Quantification of circulating steroids in individual zebrafish using stacking to achieve nanomolar detection limits with capillary electrophoresis and UV-visible absorbance detection.
Quantification of circulating steroids in individual zebrafish using stacking to achieve nanomolar detection limits with capillary electrophoresis and UV-visible absorbance detection.
复制标题
使用堆叠对单个斑马鱼中的循环类固醇进行定量，以通过毛细管电泳和紫外-可见光吸光度检测达到纳摩尔检测限。
DOI：
10.1007/s00216-015-8785-0
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发表时间：
2015
影响因子：
4.3
通讯作者：
Holland,LisaA
中科院分区：
文献类型：
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作者：
Nyakubaya,VincentT;Durney,BrandonC;Ellington,MarriahCG;Kantes,AmberD;Reed,PaigeA;Walter,ShaylynE;Stueckle,JenniferRipley;Holland,LisaA
毛细管电泳和紫外-可见吸收检测与样品堆积，以达到检测限范围为0.2至2 ng/mL（0.8至6 nM）的类固醇。使用带负电荷的环糊精类固醇载体分子在重构样品和分离电解质之间的离散pH界面处实现堆叠。然后使用毛细管电泳在5分钟内分离类固醇，该毛细管电泳通过十二烷基硫酸钠和环糊精结合二级平衡。在总循环血容量为5 μL或更少的个体雌性鱼中证明了在有限样品体积中测量多种类固醇的方法的有效性。在使用商业提取柱开发的样品处理方法之后，血浆中的类固醇回收率范围为81 - 109%，其中17α，20 β-二羟基-孕甾-4-烯-3-酮、睾酮、11-酮睾酮、雌酮、17β-雌二醇和17α-乙炔雌二醇。当应用于生殖活跃的雌性斑马鱼时，由于暴露于不同的溶剂和17β-雌二醇，检测到循环类固醇水平的变化。图形摘要类固醇是在暴露于化学物质的个体斑马鱼中测量的
Capillary electrophoresis and UV-visible absorbance detection are used with sample stacking to achieve detection limits ranging from 0.2 to 2 ng/mL (0.8 to 6 nM) for steroids. Stacking is accomplished using negatively charged cyclodextrin steroid-carrier molecules at a discrete pH interface between the reconstituted sample and the separation electrolyte. Steroids are then separated in under 5 min using capillary electrophoresis that incorporates secondary equilibria via sodium dodecyl sulfate and cyclodextrin. The effectiveness of the method for measurements of multiple steroids in limited sample volumes is demonstrated in individual female fish with total circulating blood volumes of 5 μL or less. Steroid recoveries from plasma following a sample processing method developed with commercial extraction cartridges range from 81 to 109 % for 17α,20β-dihydroxy-pregn-4-en-3-one, testosterone, 11-ketotestosterone, estrone, 17β-estradiol, and 17α-ethinyl estradiol. When applied to reproductively active female zebrafish, changes were detected in the levels of circulating steroids as a result of exposure to different solvents and 17β-estradiol.Graphical abstractSteroids are measured in individual zebrafish subject to chemical exposure