US3953295A
Expired / Lapsed
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Reagent formulations for glucose assay
Assignee: Mallinckrodt Inc
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Filing: 1974-05-06
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Granted: 1976-04-27
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Expires: 1994-05-06
Abstract
Abstract Glucose is assayed in biological specimens using granular, water soluble, substantially anhydrous, storage-stable reagent formulations containing enzymes, a chromogen and/or a buffer and a nitrogen - containing polyoxyalkylene nonionic surfactant obtained by the sequential reaction of ethylenediamine with propylene oxide and ethylene oxide in the presence of a catalyst. The surfactant contains polyoxypropylene chains having an average molecular weight of between about 750 and about 6,750, and polyoxyethylene chains constituting between about 10 and about 80 weight percent. The surfactant has an advantageous effect on granulation, dissolution and storage stability of the reagent formulations and is effective for solubilizing protein such as enzymes.
Inventors
- 👤 Alexander A. Monte
- 👤 Ching Chiang
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-26 13:02
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/54; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/72; G01N33/721
Claims Analysis
Total Claims: 19
Claim 1
Independent Claim
88 words
An enzyme reagent formulation for use in assaying a biological specimen for glucose comprising a granular, water-soluble, substantially anhydrous, storage-stable mixture containing glucose oxidase, peroxidase, gum arabic, mannitol, bovine serum albumin and a nitrogen-containing polyoxyalkylene nonionic surfactant obtained by the sequential reaction of ethylenediamine with propylene oxide and ethylene oxide in the presence of a catalyst, said surfactant containing polyoxypropylene chains having an average molecular weight of between about 750 and about 6,750 and polyoxyethylene chains constituting between about 10 and about 80 weight percent of said surfactant.
Claim 2
Dependent Claim
88 words
An enzyme reagent formulation for use in assaying a biological specimen for glucose comprising a granular, water-soluble, substantially anhydrous, storage-stable mixture containing glucose oxidase, peroxidase, gum arabic, mannitol, bovine serum albumin and a nitrogen-containing polyoxyalkylene nonionic surfactant obtained by the sequential reaction of ethylenediamine with propylene oxide and ethylene oxide in the presence of a catalyst, said surfactant containing polyoxypropylene chains having an average molecular weight of between about 750 and about 6,750 and polyoxyethylene chains constituting between about 10 and about 80 weight percent of said surfactant.
Claim 3
Dependent Claim
32 words
A reagent formulation as set forth in claim 1 wherein the nitrogen-containing polyoxyalkylene nonionic surfactant is solid and the polyoxypropylene chains thereof have an average molecular weight of less than abou 4,000.
Claim 4
Dependent Claim
39 words
A reagent formulation as set forth in clam 2 wherein the polyoxypropylene chains of the nitrogen-containing polyoxyalkylene nonionic surfactant have an average molecular weight of about 2,750 and the weight percentage of the polyoxyethylene units thereof is about 70%.
Claim 5
Dependent Claim
39 words
A reagent formulation as set forth in clam 2 wherein the polyoxypropylene chains of the nitrogen-containing polyoxyalkylene nonionic surfactant have an average molecular weight of about 2,750 and the weight percentage of the polyoxyethylene units thereof is about 70%.
Claim 6
Independent Claim
112 words
A method of preparing a granular, water-soluble, substantially anhydrous, storage-stable enzyme reagent formulation for use in assaying a biological specimen for glucose which comprises preparing a mixture of glucose oxidase, gum arabic, mannitol, bovine serum albumin, a solid nitrogen-containing polyoxyalkylene nonionic surfactant obtained by the sequential reaction of ethylene diamine with propylene oxide and ethylene oxide in the presence of a catalyst, said surfactant containing polyoxypropylene chains having an average molecular weight of between about 750 and 6,750, and polyoxyethylene chains constituting between about 10 and about 80 weight percent of said surfactant, and a solvent for the surfactant; and removing the solvent to form a substantially anhydrous, free-flowing, water-soluble, granular solid.
Claim 7
Independent Claim
112 words
A method of preparing a granular, water-soluble, substantially anhydrous, storage-stable enzyme reagent formulation for use in assaying a biological specimen for glucose which comprises preparing a mixture of glucose oxidase, gum arabic, mannitol, bovine serum albumin, a solid nitrogen-containing polyoxyalkylene nonionic surfactant obtained by the sequential reaction of ethylene diamine with propylene oxide and ethylene oxide in the presence of a catalyst, said surfactant containing polyoxypropylene chains having an average molecular weight of between about 750 and 6,750, and polyoxyethylene chains constituting between about 10 and about 80 weight percent of said surfactant, and a solvent for the surfactant; and removing the solvent to form a substantially anhydrous, free-flowing, water-soluble, granular solid.
Claim 8
Dependent Claim
31 words
A method as set forth in claim 4 wherein the nitrogen-containing polyoxyalkylene nonionic surfactant is solid and the polyoxypropylene chains thereof have an average molecular weight of less than about 4,000.
Claim 9
Dependent Claim
38 words
A method as set forth in claim 5 wherein the polyoxypropylene chains of the nitrogen-containing polyoxyalkylene nonionic surfactant have an average molecular weight of about 2,750 and the weight percentage of the polyoxyethylene units thereof is about 70%.
Claim 10
Dependent Claim
23 words
A method as set forth in claim 6 wherein the mixture is screened in wet form prior to the removal of the solvent.
Claim 11
Dependent Claim
19 words
The method as set forth in claim 7 wherein the formulation is again screened after the solvent is removed.
Claim 12
Dependent Claim
18 words
A method as set forth in claim 8 wherein the solvent is removed by drying under reduced pressure.
Claim 13
Independent Claim
153 words
A method of preparing a granular water-soluble, free-flowing, substantially anhydrous, storage-stable enzyme reagent formulation for use in assaying a biological specimen for glucose which comprises: (1) preparing a first mixture containing: glucose oxidase, gum arabic, mannitol, bovine serum albumin, a nitrogen-containing polyoxyalkylene nonionic surfactant obtained by the sequential reaction of ethylene diamine with propylene oxide and ethylene oxide in the presence of a catalyst, the surfactant containing polyoxypropylene chains having an average molecular weight of between about 750 and 6,750, and polyoxyethylene chains constituting between about 10 and about 80 weight percent of the surfactant, and a solvent for the surfactant; (2) removing the solvent to form a substantially anhydrous, free-flowing, water-soluble, granular first subformulation; (3) preparing a second mixture containing peroxidase and mannitol, the second mixture constituting a second subformulation; and (4) blending the first and second subformulations to provide a substantially anhydrous, free-flowing, water-soluble, granular solid constituting the enzyme reagent formulation.
Claim 14
Independent Claim
153 words
A method of preparing a granular water-soluble, free-flowing, substantially anhydrous, storage-stable enzyme reagent formulation for use in assaying a biological specimen for glucose which comprises: (1) preparing a first mixture containing: glucose oxidase, gum arabic, mannitol, bovine serum albumin, a nitrogen-containing polyoxyalkylene nonionic surfactant obtained by the sequential reaction of ethylene diamine with propylene oxide and ethylene oxide in the presence of a catalyst, the surfactant containing polyoxypropylene chains having an average molecular weight of between about 750 and 6,750, and polyoxyethylene chains constituting between about 10 and about 80 weight percent of the surfactant, and a solvent for the surfactant; (2) removing the solvent to form a substantially anhydrous, free-flowing, water-soluble, granular first subformulation; (3) preparing a second mixture containing peroxidase and mannitol, the second mixture constituting a second subformulation; and (4) blending the first and second subformulations to provide a substantially anhydrous, free-flowing, water-soluble, granular solid constituting the enzyme reagent formulation.
Claim 15
Dependent Claim
31 words
A method as set forth in claim 10 wherein the nitrogen-containing polyoxyalkylene nonionic surfactant is solid and the polyoxypropylene chains thereof have an average molecular weight of less than about 4,000.
Claim 16
Dependent Claim
38 words
A method as set forth in claim 11 wherein the polyoxypropylene chains of the nitrogen-containing polyoxyalkylene nonionic surfactant have an average molecular weight of about 2,750 and the weight percentage of the polyoxyethylene units thereof is about 70%.
Claim 17
Dependent Claim
166 words
In a method of assaying a biological specimen for glucose utilizing a first liquid reagent containing o-dianisidine and a second liquid reagent containing glucose oxidase, peroxidase and buffering agents, the improvement which comprises preparing the first liquid reagent by dissolving in water a granular, water-soluble, substantially anhydrous, storage-stable reagent formulation containing o-dianisidine dihydrochloride and a nitrogen-containing polyoxyalkylene nonionic surfactant obtained by the sequential reaction of ethylenediamine with propylene oxide and ethylene oxide in the presence of a catalyst, the surfactant containing polyoxypropylene chains having an average molecular weight of between about 750 to about 6,750 and polyoxyethylene chains constituting between about 10 and about 80 weight percent of the surfactant and preparing the second liquid reagent by dissolving in water a second granular, water-soluble, substantially anhydrous, storage-stable reagent formulation containing glucose oxidase, peroxidase, gum arabic, mannitol, bovine serum albumin and the nitrogen-containing polyoxyalkylene nonionic surfactant and a third granular, water-soluble, substantially anhydrous, storage-stable reagent formulation containing a phosphate buffer component and the nitrogen-containing polyoxyalkylene nonionic surfactant.
Claim 18
Dependent Claim
166 words
In a method of assaying a biological specimen for glucose utilizing a first liquid reagent containing o-dianisidine and a second liquid reagent containing glucose oxidase, peroxidase and buffering agents, the improvement which comprises preparing the first liquid reagent by dissolving in water a granular, water-soluble, substantially anhydrous, storage-stable reagent formulation containing o-dianisidine dihydrochloride and a nitrogen-containing polyoxyalkylene nonionic surfactant obtained by the sequential reaction of ethylenediamine with propylene oxide and ethylene oxide in the presence of a catalyst, the surfactant containing polyoxypropylene chains having an average molecular weight of between about 750 to about 6,750 and polyoxyethylene chains constituting between about 10 and about 80 weight percent of the surfactant and preparing the second liquid reagent by dissolving in water a second granular, water-soluble, substantially anhydrous, storage-stable reagent formulation containing glucose oxidase, peroxidase, gum arabic, mannitol, bovine serum albumin and the nitrogen-containing polyoxyalkylene nonionic surfactant and a third granular, water-soluble, substantially anhydrous, storage-stable reagent formulation containing a phosphate buffer component and the nitrogen-containing polyoxyalkylene nonionic surfactant.
Claim 19
Dependent Claim
38 words
A method as set forth in claim 13 wherein the polyoxypropylene chains of the nitrogen-containing polyoxyalkylene nonionic surfactant have an average molecular weight of about 2,750 and the weight percentage of the polyoxyethylene units thereof is about 70%.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1979-04-27 | 1979-10-27 | 1980-04-27 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1983-04-27 | 1983-10-27 | 1984-04-27 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1987-04-27 | 1987-10-27 | 1988-04-27 | $7700.00 | Window Closed / Lapsed |