Nishizawa Chiemi, Yamaura Yoshio
Nagano Research Institute for Health and Pollution.
Chudoku Kenkyu. 2003 Oct;16(4):441-5.
High-performance liquid chromatographic (HPLC) assay has been developed for the simultaneous determination of alpha-amanitin, beta-amanitin and phalloidin in serum. Three toxins were extracted by reflux in a water bath at 80 degrees C for one hour and purified by Sep-Pak Plus tC18 cartridges. The HPLC assay was performed under gradient conditions using Develosil RP AQUEOUS column. The moble phase consisted with a mixture of acetonitorile containing 0.01 M ammonium acetate(pH 5.0). The column effluence was monitored at 295 nm, 302 nm and 230 nm for 35 min. Detection limit of three toxins in serum were 0.2 microgram/ml respectively. High recovery yields in the range of 81.5-88.1% for toxins were obtained by using this method.
已开发出高效液相色谱(HPLC)分析法,用于同时测定血清中的α-鹅膏毒肽、β-鹅膏毒肽和鬼笔环肽。三种毒素在80℃水浴中回流提取1小时,并用Sep-Pak Plus tC18柱进行纯化。HPLC分析在梯度条件下使用Develosil RP AQUEOUS柱进行。流动相由含有0.01 M醋酸铵(pH 5.0)的乙腈混合物组成。在295 nm、302 nm和230 nm处监测柱流出物35分钟。血清中三种毒素的检测限分别为0.2微克/毫升。使用该方法可获得81.5-88.1%范围内的高回收率。