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US4436820A
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Method and apparatus for glycosylated hemoglobin separating and measuring fractions
Assignee: Individual
•
Filing: 1982-02-01
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Granted: 1984-03-13
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Expires: 2002-02-01
Abstract
Abstract Method and apparatus for separating and measuring the amount of the glycosylated hemoglobin fractions in a blood sample. The method and apparatus provide a means for lysing sample red blood cells and, in the same container, separating the glycosylated hemoglobin fractions from the other residual hemoglobin fractions through the utilization of a polymeric material.
Inventors
- 👤 Paul Reiter
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-08 22:24
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/72; G01N33/721; G01N33/726; G01N33/723; Y; Y10; Y10T; Y10T436/00; Y10T436/25; Y10T436/25375; Y10T436/255
Claims Analysis
Total Claims: 40
Claim 1
Independent Claim
126 words
An apparatus for separating glycosylated hemoglobin fractions from a sample of whole blood comprising: a. a container for holding said blood sample; said container being formed with one open end and one closed end; b. an ion exchange resin disposed within said container and substantially adjacent to said closed end; c. a blood cell lysing compound disposed within said container and substantially adjacent to said closed end; and d. a cap, said cap being removably attachable to said open end and forming a sealed container when attached; said cap being formed with a substantially conical shape and having an opening at the apex of the cone; said conical cap depending into said container and with the interior of said conical cap containing a viscous polymeric material.
Claim 2
Independent Claim
126 words
An apparatus for separating glycosylated hemoglobin fractions from a sample of whole blood comprising: a. a container for holding said blood sample; said container being formed with one open end and one closed end; b. an ion exchange resin disposed within said container and substantially adjacent to said closed end; c. a blood cell lysing compound disposed within said container and substantially adjacent to said closed end; and d. a cap, said cap being removably attachable to said open end and forming a sealed container when attached; said cap being formed with a substantially conical shape and having an opening at the apex of the cone; said conical cap depending into said container and with the interior of said conical cap containing a viscous polymeric material.
Claim 3
Dependent Claim
16 words
The apparatus of claim 1, wherein the viscous polymeric material comprises a silicone type of material.
Claim 4
Dependent Claim
32 words
The apparatus of claim 1, wherein the diameter of the cap adjacent to the base of the cone is substantially similar to that of the diameter of the inside of the cylinder.
Claim 5
Dependent Claim
10 words
The apparatus of claim 1, wherein the container is cylindrical.
Claim 6
Dependent Claim
12 words
The apparatus of claim 4, wherein the container is a test tube.
Claim 7
Dependent Claim
24 words
The apparatus of claim 1, wherein the cation exchange resin is mixed with the blood lysing compound to form a mixture in the container.
Claim 8
Dependent Claim
24 words
The apparatus of claim 6, wherein the volume ratio of the volume mixture of cation exchange resin to blood lysing compound is about 1:3.
Claim 9
Dependent Claim
15 words
The apparatus of claim 1, wherein the ion exchange resin is a cation exchange resin.
Claim 10
Dependent Claim
14 words
The apparatus of claim 8, wherein the cation exchange resin is an acrylic type.
Claim 11
Dependent Claim
19 words
The apparatus of claim 8, wherein the cation exchange resin has a pH of about 6 to about 7.
Claim 12
Dependent Claim
16 words
The apparatus of claim 8, wherein the cation exchange resin has a 200-400 wet mesh range.
Claim 13
Dependent Claim
16 words
The apparatus of claim 1, wherein the blood cell lysing compound comprises a elution lysing buffer.
Claim 14
Dependent Claim
18 words
The apparatus of claim 12, wherein the elution lysing buffer comprises sodium phosphate, potassium cyanide and a surfactant.
Claim 15
Dependent Claim
12 words
The apparatus of claim 12, wherein the surfactant is a hemolysing agent.
Claim 16
Dependent Claim
12 words
The apparatus of claim 12, wherein the surfactant is octyl phenoxy polyethoxyethanol.
Claim 17
Independent Claim
187 words
A method for separating glycosylated hemoglobin from a sample of whole blood fluid comprising: a. providing a quantity of a cation exchange resin to a container; b. providing a quantity of a blood cell lysing composition to said container; c. providing a quantity of blood sample to said container; d. agitating said container for a period of time; e. providing said container with a quantity of a viscous polymeric material during agitation; and f. providing a settling period for said container; whereby during agitation the blood cells of said sample are lysed by the blood cell lysing composition, glycosylated hemoglobin is released from said lysed blood cells and goes into suspension in the blood sample fluid, the remaining hemoglobin from said lysed blood cells binds with the cation exchange resin suspended in the blood sample fluid and settles to the bottom of said container with the other blood cell fragments during the settling period, and the glycosylated hemoglobin remains separately in suspension in the blood sample fluid, said viscous polymeric material providing a permanent barrier between settled solids and the glycosylated hemoglobin in the blood sample fluid.
Claim 18
Independent Claim
187 words
A method for separating glycosylated hemoglobin from a sample of whole blood fluid comprising: a. providing a quantity of a cation exchange resin to a container; b. providing a quantity of a blood cell lysing composition to said container; c. providing a quantity of blood sample to said container; d. agitating said container for a period of time; e. providing said container with a quantity of a viscous polymeric material during agitation; and f. providing a settling period for said container; whereby during agitation the blood cells of said sample are lysed by the blood cell lysing composition, glycosylated hemoglobin is released from said lysed blood cells and goes into suspension in the blood sample fluid, the remaining hemoglobin from said lysed blood cells binds with the cation exchange resin suspended in the blood sample fluid and settles to the bottom of said container with the other blood cell fragments during the settling period, and the glycosylated hemoglobin remains separately in suspension in the blood sample fluid, said viscous polymeric material providing a permanent barrier between settled solids and the glycosylated hemoglobin in the blood sample fluid.
Claim 19
Dependent Claim
18 words
The method of claim 16, wherein the container is agitated for about 10 minutes at about 23° C.
Claim 20
Dependent Claim
16 words
The method of claim 16, wherein the viscous polymeric material is a silicone type of material.
Claim 21
Dependent Claim
30 words
The method of claim 16, wherein the container is provided with a volume ratio of cation exchange resin to blood cell lysing composition to whole blood fluid of about 1:3:10.
Claim 22
Dependent Claim
11 words
The method of claim 19, wherein the container is a test-tube.
Claim 23
Dependent Claim
22 words
The method of claim 16, wherein the container is exposed to centrifugal force to decrease the time needed for the settling period.
Claim 24
Dependent Claim
19 words
The method of claim 21, wherein the container is exposed to about 10 minutes of centrifugation at high speeds.
Claim 25
Dependent Claim
16 words
The method of claim 22, wherein the viscous polymeric material is a silicone type of material.
Claim 26
Dependent Claim
16 words
The method of claim 16, wherein the blood cell lysing composition is an elution lysing buffer.
Claim 27
Dependent Claim
18 words
The method of claim 24, wherein the elution lysing buffer comprises sodium phosphate, potassium cyanide and a surfactant.
Claim 28
Dependent Claim
12 words
The method of claim 25, wherein the surfactant is a hemolysing agent.
Claim 29
Dependent Claim
12 words
The method of claim 25, wherein the surfactant is octyl phenoxy polyethoxyethanol.
Claim 30
Independent Claim
184 words
A method for measuring the amount of glycosylated hemoglobin out of the total hemoglobin in a blood sample comprising: a. adding a cation exchange resin and a blood cell lysing composition to a first container; b. adding a first blood sample to the first container; c. agitating the mixture of step b for a period of time; d. after agitation, exposing the container to centrifugal force for a period of time to form supernatant and accretion solids; e. providing a viscous polymeric material which forms a separation layer between the supernatant and accretion solids during exposure of said centrifugal force; f. measuring the supernatant with a spectrophotometer to obtain a first measurement; g. adding a second blood sample to a quantity of water in a second container; h. measuring the mixture of step g, with a spectrophotometer to obtain a second measurement; and i. measuring the difference between the first measurement and the second measurement whereby the ratio of said first measurement to said second measurement is indicative of the relative amounts of glycosylated hemoglobin out of the total hemoglobin of the blood sample.
Claim 31
Independent Claim
184 words
A method for measuring the amount of glycosylated hemoglobin out of the total hemoglobin in a blood sample comprising: a. adding a cation exchange resin and a blood cell lysing composition to a first container; b. adding a first blood sample to the first container; c. agitating the mixture of step b for a period of time; d. after agitation, exposing the container to centrifugal force for a period of time to form supernatant and accretion solids; e. providing a viscous polymeric material which forms a separation layer between the supernatant and accretion solids during exposure of said centrifugal force; f. measuring the supernatant with a spectrophotometer to obtain a first measurement; g. adding a second blood sample to a quantity of water in a second container; h. measuring the mixture of step g, with a spectrophotometer to obtain a second measurement; and i. measuring the difference between the first measurement and the second measurement whereby the ratio of said first measurement to said second measurement is indicative of the relative amounts of glycosylated hemoglobin out of the total hemoglobin of the blood sample.
Claim 32
Dependent Claim
14 words
The method of claim 28, wherein the mixture is agitated for about 10 minutes.
Claim 33
Dependent Claim
17 words
The method of claim 28, wherein the mixture is centrifuged at high speeds for about 10 minutes.
Claim 34
Dependent Claim
17 words
The method of claim 28, wherein the supernatant is measured in a spectrophotometer at about 415 nm.
Claim 35
Dependent Claim
16 words
The method of claim 28, wherein the quantity of the second blood sample is about 5μ.
Claim 36
Dependent Claim
17 words
The method of claim 28, wherein the quantity of water in step g is about 20 ml.
Claim 37
Dependent Claim
41 words
The method of claim 28, wherein the ratio of the spectrophotometric reading of the first blood sample to the spectrophotometric reading of the second blood sample multiplied by 6.66±0.01 equals the percentage of glycosylated hemoglobin out of the total hemoglobin present.
Claim 38
Dependent Claim
16 words
The method of claim 28, wherein the viscous polymeric material is a silicone type of material.
Claim 39
Dependent Claim
15 words
The method of claim 28, wherein the quantity of first blood sample is about 5μ.
Claim 40
Dependent Claim
24 words
The method of claim 36, wherein the total quantity of the cation exchange resin and a blood cell lysing composition is about 4 ml.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1987-03-13 | 1987-09-12 | 1988-03-13 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-03-13 | 1991-09-12 | 1992-03-13 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-03-13 | 1995-09-12 | 1996-03-13 | $7700.00 | Window Closed / Lapsed |