{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T16:15:00Z","timestamp":1792080900539,"version":"4.4.0"},"reference-count":26,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2025,11,15]],"date-time":"2025-11-15T00:00:00Z","timestamp":1763164800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2025,11,15]],"date-time":"2025-11-15T00:00:00Z","timestamp":1763164800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100009827","name":"Alexandria University","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100009827","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["BMC Microbiol"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Method suitability for microbial limit tests during microbiological quality control (QC) can be a complicated process that requires multiple steps of optimization and plays a central role in ensuring reliable QC results. When antimicrobial activity of a given product cannot be neutralized, it is assumed that the inhibited microorganism is not present in the product, as specified by the U.S. pharmacopeia. This can potentially lead to contaminants that can multiply during storage or use, resulting in potential health risks or even death.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Method suitability testing was performed on a total of 133 pharmaceutical finished products as part of microbiological QC testing performed prior to product marketing. For total microbial count, depending on the pharmaceutical dosage form, testing included sequential trials of different dilution factors up to 1:200, the addition of 1\u20135% tween (polysorbate) 80, 0.7% lecithin, and\/or filtration using different membrane filter types until the optimal method was selected for each product. Method suitability for microbial limit tests as well as for absence of specific pathogens was verified using a range of bacteria and fungi including\n                      <jats:italic>Burkholderia cepacia<\/jats:italic>\n                      .\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>There was an acceptable microbial recovery of at least 84% for all standard strains with all neutralization methods, demonstrating minimal to no toxicity. Forty of 133 finished products required multiple steps of optimization. Of these, 18 were neutralized through 1:10 dilution with diluent warming. Another 8 had no inherent antimicrobial activity to their API and were neutralized through dilution and the addition of tween 80. Neutralization of the remaining 13 products (mostly antimicrobial drugs) was achieved through variations of different dilution factors and filtration with different membrane filter types with multiple rinsing steps.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      Our results provide detailed protocols that can help guide method suitability testing of finished pharmaceutical products for which neutralization proves challenging. We also include testing for\n                      <jats:italic>B. cepacia<\/jats:italic>\n                      complex in aqueous dosage form, often overlooked in the QC process. Since the current QC practice mostly assumes that these products do not require further testing, similar studies are especially needed to ensure the safety of finished pharmaceutical products during marketing, storage, and use.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s12866-025-04456-9","type":"journal-article","created":{"date-parts":[[2025,11,15]],"date-time":"2025-11-15T08:27:16Z","timestamp":1763195236000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Optimizing neutralization strategies for microbial testing of non-sterile pharmaceutical finished products with challenging method suitability"],"prefix":"10.1186","volume":"25","author":[{"given":"Ahmed","family":"Eldemerdash","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Radwa","family":"Ewaisha","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mustafa","family":"Alseqely","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0418-456X","authenticated-orcid":false,"given":"Michael G.","family":"Shehat","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,11,15]]},"reference":[{"key":"4456_CR1","unstructured":"World Health Organization (WHO). WHO Expert Committee on Specifications for Pharmaceutical Preparations: fifty-eight report 2025. Available from: https:\/\/www.who.int\/publications\/i\/item\/9789240108264. Updated April 15, 2025; cited 2025 June 23, 2025."},{"key":"4456_CR2","doi-asserted-by":"publisher","unstructured":"United States Pharmacopeia (USP). < Chapter 1111> Microbiological examination of nonsterile products: Acceptance criteria for pharmaceutical preparations and substances for pharmaceutical use. In: USP-NF Online. Rockville, MD: United States Pharmacopeial Convention 2013.https:\/\/doi.org\/10.31003\/USPNF_M99830_01_01. Accessed\u00a05 January 2024.","DOI":"10.31003\/USPNF_M99830_01_01"},{"key":"4456_CR3","doi-asserted-by":"publisher","unstructured":"United States Pharmacopeia (USP). < Chapter 61\u2009>\u2009Microbiological examination of nonsterile products: Microbial enumeration tests. In: USP-NF Online. Rockville, MD: United States Pharmacopeial Convention 2013. https:\/\/doi.org\/10.31003\/USPNF_M98800_01_01. Accessed 5 January 2024.","DOI":"10.31003\/USPNF_M98800_01_01"},{"key":"4456_CR4","doi-asserted-by":"publisher","unstructured":"Jimenez L. Microbial limits. In L. Jiminez (Ed.), Microbial contamination control in the pharmaceutical industry. CRC Press; 1st ed.,\u00a02004. pp. 15-44. https:\/\/doi.org\/10.1201\/9780203026267.","DOI":"10.1201\/9780203026267"},{"key":"4456_CR5","doi-asserted-by":"publisher","unstructured":"United States Pharmacopeia (USP). <62\u2009>\u2009microbiological examination of nonsterile products: Tests for specified microorganisms. In: USP-NF Online. Rockville, MD: United States Pharmacopeial Convention 2013.https:\/\/doi.org\/10.31003\/USPNF_M98802_01_01. Accessed 5 January 2024.","DOI":"10.31003\/USPNF_M98802_01_01"},{"key":"4456_CR6","doi-asserted-by":"crossref","unstructured":"Ratajczak M, et al. 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Microbial contamination of nonsterile pharmaceuticals in public hospital settings.Ther Clin Risk Manag. 2010;6:443\u20138.","DOI":"10.2147\/TCRM.S12253"},{"key":"4456_CR11","doi-asserted-by":"crossref","unstructured":"Myemba DT, et al.Microbiological Quality of Selected Local and Imported Non-Sterile Pharmaceutical Products in Dar es Salaam, Tanzania. Infect Drug Resist. 2022;15:2021\u201334.","DOI":"10.2147\/IDR.S355331"},{"key":"4456_CR12","unstructured":"Food and Drug Administration (FDA) Office of Regulatory Affairs. Pharmaceutical microbiology manual. 2020 . Accessed\u00a06 June 2025. Available from: https:\/\/www.fda.gov\/media\/88801\/download"},{"key":"4456_CR13","unstructured":"The GMP Journal. Microbiological Suitability Tests of Non-Sterile Preparations. 2022. Available from: https:\/\/www.gmp-journal.com\/current-articles\/details\/microbiological-suitability-tests-of-non-sterile-preparations.html. 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