Sundström Ingela, Andrén Per E, Westerlund Douglas
Analytical Pharmaceutical Chemistry, Uppsala University, Biomedical Centre, Box 574, SE-751 23 Uppsala, Sweden; Department of Pharmaceutical Biosciences, Medical Mass Spectrometry, Uppsala University, Biomedical Centre, Box 583, SE-751 23 Uppsala, Sweden.
J Chromatogr A. 2008 May 2;1189(1-2):503-13. doi: 10.1016/j.chroma.2008.02.043. Epub 2008 Feb 16.
Methodologies for identification of ketobemidone metabolites in microdialysate samples utilizing coupled-column capillary liquid chromatography-electrospray quadrupole time-of-flight tandem mass spectrometry are presented. Two different methods were developed to efficiently analyze the metabolites norketobemidone, ketobemidone N-oxide and hydroxyketobemidone, respectively. Both methods include on-line desalting and trapping of the analytes on micro-solid-phase extraction columns with different retention mechanisms. Norketobemidone and ketobemidone N-oxide were trapped on a C18 column and then eluted by back-flush followed by isocratic separation on a fluorinated reversed-phase type silica gel column (Fluofix). Retention equations are proposed for the chromatographic observations made on the Fluofix column. Hydroxyketobemidone was trapped on a phenylboronic acid column by complex formation at basic pH and then eluted at acidic pH directly into to the mass spectrometer. Oxidation of hydroxyketobemidone to its corresponding quinone was also observed. The methods were successfully used to analyze synthetic ketobemidone metabolites in dilute low-volume microdialysis samples.
本文介绍了利用联用柱毛细管液相色谱-电喷雾四极杆飞行时间串联质谱法鉴定微透析样品中凯托米酮代谢物的方法。开发了两种不同的方法，分别用于高效分析去甲凯托米酮、凯托米酮N-氧化物和羟基凯托米酮等代谢物。两种方法均包括在线脱盐以及通过具有不同保留机制的微固相萃取柱对分析物进行捕集。去甲凯托米酮和凯托米酮N-氧化物被捕集在C18柱上，然后通过反冲洗脱，随后在氟化反相硅胶柱（Fluofix）上进行等度分离。针对在Fluofix柱上的色谱观测结果提出了保留方程。羟基凯托米酮在碱性pH下通过形成络合物被捕集在苯基硼酸柱上，然后在酸性pH下洗脱，直接进入质谱仪。还观察到羟基凯托米酮氧化为其相应的醌。这些方法成功用于分析稀释的小体积微透析样品中的合成凯托米酮代谢物。
