Hjemdahl P
Acta Physiol Scand Suppl. 1984;527:43-54.
Thirty-four laboratories performed altogether 41 plasma catecholamine assays on each of four samples. Various radioenzymatic assays, high performance liquid chromatographic (HPLC) assays with electrochemical detection and fluorimetric assays were used. There was reasonable agreement that the levels of adrenaline and noradrenaline in a basal plasma pool were about 0.2 and 1.8 nM, respectively. The levels in a pool of plasma obtained after exercise were about 0.7 and 11 nM, respectively. The study, however, revealed a sometimes considerable variability between methods as well as between laboratories using the same method. Results from duplicate determinations of noradrenaline suggest frequent problems with intra-laboratory reproducibility. Results concerning the recoveries of 0.7 or 3.0 nM adrenaline or 2.0 nM noradrenaline (added to the basal plasma pool) showed a rather frequent need for improved precision. Fluorimetric assays gave unacceptable results. Plasma free dopamine measurements showed a basal level of 0.1-0.2 nM with most HPLC assays and a tendency towards higher levels and greater scatter with radioenzymatic methods. On the whole, reverse phase HPLC methods and an inhomogeneous group of single-isotope derivative radioenzymatic assays showed the largest variability. Less variability was found with the radioenzymatic assay of Peuler & Johnson (1977), provided that a few obviously erroneous results were excluded. The smallest variability was found with microparticulate cation exchange HPLC. It is concluded that plasma catecholamine assays would benefit from better standardization and a continuous quality control. Problems associated with validation of new assays, as well as modifications of old assays are discussed.
34个实验室对4个样本中的每一个都总共进行了41次血浆儿茶酚胺测定。使用了各种放射酶测定法、带电化学检测的高效液相色谱(HPLC)测定法和荧光测定法。大家普遍认同,基础血浆库中肾上腺素和去甲肾上腺素的水平分别约为0.2和1.8 nM。运动后获得的血浆库中,其水平分别约为0.7和11 nM。然而,该研究显示,不同方法之间以及使用相同方法的不同实验室之间有时存在相当大的差异。去甲肾上腺素重复测定的结果表明,实验室内的可重复性经常出现问题。关于向基础血浆库中添加0.7或3.0 nM肾上腺素或2.0 nM去甲肾上腺素后的回收率结果显示,相当频繁地需要提高精密度。荧光测定法给出的结果不可接受。大多数HPLC测定法显示血浆游离多巴胺测量的基础水平为0.1 - 0.2 nM,而放射酶法显示其有升高和更大离散度的趋势。总体而言,反相HPLC方法和一组不均匀的单同位素衍生物放射酶测定法显示出最大的差异。如果排除一些明显错误的结果,Peuler和Johnson(1977年)的放射酶测定法的差异较小。微粒阳离子交换HPLC的差异最小。结论是,血浆儿茶酚胺测定将受益于更好的标准化和持续的质量控制。讨论了与新测定法验证以及旧测定法修改相关的问题。