Column-Chromatography of TSH Preparation

Abstract

Studing on biological actions of TSH employing Pretiron (A.G. Schering) as this source, we found that the administration of this preparation has induced the excessive ovarian responses in mature female rabbits, such as multiple ovulations and corpora lutea formation-Friedman's Reaction.The data suggests that it has a relatively high gonadotropic potency or surely it is contaminated with gonadotropic hormones. So we tried to estimate the gonadotropic potency of this preparation and, moreover desirably, to separate gonadotropic substances from this.We used both mouse uterine weight test and Junkmann-Schoeller's method for gonadotropin and for TSH estimation respectively.It has found that Pretiron had relatively high gonadotropic potency ; 10-J.S. unit of it showed at least I m.u.u. of gonadotropic activity.Then, in order to separate gonadotropic fraction from TSH preparation, we applied Condliffe's column-chromatography method, employing IRC-50 as adsorbent.We used 0.2M-Na-phosphate-buffer solution for the first elution, and for the second 1.0-M-NaCl-solution. The former fraction we named Fraction-I, the later Fraction-II.The biological assay of Fraction-I appeared to be mostly specific for the gonadotropic hormone, but for the thyrotropic activity we could recognize nothing in it.On the other hand Fraction-II showed biologically high thyrotropic activity but not any gonadotropic potencies.For these reasons we can say that this column-chromatography method is very usefull to separate gonadotropic substance from TSH Preparation, namely to purify the TSH preparation for biological applications.