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Herein, semi\u2010preparative separation method of diclazuril enantiomers has been developed through normal\u2010phase high\u2010performance liquid chromatography. Effects of chiral stationary phases, alcoholic modifiers, and column temperature on separation of diclazuril were discussed in detail. Both the single\u2010urea\u2010bound 4\u2010chlorophenylcarbamoylated \u03b2\u2010cyclodextrin and amylose tris(3,5\u2010dimethylphenylcarbamate)\u2010coated chiral stationary phases showed strong ability in separation of diclazuril by using <jats:italic>n<\/jats:italic>\u2010hexane\u2013trifluoroacetic acid\u2013ethanol. Then, semi\u2010preparative separation of diclazuril was carried out through stacked injection, and the \"enantiomeric excess\" purities of two fractions were over 98%. Next, the electronic circular dichroism profiles of these two fractions in ethanol solution displayed the mirror image of each other in the range 360\u2013200\u00a0nm. Moreover, effects of acidic\/basic additive, time, and temperature on racemization of diclazuril enantiomers in ethanol solution have been studied in detail through normal\u2010phase high\u2010performance liquid chromatography. Racemization of diclazuril enantiomers was remarkably accelerated through adding triethylamine at high temperature. We envision that this systematic investigation of diclazuril at an enantiomeric level would provide valuable information in future studies involving enantioselective bioactive, metabolic, and toxicological activities.<\/jats:p>","DOI":"10.1002\/jssc.201901201","type":"journal-article","created":{"date-parts":[[2020,1,7]],"date-time":"2020-01-07T07:34:24Z","timestamp":1578382464000},"page":"1240-1247","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["HPLC semi\u2010preparative separation of diclazuril enantiomers and racemization in solution"],"prefix":"10.1002","volume":"43","author":[{"given":"Zhan","family":"Huang","sequence":"first","affiliation":[{"name":"School of Chemistry South China Normal University  Guangzhou P. R. China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dong","family":"Guo","sequence":"additional","affiliation":[{"name":"School of Chemistry South China Normal University  Guangzhou P. R. China"},{"name":"Guangzhou Research &amp; Creativity Biotechnology Co. Ltd.  Guangzhou P. R. China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2986-8551","authenticated-orcid":false,"given":"Jun","family":"Fan","sequence":"additional","affiliation":[{"name":"School of Chemistry South China Normal University  Guangzhou P. R. China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yujing","family":"Zhong","sequence":"additional","affiliation":[{"name":"School of Chemistry South China Normal University  Guangzhou P. R. 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