Li Xiaoguang Sunny, Li Shu, Wynveen Paul, Mork Kathy, Kellermann Gottfried
Pharmasan Labs, Inc, 373 280th Street, Osceola, WI, 54020, USA,
Anal Bioanal Chem. 2014 Nov;406(28):7287-97. doi: 10.1007/s00216-014-8120-1. Epub 2014 Sep 17.
Catecholamines are a class of biogenic amines that play an important role as neurotransmitters and hormones. We developed and validated a rapid, specific and sensitive LC-MS/MS method for quantitative determination of catecholamines in human urine. Linearity, specificity, sensitivity, precision, accuracy, matrix effect, carryover, analyte stability, method comparison and reference range were evaluated. The catecholamine measurements were not affected by 35 structurally-related drugs and metabolites. The outstanding specificity was achieved by use of a specific diphenylborate-based solid phase extraction and subsequent selective LC-MS/MS analysis. Excellent sensitivity, accuracy and precision (average intra-assay variations <2.9 % and inter-assay variations <4.6 %) were obtained. The method was successfully applied in the study of day-to-day biological within- and between-subject variations of 25 healthy people under free-living conditions over three consecutive days. We observed that catecholamine excretions for second morning sampling had least day-to-day within-subject variation and excellent reproducibility. This work is one of the rare studies on these topics and represents the first utilization of advanced LC-MS/MS technology. Additionally, we found significant correlations between spot and conventional 24 h collections of human urine (n = 22, r > 0.853, p < 0.0001). These findings suggest that determining the catecholamine concentrations in the second morning urine sample presents accurate, convenient and reliable measurement of catecholamine excretions. In addition, consistent and significant diurnal variations for norepinephrine and epinephrine excretions were observed during the three-day period, while dopamine did not exhibit a diurnal rhythm. The LC-MS/MS method presented here is rapid, sensitive and specific, which could be an advantage in clinical laboratories.
儿茶酚胺是一类生物胺，作为神经递质和激素发挥着重要作用。我们开发并验证了一种快速、特异且灵敏的液相色谱-串联质谱法（LC-MS/MS），用于定量测定人尿液中的儿茶酚胺。评估了线性、特异性、灵敏度、精密度、准确度、基质效应、残留、分析物稳定性、方法比较和参考范围。儿茶酚胺的测量不受35种结构相关药物和代谢物的影响。通过使用基于特定二苯硼酸盐的固相萃取和随后的选择性LC-MS/MS分析，实现了出色的特异性。获得了优异的灵敏度、准确度和精密度（平均批内变异<2.9%，批间变异<4.6%）。该方法成功应用于25名健康人在自由生活条件下连续三天的日常生物体内和个体间变异研究。我们观察到，第二次晨尿采样的儿茶酚胺排泄量在个体内的日常变异最小，且具有出色的重现性。这项工作是关于这些主题的罕见研究之一，代表了先进LC-MS/MS技术的首次应用。此外，我们发现人尿液的即时和传统24小时收集之间存在显著相关性（n = 22，r > 0.853，p < 0.0001）。这些发现表明，测定第二次晨尿样本中的儿茶酚胺浓度可提供准确、便捷且可靠的儿茶酚胺排泄量测量。此外，在三天期间观察到去甲肾上腺素和肾上腺素排泄量存在一致且显著的昼夜变化，而多巴胺未表现出昼夜节律。本文介绍的LC-MS/MS方法快速、灵敏且特异，这在临床实验室中可能是一个优势。
