著者

福留 厚

出版者

昭和大学学士会

雑誌

昭和医学会雑誌 (ISSN:00374342)

巻号頁・発行日

vol.37, no.5, pp.425-435, 1977-10-28 (Released:2010-09-09)

参考文献数

80

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Fot the determination of lactose by galactose-oxidase peroxidase (GOP) method, it is necessary to measure the amount of galactose liberated after the hydrolysis of lactose.(1) Hydrolysis of lactose-Lactose was hydrolyzed with 0.2% H2SO4or with living cells of Escherichia coli3-MT (a mutbile-type variant which decomposes lactose but not galactose) . By H2SO4method, the calculated dose of galactose was liberated after 2 hours. By MT method, free galactose increased rapidly after 2 hours and reached maximum after 3 hours. The volume of lactose solution was diluted to 1: 4 in the procedures of acid hydrolysis and neutralization, but not in MT method.(2) Determination of galactose liberated from lactose after hydrolysis- (i) Free galactose in the H2SO4hydrolysate of standard solution of lactose was measured by GOP or a cup-plate method using M (galactose-sensitive mutant of enteric bacteria) as the test organism. The amount of galactose was measurable in the range of 25-200γ/2 ml in the case of GOP method, and 1.25-10γ/0.1 ml in the case of M-cup method. (ii) Galactose in the MT hydrolysate was measured by GOP or M-cup method. The amount of galactose could be estimated in the range of 25-100 γ/2 ml by GOP method, but the volume of MT hydrolysate was diluted to 1: 8 because of the necessity of deproteinizing. On the contrary, the amount of 1.25-10γ/0.1 ml could be estimated by M-cup method without the need of deproteinizing.(3) Determination of lactose in the biological material- (i) Free galactose in the H2SO4 hydrolysate of milk was measured by GOP or M-cup method. Although deproteinizing of the hydrolysate was necessary, the determination of galactose in the range of 25-100γ/2 ml was possible by GOP method. By M-cup method, 1.25-10γ/0.1 ml of galactose could be determined without the need of deproteinizing. (ii) The amount of galactose in the MT hydrolysate of milk was measured by GOP or M-cup method. While the estimation of 25-100γ/2 ml of galactose was possible by GOP method after deproteinizing, 1.25-10 γ/0.1 ml of galactose could be estimated by M-cup method without deproteinizing.From these experimental results, the combination of MT hydrolysis and M-cup method proved to be most sensitive for the determination of lactose in the biological material.