved [1]. Based on these findings we created a smartphone-based, electrochemical, immunomagnetic assay for the detection of ochratoxin A and ergometrine in real samples. Therefore, a competitive assay was performed on magnetic beads using HRP and TMB/H2O2 to generate the signal. Enzymatically oxidized TMB was quantified after addition of H2SO4 by amperometry with screen-printed gold electrodes in a custom-made wall-jet flow cell. The results are in good correlation with the established photometric detection method, providing a solid basis for sensing of further analytes in HRP-based assays using the newly developed miniaturized smartphone-based, electrochemical, immunomagnetic assay. T2 - Electrochemistry 2022 CY - Berlin, Germany DA - 28.09.2022 KW - Cylic Voltammetry KW - Immunoassay KW - TMB KW - Amperometry PY - 2022 N1 - Geburtsname von Riedel, Soraya: Höfs, S. - Birth name of Riedel, Soraya: Höfs, S. AN - OPUS4-55876 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Singh, Chandan T1 - Functionalized Titanium Carbide Nanosheets based Biosensor for Detection of the SARS-CoV-2 Nucleocapsid Protein N2 - MXenes are a new family of two-dimensional (2D) transition metal carbides, carbonitrides, and nitrides discovered in 2011. Among many reported family members, titanium carbide is the most widely studied and explored due to the optimized synthesis conditions and promising characteristics like good mechanical strength, solution processability, and excellent conductivity. Here, we report the development of an electrochemical biosensor involving the amine-functionalized Few-Layered-Titanium Carbide Nanosheets and monoclonal antibodies against the SARS-CoV-2 nucleocapsid protein (anti-SARS-CoV-2 mAb) to design a point-of-care device for detection of the SARS-CoV-2 nucleocapsid protein (SARS-CoV-2 NP) antigen. T2 - SALSA's Make and Measure 2022 CY - Berlin, Germany DA - 15.09.2022 KW - MXenes KW - Titanium Carbide Nanosheets KW - Biosensors KW - Electrochemistry PY - 2022 N1 - Geburtsname von Riedel, Soraya: Höfs, S. - Birth name of Riedel, Soraya: Höfs, S. AN - OPUS4-55856 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - GEN A1 - Tchipilov, Teodor A1 - Raysyan, Anna A1 - Weller, Michael G. T1 - Methods for the quantification of particle-bound protein – Application to reagents for lateral-flow immunoassays (LFIA) N2 - Protein immobilization for the functionalization of particles is used in various applications, including biosensors, lateral-flow immunoassays (LFIA), bead-based assays, and others. Common methods for the quantification of bound protein are measuring protein in the supernatant before and after coating and calculating the difference. This popular approach has the potential for a significant overestimation of the amount of immobilized protein since layers not directly bound to the surface (soft protein corona) are usually lost during washing and handling. Only the layer directly bound to the surface (hard corona) can be used in subsequent assays. A simplified amino acid analysis method based on acidic hydrolysis and RP-HPLC-FLD of tyrosine and phenylalanine (aromatic amino acid analysis, AAAA) is proposed to directly quantify protein bound to the surface of gold nano- and latex microparticles. The results are compared with indirect methods such as colorimetric protein assays, such as Bradford, bicinchoninic acid (BCA), as well as AAAA of the supernatant. For both particle types, these indirect quantification techniques show a protein overestimation of up to 1700% compared to the direct AAAA measurements. In addition, protein coating on latex particles was performed both passively through adsorption and covalently through EDC/sulfo-NHS chemistry. Our results showed no difference between the immobilization methodologies. This finding suggests that usual protein determination methods are no unambiguous proof of a covalent conjugation on particles or beads. KW - Soft protein corona KW - Hard protein corona KW - Gold particles KW - Nanoparticles KW - Mikroparticles KW - Antibody KW - Bioconjugation KW - Protein quantification KW - Supernatant KW - Sodium chloride method KW - Covalent conjugation KW - Latex particles KW - Lateral flow immunoassays PY - 2022 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-545365 DO - https://doi.org/10.20944/preprints202203.0332.v1 SP - 1 EP - 8 PB - MDPI CY - Basel AN - OPUS4-54536 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -