{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,23]],"date-time":"2026-08-23T16:50:53Z","timestamp":1787503853914,"version":"build-2736575974"},"reference-count":48,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2021,3,12]],"date-time":"2021-03-12T00:00:00Z","timestamp":1615507200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002347","name":"Bundesministerium f\u00fcr Bildung und Forschung","doi-asserted-by":"publisher","award":["FKZ 01DQ18003A"],"award-info":[{"award-number":["FKZ 01DQ18003A"]}],"id":[{"id":"10.13039\/501100002347","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100018761","name":"Indo-German Science and Technology Centre","doi-asserted-by":"publisher","award":["FKZ 01DQ18003A"],"award-info":[{"award-number":["FKZ 01DQ18003A"]}],"id":[{"id":"10.13039\/501100018761","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Antibiotics"],"abstract":"<jats:p>Contamination of waters with pharmaceuticals is an alarming problem as it may support the evolution of antimicrobial resistance. Therefore, fast and cost-effective analytical methods for potential on-site analysis are desired in order to control the water quality and assure the safety of its use as a source of drinking water. Antibody-based methods, such as the enzyme-linked immunosorbent assay (ELISA), can be helpful in this regard but can also have certain pitfalls in store, depending on the analyte. As shown here for the class of \u03b2-lactam antibiotics, hydrolysis of the \u03b2-lactam ring is a key factor in the immunochemical analysis as it influences antibody recognition. With the antibody used in this study, the limit of detection (LOD) in the immunoassay could be significantly reduced by hydrolysis for the five tested penicillins, with the lowest LOD for carbenicillin (0.2 nmol\/L) and the greatest impact on penicillins G and V (reduction by 85%). In addition to enhanced quantification, our strategy also provides access to information about the degree of hydrolysis in water samples as shown for the most abundant penicillin amoxicillin.<\/jats:p>","DOI":"10.3390\/antibiotics10030298","type":"journal-article","created":{"date-parts":[[2021,3,12]],"date-time":"2021-03-12T11:56:55Z","timestamp":1615550215000},"page":"298","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Pitfalls in the Immunochemical Determination of \u03b2-Lactam Antibiotics in Water"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2954-6854","authenticated-orcid":false,"given":"Alexander","family":"Ecke","sequence":"first","affiliation":[{"name":"BAM Federal Institute for Materials Research and Testing, 12205 Berlin, Germany"},{"name":"Department of Chemistry, Humboldt-Universit\u00e4t zu Berlin, 12489 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2228-1248","authenticated-orcid":false,"given":"Rudolf J.","family":"Schneider","sequence":"additional","affiliation":[{"name":"BAM Federal Institute for Materials Research and Testing, 12205 Berlin, Germany"},{"name":"Faculty III Process Sciences, Technische Universit\u00e4t Berlin, 10623 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,3,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"3245","DOI":"10.1016\/S0043-1354(98)00099-2","article-title":"Occurrence of drugs in German sewage treatment plants and rivers","volume":"32","author":"Ternes","year":"1998","journal-title":"Water Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1016\/S0378-4274(02)00041-3","article-title":"Occurrence, fate, and removal of pharmaceutical residues in the aquatic environment: A review of recent research data","volume":"131","author":"Heberer","year":"2002","journal-title":"Toxicol. Lett."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"320","DOI":"10.1016\/j.watres.2019.06.039","article-title":"Antibiotic resistance genes in treated wastewater and in the receiving water bodies: A pan-European survey of urban settings","volume":"162","author":"Cacace","year":"2019","journal-title":"Water Res."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Amador, P., Fernandes, R., Prud\u00eancio, C., and Duarte, I. (2019). Prevalence of Antibiotic Resistance Genes in Multidrug-Resistant Enterobacteriaceae on Portuguese Livestock Manure. Antibiotics, 8.","DOI":"10.3390\/antibiotics8010023"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1016\/S1473-3099(18)30605-4","article-title":"Attributable deaths and disability-adjusted life-years caused by infections with antibiotic-resistant bacteria in the EU and the European Economic Area in 2015: A population-level modelling analysis","volume":"19","author":"Cassini","year":"2019","journal-title":"Lancet Infect. Dis."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"M\u00fcller, H., Sib, E., Gajdiss, M., Klanke, U., Lenz-Plet, F., Barabasch, V., Albert, C., Schallenberg, A., Timm, C., and Zacharias, N. (2018). Dissemination of multi-resistant Gram-negative bacteria into German wastewater and surface waters. FEMS Microbiol. Ecol., 94.","DOI":"10.1093\/femsec\/fiy057"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1016\/j.watres.2014.02.003","article-title":"Insights into the relationship between antimicrobial residues and bacterial populations in a hospital-urban wastewater treatment plant system","volume":"54","author":"Varela","year":"2014","journal-title":"Water Res."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Singer, A.C., Shaw, H., Rhodes, V., and Hart, A. (2016). Review of Antimicrobial Resistance in the Environment and Its Relevance to Environmental Regulators. Front. Microbiol., 7.","DOI":"10.3389\/fmicb.2016.01728"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Kraemer, S.A., Ramachandran, A., and Perron, G.G. (2019). Antibiotic Pollution in the Environment: From Microbial Ecology to Public Policy. Microorganisms, 7.","DOI":"10.3390\/microorganisms7060180"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1441","DOI":"10.1093\/jac\/dkn386","article-title":"Outpatient antibiotic use in the four administrations of the UK: Cross-sectional and longitudinal analysis","volume":"62","author":"Davey","year":"2008","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Fuentes, A.V., Pineda, M.D., and Venkata, K.C.N. (2018). Comprehension of Top 200 Prescribed Drugs in the US as a Resource for Pharmacy Teaching, Training and Practice. Pharmacy, 6.","DOI":"10.3390\/pharmacy6020043"},{"key":"ref_12","unstructured":"(2020, February 10). The Top 200 Drugs of 2020. Available online: https:\/\/clincalc.com\/DrugStats\/."},{"key":"ref_13","unstructured":"(2020, February 10). Center for Financing, Access and Cost Trends, Agency for Healthcare Research and Quality, Medical Expenditure Panel Survey, 1996\u20132017, Available online: https:\/\/www.meps.ahrq.gov\/mepstrends\/hc_pmed\/."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1016\/j.ijheh.2018.12.013","article-title":"The occurrence of antimicrobial substances in toilet, sink and shower drainpipes of clinical units: A neglected source of antibiotic residues","volume":"222","author":"Voigt","year":"2019","journal-title":"Int. J. Hyg. Environ. Health"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"6832","DOI":"10.1021\/es049509a","article-title":"Antibiotics in the Environment: Occurrence in Italian STPs, Fate, and Preliminary Assessment on Algal Toxicity of Amoxicillin","volume":"38","author":"Andreozzi","year":"2004","journal-title":"Environ. Sci. Technol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2711","DOI":"10.1016\/j.scitotenv.2008.11.059","article-title":"The occurrence of antibiotics in an urban watershed: From wastewater to drinking water","volume":"407","author":"Watkinson","year":"2009","journal-title":"Sci. Total Environ."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1016\/j.ecoenv.2016.11.014","article-title":"A review of the occurrence of pharmaceuticals and personal care products in Indian water bodies","volume":"137","author":"Balakrishna","year":"2017","journal-title":"Ecotoxicol. Environ. Saf."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1002\/aheh.200390014","article-title":"Determination of Antibiotic Residues in Manure, Soil, and Surface Waters","volume":"31","author":"Christian","year":"2003","journal-title":"Acta Hydrochim. Hydrobiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"3498","DOI":"10.1016\/j.watres.2008.04.026","article-title":"The occurrence of pharmaceuticals, personal care products, endocrine disruptors and illicit drugs in surface water in South Wales, UK","volume":"42","author":"Dinsdale","year":"2008","journal-title":"Water Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1961","DOI":"10.1021\/ed500482j","article-title":"Development and Validation of a Fast Procedure To Analyze Amoxicillin in River Waters by Direct-Injection LC\u2013MS\/MS","volume":"91","author":"Homem","year":"2014","journal-title":"J. Chem. Educ."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1007\/s00244-010-9550-1","article-title":"Determination of Antibiotics in Brazilian Surface Waters Using Liquid Chromatography\u2013Electrospray Tandem Mass Spectrometry","volume":"60","author":"Locatelli","year":"2011","journal-title":"Arch. Environ. Contam. Toxicol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Larramendy, M.L., and Soloneski, S. (2016). Amoxicillin in the aquatic environment, its fate and environmental risk. Environmental Health Risk\u2014Hazardous Factors to Living Species, IntechOpen.","DOI":"10.5772\/61472"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"880","DOI":"10.1016\/j.watres.2015.11.028","article-title":"New hydrolysis products of the beta-lactam antibiotic amoxicillin, their pH-dependent formation and search in municipal wastewater","volume":"88","author":"Hirte","year":"2016","journal-title":"Water Res."},{"key":"ref_24","first-page":"1684","article-title":"Degradation of \u03b2-lactam antibiotics","volume":"87","author":"Deshpande","year":"2004","journal-title":"Curr. Sci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"985","DOI":"10.1016\/j.chemosphere.2013.01.095","article-title":"Amoxicillin-degradation products formed under controlled environmental conditions: Identification and determination in the aquatic environment","volume":"91","author":"Gozlan","year":"2013","journal-title":"Chemosphere"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1670","DOI":"10.1016\/j.jasms.2005.06.002","article-title":"Structure Elucidation of Degradation Products of the Antibiotic Amoxicillin with Ion Trap MSn and Accurate Mass Determination by ESI TOF","volume":"16","author":"Moritz","year":"2005","journal-title":"J. Am. Soc. Mass Spectrom."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"731","DOI":"10.1002\/rcm.4902","article-title":"Behavior of amoxicillin in wastewater and river water: Identification of its main transformation products by liquid chromatography\/electrospray quadrupole time-of-flight mass spectrometry","volume":"25","author":"Heinzen","year":"2011","journal-title":"Rapid Commun. Mass Spectrom."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1016\/j.jhazmat.2017.11.050","article-title":"Variations in the fate and risk analysis of amoxicillin and its degradation products during pig manure aerobic composting","volume":"346","author":"Liu","year":"2018","journal-title":"J. Hazard. Mater."},{"key":"#cr-split#-ref_29.1","unstructured":"European Commission (2018). Commission Implementing Decision"},{"key":"#cr-split#-ref_29.2","unstructured":"(EU) 2018\/840 of 5 June 2018 establishing a watch list for Union-wide monitoring in the field of water policy pursuant to Directive 2008\/105\/EC of the European Parliament and of the Council and repealing Commission Implementing Decision"},{"key":"#cr-split#-ref_29.3","unstructured":"(EU) 2015\/495. Off. J. Eur. Union, L141, 9-12."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1016\/S0021-9673(01)01154-2","article-title":"Determination of drugs in surface water and wastewater samples by liquid chromatography\u2013mass spectrometry: Methods and preliminary results including toxicity studies with Vibrio fischeri","volume":"938","author":"Ferrer","year":"2001","journal-title":"J. Chromatogr. A"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/S0021-9673(00)01187-0","article-title":"Determination of drug residues in water by the combination of liquid chromatography or capillary electrophoresis with electrospray mass spectrometry","volume":"910","author":"Ahrer","year":"2001","journal-title":"J. Chromatogr. A"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"859","DOI":"10.1016\/j.chemosphere.2010.08.011","article-title":"Analysis of llicit and illicit drugs in waste, surface and lake water samples using large volume direct injection high performance liquid chromatography\u2014Electrospray tandem mass spectrometry (HPLC\u2013MS\/MS)","volume":"81","author":"Berset","year":"2010","journal-title":"Chemosphere"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1748","DOI":"10.1016\/j.chroma.2010.01.046","article-title":"Determination of drugs of abuse in water by solid-phase extraction, derivatisation and gas chromatography\u2013ion trap-tandem mass spectrometry","volume":"1217","author":"Quintana","year":"2010","journal-title":"J. Chromatogr. A"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"678","DOI":"10.1016\/j.talanta.2006.05.024","article-title":"Development of a multi-residue analytical methodology based on liquid chromatography\u2013tandem mass spectrometry (LC\u2013MS\/MS) for screening and trace level determination of pharmaceuticals in surface and wastewaters","volume":"70","author":"Gros","year":"2006","journal-title":"Talanta"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1545","DOI":"10.1128\/AAC.36.7.1545","article-title":"Use of an enzyme-linked immunosorbent assay to assess penetration of amoxicillin into lung secretions","volume":"36","author":"Hill","year":"1992","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1016\/0300-483X(94)02983-2","article-title":"Epitope mapping of \u03b2-lactam antibiotics with the use of monoclonal antibodies","volume":"97","author":"Mayorga","year":"1995","journal-title":"Toxicology"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"577","DOI":"10.3382\/ps.2007-00167","article-title":"Development of Amoxicillin Enzyme-Linked Immunosorbent Assay and Measurements of Tissue Amoxicillin Concentrations in a Pigeon Microdialysis Model","volume":"87","author":"Yeh","year":"2008","journal-title":"Poult. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1080\/09540100120075817","article-title":"Highly Sensitive Enzyme Immunoassays for the Detection of \u03b2-Lactam Antibiotics","volume":"13","author":"Grubelnik","year":"2001","journal-title":"Food Agric. Immunol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"21","DOI":"10.13005\/ojc\/360103","article-title":"Immune Recognition of Closed and Open Lactam Rings and Their Influence on Immunoassays of Ampicillin Antibiotics","volume":"36","author":"Komova","year":"2020","journal-title":"Orient. J. Chem."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1093\/jac\/28.3.357","article-title":"Anti-ampicillin monoclonal antibodies and their cross-reactivities to various \u03b2-lactams","volume":"28","author":"Nagakura","year":"1991","journal-title":"J. Antimicrob. Chemoth."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"8171","DOI":"10.1021\/jf101243z","article-title":"Development of an Enzyme-Linked Immunosorbent Assay To Detect Benzylpenicilloic Acid, a Degradation Product of Penicillin G in Adulterated Milk","volume":"58","author":"Zhang","year":"2010","journal-title":"J. Agric. Food Chem."},{"key":"ref_42","unstructured":"Hunter, W.M., and Corrie, J.E.T. (1983). The precision profile: Its use in assay design, assessment and quality control. Immunoassays for Clinical Chemistry, Churchill Livingstone. [2nd ed.]."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1725","DOI":"10.1039\/P29800001725","article-title":"Metal-ion catalysed hydrolysis of some \u03b2-lactam antibiotics","volume":"2","author":"Gensmantel","year":"1980","journal-title":"J. Chem. Soc. Perkin Trans."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1197","DOI":"10.1002\/jps.2600670903","article-title":"Preparation and immunological cross-reactions of penicilloic and penilloic acids","volume":"67","author":"Munro","year":"1978","journal-title":"J. Pharm. Sci."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/0378-5173(79)90044-9","article-title":"Piperazinedione formation from reaction of ampicillin with carbohydrates and alcohols in aqueous solution","volume":"3","author":"Bundgaard","year":"1979","journal-title":"Int. J. Pharm."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Frenzel, A., Hust, M., and Schirrmann, T. (2013). Expression of Recombinant Antibodies. Front. Immunol., 4.","DOI":"10.3389\/fimmu.2013.00217"}],"container-title":["Antibiotics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-6382\/10\/3\/298\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:35:03Z","timestamp":1760160903000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-6382\/10\/3\/298"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,3,12]]},"references-count":48,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2021,3]]}},"alternative-id":["antibiotics10030298"],"URL":"https:\/\/doi.org\/10.3390\/antibiotics10030298","relation":{},"ISSN":["2079-6382"],"issn-type":[{"value":"2079-6382","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,3,12]]}}}