{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T04:24:24Z","timestamp":1782275064347,"version":"3.54.5"},"reference-count":22,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2013,3,25]],"date-time":"2013-03-25T00:00:00Z","timestamp":1364169600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomedical Chromatography"],"published-print":{"date-parts":[[2013,7]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>22\u2010[<jats:italic>N<\/jats:italic>(\u22127\u2010Nitrobenz\u20102\u2010oxa\u20101,3\u2010diazol\u20104\u2010yl)amino]\u201023,24\u2010bisnor\u20105\u2010cholen\u20103<jats:italic>\u03b2<\/jats:italic>\u2010ol (NBD\u2010cholesterol), a fluorescent cholesterol analog, was an extragenous cholesterol tracer used to study cholesterol absorption and metabolism in cultured cells. In order to measure free intracellular cholesterol and its esters, a precise and sensitive method employing high\u2010performance liquid chromatography\/fluorescence detection (HPLC\u2010FLD) was developed for the first time. Method validation showed a limit of detection at 30\u2009ng\/mL. The calibration curve was linear within the range of 0.0625\u201310.0\u2009\u00b5g\/mL (<jats:italic>r<\/jats:italic><jats:sup>2<\/jats:sup>\u2009=\u20090.999). Accuracy and precision were highlighted by good recovery and low variations. Apart from NBD\u2010cholesteryl oleate, two additional cellular metabolites of NBD\u2010cholesterol, probably an isomer and an oxidation product, were determined in the lipid extracts of Caco\u20102 human colon adenocarcinoma cells according to mass spectrometry. In AC29 mouse malignant mesothelioma cells overexpressing acyl\u2010CoA:cholesterol acyltransferase\u20101 (ACAT1) or ACAT2, only the oxidized metabolite was detected. Using the newly developed method, YIC\u2010C8\u2010434, a known ACAT inhibitor, was shown to inhibit ACAT activity in Caco\u20102 cells, as well as in AC29\/ACAT1 or AC29\/ACAT2 cells. In conclusion, the sensitive and specific HPLC\u2010FLD method is a powerful tool for simultaneous quantification of intracellular NBD\u2010cholesterol and its oleoyl\u2010ester. Copyright \u00a9 2013 John Wiley &amp; Sons, Ltd.<\/jats:p>","DOI":"10.1002\/bmc.2881","type":"journal-article","created":{"date-parts":[[2013,3,25]],"date-time":"2013-03-25T00:56:37Z","timestamp":1364172997000},"page":"910-915","source":"Crossref","is-referenced-by-count":6,"title":["HPLC\u2010FLD determination of NBD\u2010cholesterol, its ester and other metabolites in cellular lipid extracts"],"prefix":"10.1002","volume":"27","author":[{"given":"Xiu\u2010zhen","family":"Cao","sequence":"first","affiliation":[{"name":"Unilever R&amp;D Shanghai  66 Lin Xin Road Shanghai 200335 People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ting\u2010yan","family":"Mi","sequence":"additional","affiliation":[{"name":"Unilever R&amp;D Shanghai  66 Lin Xin Road Shanghai 200335 People's Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lin","family":"Li","sequence":"additional","affiliation":[{"name":"Unilever R&amp;D Shanghai  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