Abstract
A radioimmunoassay (RIA) using a 125I-tracer is described for measurement of serotonin (5-hydroxytryptamine, 5-HT) in human platelet-rich plasma (PRP). Antisera were raised against 5-HT-succinamate conjugated to bovine albumin and, to improve assay sensitivity, the analyte was made chemically similar to the immunogen by conversion to N-acetylserotonin prior to assay, using the specific amino reagent N-acetoxysuccinimide. The assay shows good correlation with a high-pressure liquid chromatography (HPLC) reference method (5-HT RIA = 1.007 × 5HT HPLC + 29.3, r = 0.936, p < 0.001, n = 40), indicating that no significant cross-reactions were detected. Samples of PRP are diluted 1 20 to fall within the working range (80–15% B/ B o ) of the assay, which is 4.75–325 nmol/1, (0.95–65.0 pmol/ tube), corresponding to 95–6500 nmol/1 in PRP. Intra- and interassay coefficients of variation were 5.0–10.5% and 12.0–21.2% respectively for serotonin concentrations of 250–2500 nmol/l added to platelet-poor plasma. With this improved assay, it is possible to analyse up to 100 samples/ day, compared with 10–20 samples/ day by HPLC.
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