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An enzyme immunoassay for 6‐sulphatoxy‐melatonin in human urine
Author(s) -
PenistonBird John F.,
Di WeiLi,
Street Catherine A.,
Edwards Ray,
Little John A.,
Silman Robert E.
Publication year - 1996
Publication title -
journal of pineal research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.881
H-Index - 131
eISSN - 1600-079X
pISSN - 0742-3098
DOI - 10.1111/j.1600-079x.1996.tb00239.x
Subject(s) - radioimmunoassay , melatonin , immunoassay , urine , chemistry , bovine serum albumin , chromatography , coefficient of variation , conjugate , horseradish peroxidase , enzyme , endocrinology , antibody , biology , biochemistry , immunology , mathematics , mathematical analysis
We describe a newly developed enzyme immunoassay (EIA) for the determination of 6‐sulphatoxy‐melatonin (aMT6s) in human urine, using a aMT6s‐bovine serum albumin‐horseradish peroxidase (aMT6s‐BSA‐HRP) conjugate as the enzyme label. The assay incorporates a highly specific antibody raised in rabbits. The EIA has a sensitivity of 2 pg/well (40 pg/ml) with intraassay coefficients of variation of 2.3–6.1% in the range of the assay. The material with the highest level of cross‐reactivity was N‐acetyl serotonin sulphate, with a relative potency of 0.000078%. One hundred thirty‐four urine samples from children and adults at different time points were assayed and the results compared with those from an established radioimmunoassay (RIA) and with a newly developed RIA using the same antibody as the EIA. The correlation coefficient, r , comparing the two RIA's was 0.9869, and the regression equation was log (kit) = 0.9340 log (new) + 0.1213. The correlation coefficient, r , comparing the EIA with the newly developed RIA, was 0.9686, and regression equation log (new) = 0.9674 log (EIA) + 0.0600. The EIA for the measurement of aMT6s in urine represents a new approach in the investigation of pineal function.