The assay of the catecholamine content of small volumes of human plasma.
Plasma catecholamines are routinely measured using high-performance liquid chromatography (HPLC) with electrochemical detection. Most of the present assays require sample volumes of at least 500 microL and are complex and labour-intensive procedures, or require large capital investment to reduce the sample size. This paper describes a liquid/liquid plasma catecholamine extraction procedure, HPLC separation and electrochemical detection method which is simple, sensitive and reproducible. The resting catecholamine concentration of 50 microL adult human plasma can be assayed using standard electrochemical detection. The limits of detection were 0.1 fmol injected onto the column for each catecholamine. This method allows the routine assay of plasma catecholamine concentrations within the normal adult ranges in both 500 and 50 microL samples. The within assay coefficient of variation (CV) for noradrenaline (NA) was 1.2% in 500 microL plasma and 1.9% for 50 microL plasma, corresponding values for adrenaline (A) were 8.5 and 6.6%. The between assay CVs were 3.9 and 7.8% for NA, and 9.9 and 5.7% for A.
Forster
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CD
School of Biomedical Sciences, Nottingham University Medical School, Queen's Medical Centre, UK.
Macdonald
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IA
D000143:Acids; D000328:Adult; D002395:Catecholamines / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004563:Electrochemistry; D006801:Humans; D007202:Indicators and Reagents; D012015:Reference Standards; D012016:Reference Values; D015203:Reproducibility of Results
D000143:Acids; D002395:Catecholamines; D007202:Indicators and Reagents