{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T13:08:33Z","timestamp":1788872913482,"version":"build-2803163510"},"reference-count":19,"publisher":"Springer Science and Business Media LLC","license":[{"start":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T00:00:00Z","timestamp":1788825600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T00:00:00Z","timestamp":1788825600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100005416","name":"Norges Forskningsr\u00e5d","doi-asserted-by":"publisher","award":["105303"],"award-info":[{"award-number":["105303"]}],"id":[{"id":"10.13039\/501100005416","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005366","name":"University of Oslo","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100005366","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Anal Bioanal Chem"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Sample preparation in bioanalysis can require significant numbers of (manual) steps and consumables. Such procedures can be bottlenecks regarding cost, throughput, and greenness. This is also the case for analysis of new approach methodologies (NAMs) such as organoids and organ-on-a-chip systems. In this context, approaches tailored for measuring drugs in NAM-related cell culture media (CCM) are being developed. For mass spectrometry-based analysis, automated filtration\/filter backflush solid-phase extraction liquid chromatography AFFL-SPE-LC is utilized. Case studies have shown that the \u201cAFFL\u201d platform allows for vast reductions in sample preparation efforts, as the \u201cself-cleaning\u201d filtration features and SPE conditions remove potentially clogging\/contaminating materials from samples of biological origin, e.g., the salt and protein content in CCM. Here, broader demonstrations of the AFFL platform\u2019s traits for CCM analysis are provided, employing an extended panel of hydrophobic small-molecule drugs. The investigated AFFL platform shows reproducible chromatographic performance, as well as fit-for-purpose inter-matrix and inter-column robustness. Hundred-scale injections can be performed with satisfactory repeatability (e.g., retention time RSDs\u2009&lt;\u20091%), with limited interference from the various CCM matrices investigated. Practical NAM applications are also demonstrated. The approach has clear advantages in greenness, obtaining an AGREE prep score of 0.71, contrasting the\u2009&lt;\u20090.55 scores of more conventional\/commercial approaches. The plastic consumable usage of our approach is 28\u00a0g\/100 samples, a nearly 20-fold improvement over a previously established benchmark (500\u00a0g\/100 samples). Taken together, these results demonstrate that AFFL provides a robust, low-waste, and scalable LC-MS-based workflow for chemical analysis of NAM-derived samples.<\/jats:p>","DOI":"10.1007\/s00216-026-06775-w","type":"journal-article","created":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T12:29:21Z","timestamp":1788870561000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Expanded evaluation of a robust on-line sample cleanup-LC-MS platform suited for new approach methodologies (NAM)"],"prefix":"10.1007","author":[{"given":"Marcus","family":"Kamande","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julie","family":"Bekkhus","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stian","family":"Kogler","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aysan","family":"Safavi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mads T. S.","family":"Gisle Johnsen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrik","family":"Appelblad","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steven Ray","family":"Wilson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hanne","family":"R\u00f8berg-Larsen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,9,8]]},"reference":[{"key":"6775_CR1","doi-asserted-by":"publisher","unstructured":"Kogler S, K\u00f8murcu KS, Olsen C, Shoji JY, Skottvoll FS, Krauss S, Wilson SR, R\u00f8berg-Larsen H. Organoids, organ-on-a-chip, separation science and mass spectrometry: an update. TrAC Trends Anal Chem. 2023;161:116996. https:\/\/doi.org\/10.1016\/j.trac.2023.116996.","DOI":"10.1016\/j.trac.2023.116996"},{"issue":"1","key":"6775_CR2","doi-asserted-by":"publisher","DOI":"10.1038\/s43586-022-00118-6","volume":"2","author":"CM Leung","year":"2022","unstructured":"Leung CM, de Haan P, Ronaldson-Bouchard K, Kim GA, Ko J, Rho HS, et al. A guide to the organ-on-a-chip. Nat Rev Methods Primer. 2022;2(1):33. https:\/\/doi.org\/10.1038\/s43586-022-00118-6.","journal-title":"Nat Rev Methods Primer"},{"key":"6775_CR3","doi-asserted-by":"publisher","unstructured":"Zhao Z, Chen X, Dowbaj AM, Sljukic A, Bratlie K. Lin L,\u00a0Fong EL, Balachander GM, Chen Z, Soragni A, Huch M.\u00a0Organoids. Nat Rev Methods Primer. 2022;2(1):94. https:\/\/doi.org\/10.1038\/s43586-022-00174-y.","DOI":"10.1038\/s43586-022-00174-y"},{"issue":"3","key":"6775_CR4","doi-asserted-by":"publisher","first-page":"204","DOI":"10.1038\/s41573-025-01317-y","volume":"25","author":"D Wang","year":"2026","unstructured":"Wang D, Villenave R, Stokar-Regenscheit N, Clevers H. Human organoids as 3D in vitro platforms for drug discovery: opportunities and challenges. Nat Rev Drug Discov. 2026;25(3):204\u201326. https:\/\/doi.org\/10.1038\/s41573-025-01317-y.","journal-title":"Nat Rev Drug Discov"},{"key":"6775_CR5","doi-asserted-by":"publisher","DOI":"10.3389\/fddsv.2024.1355044","author":"T Hartung","year":"2024","unstructured":"Hartung T. The (misleading) role of animal models in drug development. Front Drug Discov. 2024. https:\/\/doi.org\/10.3389\/fddsv.2024.1355044.","journal-title":"Front Drug Discov"},{"key":"6775_CR6","doi-asserted-by":"publisher","unstructured":"FDA pushes to replace animal testing. Nat Biotechnol. 2025;43(5):655\u2013655. https:\/\/doi.org\/10.1038\/s41587-025-02690-0.","DOI":"10.1038\/s41587-025-02690-0"},{"issue":"29","key":"6775_CR7","doi-asserted-by":"publisher","first-page":"12129","DOI":"10.1021\/acs.analchem.4c02246","volume":"96","author":"S Kogler","year":"2024","unstructured":"Kogler S, Pedersen GM, Mart\u00ednez-Ram\u00edrez F, Aizenshtadt A, Busek M, Krauss SJK, et al. An FDA-validated, self-cleaning liquid chromatography\u2013mass spectrometry system for determining small-molecule drugs and metabolites in organoid\/organ-on-chip medium. Anal Chem. 2024;96(29):12129\u201338. https:\/\/doi.org\/10.1021\/acs.analchem.4c02246.","journal-title":"Anal Chem"},{"issue":"9","key":"6775_CR8","doi-asserted-by":"publisher","first-page":"4923","DOI":"10.1021\/acs.analchem.4c04994","volume":"97","author":"S Kogler","year":"2025","unstructured":"Kogler S, Skottvoll FS, Hru\u0161kov\u00e1 H, Rise F, Aizenshtadt A, Krauss S, et al. Electromembrane extraction provides unprecedented selectivity for drugs in cell culture media used in organoid and organ-on-chip systems. Anal Chem. 2025;97(9):4923\u201331. https:\/\/doi.org\/10.1021\/acs.analchem.4c04994.","journal-title":"Anal Chem"},{"issue":"9","key":"6775_CR9","doi-asserted-by":"publisher","first-page":"2005","DOI":"10.1038\/s12276-023-01074-1","volume":"55","author":"SP Harrison","year":"2023","unstructured":"Harrison SP, Siller R, Tanaka Y, Chollet ME, de la Morena-Barrio ME, Xiang Y. Scalable production of tissue-like vascularized liver organoids from human PSCs. Exp Mol Med. 2023;55(9):2005\u201324. https:\/\/doi.org\/10.1038\/s12276-023-01074-1.","journal-title":"Exp Mol Med"},{"key":"6775_CR10","doi-asserted-by":"publisher","first-page":"120","DOI":"10.1016\/j.jpba.2013.05.006","volume":"87","author":"M Rogeberg","year":"2014","unstructured":"Rogeberg M, Malerod H, Roberg-Larsen H, Aass C, Wilson SR. On-line solid phase extraction\u2013liquid chromatography, with emphasis on modern bioanalysis and miniaturized systems. J Pharm Biomed Anal. 2014;87:120\u20139. https:\/\/doi.org\/10.1016\/j.jpba.2013.05.006.","journal-title":"J Pharm Biomed Anal"},{"key":"6775_CR11","doi-asserted-by":"publisher","DOI":"10.1016\/j.microc.2023.108395","volume":"187","author":"M\u00c1 de la Serna Calleja","year":"2023","unstructured":"de la Serna Calleja M\u00c1, Bolado S, Jim\u00e9nez JJ, L\u00f3pez-Serna R. Performance critical comparison of offline SPE, online SPE, and direct injection for the determination of CECs in complex liquid environmental matrices. Microchem J. 2023;187:108395. https:\/\/doi.org\/10.1016\/j.microc.2023.108395.","journal-title":"Microchem J"},{"key":"6775_CR12","doi-asserted-by":"publisher","DOI":"10.1016\/j.trac.2023.116976","volume":"160","author":"L Chen","year":"2023","unstructured":"Chen L, Yan X, Zhou X, Peng P, Sun Q, Zhao F. Advances in the on-line solid-phase extraction-liquid chromatography-mass spectrometry analysis of emerging organic contaminants. TrAC Trends Anal Chem. 2023;160:116976. https:\/\/doi.org\/10.1016\/j.trac.2023.116976.","journal-title":"TrAC Trends Anal Chem"},{"issue":"1","key":"6775_CR13","doi-asserted-by":"publisher","DOI":"10.1038\/srep31693","volume":"6","author":"OK Brandtzaeg","year":"2016","unstructured":"Brandtzaeg OK, Johnsen E, Roberg-Larsen H, Seip KF, MacLean EL, Gesquiere LR. Proteomics tools reveal startlingly high amounts of oxytocin in plasma and serum. Sci Rep. 2016;6(1):31693. https:\/\/doi.org\/10.1038\/srep31693.","journal-title":"Sci Rep"},{"issue":"21","key":"6775_CR14","doi-asserted-by":"publisher","first-page":"3020","DOI":"10.1002\/jssc.201100553","volume":"34","author":"KO Svendsen","year":"2011","unstructured":"Svendsen KO, Larsen HR, Pedersen SA, Brenna I, Lundanes E, Wilson SR. Automatic filtration and filter flush for robust online solid-phase extraction liquid chromatography. J Sep Sci. 2011;34(21):3020\u20132. https:\/\/doi.org\/10.1002\/jssc.201100553.","journal-title":"J Sep Sci"},{"issue":"16","key":"6775_CR15","doi-asserted-by":"publisher","first-page":"11899","DOI":"10.1021\/acs.analchem.5c08225","volume":"98","author":"L Noreng","year":"2026","unstructured":"Noreng L, \u00d8iestad \u00c5ML, Hansen FA, R\u00f8berg-Larsen H, Wilson SR, \u00d8iestad EL. Toward Sustainable Clinical Analysis: Benchmarking Plastic Use in LC\u2013MS Sample Preparation \u2013 Exemplified by Ketamine Analogs in Whole Blood. Anal Chem. 2026;98(16):11899\u2013909. https:\/\/doi.org\/10.1021\/acs.analchem.5c08225.","journal-title":"Anal Chem"},{"issue":"8","key":"6775_CR16","doi-asserted-by":"publisher","first-page":"3544","DOI":"10.1021\/acs.analchem.3c05431","volume":"96","author":"CR Canez","year":"2024","unstructured":"Canez CR, Li L. Studies of labware contamination during lipid extraction in mass spectrometry-based lipidome analysis. Anal Chem. 2024;96(8):3544\u201352. https:\/\/doi.org\/10.1021\/acs.analchem.3c05431.","journal-title":"Anal Chem"},{"issue":"7583","key":"6775_CR17","doi-asserted-by":"publisher","first-page":"479","DOI":"10.1038\/528479c","volume":"528","author":"MA Urbina","year":"2015","unstructured":"Urbina MA, Watts AJR, Reardon EE. Labs should cut plastic waste too. Nature. 2015;528(7583):479\u2013479. https:\/\/doi.org\/10.1038\/528479c.","journal-title":"Nature"},{"key":"6775_CR18","doi-asserted-by":"publisher","DOI":"10.1016\/j.trac.2022.116553","volume":"149","author":"W Wojnowski","year":"2022","unstructured":"Wojnowski W, Tobiszewski M, Pena-Pereira F, Psillakis E. AGREEprep \u2013 Analytical greenness metric for sample preparation. TrAC Trends Anal Chem. 2022;149:116553. https:\/\/doi.org\/10.1016\/j.trac.2022.116553.","journal-title":"TrAC Trends Anal Chem"},{"issue":"9","key":"6775_CR19","doi-asserted-by":"publisher","first-page":"6792","DOI":"10.1021\/acs.analchem.5c07140","volume":"98","author":"KS K\u00f8murcu","year":"2026","unstructured":"K\u00f8murcu KS, Zawadzka ME, Meszka I, Aizenshtadt A, Hru\u0161kov\u00e1 H, Aakervik LE. A validated mass spectrometry platform for oxysterol analysis of single human gastruloids and liver organoids. Anal Chem. 2026;98(9):6792\u2013804. https:\/\/doi.org\/10.1021\/acs.analchem.5c07140.","journal-title":"Anal Chem"}],"container-title":["Analytical and Bioanalytical Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s00216-026-06775-w.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1007\/s00216-026-06775-w","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s00216-026-06775-w.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T12:29:22Z","timestamp":1788870562000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1007\/s00216-026-06775-w"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,8]]},"references-count":19,"alternative-id":["6775"],"URL":"https:\/\/doi.org\/10.1007\/s00216-026-06775-w","relation":{},"ISSN":["1618-2642","1618-2650"],"issn-type":[{"value":"1618-2642","type":"print"},{"value":"1618-2650","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,8]]},"assertion":[{"value":"2 June 2026","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"19 August 2026","order":2,"name":"revised","label":"Revised","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"20 August 2026","order":3,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"8 September 2026","order":4,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"PA is employed by Merck Life Science AS in Norway and holds a role within the global Merck Life Science Chemistry R&D organization. The remaining authors declare no conflicts of interest.","order":1,"name":"Ethics","label":"Conflict of interest","group":{"name":"EthicsHeading","label":"Declarations"}}]}}