Enzyme electrode and method for dextran analysis

• US 4,552,840 A
• Filed: 12/02/1982
• Issued: 11/12/1985
• Est. Priority Date: 12/02/1982
• Status: Expired due to Fees

• Current Assignee: California and Hawaiian Sugar Company
• Original Assignee: California and Hawaiian Sugar Company
• Inventors: Riffer, Richard
• Primary Examiner(s): Wiseman, Thomas G.
• Assistant Examiner(s): Huleatt, Jayme A.
• Application Number:
  US06/433,255
• Time in Patent Office: 1,076 Days
• Field of Search: 435/14, 435/18, 435/22, 435/27, 435/28, 435/175, 435/177, 435/179, 435/181, 435/188, 435/190, 435/200, 435/211, 435/288, 435/291, 435/808, 435/817, 204/403, 204/405, 204/1 E, 204/1 P
• US Class Current: 205/778
• CPC Class Codes: C12M 21/18 Apparatus specially designed for the use of free, immobilized or carrier-bound enzymes
• CPC Class Codes: C12M 29/04 Filters; Permeable or porous membranes or plates, e.g. dialysis
• CPC Class Codes: C12M 41/32 of substances in solution
• CPC Class Codes: C12Q 1/005 involving specific analytes or enzymes including groups of enzymes, e.g. oxydases; C12Q1/004 takes precedence
• CPC Class Codes: Y10S 435/817 Enzyme or microbe electrode

Abstract

A method for the potentiometric determination of a dextran solution is provided wherein the dextran is enzymatically hydrolyzed to glucose, the glucose is oxidized to form hydrogen peroxide and the hydrogen peroxide is measured utilizing a redox electrode. A novel electrode having a plurality of enzyme impregnated layers is provided for converting the dextran to glucose.

Claims

1. An electrode for the potentiometric determination of dextran in a solution comprising a platinum redox electrode, a first cellulose absorbent sheath surrounding the electrode with immobilized glucose oxidase, a second absorbent sheath thereover with immobilized enzymes selected from the group consistinng of α

-glucosidase and isomaltase, with the provision that a means for excluding compounds having a molecular weight of 14,000 or greater surrounding said second sheath is included when α

-glucosidase is selected and when the solution contains starch and an outer absorbent sheath with immobilized dextranase surrounding said means wherein the amounts of glucose oxidase, α

-glucosidase or isomaltase and dextranase are sufficient to determine the amount of dextran.

2. The electrode of claim 1 wherein said means is a dialysis membrane.

3. The electrode of claim 1 wherein α

-glucosidase is selected.

4. The electrode of claim 1 wherein isomaltase is selected.

5. The electrode of claim 1 wherein each sheath is made from a cellulose filter paper sheet.

6. The electrode of claim 5 wherein each enzyme is immobilized on cellulose by covalent bonding utilizing a bifunctional reagent.

7. The electrode of claim 6 wherein the bifunctional reagent is sym-triazine trichloride.

8. The electrode of claim 1 wherein the dextranase is mold dextranase.

9. A method for the potentiometric determination of a dextran in a solution comprising immersing into a sample, an electrode containing a platinum redox electrode, a first cellulose absorbent sheath surrounding the electrode with immobilized glucose oxidase, a second absorbent sheath thereover with immobilized enzymes selected from the group consisting of α

-glucosidase and isomaltase, with the provision that a means for excluding compounds having a molecular weight of 14,000 or greater surrounding said second sheath is included when α

-glucosidase is selected and when the solution contains starch and an outer absorbent sheath with immobilized dextranase surrounding said means wherein the amounts of glucose oxidase, α

-glucosidase or isomaltase and dextranase are sufficient to determine the amount of dextran, incubating and measuring the voltage which correlates with the amount of dextran present.

10. The method of claim 9 wherein α

-glucosidase is selected.

11. The method of claim 9 wherein isomaltase is selected.

12. The method of claim 9 wherein said means is a dialysis membrane.