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Clinical Chemistry 29: 1937-1940, 1983;
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Clinical Chemistry, Vol 29, 1937-1940, Copyright © 1983 by American Association for Clinical Chemistry

Separation of bound and free ligand by ethacridine (Rivanol) in thyroxin radioimmunoassay

V Bhupal and RS Mani

In this simple and low-cost radioimmunoassay (RIA) procedure for thyroxin (T4), ethacridine (6,9-diamino-2-ethoxyacridine lactate; Rivanol) is used to separate antibody-bound ligand and free ligand. This acridine dye precipitates free T4 not bound to antibody. At pH 8.6, it precipitates more than 90% of 125I-labeled T4. Precipitation is rapid and is constant over the concentration range of 1 to 3 g of ethacridine per liter; time of addition and temperature are not critical. The inherent yellow color of the dye gives it the advantage of a color-coded reagent. Analysis of 65 serum samples by the proposed method and by a procedure involving polyethylene glycol gave similar results: correlation coefficient, 0.988; slope, 0.981; y-intercept, - 1.42. Within- and between-assay variations (CV) were less than 5 and 6%, respectively.




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Copyright © 1983 by the American Association for Clinical Chemistry.