Hamilton C L, Kirkwood J A, Carter G, Williams R S
Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285.
J Chromatogr. 1993 Apr 2;613(2):365-70. doi: 10.1016/0378-4347(93)80156-x.
A reversed-phase HPLC method is reported for the determination of a new M1 agonist, LY246708, in human plasma. The compound and an internal standard were extracted from plasma with hexane at basic pH. The organic extract was evaporated to dryness and the residue was reconstituted with mobile phase [0.5% diethylamine (pH 3, adjusted with phosphoric acid)-acetonitrile (70:30, v/v)]. The analytes were separated from endogenous substances on a Zorbax CN column; the effluent was monitored by measuring its absorbance at 296 nm. The limit of quantification was determined at 1.5 ng/ml and the response was linear from 1.5 to 20 ng/ml. Validation studies showed the method to be both repeatable and reproducible. Its robustness was demonstrated by transfer between analytical laboratories and continued use in support of pharmacokinetic studies and therapeutic monitoring of the compound.
报道了一种反相高效液相色谱法，用于测定人血浆中一种新型M1激动剂LY246708。该化合物和内标在碱性pH条件下用己烷从血浆中萃取。有机萃取液蒸发至干，残渣用流动相[0.5%二乙胺（pH 3，用磷酸调节）-乙腈（70:30，v/v）]复溶。分析物在Zorbax CN柱上与内源性物质分离；通过测量流出物在296 nm处的吸光度进行监测。定量限为1.5 ng/ml，响应在1.5至20 ng/ml范围内呈线性。验证研究表明该方法具有可重复性和再现性。通过在分析实验室之间转移并持续用于支持该化合物的药代动力学研究和治疗监测，证明了其稳健性。
