1. Carring out the determination of sugars after H
AGEDORN and J
ENSEN but by en'arging the original scale by ten times, reported H
ANES that in the case of glucose the relation between the concentration of sugar and the amount of ferricyanide consumed is represented by two straight lines which cross each other at the point corresponding to about 2mg./5 c.c. of sugar. But after repeated experiments the present author found that the relation can be expressed by one sfraight line in whole range of concentration (0•5-3•5mg./5 c.c.) and this also hold good of fructose and xylose, where 1 c.c. of N/100 thiosulfate is equivalent to 0•336mg. of glucose, 0•338mg. of fructose and 0•305mg. of xylose.
2. Influence on this method of some substances, often met with in a biochemical research, was examined and the procedure to estimate sugars in the presence of protein, pepton, creatin or creatinin as an interfering substance was given. Urea which is possible to cause a large error in copper method, had no effect on this estimation.
3. The amount of ferrieyanide (
T ?? ) reduced by glucose of a certain concentration can be given by the equation
Tt=
T-(
T1e-k1t+
T2e-k2t),
where
t is time of heating,
T is the value of
Tt when
t=∞,
T1+
T2=
T, and
k1 and
k2 are constanst. Hence this reaction can be said to be composed of two monomolecular reactions. Then
T1 and
T2 are respectively the limiting amounts of ferricyanide consumed in these reactions. As shown in Table 5, which gives the values of these terms for various concentration of sugar,
T1 is proportional to the amount of sugar,
T2 and
k1 are nearly constant while
k2 increases slightly with sugar concentration. These facts lead us to the assumption that the one of these two reactions, which has velocity constant
k1, is oxidation of glucose and the other of which the velocity constant is
k2, an autodecomposition of ferricyanide caused by the presence of glucose.
The actual consumption of ferricyanide amounts three times as much as that expected when glucose is oxidized only to gluconic acid.
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