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US4049673A
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Preparation of ferrous hemoglobin and enzymatic digestion products thereof active for absorption of carbon monoxide
Assignee: Individual
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Filing: 1972-09-25
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Granted: 1977-09-20
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Expires: 1992-09-25
Abstract
Abstract Ferrous hemoglobin in a form suitable for absorption of carbon monoxide is produced from a hemoglobin solution, buffered to prevent denaturation of the hemoglobin, and containing an effective reductant. The quantity of reductant present is at least sufficient to counteract the effect of oxygen in such air as comes in contact with the hemoglobin. Enzymatic digestion products of hemoglobin, also active for the absorption of carbon monoxide are produced by controlled decomposition of hemoglobin. These products are lower in equivalent weight than is hemoglobin so that the quantity needed for absorbing a given quantity of carbon monoxide is reduced.
Inventors
- 👤 Israel Herbert Scheinberg
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-03 13:59
CPC Classifications
A; A61; A61K; A61K31/00; A61K31/33; A61K31/395; A61K31/495; A61K31/50; A24; A24D; A24D3/00; A24D3/06; A24D3/14; B; B01; B01D; B01D39/00; B01D39/02; B01D39/04; B01D53/00; B01D53/02
Claims Analysis
Total Claims: 22
Claim 1
Independent Claim
98 words
Method of producing ferrous hemoglobin from an aqueous solution of hemoglobin wherein said solution is to be exposed to a limited quantity of oxygen during production of said ferrous hemoglobin, comprising the steps of adding during agitation of said solution a buffer under conditions such as to maintain said solution at a pH between about 6.0 and 8.5 both locally and overall, and during addition of reductant and adding to said solution a quantity of an effective reductant sufficient to substantially prevent the inactivation of said hemoglobin for the absorption of CO by said limited quantity of oxygen.
Claim 2
Dependent Claim
98 words
Method of producing ferrous hemoglobin from an aqueous solution of hemoglobin wherein said solution is to be exposed to a limited quantity of oxygen during production of said ferrous hemoglobin, comprising the steps of adding during agitation of said solution a buffer under conditions such as to maintain said solution at a pH between about 6.0 and 8.5 both locally and overall, and during addition of reductant and adding to said solution a quantity of an effective reductant sufficient to substantially prevent the inactivation of said hemoglobin for the absorption of CO by said limited quantity of oxygen.
Claim 3
Dependent Claim
19 words
Method as defined in claim 1, wherein said buffer is a phosphate buffer or a tris chydroxymethyl aminoethone buffer.
Claim 4
Dependent Claim
23 words
Method as defined in claim 1, wherein said reductant is selected from the group consisting of ascorbic acid, methylene white and dithionite ion.
Claim 5
Dependent Claim
36 words
Method as defined in claim 1, further comprising the step of adding buffer together with said reductant to bring the pH of said solution toward the center of the pH range of about 6.0 to 8.5.
Claim 6
Dependent Claim
45 words
Method as defined in claim 1, wherein the quantity of effective reductant added is substantially sufficient to prevent the inactivation of said hemoglobin for absorption of CO by oxygen to which it is expected that said hemoglobin will be exposed during and subsequent to production.
Claim 7
Dependent Claim
39 words
Method as defined in claim 1, wherein the ratio of effective reductant to the quantity of hemoglobin present, expressed in equivalents with respect to the ferrous iron content of said hemoglobin, is in the range of 1:1 to 5:1.
Claim 8
Dependent Claim
34 words
Method as defined in claim 1, further comprising the step of drying said solution by one of the processes, freeze-drying, drum-drying and spray-drying, and placing in proximity to a desiccant, thereby producing amorphous hemoglobin.
Claim 9
Dependent Claim
92 words
The process as defined in claim 1, further comprising the steps of adding a quantity of ammonium sulfate to said solution, monitoring the pH of said solution during addition of said ammonium sulfate and adding buffer as necessary to keep the pH from dropping below 6.0, the quantity of added ammonium sulfate being from 45% to 55% of that necessary to saturate said solution, chilling said solution to -15° C. to 0° C., allowing said solution to stand until crystallization of said ferrous hemoglobin is complete and separating off the crystals formed.
Claim 10
Dependent Claim
56 words
The process as defined in claim 8, further comprising the steps of redissolving said crystals in a solution containing effective reductant and buffered to a pH between about 7.5 and 8.5, adjusting the pH to about 7.0 and cooling said solution to between 0° C. and 10° C. to crystallize out said hemoglobin in purified form.
Claim 11
Dependent Claim
52 words
The process as defined in claim 8, further comprising the steps of adding ice-cold ethanol in quantity which is about 25% of the weight of said solution, cooling said solution to between -15° C. to 0° C., allowing said solution to stand until said crystallization is complete and separating off said crystals.
Claim 12
Dependent Claim
56 words
The process as defined in claim 10, further comprising the steps of redissolving said crystals in a solution containing effective reductant and buffered to a pH between about 7.5 and 8.5, adjusting the pH to about 7.0 and cooling said solution to between 0° C. and 10° C. to crystallize out said hemoglobin in purified form.
Claim 13
Dependent Claim
40 words
Method as defined in claim 1, wherein the hemoglobin in said aqueous hemoglobin solution is a component of the blood of a living creature, said living creature being of a type which utilizes ferrous hemoglobin for the transport of oxygen.
Claim 14
Dependent Claim
48 words
Method as defined in claim 12, wherein said blood is collected in an anti-coagulant, red cells therein are separated by centrifugation, the red cells are washed with a solution containing about 0.85% sodium chloride, the red cells are lysed, and red cell ghosts are separated off by centrifugation.
Claim 15
Dependent Claim
46 words
Method as defined in claim 12, wherein said blood is defibrinated, the red cells are separated by centrifugation, the red cells are washed with a solution containing about 0.85% sodium chloride, the red cells are lysed, and the red cell ghosts are separated off by centrifugation.
Claim 16
Dependent Claim
19 words
Method as defined in claim 13, wherein said lysing is effected by suspending said red cells in distilled water.
Claim 17
Dependent Claim
15 words
The product comprising amorphous hemoglobin, buffer, and reductant produced by the method of claim 7.
Claim 18
Dependent Claim
15 words
A product comprising crystalline hemoglobin, buffer and reductant produced by the method of claim 8.
Claim 19
Dependent Claim
47 words
The process as defined in claim 8 wherein the pH of said solution is maintained at an acid pH at least as high as 6.0 during said crystallization step with the objective of obtaining hemoglobin suitable for absorption of negatively charged organic volatiles in addition to CO.
Claim 20
Dependent Claim
43 words
The process as defined in claim 8 wherein the pH is maintained at an alkaline pH with 8.5 as the upper limit during said crystallization step with the objective of obtaining hemoglobin suitable for absorption of positively charged volatiles in addition to CO.
Claim 21
Dependent Claim
12 words
A product comprising crystalline hemoglobin made by the method of claim 18.
Claim 22
Dependent Claim
12 words
A product comprising crystalline hemoglobin made by the method of claim 19.
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) | 1980-09-19 | 1981-03-21 | 1981-09-20 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1984-09-19 | 1985-03-21 | 1985-09-20 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1988-09-19 | 1989-03-21 | 1989-09-20 | $7700.00 | Window Closed / Lapsed |