{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T03:16:22Z","timestamp":1788837382997,"version":"build-2803163510"},"reference-count":32,"publisher":"Wiley","issue":"17","license":[{"start":{"date-parts":[[2018,8,5]],"date-time":"2018-08-05T00:00:00Z","timestamp":1533427200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"US Department of Agriculture, Agricultural Research Service (USDA-ARS)"}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Rapid Comm Mass Spectrometry"],"published-print":{"date-parts":[[2018,9,15]]},"abstract":"<jats:sec>\n                    <jats:title>Rationale<\/jats:title>\n                    <jats:p>The occurrence of antibiotic\u2010resistant bacteria is a worldwide issue that has the potential, if not addressed, to eliminate classes of antibiotics that have extended life expectancy in the last century. An approach to confront this threat is the development of technologies that greatly accelerate the detection of antibiotic resistance to minimize unnecessary treatment involving antibiotics. Development of an analytical method for rapid detection of aminoglycoside resistance using liquid chromatography\/mass spectrometry (LC\/MS) has not been reported in the literature and is described here.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      A strain of\n                      <jats:styled-content style=\"fixed-case\">\n                        <jats:italic>Escherichia coli<\/jats:italic>\n                      <\/jats:styled-content>\n                      carrying a plasmid encoding an aminoglycoside\u2010modifide enzyme (N\u2010acetyltransferase) was incubated with kanamycin, an aminoglycoside. The antibiotic and its modified form were observed using LC\/MS. An ABSciex QTrap 6500+ was used for kinetic and quantitative analysis and high\u2010resolution structural elucidation was performed using a Thermo Fisher Q\u2010Exactive hybrid quadrupole\u2010orbitrap mass spectrometer.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Detection of kanamycin modification was achieved in less than an hour of incubation. Calibration curves for both modified and unmodified kanamycin from 0.5 to 50 \u03bcg mL\n                      <jats:sup>\u22121<\/jats:sup>\n                      were obtained. Generation and depletion of modified and unmodified kanamycin as a function of time were performed. High\u2010resolution mass spectrometry was employed for confirmation and structural elucidation of the novel precursor and product ion biomarkers with high mass accuracy (\u22647 ppm).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>A newly developed analytical method is able to determine bacterial resistance to aminoglycosides (via acetylation of kanamycin), qualitatively and quantitatively, within 30\u00a0minutes and 6\u00a0hours of incubation with kanamycin, respectively. High\u2010resolution data support the placement of an acetyl group on kanamycin confirming aminoglycoside resistance and its mechanism. Quantification was achieved for both forms of the antibiotic 50\u2010 to 100\u2010fold lower than the minimum inhibitory concentration for the resistant bacteria and can be used to replace conventional antimicrobial susceptibility tests.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/rcm.8160","type":"journal-article","created":{"date-parts":[[2018,5,21]],"date-time":"2018-05-21T01:58:43Z","timestamp":1526867923000},"page":"1549-1556","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["Detection of acetyltransferase modification of kanamycin, an aminoglycoside antibiotic, in bacteria using ultrahigh\u2010performance liquid chromatography tandem mass spectrometry"],"prefix":"10.1002","volume":"32","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0789-5471","authenticated-orcid":false,"given":"Johnny J.","family":"Perez","sequence":"first","affiliation":[{"name":"Residue Chemistry and Predictive Microbiology Research Unit US Department of Agriculture  600 East Mermaid Lane Wyndmoor PA 19038 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chin\u2010Yi","family":"Chen","sequence":"additional","affiliation":[{"name":"Molecular Characterization of Foodborne Pathogens Research Unit US Department of Agriculture, Agricultural Research Service, Eastern Regional Research Center  600 East Mermaid Lane Wyndmoor PA 19038 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2018,8,5]]},"reference":[{"key":"e_1_2_6_2_1","first-page":"277","volume":"40","author":"Ventola CL","year":"2015","journal-title":"Pharm 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