Ganhao M F, Hattingh J, Hurwitz M L, Pitts N I
Department of General Physiology, Dental School, University of Witwatersrand, Johannesburg, South Africa.
J Chromatogr. 1991 Mar 8;564(1):55-66. doi: 10.1016/0378-4347(91)80069-o.
The modified extraction method for catecholamines described in this study is reproducible, simple, rapid, economical and relatively hazard-free. This method is based on the principle that plasma catecholamines are selectively adsorbed on acid-washed alumina at pH 8.6 and then eluted at a pH between 1.0 and 2.0. No statistically significant differences were obtained by using either 0.5 or 1.0 ml of plasma with 0.5 or 1.0 ml of Tris buffer. A 15-min mixing time during the adsorption and desorption steps was found to be practical, but any standardized time up to 1 h can be used. If the washing step was omitted, the catecholamines could not be eluted from the acid-washed alumina. To prevent dilution, the alumina had to be centrifuged and not aspirated to dryness after the washing step. An amount of 50 mg of WA-4 alumina was found to be the most practical in this study. Extracted or unextracted plasma as well as catecholamine standards were stable for four months at -20 degrees C.
本研究中描述的儿茶酚胺改良提取方法具有可重复性、简单、快速、经济且相对无危害的特点。该方法基于以下原理：血浆儿茶酚胺在pH 8.6时被选择性吸附在酸洗过的氧化铝上，然后在pH 1.0至2.0之间洗脱。使用0.5或1.0毫升血浆与0.5或1.0毫升Tris缓冲液时，未获得统计学上的显著差异。发现在吸附和解吸步骤中15分钟的混合时间是可行的，但也可以使用长达1小时的任何标准化时间。如果省略洗涤步骤，儿茶酚胺就无法从酸洗过的氧化铝上洗脱下来。为防止稀释，洗涤步骤后必须对氧化铝进行离心，而不是抽吸至干燥。在本研究中发现50毫克WA - 4氧化铝的量最为实用。提取或未提取的血浆以及儿茶酚胺标准品在-20℃下可稳定保存四个月。
