US4892815A
Expired / Lapsed
US
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Process and reagent for the specific determination of the cholesterol of the HDL fraction
Assignee: Roche Diagnostics GmbH
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Filing: 1987-10-06
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Granted: 1990-01-09
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Expires: 2007-10-06
Abstract
Abstract The present invention provides a process for the specific determination of the cholesterol of the HDL fraction in the presence of the LDL fraction of serum lipoproteins. Pancreatic cholesterol esterase is used to liberate cholesterol, and the liberated cholesterol then reacts with cholesterol oxidase and oxygen to form hydrogen peroxide. The kinetics of either of hydrogen peroxide formation or oxygen consumption is measured within 2 to 15 minutes after the start of the reaction between cholesterol and the oxidase. The temperature is maintained within a range of 20 DEG C., during a predetermined time interval. Specific concentrations of reactants are maintained in the reaction solution, i.e., from 0.05 to 30 U/ml pancreatic cholesterol esterase; from 0.1 to 50 U/ml cholesterol oxidase; from 1.0 to 20 mMole/liter of a tenside of the bile acid group, and 0.1 to 10 g/liter of a non-ionic detergent. The pH is kept within a range of 5 to 9. In addition, a reagent is provided which is used for specific determination of HDL fraction cholesterol in the presence of the LDL fraction of serum lipoproteins. The reagent contains pancreatic esterase (0.05 to 30 U/ml); cholesterol oxidase (0.1 to 50 U/ml) a tenside of the bile acid grop (1.5 to 8 mMole/liter) and a non-ionic detergent (0.1 to 10 g/liter), all concentrations referring to the dilution used in the test. The reagent also contains a buffer at a pH of from 5 to 9, and a system for photometric determination of hydrogen peroxide.
Inventors
- 👤 Lorenz Kerscher
- 👤 Brigitte Pautz
- 👤 Gisela Trunk
- 👤 Joachim Ziegenhorn
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-08 15:19
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/60; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/92; G01N2800/00; G01N2800/04; G01N2800/044; Y; Y10; Y10S; Y10S435/00; Y10S435/805; Y10S435/962; Y10S436/00; Y10S436/807; Y10S436/81; Y10S436/824
Claims Analysis
Total Claims: 19
Claim 1
Independent Claim
147 words
Process for specific determination of HDL cholesterol in a serum lipoprotein containing sample which contains LDL comprising adding to said sample pancreatic cholesterol esterase to liberate cholesterol from cholesterol esters, cholesterol oxidase and oxygen to oxidize said liberated cholesterol with the formation of hydrogen peroxide and kinetically measuring one of hydrogen peroxide formation or oxygen consumption, within 2-15 minutes thereafter as a measurement of HDL cholesterol, said measuring and reaction taking place at a temperature of from 20° C to 40° C during a predetermined time interval during which measurement one maintains a pancreatic cholesterol esterase concentration of from 0.05 to 30 U/ml, a cholesterol oxidase concentration of from 0.1 to 50 U/ml, a tenside of a bile acid group at a concentration of from 1.0 to 20 mMole/liter, concentration of a non-ionic detergent of from 0.1 to 10g/liter and a pH of from 5 to 9.
Claim 2
Dependent Claim
147 words
Process for specific determination of HDL cholesterol in a serum lipoprotein containing sample which contains LDL comprising adding to said sample pancreatic cholesterol esterase to liberate cholesterol from cholesterol esters, cholesterol oxidase and oxygen to oxidize said liberated cholesterol with the formation of hydrogen peroxide and kinetically measuring one of hydrogen peroxide formation or oxygen consumption, within 2-15 minutes thereafter as a measurement of HDL cholesterol, said measuring and reaction taking place at a temperature of from 20° C to 40° C during a predetermined time interval during which measurement one maintains a pancreatic cholesterol esterase concentration of from 0.05 to 30 U/ml, a cholesterol oxidase concentration of from 0.1 to 50 U/ml, a tenside of a bile acid group at a concentration of from 1.0 to 20 mMole/liter, concentration of a non-ionic detergent of from 0.1 to 10g/liter and a pH of from 5 to 9.
Claim 3
Dependent Claim
29 words
Process according to claim 1, comprising adding the non-ionic detergent after the addition of the oxidase to said sample but from 1 to 14 minutes before said kinetic measurement.
Claim 4
Dependent Claim
22 words
Process according to claim 1, comprising carrying out the kinetic measurement within 3 to 5 minutes after addition of the cholesterol oxidase.
Claim 5
Dependent Claim
18 words
Process according to claim 1, comprising maintaining a pancreatic cholesterol esterase concentration of from 0.05 to 10 U/ml.
Claim 6
Dependent Claim
17 words
Process according to claim 1 comprising maintaining a cholesterol oxidase concentration of from 1 to 30 U/ml.
Claim 7
Dependent Claim
17 words
Process according to claim 1 comprising maintaining a concentration of tenside of from 1.5 to 8 mMole/liter.
Claim 8
Dependent Claim
17 words
Process according to claim 1 comprising maintaining a concentration of non-ionic detergent of from 0.4 to 4g/liter.
Claim 9
Dependent Claim
20 words
Process according to claim 1, comprising carrying out said measurement at a temperature of from 25° C to 37° C.
Claim 10
Dependent Claim
11 words
Process according to claim 1, wherein said tenside is sodium cholate.
Claim 11
Dependent Claim
16 words
Process according to claim 1 wherein said non-ionic detergent is a polyethylene oxide group containing detergent.
Claim 12
Dependent Claim
24 words
Process according to claim 1, further comprising carrying out said process in the presence of at least one of anti-LDL-antibodies and antiapolipoprotein B antibodies.
Claim 13
Dependent Claim
25 words
Process according to claim 1, further comprising determining LDL cholesterol within 2 minutes after addition of the cholesterol oxidase by determining hydrogen peroxide formation kinetically.
Claim 14
Dependent Claim
54 words
Process according to claim 1, further comprising adding an amount of at least one of detergent and pancreatic cholesterol esterase sufficient to oxidise total cholesterol contained in the sample to form hydrogen peroxide and determining an end value of a color forming reaction as a measure of the total cholesterol content of the sample.
Claim 15
Dependent Claim
102 words
Reagent for specific determination of HDL cholesterol in a serum lipoprotein containing sample which contains LDL comprising pancreatic cholesterol esterase, cholesterol oxidase, a tenside of a bile acid group, a non-ionic detergent, a buffer at pH from 5 to 9, and a system for photometric determination of hydrogen peroxide, wherein said reagent contains cholesterol esterase in a concentration of from 0.05 to 30 U/ml., cholesterol oxidase in a concentration of from 0.1 to 50 U/ml., tenside of the bile acid group in a concentration of from 1.5 to 8 mMole/liter and non-ionic detergent in a concentration of from 0.1 to 10 g/liter.
Claim 16
Dependent Claim
102 words
Reagent for specific determination of HDL cholesterol in a serum lipoprotein containing sample which contains LDL comprising pancreatic cholesterol esterase, cholesterol oxidase, a tenside of a bile acid group, a non-ionic detergent, a buffer at pH from 5 to 9, and a system for photometric determination of hydrogen peroxide, wherein said reagent contains cholesterol esterase in a concentration of from 0.05 to 30 U/ml., cholesterol oxidase in a concentration of from 0.1 to 50 U/ml., tenside of the bile acid group in a concentration of from 1.5 to 8 mMole/liter and non-ionic detergent in a concentration of from 0.1 to 10 g/liter.
Claim 17
Dependent Claim
19 words
Reagent according to claim 14 further comprising at least one of antibodies against LDL and antibodies against apolipoprotein B.
Claim 18
Dependent Claim
16 words
Reagent according to claim 14 wherein said reagent is impregnated in or on a carrier material.
Claim 19
Dependent Claim
16 words
Reagent according to claim 16, wherein the carrier material is an absorbent, swellable or film-forming material.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1993-01-08 | 1993-07-10 | 1994-01-09 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1997-01-08 | 1997-07-10 | 1998-01-09 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2001-01-08 | 2001-07-10 | 2002-01-09 | $7700.00 | Window Closed / Lapsed |