Husseini H, Mitrovic V, Schlepper M
Max Planck Institute for Physiological and Clinical Research, Kerckhoff-Klinik GmbH, Bad Nauheim, Germany.
J Chromatogr. 1993 Oct 22;620(1):164-8. doi: 10.1016/0378-4347(93)80066-d.
A sensitive and specific high-performance liquid chromatographic method with electrochemical detection was developed for measuring dobutamine in human plasma samples. Following an external standard method, 0.1 ml of EDTA-glutathione plasma was diluted on ice with 0.2 ml of a 5% trichloracetic acid solution. The mixture was centrifuged, filtered, and 30 microliters were injected. Assessment was done by electrochemical detection. The assay was linear from 1 to 400 ng/ml plasma. For determination of dobutamine we also used a liquid-liquid extraction method routinely applied for plasma catecholamines. Liquid-liquid extraction requires application of 100-1000 microliters of plasma. The standard curve was linear from 0.1 to 600 ng/ml. Absolute recovery of dobutamine was 90 +/- 3% with the liquid-liquid extraction procedure and 91 +/- 3% with the protein precipitation method. For both methods dobutamine was separated on Nova-Pak C18 columns. The mobile phase used was 0.1 molar phosphate buffer-acetonitrile (80:20, v/v) with 1-octanesulfonic acid and triethylamine as ion-pair reagents. The pH was adjusted to 2.7.
建立了一种灵敏且特异的高效液相色谱电化学检测法，用于测定人血浆样本中的多巴酚丁胺。采用外标法，将0.1 ml乙二胺四乙酸 - 谷胱甘肽血浆在冰上用0.2 ml 5%三氯乙酸溶液稀释。混合物离心、过滤后，进样30微升。通过电化学检测进行评估。该测定法在血浆浓度1至400 ng/ml范围内呈线性。为测定多巴酚丁胺，我们还使用了一种常用于血浆儿茶酚胺的液 - 液萃取法。液 - 液萃取需要使用100 - 1000微升血浆。标准曲线在0.1至600 ng/ml范围内呈线性。液 - 液萃取法中多巴酚丁胺的绝对回收率为90±3%，蛋白质沉淀法为91±3%。两种方法中多巴酚丁胺均在Nova - Pak C18柱上分离。所用流动相为0.1摩尔磷酸盐缓冲液 - 乙腈（80:20，v/v），含1 - 辛烷磺酸和三乙胺作为离子对试剂。将pH调至2.7。
