Hurst P R, Whelpton R
Department of Pharmacology, London Hospital Medical College, UK.
Biomed Chromatogr. 1989 Sep;3(5):226-32. doi: 10.1002/bmc.1130030511.
A simple, selective and very sensitive assay is described for the quantification of physostigmine in blood, plasma and urine. The most appropriate solid phase column was selected after a systematic investigation of nine types of phase. The conditions for solid phase extraction were optimized using [3H]physostigmine so that the overall recoveries were greater than 90%. Physostigmine was retained on alkaline treated cyanopropyl columns and eluted into the minimum volume of methanol, obviating the need for an evaporation step. Extracted samples were quantified by HPLC with a three electrode coulometric detection system. The limit of detection was 50 pg/mL for a 0.5 mL plasma sample. The precision (CV) for 0.5 mL plasma samples containing 50 pg was 8.1%. Application of the method to plasma, blood and urine samples is presented.
本文描述了一种简单、选择性强且灵敏度高的方法，用于定量测定血液、血浆和尿液中的毒扁豆碱。在对九种类型的固定相进行系统研究后，选择了最合适的固相柱。使用[3H]毒扁豆碱对固相萃取条件进行了优化，使总回收率大于90%。毒扁豆碱保留在经碱处理的氰丙基柱上，并用最小体积的甲醇洗脱，无需蒸发步骤。通过带有三电极库仑检测系统的高效液相色谱法对提取的样品进行定量。对于0.5 mL血浆样品，检测限为50 pg/mL。含50 pg的0.5 mL血浆样品的精密度（CV）为8.1%。本文还介绍了该方法在血浆、血液和尿液样品中的应用。
