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Assay of human plasma catecholamines.

机译:人血浆儿茶酚胺的测定。

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摘要

Concentrations of catecholamines (CA) in plasma reflect, to a significant extent, the activity level of the sympathetic nervous system but a sensitive means to accurately quantify the CA on a routine basis had not previously been readily available.;A number of unique features have been incorporated into this assay to achieve and maintain its sensitivity. These include a novel method of cleaning the HPLC-ECD apparatus on a daily basis with no attendant loss of sensitivity and with a minimal re-equilibration period (i.e., less than one-half hour). Additionally, a novel method of polishing the glassy carbon working electrode (WE) is described. This polishing technique restores the sensitivity of the WE to its original status (as determined when the WE is new). Recommendations are made regarding the details essential to the operation of the assay on a regular basis and to the continued preservation of the high level of sensitivity of the assay for routine use.;The performance of this CA assay was tested under the constraints of a clinical setting in an effort to judge the ability of the assay to perform under these conditions and in an attempt to answer a clinical question: are plasma CA concentrations elevated in patients who have suffered a subarachnoid hemorrhage due to rupture of a cerebral artery aneurysm? Based on the data collected in the present instance, no firm conclusions could be drawn about the contribution of plasma CA levels to the development of clinical complications in this group of patients.;The present work has involved the development of a reliable, sensitive and reproducible human plasma CA assay. An efficient boric acid extraction method was used to extract the CA from plasma; the CA were subsequently separated and detected by high pressure liquid chromatography (HPLC) and (amperometric) electrochemical detection (ECD), respectively. An on-line, dedicated integrator was used to obtain and record an objective assessment of the peak heights of the analytes on the chromatogram. The entire assay technique described herein has been optimized to accommodate the operation of the ECD at the high sensitivity level (namely, 1 nanoamp full scale deflection of the ECD response) required by the low levels of endogenous CA found in resting human plasma.
机译:血浆中儿茶酚胺(CA)的浓度在很大程度上反映了交感神经系统的活动水平,但以前尚无法获得一种以常规方式准确定量CA的灵敏方法。许多独特的功能被并入该测定中以实现并维持其灵敏度。这些方法包括每天清洁HPLC-ECD设备的新方法,且不会伴随灵敏度的损失,并且重新平衡的时间最少(不到半小时)。另外,描述了抛光玻璃碳工作电极(WE)的新方法。这种抛光技术将WE的灵敏度恢复到其原始状态(由WE新建时确定)。提出了有关定期进行测定操作以及对于常规使用的测定方法的高灵敏度持续保持必不可少的细节的建议。该CA测定法的性能在临床条件下进行了测试为了判断该试验在这些条件下的执行能力并试图回答一个临床问题而设置:在由于脑动脉瘤破裂而遭受蛛网膜下腔出血的患者中血浆CA浓度是否升高?根据目前的数据,尚不能就血浆CA水平对这组患者的临床并发症的发展做出确切的结论。;目前的工作涉及可靠,敏感和可重复的开发。人血浆CA分析。一种有效的硼酸提取方法用于从血浆中提取CA。随后将CA分离并分别通过高压液相色谱(HPLC)和(安培)电化学检测(ECD)检测。在线专用积分仪用于获得并记录色谱图上分析物峰高的客观评估。已经对本文所述的整个测定技术进行了优化,以适应在静息人血浆中发现的低水平内源性CA所需的高灵敏度水平(即ECD响应的1纳安全刻度偏转)下的ECD操作。

著录项

  • 作者

    Damjanovic, Dragana.;

  • 作者单位

    University of Alberta (Canada).;

  • 授予单位 University of Alberta (Canada).;
  • 学科 Medicine.
  • 学位 Ph.D.
  • 年度 1989
  • 页码 1 p.
  • 总页数 1
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 老年病学 ;
  • 关键词

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