US3819488A
Expired / Lapsed
US
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Diagnostic compositions for glutamic oxalic transaminase (got) and glutamic pyruvic transaminase (gpt) determinations
Assignee: Bayer Corp
•
Filing: 1971-07-20
•
Granted: 1974-06-25
•
Expires: 1991-07-20
Abstract
Abstract Stable, lyophilized diagnostic compositions are disclosed effective in determining glutamic oxalic transaminase (GOT) and glutamic pyruvic transaminase (GPT) comprising malate dehydrogenase (for GOT) or lactate dehydrogenase (for GPT), a pyridine nucleotide and a buffer having a sodium content of less than about 600 meq./1. capable of regulating the pH of the composition in reconstituted form. Also disclosed are the aqueous reconstituted products, the method of determining the activity of GOT and GPT in a sample using the subject compositions and the process for making the disclosed diagnostic compositions.
Inventors
- 👤 R Rush
- 👤 L Leon
- 👤 Lea A De
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-06 16:30
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/48; C12Q1/52; C12Q1/26; C12Q1/32
Claims Analysis
Total Claims: 45
Claim 1
Independent Claim
31 words
The composition as claimed in claim 1 wherein said buffer is potassium phosphate present in an amount to provide a concentration of from about 0.05 M to 0.4 M upon reconstitution.
Claim 2
Dependent Claim
13 words
The composition as claimed in claim 1 wherein said pyridine nucleotide is NADH.
Claim 3
Dependent Claim
16 words
The composition as claimed in claim 1 wherein said activity of MDH is about 3 units/ml.
Claim 4
Dependent Claim
13 words
The composition as claimed in claim 1 wherein said optical density is 1.8.
Claim 5
Dependent Claim
13 words
The composition as claimed in claim 2 wherein said concentration is 0.1 M.
Claim 6
Dependent Claim
37 words
The composition as claimed in claim 1 wherein said buffer is a mixture of potassium phosphate and sodium phosphate and albumin is included in an amount to provide a concentration upon reconstitution of about 0.15 g./100 ml.
Claim 7
Dependent Claim
24 words
A stable aqueous solution having a pH in the range from 6.0 to 8.0 which comprises the composition of claim 1 dissolved in water.
Claim 8
Dependent Claim
143 words
A stable, lyophilized diagnostic composition effective in determining glutamic pyruvic transaminase (GPT) in a sample containing said enzyme which consists of lactate dehydrogenase (LDH) present in an amount to provide an activity upon reconstitution of from 8 units/ml to about 80 units/ml; a pyridine nucleotide selected from the group consisting of reduced nicotinamide adenine dinucleotide (NADH) and reduced nicotinamide adenine dinucleotide phosphate (NADPH) said pyridine nucleotide present in an amount to obtain an optical density upon reconstitution in the range of from 2.1 to 1.0 spectrophotometrically determined at 340 nm using a 10 mm light path; and a buffer selected from the group consisting of potassium phosphate and a mixture of potassium phosphate and sodium phosphate, said mixture having a sodium content less than 600 meq./1 upon reconstitution, which regulates the pH of the reconstituted product in the range from 6 to 8.
Claim 9
Dependent Claim
31 words
The composition as claimed in claim 9 wherein said buffer is potassium phosphate present in an amount to provide a concentration of from about 0.05 M to 0.4 M upon reconstitution.
Claim 10
Dependent Claim
13 words
The composition as claimed in claim 9 wherein said pyridine nucleotide is NADH.
Claim 11
Dependent Claim
16 words
The composition as claimed in claim 9 wherein said activity of LDH is about 40 units/ml.
Claim 12
Dependent Claim
13 words
The composition as claimed in claim 9 wherein said optical density is 1.8.
Claim 13
Dependent Claim
13 words
The composition as claimed in claim 10 wherein said concentration is 0.1 M.
Claim 14
Dependent Claim
37 words
The composition as claimed in claim 9 wherein said buffer is a mixture of potassium phosphate and sodium phosphate and albumin is included in an amount to provide a concentration upon reconstitution of about 0.15 g./100 ml.
Claim 15
Dependent Claim
24 words
A stable aqueous solution having a pH in the range from 6.0 to 8.0 which comprises the composition of claim 9 dissolved in water.
Claim 16
Dependent Claim
56 words
In a method for quantitatively determining the activity of glutamic oxalic transaminase (GOT) in a sample by mixing said sample and a substrate and analyzing for GOT; the improvement which comprises combining the stabilized composition as claimed in claim 1 with the reaction product from said mixing step and photometrically measuring the decrease in NADH concentration.
Claim 17
Dependent Claim
24 words
The method of claim 17 including the further step of passing the reaction product through a dialysis membrane prior to combining with said composition.
Claim 18
Dependent Claim
19 words
The method of claim 17 including the further step of heating the reacTion mixture of said sample and substrate.
Claim 19
Dependent Claim
17 words
The method of claim 17 wherein said substrate is an aspartic acid - Alpha -ketoglutaric acid mixture.
Claim 20
Dependent Claim
20 words
The method of claim 17 wherein said composition buffer regulates the reconstituted product in the range from 6 to 8.
Claim 21
Dependent Claim
11 words
The method of claim 21 wherein said buffer is potassium phosphate.
Claim 22
Dependent Claim
17 words
The method of claim 21 wherein said buffer is a mixture of potassium phosphate and sodium phosphate.
Claim 23
Dependent Claim
15 words
The method of claim 22 wherein said composition comprises malate dehydrogenase, NADH and potassium phosphate.
Claim 24
Dependent Claim
26 words
The method of claim 17 wherein said composition is in the form of an aqueous solution having a pH in the range from 6.0 to 8.0.
Claim 25
Dependent Claim
56 words
In a method for quantitatively determining the activity of glutamic pyruvic transaminase (GPT) in a sample by mixing said sample and a substrate and analyzing for GPT, the improvement which comprises combining the stabilized composition as claimed in claim 9 with the reaction product from said mixing step and photometrically measuring the decrease in NADH concentration.
Claim 26
Dependent Claim
30 words
The method of claim 26 including the further step of passing the reaction product of said sample and said substrate through a dialysis membrane prior to combining with said composition.
Claim 27
Dependent Claim
18 words
The method of claim 26 including the further step of heating the mixture of said sample and substrate.
Claim 28
Dependent Claim
16 words
The method of claim 26 wherein said substrate is an alanine - Alpha -ketoglutaric acid mixture.
Claim 29
Dependent Claim
20 words
The method of claim 26 wherein said composition buffer regulates the reconstituted product in the range from 6 to 8.
Claim 30
Dependent Claim
11 words
The method of claim 30 wherein said buffer is potassium phosphate.
Claim 31
Dependent Claim
17 words
The method of claim 30 wherein said buffer is a mixture of potassium phosphate and sodium phosphate.
Claim 32
Dependent Claim
15 words
The method of claim 31 wherein said composition comprises lactate dehydrogenase, NADH and potassium phosphate.
Claim 33
Dependent Claim
26 words
The method of claim 26 wherein said composition is in the form of an aqueous solution having a pH in the range from 6.0 to 8.0.
Claim 34
Dependent Claim
110 words
A process for the preparation of a stable, lyophilized diagnostic composition as claimed in claim 1 which comprises admixing malate dehydrogenase, a pyridine nucleotide selected from the group consisting of reduced nicotinamide adenine dinucleotide (NADH) and nicotinamide adenine dinucleotide phosphate (NADPH); and a buffer selected from the group consisting of potassium phosphate and a mixture of potassium phosphate and sodium phosphate which regulates the pH of the reconstituted product in the range from 6 to 8; and lyophilizing the resulting aqueous product which upon reconstitution is characterized by an optical density within the range from 2.1 to 1.0 and an MDH activity of from 0.5 units/ml. to about 10 units/ml.
Claim 35
Dependent Claim
28 words
The process of claim 35 wherein said buffer is potassium phosphate added as an 0.5 M aqueous solution whose concentration upon reconstitution is from 0.05 to 0.4 M.
Claim 36
Dependent Claim
31 words
The process of claim 35 wherein said pyridine nucleotide is NADH and the reconstituted lyophilized product is characterized by an optical density of 1.8 and an MDH activity of 3 units/ml.
Claim 37
Dependent Claim
14 words
The process of claim 36 wherein said potassium phosphate concentration is about 0.1 M.
Claim 38
Dependent Claim
17 words
The process of claim 35 wherein said buffer is a mixture of potassium phosphate and sodium phosphate.
Claim 39
Dependent Claim
25 words
The process of claim 39 wherein albumin is added in an amount to provide a concentration of about 0.15 g./100 ml of reconstituted lyophilized product.
Claim 40
Dependent Claim
111 words
A process for the preparation of a stable, lyophilized diagnostic composition as claimed in claim 9 which comprises admixing lactate dehydrogenase; a pyridine nucleotide selected from the group consisting of reduced nicotinamide adenine dinucleotide (NADH) and nicotinamide adenine dinucleotide phosphate (NADPH); and a buffer selected from the group consisting of potassium pHosphate and a mixture of potassium phosphate and sodium phosphate which regulates the pH of the reconstituted product in the range from 6 to 8; and lyophilizing the resulting aqueous product which upon reconstitution is characterized by an optical density within the range from 2.1 to 1.0 acid and an LDH activity of from 8 units/ml to about 80 units/ml.
Claim 41
Dependent Claim
29 words
The process of claim 41 wherein said buffer is potassium phosphate added as an 0.5 M aqueous solution whose concentration upon reconstitution is from 0.05 M to 0.4 M.
Claim 42
Dependent Claim
31 words
The process of claim 41 wherein said pyridine nucleotide is NADH and the reconstituted lyophilized product is characterized by an optical density of 1.8 and an LDH activity of 40 units/ml.
Claim 43
Dependent Claim
20 words
The process of claim 41 wherein said buffer is potassium phosphate and said potassium phosphate concentration is about 0.1 M.
Claim 44
Dependent Claim
17 words
The process of claim 41 wherein said buffer is a mixture of potassium phosphate and sodium phosphate.
Claim 45
Dependent Claim
25 words
The process of claim 45 wherein albumin is added in an amount to provide a concentration of about 0.15 g./100 ml of reconstituted lyophilized product.
References Cited (Backward Citations)
US3413198A
Cited
GB1153109A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1977-06-24 | 1977-12-24 | 1978-06-25 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1981-06-24 | 1981-12-24 | 1982-06-25 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1985-06-24 | 1985-12-24 | 1986-06-25 | $7700.00 | Window Closed / Lapsed |