Clinical Pharmacology/Psychopharmacology, Department of Psychiatry and Psychotherapy, University of Regensburg, Universitätsstrasse 84, 93053 Regensburg, Germany.
J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Jan 15;881-882:1-11. doi: 10.1016/j.jchromb.2011.10.027. Epub 2011 Nov 10.
A novel, simple, specific and sensitive high performance liquid chromatography (HPLC) assay for the detection and quantification of donepezil in serum of demented patients has been developed and validated. The analytical procedure involves an offline serum preextraction using solid phase extraction (SPE) cartridges (Oasis® HLB, Waters Co). The chromatographic analyses were performed on a Dionex HPLC system with a Phenomenex Luna Phenyl-Hexyl analytical column, and a mobile phase with the two components 0.02 mol/l phosphate buffer and acetonitrile. The flow rate was 0.4 ml/min. For the detection of donepezil three different UV wavelengths were used as an interference-control check. Interference tests between donepezil and 100 of the most commonly used concomitant medications allow quantification of donepezil under the polypharmaceutical conditions of the daily clinical routine. The retention time for donepezil was 12.1 min. The method was validated according to the guidelines of the Society of Toxicology and Forensic Chemistry (GTFCh): The calibration curve was linear over a concentration range from 5 to 160 ng/ml (n=8/r²>0.999). No endogenous compounds were found to interfere with the analyte, which was shown by retention times for the comedication most often prescribed to demented patients. The method had an accuracy of >85%. Intra- and inter-assay coefficients of variation were <6% and <8%, respectively, at three different concentrations. The limit of quantification (LOQ) and the limit of detection (LOD) were found to be 6.1 and 1.7 ng/ml for donepezil. Application of the method to patient serum samples discovered that concentrations suggested as "therapeutic" in the literature may only be reached either by high, off-label dosages or by utilization of inhibitory metabolic effects of the comedication.
一种新颖、简单、特异、灵敏的高效液相色谱（HPLC）法已被建立并验证，用于检测和定量痴呆患者血清中的多奈哌齐。分析程序涉及使用固相萃取（SPE）小柱（Oasis®HLB，Waters Co.）进行离线血清预提取。色谱分析在 Dionex HPLC 系统上进行，采用 Phenomenex Luna Phenyl-Hexyl 分析柱，流动相由两种成分 0.02 mol/l 磷酸盐缓冲液和乙腈组成。流速为 0.4 ml/min。为了检测多奈哌齐，使用了三个不同的紫外波长作为干扰控制检查。多奈哌齐与 100 种最常用伴随药物之间的干扰试验允许在日常临床常规的多药治疗条件下定量多奈哌齐。多奈哌齐的保留时间为 12.1 min。该方法按照毒理学和法医化学学会（GTFCh）的指南进行了验证：校准曲线在 5 至 160 ng/ml 浓度范围内呈线性（n=8/r²>0.999）。没有发现内源性化合物干扰分析物，这是通过保留时间证明的，对于最常开给痴呆患者的伴随药物。该方法的准确度>85%。在三个不同浓度下，日内和日间变异系数分别<6%和<8%。多奈哌齐的定量限（LOQ）和检测限（LOD）分别为 6.1 和 1.7 ng/ml。该方法应用于患者血清样本，发现文献中建议的“治疗”浓度可能仅通过高剂量、超说明书剂量或利用伴随药物的抑制代谢作用来达到。
