Said R, Robinet D, Barbier C, Sartre J, Huguet C
Laboratoire Cerba, Pontoise, France.
J Chromatogr. 1990 Aug 24;530(1):11-8. doi: 10.1016/s0378-4347(00)82297-5.
A totally automated and reliable high-performance liquid chromatographic method is described for the routine determination of free catecholamines (norepinephrine, epinephrine and dopamine) in urine. The catecholamines were isolated from urine samples using small alumina columns. A standard automated method for pH adjustment of urine before the extraction step has been developed. The extraction was performed on an ASPEC (Automatic Sample Preparation with Extraction Columns, Gilson). The eluate was collected in a separate tube and then automatically injected into the chromatographic column. The catecholamines were separated by reversed-phase ion-pair liquid chromatography and quantified by fluorescence detection. No manual intervention was required during the extraction and separation procedure. One sample may be run every 15 min, ca. 96 samples in 24 h. Analytical recoveries for all three catecholamines are 63-87%, and the detection limits are 0.01, 0.01, and 0.03 microM for norepinephrine, epinephrine and dopamine, respectively, which is highly satisfactory for urine. Day-to-day coefficients of variation were less than 10%.
本文描述了一种完全自动化且可靠的高效液相色谱法，用于尿液中游离儿茶酚胺（去甲肾上腺素、肾上腺素和多巴胺）的常规测定。使用小氧化铝柱从尿液样本中分离儿茶酚胺。已开发出一种在萃取步骤前对尿液进行pH调节的标准自动化方法。萃取在ASPEC（带萃取柱的自动样品制备仪，吉尔森公司）上进行。洗脱液收集在单独的试管中，然后自动注入色谱柱。儿茶酚胺通过反相离子对液相色谱法分离，并通过荧光检测进行定量。在萃取和分离过程中无需人工干预。每15分钟可运行一个样品，24小时内约可分析96个样品。三种儿茶酚胺的分析回收率为63 - 87%，去甲肾上腺素、肾上腺素和多巴胺的检测限分别为0.01、0.01和0.03微摩尔，这对于尿液检测来说非常令人满意。日常变异系数小于10%。
