甲状腺刺戟ホルモンの生物学的測定法, 抽出法並びに化学的性状の研究

抄録

Investigating the basic factor of bioassay utilizing the P³² uptake of one-day-old chick thyroids, we found that the uptake of P³² by thyroid, reached a maximal level in 3 hours, not only in the untreated chicks but also in the chicks which had been subcutaneously injected with TSH. In the case of subcutaneous injections of TSH, the maximal level was obtained between 12-18 hours after injection, and in the intracardially injected with TSH, the response reached the maximal level at around 6 hours after TSH administration.
By comparing Lambergs' subcutaneous and Greenspan's intracardiac injections of TSH for its bioassay utilizing the P³², we recognized that the Greenspan's method is more appropriate than the Lamberg's with regard to the minimal detectable dose, which was 0.01 J.S.u. in the former while 0.1 J.S.u. in the latter, and the precision index also indicated the former's superiority, that is, 0.31 in Greenspan's and 0.64 in Lamberg's method.
As to the extraction method of TSH from blood, there is a classical Fellinger's procedure. Using Fellinger's method and Greenspan's intracardiac method, the recovery test was made and showed 24-29% of recovery rate. Substituting the method of centrifugation for filtration, in order to separate the precipitate produced with addition of diluted acetone, the author obtained a satisfactory recovery rate of 53-58%.
The TSH activity of each fraction was exemined : about 18% of the added TSH was transfered to the first precipitate and 0.5% of it was contained in the second supernant. Therefore it was apparent that 30% undetectably disappeared during the course of the procedure.
The TSH activities of the blood were examined by the procedure described.
The chemical analysis of TSH was then carried out by papergraphy, and the author found that the Amidoblack 10 B was most satisfactory for the staining and that the best result was obtained when TSH was dissolved in physiological saline. The author also found that at least 4 components of Ninhydrin positive were separated when the filter paper was developed with the solvent butanol-glacial acetic acid-water.
Furthermore the author recognized the glucosamine as a suger component of TSH.