TY - CONF A1 - Raysyan, Anna T1 - Determination of mycotoxins in food and pharmaceuticals by immunochemical analysis N2 - Mycotoxins are toxic secondary metabolites produced by filamentous fungi. The contamination of mycotoxins in food and feed products has been recognized to be an important safety issue for human and animal health. In the context of food safety, medical herbs nowadays are also considered as potential source of mycotoxins. Considering that mycotoxins are generally stable compounds and could not be destroyed completely during food-processing operations, prevention of contamination is identified as main issue. Currently, biochemical, and immunochemical methods based on the use of antibodies as a specific recognition bioreagents are increasingly found to be applied in screening. Along with good common enzyme linked immunosorbent assay (ELISA), more and more widely used method is fluorescence polarization immunoassay (FPIA). These methods help to monitor of many toxic substances in large number of samples and carried out quickly, easily, cheaply, and give good results in quantifying the one or more substances. That is why the use of immunochemical methods of analysis, such as ELISA and FPIA, is the most promising for solving this problem. Thus, the aim of our work is to optimize the determination of mycotoxins by ELISA and FPIA in medical herbs. We selected antibodies and a tracer for the FPIA, then we constructed obtained a calibration curve and determined the sensitivity of this method for the samples in aqueous media. Then, the optimal sample preparation was elaborated: for spiked nuts samples were selected and prepared and a calibration curve was obtained with respect to the matrix effect. After that, the analysis of spiked nuts samples was performed by ELISA and FPIA and the sensitivity of both methods was compared. T2 - Global Food Science Student Competition CY - Jiangnan University, Wuxi, China DA - 14.11.2018 KW - Mycotoxins KW - FPIA KW - ELISA PY - 2018 AN - OPUS4-50048 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Raysyan, Anna A1 - Eremin, S. A. A1 - Beloglazova, N. V. A1 - De Saeger, S. A1 - Gravel, I. V. T1 - Immunochemical approaches for detection of aflatoxin B1 in herbal medicines N2 - Introduction Aflatoxin B1 (AFB1) is a toxic low‐molecular‐weight secondary metabolite of Aspergillus flavus and A. parasiticus. AFB1 was classified as a Group I carcinogen by the World Health Organisation for Research on Cancer in 1993. AFB1 is an unavoidable natural contaminant of some herbal medicine, able to cause serious health issues for humans consuming the related medicine. Objective Therefore, this study aimed to develop an efficient fluorescence polarisation immunoassay (FPIA) and a rapid, low‐cost, and easy‐to‐use membrane‐based flow‐through immunoassay (MBA) for determination of AFB1 in herbal medicine Origanum vulgare L., Rubus idaeus L., Urtica dioica L. and Sorbus aucuparia L. Results A cut‐off level of the developed MBA was 0.8 ppb. Validation of the developed test was performed with blank and spiked samples. Using three naturally contaminated or three artificially spiked samples. The FPIA showed a linear working range of 8.6 to 64 ppb, and a half maximal inhibitory concentration (IC50) of 24 ppb. Conclusion The results were in good correlation with the enzymelinked immunosorbent assay (ELISA) results (the IC50 0.1 ppb). Both the sample preparation and analysis are simple, cost‐effective and easy to perform on‐site in non‐laboratory environments. Liquid chromatography with tandem mass spectrometry (LC‐MS/MS) was used as a confirmatory technique. KW - Fluorescence Polarisation Immunoassay KW - membrane-based flow-through immunoassay KW - Mycotoxins KW - Mykotoxine KW - Schnelltest PY - 2020 DO - https://doi.org/10.1002/pca.2931 SN - 1099-1565 VL - 31 IS - 5 SP - 662 EP - 669 PB - Wiley Online Libary CY - Hoboken, NJ, USA AN - OPUS4-51341 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Raysyan, Anna A1 - Galvidis, I. A. A1 - Schneider, Rudolf A1 - Eremin, S. A. A1 - Burkin, M. A. T1 - Development of a latex particles-based lateral flow immunoassay for group determination of macrolide antibiotics in breast milk N2 - A lateral flow immunoassay (LFIA) using latex particles labeled with antibody to BSAclarithromycin (CLA) was developed for the rapid simultaneous group determination of six macrolide antibiotics. Optimization of antigen spotting on the membrane and latex probe loading allowed improving visual detectability (vLOD) 100 times, which was 1, 1, 10, 10, 50, and 1000 ng/mL for CLA, roxithromycin, erythromycin, dirithromycin, azithromycin, and oleandomycin in buffer, respectively. The calculated limits of instrumental detection (cLOD) were respectively 0.12, 0.15, 1.4, 2.1, 2.4, and 3.3 ng/mL. To avoid a strong influence of breast milk of a very diverse and variable composition, a sample pretreatment is proposed. The six macrolides mentioned can be visually detected in breast milk after 20 minutes pretreatment at concentrations of 10-1000 ng / ml or instrumentally with cLOD of 4.0, 2.5, 30, 42, 42 and 180 ng/mL. The recovery rate from the spiked samples carried out using a strip scanner device ranged from 71% to 110%, and precision expressed as relative standard deviation was between 3-14%. The described rapid on-site diagnostic assay format can be useful for monitoring the content of antibiotics in breast milk during macrolide treatment to ensure safe breastfeeding of infants. KW - Clarithromycin KW - Roxithromycin KW - Erythromycin KW - Dirithromycin KW - Azithromycin KW - Oleandomycin KW - Macrolide antibiotics KW - Makrolide KW - Antibiotika PY - 2020 DO - https://doi.org/10.1016/j.jpba.2020.113450 SN - 0731-7085 VL - 189 IS - 113450 SP - 1 EP - 9 PB - Elsevier CY - Amsterdam, NL AN - OPUS4-51343 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Raysyan, Anna A1 - Moerer, R. A1 - Coesfeld, Bianca A1 - Eremin, S. A1 - Schneider, Rudolf T1 - Fluorescence polarization immunoassay for the determination of diclofenac in wastewater N2 - Pharmacologically active compounds are often detected in wastewater and surface waters. The nonsteroidal anti-inflammatory drug diclofenac (DCF) was included in the European watch list of substances that requires its environmental monitoring in the member states. DCF may harmfully influence the ecosystem already at concentrations ≤ 1 μg L−1. The fast and easy quantification of DCF is becoming a subject of global importance. Fluorescence polarization immunoassay (FPIA) is a homogeneous mix-and-read method which does not require the immobilization of reagents. FPIA can be performed in one phase within 20–30 min, making it possible to analyse wastewater without any complicated pre-treatment. In this study, new tracermolecules with different structures, linking fluorophores to derivatives of the analyte, were synthesized, three homologous tracers based on DCF, two including a C6 spacer, and one heterologous tracer derived from 5-hydroxy-DCF. The tracer molecules were thoroughly assessed for performance. Regarding sensitivity of the FPIA, the lowest limit of detection reached was 2.0 μg L−1 with a working range up to 870 μg L−1. The method was validated for real wastewater samples against LC-MS/MS as reference method with good agreement of both methods. KW - Abwasser KW - Umweltschadstoffe KW - Antikörper KW - Antibody PY - 2020 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-518398 DO - https://doi.org/10.1007/s00216-020-03058-w SN - 1618-2642 VL - 413 IS - 4 SP - 999 EP - 107 PB - Springer CY - Heidelberg AN - OPUS4-51839 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Raysyan, Anna A1 - Schneider, Rudolf T1 - Development of a Lateral Flow Immunoassay (LFIA) to Screen for the Release of the Endocrine Disruptor Bisphenol A from Polymer Materials and Products N2 - One of the most important chemicals used in the production of polymer plastics and coatings is bisphenol A. However, despite the large number of studies on the toxicity and hormonal activity of BPA, there are still open questions and thus considerable media attention regarding BPA toxicity. Hence, it is necessary to develop a sensitive, simple, cost-efficient, specific, portable, and rapid method for monitoring bisphenol A and for high sample throughput and on-site screening analysis. Lateral flow immunoassays have potential as rapid tests for on-site screening. To meet sensitivity criteria, they must be carefully optimized. A latex microparticle-based LFIA for detection of BPA was developed. The sensitivity of the assay was improved by non-contact printing of spot grids as the control and test lines with careful parameter optimization. Results of the test could be visually evaluated within 10 min with a visual cut-off of 10 µg/L (vLOD). Alternatively, photographs were taken, and image analysis performed to set up a calibration, which allowed for a calculated limit of detection (cLOD) of 0.14 µg/L. The method was validated for thermal paper samples against ELISA and LC–MS/MS as reference methods, showing good agreement with both methods KW - Endokrine Disruptoren KW - Polymere KW - Schnelltest KW - Endocrine disruptor KW - Bisphenol A KW - LFIA KW - Immunoassay PY - 2021 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-540492 DO - https://doi.org/10.3390/bios11070231 VL - 11 IS - 7 SP - 1 EP - 11 PB - MDPI CY - Basel AN - OPUS4-54049 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Raysyan, Anna A1 - Zwigart, S. A1 - Eremin, S. A. A1 - Schneider, Rudolf T1 - BPA Endocrine Disruptor Detection at the Cutting Edge: FPIA and ELISA Immunoassays N2 - BPA is a chemical commonly used in the production of polymer-based materials that can have detrimental effects on the thyroid gland and impact human reproductive health. Various expensive methods, such as liquid and gas chromatography, have been suggested for detecting BPA. The fluorescence polarization immunoassay (FPIA) is an inexpensive and efficient homogeneous mix-and-read method that allows for high-throughput screening. FPIA offers high specificity and sensitivity and can be carried out in a single phase within a timeframe of 20–30 min. In this study, new tracer molecules were designed that linked the fluorescein fluorophore with and without a spacer to the bisphenol A moiety. To assess the influence of the C6 spacer on the sensitivity of an assay based on the respective antibody, hapten–protein conjugates were synthesized and assessed for performance in an ELISA setup, and this resulted in a highly sensitive assay with a detection limit of 0.05 µg/L. The lowest limit of detection was reached by employing the spacer derivate in the FPIA and was 1.0 µg/L, working range from 2 to 155 µg/L. The validation of the methods was conducted using actual samples compared to LC–MS/MS, which served as the reference method. The FPIA and ELISA both demonstrated satisfactory concordance. KW - Fluorescence Polarization KW - Enzyme-Linked Immunosorbent Assay KW - Endokriner Disruptor KW - Bisphenol A KW - Freisetzung PY - 2023 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-579497 DO - https://doi.org/10.3390/bios13060664 VL - 13 IS - 6 SP - 1 EP - 12 PB - MDPI CY - Basel AN - OPUS4-57949 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Raysyan, Anna T1 - Fluorescence polarization immunoassay for determination of diclofenac in wastewater N2 - Pharmacologically active compounds are often detected in wastewater and surface waters. The nonsteroidal anti-inflammatory drug diclofenac was included in the watch list of substances in the EU that requires their environmental monitoring in the member states. DCF may harmfully influence the ecosystem and the natural organization already at concentrations of ≤1 μg/l. The fast and easy quantification of DCF is becoming a subject of global importance. Fluorescence polarization immunoassay (FPIA) is a homogeneous mix-and-read method, which does not require the immobilization of reagents, they can be performed in one phase, making it possible to analyze wastewater without any complicated pre-treatment, performed within 20–30 min. T2 - Potsdam days on Bioanalysis 2017 CY - Potsdam, Germany DA - 23.11.2017 KW - Diclofenac KW - DCF KW - FPIA KW - Wastewater PY - 2017 AN - OPUS4-43127 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Raysyan, Anna T1 - Development of a quick test to screen for bisphenol A release from polymer materials N2 - Bisphenol A (BPA) is a common industrial chemical that is widely used as a monomer or additive in the production of polymer materials. This substance causes impairment of the childbearing function, acceleration of sexual aging, negatively impacts the brain, and also contributes to the development of certain oncological diseases, as well as increasing the risk of a fetus developing the Down syndrome. But the biggest downside of bisphenol is the fact that it accumulates in the system during its use, as well as after. Many methods have been proposed for the determination of BPA, in particular, gas and liquid chromatography. The main disadvantages of such approaches are the high cost of the equipment, the significant duration of an analysis, and difficulties in screening a large number of samples. Immunoanalytical techniques attributes are high throughput, high specificity and sensitivity, as well as low cost and simplicity. Particularly interesting as a quick test for screenings is the lateral flow immunoassay (LFIA), a format widely in use for the detection of pregnancy. LFIA for BPA have been described before, but lack sensitivity. In this work, focuses on the conjugation (by passive adsorption) efficiency of the different monoclonal antibodies with 40 nm gold nanoparticles. The effect of factors such as pH value and concentration of antibody has been quantificationally discussed using spectra methods after adding 10% NaCl which induced gold nanoparticle aggregation. Consequently, 40 nm gold nanoparticles should be labeled with antibody under optimal pH value and optimal concentrations of antibody. It will be helpful for the application of antibody-labeled gold nanoparticles for the determination of bisphenol A in aqueous solutions (water from different sources, leachates and extracts). T2 - Adlershofer Forschungsforum 2017 CY - Berlin, Germany DA - 10.11.2017 KW - Bisphenol A KW - BPA KW - LFIA KW - ELISA KW - GNP KW - Passive absorption PY - 2017 AN - OPUS4-43125 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Raysyan, Anna T1 - Development of dipstick immunoassays for environmental and diagnostic applications missions from plastic materials N2 - Development of dipstick immunoassays for environmental analysis and integration the technology for diagnostic applications missions. T2 - Arbeitskreistreffen Immunoassays CY - Hennigsdorf, Germany DA - 26.11.2018 KW - Environmental analysis KW - LFIA KW - BPA KW - DCF PY - 2018 AN - OPUS4-46801 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Raysyan, Anna T1 - A dipstick immunoassay for the determination of Bisphenol N2 - Bisphenol A (BPA), a monomer commonly used to manufacture polycarbonate plastic products and the epoxy resins that line metal cans, has resulted in an increased human exposure because both products come in contact with food and beverages. Since BPA is an endocrine disruptor excessive exposure could affect human reproductive function. In response, various countries have adopted a range of measures, from voluntary reductions to outright bans of BPA in some products, mostly in baby bottles and containers for canned foods. Pregnancy-test like strips in combination with hand-held readers provide a simple and versatile tool to determine the presence of bisphenol A quantitatively, making it a very attractive analytical approach especially for on-site applications at critical points, such as environmental analysis, consumer protection, and materials testing. T2 - BAM-BfR Seminar CY - Berlin, Germany DA - 15.02.2018 KW - BPA KW - Dipstickassay KW - LFIA PY - 2018 AN - OPUS4-46751 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Raysyan, Anna T1 - An immunochromatographic assay for the determination of bisphenol a releases from plastic materials N2 - Background. The worldwide increase in the use of bisphenol A (BPA), a monomer commonly used to manufacture polycarbonate plastic products and the epoxy resins that line metal cans, has resulted in an increased human exposure because both products come in contact with food and beverages. Since BPA is an endocrine disruptor excessive exposure could affect human reproductive function. In response, various countries have adopted a range of measures, from voluntary reductions to outright bans of BPA in some products, mostly in baby bottles and containers for canned foods. Nonetheless, consumers would certainly appreciate a quick test for their own assessment of contaminations by bisphenol A. Methods. An immunochromatographic test strip was developed. This lateral-flow immunoassay (LFIA) is based on anti-BPA antibodies (from mouse) conjugated with gold nanoparticles as the marker, deposited in an elaborate pad. To form control and test zones, anti-mouse antibody and a BPA-BSA conjugate were spotted on the nitrocellulose membrane, respectively. Results. Negative samples are revealed by red lines both in the test and control zones, respectively, whereas positive samples produce a single red line only in the control zone. The limit of detection (LOD) was found to be 0.05 µg/l, visually detected by the naked eye within 5 minutes. In addition, the intensities at the test line can be read by dedicated lateral flow readers (opTrilyzer® and Cube by opTricon, Berlin) for a more precise and documented determination. Conclusion. Pregnancy-test like strips in combination with hand-held readers provide a simple and versatile tool to determine the presence of bisphenol A quantitatively, making it a very attractive analytical approach especially for on-site applications at critical points, such as environmental analysis, consumer protection, and materials testing. T2 - International Biotech Innovation Days CY - Senftenberg, Germany DA - 22.05.2018 KW - Immunoassay KW - BPA KW - LFIA KW - Plastic PY - 2018 AN - OPUS4-46752 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Raysyan, Anna T1 - A fast dipstick immunoassay for the determination of DCF in the breast milk N2 - Diclofenac (DCF) is a non-steroidal anti-inflammatory drug (NSAID) with analgetic, anti-inflammatory, and antipyretic properties. The mechanism of action of diclofenac, like that of other NSAIDs, involves inhibition of cyclooxygenase (COX-1 and COX-2). Another pharmacological effect is preventing of prostaglandin synthesis in vitro. Prostaglandins are mediators of inflammation, because diclofenac is an inhibitor of prostaglandin synthesis. A method has been developed to analyse for diclofenac (DCF) in the milk. T2 - BIONNALE Speed Lecture Award 2018 CY - Berlin, Germany DA - 20.06.2018 KW - Milk KW - DCF KW - Immunoassay PY - 2018 AN - OPUS4-46753 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -