LATHYROGENIC EFFECT OF INH:Report I. Quantitative Determination of Hydroxyproline

概要

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In order to make the experiments with chick embryos for the purpose of investigating about the influence on fetus by the dosage of INH, which is widely used today for the treatment of tuberculous patients, it was necessary to determine hydroxyproline quantitatively in the 1M. sodium chloride extract from the long bones of chick embryos treated with INH. The method of determination was almost the same as that of Robert E. Newman and Milan A. Logan, but somewhat changed because of availability of laboratory instruments. Procedures: The tibia and the femur were stripped of muscles, tendon and periostium, minced with scissors and extracted in 20 volumes (v/w) of 1M. cold saline buffered with phos phate (pH.7.6, ionic strength 0.02). The extract was shaked for 24 hours at 5°C., then centrifuged at 15, 000 r.p.m. for 1 hour (Model Tominaga Factory, Toyo No.71). The supernatant solution was passed through a glass filter, No.4. The viscosity was measured with the Ostwald viscosimeter. The extract was analysed for hydroproline, for hydrolysis of extracted collagen is: _??_ 1cc. of this neutralized sample was used for the determination of hydroxyproline content by Newman and Longan's method. The recovery of hydroxyproline from a bovine achilles tendon by our method was 95-100%.
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