Abstract
Determination of conjugated chemocorticoids has developed and its clinical significance has been observed. It is reported in this paper the hydrolysis of conjugated formaldehydrogenic chemocorticoids in blood and urine with spleen β-glucuronidase which prepared by Fishman's method.
For urinary conjugated chemocorticoids, the method which investigated by Cohen is advisable, but the problem has been remained obscure in the enzymatic hydrolysis. We researched the chemical equilibrium and deviced the incubation method. That is to say, after extracting of free from with chloroform, β-glucuronidase was added to the remaining fluid which was brought to pH 5.0 with acetate buffer. Then 10ml. of chloroform was added and incubated 40 hrs. at 38°C. For the incubation time, the flask was shaken several times in order to extracting the hydrolyzed free steroids to chloroform, and β-glucuronidase was supplied three times. Under this condition the hydrolyzed chemocortiroids was twice of these obtained without adding chloroform as much.
For conjugated chemocorticoids in blood, Bongiovanni reported the method of estimation of conjugated 17-hydroxycorticosteroids, but the method of formaldehydogenie chemocorticoids has not been reported. The results obtained by our method conjugated formaldehydogenie chemocorticoids was about two times of free form.
By these methods, it is estimated formaldehydogenic chemocorticoids in the patient with heart failure and revealed that the conjugates was less in the patient with liver dysfunction than with normal liver.
