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Accurate determination of its concentration is essential for quality control, therapeutic drug monitoring, and preventing toxic effects associated with overdosing or accumulation in the environment. In this work, a simple, rapid, and highly sensitive spectrofluorimetric method was developed for quantifying streptomycin in pure form, pharmaceutical vials, and human plasma using fluorescamine as a fluorogenic probe. The method is based on the reaction of fluorescamine with the primary amine group of streptomycin under mild alkaline conditions to yield a fluorescent product emitting at 482\u00a0nm (excitation 390\u00a0nm). The calibration curve was linear over the range 100\u2013600 ng\/mL, with a correlation coefficient of 0.9999, a detection limit of 8.3 ng\/mL, and a quantitation limit of 25.2 ng\/mL, ensuring sensitive and accurate determination across the studied range. The procedure demonstrated high precision (RSD\u2009&lt;\u20092%) and accuracy (recoveries 98.1\u2013101.3%). The method was successfully applied to commercial streptomycin vials (% recovery\u2009=\u200999.7\u2009\u00b1\u20090.8) and to spiked human plasma samples with negligible matrix interference. Owing to its simplicity, short analysis time (3\u00a0min), and excellent reproducibility, the proposed assay offers a cost-effective alternative to chromatographic methods for routine quality control and bioanalytical applications.<\/jats:p>\n                  <jats:p>\n                    <jats:bold>Graphical abstract<\/jats:bold>\n                  <\/jats:p>","DOI":"10.1186\/s13065-025-01708-7","type":"journal-article","created":{"date-parts":[[2026,1,17]],"date-time":"2026-01-17T17:15:44Z","timestamp":1768670144000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Turning on fluorescent probe for sensitive detection of streptomycin in pure, pharmaceutical formulations and human plasma"],"prefix":"10.1186","volume":"20","author":[{"given":"Bassant","family":"Samy","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mokhtar M.","family":"Mabrouk","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mohamed A. 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Ethical approval was obtained from the Research Ethical Committee of the Faculty of Pharmacy, Tanta University, Egypt, under approval number TR\/RE\/4\/25 p-006. The research involved the use of spiked human plasma from a healthy volunteer, without administering any drug to the participant. The volunteer was fully informed about the objectives and nature of the study, and written informed consent was obtained prior to participation.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable. The manuscript does not contain any individual person\u2019s data in any form.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"The authors declare no competing interests.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"25"}}