Rapid analysis of plural components

• US 5,175,088 A
• Filed: 11/20/1990
• Issued: 12/29/1992
• Est. Priority Date: 03/08/1983
• Status: Expired due to Fees

• Current Assignee: Oriental Yeast Company Limited (Nisshin Seifun Group Incorporated), Oriental Electronics Co., Ltd.
• Original Assignee: Oriental Yeast Company Limited (Nisshin Seifun Group Incorporated), Oriental Electronics Co., Ltd.
• Inventors: Oka, Osamu, Ohashi, Minoru, Numazawa, Kenichi, Utugi, Yoshio, Arakawa, Nobuhiko
• Primary Examiner(s): Kepplinger, Esther L.
• Assistant Examiner(s): SCHEINER, TONI R
• Application Number:
  US07/616,766
• Time in Patent Office: 770 Days
• Field of Search: 435/4, 435/10, 435/14, 435/18, 435/21, 435/22, 435/15, 435/25
• US Class Current: 435/14
• CPC Class Codes: G01N 33/14 Beverages

Abstract

A new method of rapidly analyzing plural substances in the presence of biological catalyzers is disclosed. The method is practiced by way of the steps of injecting both pH buffer solution and specimen into a reaction cell, successively adding a plurality of enzymes to induce up-take reaction of dissolved oxygen, causing the plurality of substances to be subjected to selective oxidation in the stepwise manner, obtaining a stepdown curve of dissolved oxygen by automatically recording the oxidative process by means of a dissolved oxygen sensor, qualitatively determining each of the substances with reference to the kind of added enzymes and the order of their addition and quantitatively determining the same with reference to the extent of decrease in dissolved oxygen. Typically, oxidation of the substances is carried out by way of two or three or further more steps. The reaction cell for performing the method is equipped with a dissolved oxygen sensor on the one side wall and its upper portion is air-tightly closed with a plug through which a fine bore is formed for the purpose of supply of buffer solution, specimen and enzymes.

Claims

1. A method for analyzing glucose and ethyl alcohol in a liquid containing them, which comprises the steps:

(a) adding a volume of the liquid and a pH buffer solution to a reaction cell designed to prevent air from entering the cell, said reaction cell being equipped with an oxygen sensor connected to an automatic recording device,

(b) adding an enzyme selected from the group consisting of glucose oxidase and alcohol oxidase to the liquid which catalyzes oxidation of one component in the liquid,

(c) continuously recording oxygen sensor output,

(d) adding an enzyme selected from the group consisting of glucose oxidase and alcohol oxidase which is different from the enzyme added in step (b) to the liquid which catalyzes oxidation of a second component in the liquid,

(e) continuously recording oxygen sensor output, and

(f) calculating the amount of said one component of step (b) from the decrease in oxygen sensor output in step (c) and calculating the amount of said second component present in the liquid from the decrease in oxygen sensor output in step (e).

2. A method for analyzing glucose and mono-sodium glutamate in a liquid containing them, which comprises the steps:

(a) adding a volume of the liquid and a pH buffer solution to a reaction cell designed to prevent air from entering the cell, said reaction cell being equipped with an oxygen sensor connected to an automatic recording device,

(b) adding an enzyme selected from the group consisting of glucose oxidase and glutamic acid oxidase to the liquid which catalyzes oxidation of one component in the liquid,

(c) continuously recording oxygen sensor output,

(d) adding an enzyme selected from the group consisting of glucose oxidase and glutamic acid oxidase which is different from the enzyme added in step (b) to the liquid which catalyzes oxidation of a second component in the liquid,

(e) continuously recording oxygen sensor output, and

(f) calculating the amount of said one component of step (b) from the decrease in oxygen sensor output in step (c) and calculating the amount of said second component present in the liquid from the decrease in oxygen sensor output in step (e).

3. A method for analyzing inosinic acid and glutamic acid in a liquid containing them, which comprises the steps:

(a) adding a volume of the liquid and a pH buffer solution to a reaction cell designed to prevent air from entering the cell, said reaction cell being equipped with an oxygen sensor connected to an automatic recording device,

(b) adding xanthene oxidase and an enzyme selected from the group consisting of nucleoside phosphorylase and glutamic acid oxidase to the liquid,

(c) continuously recording oxygen sensor output,

(d) adding an enzyme selected from the group consisting of nucleoside phosphorylase and glutamic acid oxidase which is different from the enzyme added in step (b) to the liquid which catalyzes oxidation of a second component in the liquid,

(e) continuously recording oxygen sensor output, and

(f) calculating the amount of said one component of step (b) from the decrease in oxygen sensor output in step (c) and calculating the amount of said second component present in the liquid from the decrease in oxygen sensor output in step (e) provided that when nucleoside phosphorylase is the added enzyme alkaline phosphatase is added together with the nucleoside phosphorylase.

4. A method for analyzing glutamic acid and glucose in a liquid containing them, which comprises the steps:

(a) adding a volume of the liquid and a pH buffer solution to a reaction cell designed to prevent air from entering the cell, said reaction cell being equipped with an oxygen sensor connected to an automatic recording device,

(b) adding an enzyme selected from the group consisting of glutamic acid oxidase and glucose oxidase to the liquid which catalyzes oxidation of one component in the liquid,

(c) continuously recording oxygen sensor output,

(d) adding an enzyme selected from the group consisting of glutamic acid oxidase and glucose oxidase which is different from the enzyme added in step (b) to the liquid which catalyzes oxidation of a second component in the liquid,

(e) continuously recording oxygen sensor output, and

(f) calculating the amount of said one component of step (b) from the decrease in oxygen sensor output in step (c) and calculating the amount of said second component present in the liquid from the decrease in oxygen sensor output in step (e).

5. A method for analyzing ascorbic acid and glucose in a liquid containing them, which comprises the steps:

(a) adding a volume of the liquid and a pH buffer solution to a reaction cell designed to prevent air from entering the cell, said reaction cell being equipped with an oxygen sensor connected to an automatic recording device,

(b) adding an enzyme selected from the group consisting of ascorbate oxidase and glucose oxidase to the liquid which catalyzes oxidation of one component in the liquid,

(c) continuously recording oxygen sensor output,

(d) adding an enzyme selected from the group consisting of ascorbate oxidase and glucose oxidase which is different from the enzyme added in step (b) to the liquid which catalyzes oxidation of a second component in the liquid,

(e) continuously recording oxygen sensor output, and

(f) calculating the amount of said one component of step (b) from the decrease in oxygen sensor output in step (c) and calculating the amount of said second component present in the liquid from the decrease in oxygen sensor output in step (e).