DETERMINATION OF PROSCILLARIDIN IN PLASMA BY RADIOIMMUNOASSAY

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The development of a sensitive and selective radioimmunoassay for determination of proscillaridin in plasma is described. Antiserum against proscillaridin was obtained from guinea pig immunized with an immunogen prepared by conjugating methylproscillaridin, 14β-hydroxy-3β-[(4-O-methyl-α-L-rhamnosyl) oxy] bufa-4,20,22-trienolide to bovine serum albumin. Methyl N-[3-[(14β-hydroxybufa-4,20,22-trienolid-3β-yl) oxy-carbonyl] propanoyl]-L-[3', 5'-^<125>I_2] diiodotyrosinate ([^<125>I] SST) was used as a radioactive ligand. [^<125>I] SST and antiserum were added to the recovered sample from plasma using a Bond Elut^[○!R] column. After overnight incubation, the antibody-bound and free [^<125>I] SST were separated using polyethylene glycol. The mean coefficients of variation of intra and interassay at four different plasma concentrations were 4.0 and 5.0%, respectively. The assay was able to determine as little as 125 pg/ml of proscillaridin in plasma by using 1.5 ml of sample. Beagle dogs were orally administered one tablet (Talusin^[○!R]) containing 0.25 mg of proscillaridin. Proscillaridin was rapidly absorbed, exhibiting plasma maximum concentration of 2.06 ng/ml at 20 min. Thereafter, the plasma level declined biphasically with half-lives of 0.6 and 25.4 h.
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