Laboratory of Pharmaco-Toxicological Analysis, Department of Pharmaceutical Sciences, University of Bologna, Via Belmeloro 6, I-40126, Bologna, Italy.
J Chromatogr A. 2011 Apr 22;1218(16):2153-9. doi: 10.1016/j.chroma.2011.01.071. Epub 2011 Feb 4.
A novel analytical approach has been developed for the determination of clozapine and its metabolites in dried blood spots on filter paper, using a chromatographic method coupled with a microextraction by packed sorbent procedure. The analytes were separated on a RP-C18 column using a mobile phase composed of 20% methanol, 16% acetonitrile and 64% aqueous phosphate buffer. Coulometric detection was used, setting the guard cell at +0.050 V, the first analytical cell at -0.200 V and the second analytical cell at +0.500 V. Clozapine and its metabolites were extracted from dried blood spots with phosphate buffer and, then, a microextraction by packed sorbent procedure for the sample clean-up was implemented obtaining good extraction yields. The calibration curve was linear over the 2.5-1000 ng mL(-1) blood concentration ranges for all the analytes. The method validation gave satisfactory results in terms of sensitivity, precision, selectivity and accuracy. The analytical method was successfully applied to dried blood spots from several psychiatric patients for therapeutic drug monitoring purpose.
已开发出一种新颖的分析方法，用于在滤纸干血斑上测定氯氮平和其代谢物，采用色谱法与填充固相微萃取程序相结合。使用由 20%甲醇、16%乙腈和 64%磷酸缓冲水溶液组成的流动相，在 RP-C18 柱上分离分析物。采用库仑检测法，将保护池设置为+0.050 V，第一分析池设置为-0.200 V，第二分析池设置为+0.500 V。氯氮平和其代谢物用磷酸盐缓冲液从干血斑中提取，然后进行固相微萃取程序的微萃取以实现样品净化，获得了良好的提取效率。校准曲线在所有分析物的 2.5-1000ngmL(-1)血浓度范围内呈线性。该方法在灵敏度、精密度、选择性和准确性方面均取得了令人满意的结果。该分析方法成功地应用于来自几位精神科患者的干血斑，用于治疗药物监测目的。
