WO1995010044B1 - A method and apparatus for detecting hemolysis in a fluid sample - Google Patents
A method and apparatus for detecting hemolysis in a fluid sampleInfo
- Publication number
- WO1995010044B1 WO1995010044B1 PCT/US1994/010588 US9410588W WO9510044B1 WO 1995010044 B1 WO1995010044 B1 WO 1995010044B1 US 9410588 W US9410588 W US 9410588W WO 9510044 B1 WO9510044 B1 WO 9510044B1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- sample
- fraction
- concentration
- whole
- hemolysis
- Prior art date
Links
- 206010018910 Haemolysis Diseases 0.000 title claims abstract 19
- 230000002949 hemolytic Effects 0.000 title claims abstract 19
- 239000012530 fluid Substances 0.000 title claims abstract 6
- 239000008280 blood Substances 0.000 claims abstract 21
- 229940037179 Potassium Ion Drugs 0.000 claims abstract 10
- 238000001514 detection method Methods 0.000 claims abstract 10
- NPYPAHLBTDXSSS-UHFFFAOYSA-N potassium ion Chemical compound [K+] NPYPAHLBTDXSSS-UHFFFAOYSA-N 0.000 claims abstract 10
- 229910001414 potassium ion Inorganic materials 0.000 claims abstract 10
- 102000001554 Hemoglobins Human genes 0.000 claims 22
- 108010054147 Hemoglobins Proteins 0.000 claims 22
- 239000000463 material Substances 0.000 claims 21
- 210000003743 Erythrocytes Anatomy 0.000 claims 17
- 210000003324 RBC Anatomy 0.000 claims 17
- 238000000926 separation method Methods 0.000 claims 15
- 210000002381 Plasma Anatomy 0.000 claims 9
- 235000019642 color hue Nutrition 0.000 claims 9
- 210000004369 Blood Anatomy 0.000 claims 8
- 239000012491 analyte Substances 0.000 claims 7
- 230000000694 effects Effects 0.000 claims 6
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims 6
- 229910052700 potassium Inorganic materials 0.000 claims 6
- 239000011591 potassium Substances 0.000 claims 6
- 239000003153 chemical reaction reagent Substances 0.000 claims 3
- 235000019571 color Nutrition 0.000 claims 2
- YRNWIFYIFSBPAU-UHFFFAOYSA-N 4-[4-(dimethylamino)phenyl]-N,N-dimethylaniline Chemical compound C1=CC(N(C)C)=CC=C1C1=CC=C(N(C)C)C=C1 YRNWIFYIFSBPAU-UHFFFAOYSA-N 0.000 claims 1
- 102000013563 Acid Phosphatase Human genes 0.000 claims 1
- 108010051457 Acid Phosphatase Proteins 0.000 claims 1
- 229920003043 Cellulose fiber Polymers 0.000 claims 1
- 102000003855 L-lactate dehydrogenases Human genes 0.000 claims 1
- 108091000084 L-lactate dehydrogenases Proteins 0.000 claims 1
- 210000002700 Urine Anatomy 0.000 claims 1
- SBUBPFHJZHQNNT-UHFFFAOYSA-L [O-]O.[O-]O.CC(C)C1=CC=CC=C1C(C)C Chemical compound [O-]O.[O-]O.CC(C)C1=CC=CC=C1C(C)C SBUBPFHJZHQNNT-UHFFFAOYSA-L 0.000 claims 1
- 238000004159 blood analysis Methods 0.000 claims 1
- 230000001413 cellular Effects 0.000 claims 1
- 239000002131 composite material Substances 0.000 claims 1
- 230000001809 detectable Effects 0.000 claims 1
- 239000000835 fiber Substances 0.000 claims 1
- 239000003365 glass fiber Substances 0.000 claims 1
- 150000002500 ions Chemical class 0.000 claims 1
- RNVCVTLRINQCPJ-UHFFFAOYSA-N o-toluidine Chemical compound CC1=CC=CC=C1N RNVCVTLRINQCPJ-UHFFFAOYSA-N 0.000 claims 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 claims 1
- 229910052710 silicon Inorganic materials 0.000 claims 1
- 239000010703 silicon Substances 0.000 claims 1
- 229920002994 synthetic fiber Polymers 0.000 claims 1
- 239000004758 synthetic textile Substances 0.000 claims 1
Abstract
The present invention concerns a method of detecting hemolysis in a whole-blood sample, a method of determining an elevation in the potassium ion concentration of a whole-blood sample, an apparatus for detecting hemolysis and/or determining an elevation in the potassium ion concentration in a fluid sample, an apparatus for detecting hemolysis and/or determining an elevation in the potassium ion concentration in a whole-blood sample, and a single-use cartridge containing a plurality of microfabricated biosensors which further contains a hemolysis detection unit.
Claims
1. A method of detecting hemolysis in a whole-blood sample comprising:
(a) contacting a whole-blood sample comprising intact red blood cells with a dry separation material having a site where said whole blood sample is introduced and a remainder, said material having physical characteristics effective to separate by wicking action a fraction, which contains extracellular hemoglobin that may be present in said sample, from the intact red blood cells;
(b) allowing said sample to remain in contact with said material for a period of time sufficient to effect said separation; and
(c) detecting at a location in said remainder the presence of extracellular hemoglobin in said fraction.
2. The method of claim 1 in which said fraction includes plasma.
3. The method of claim 1 in which said detecting step comprises visually inspecting said fraction for the presence of a color hue.
4. The method of claim 3 which further comprises comparing said color hue to a colored chart displaying a variety of color hues, each of said color hues indicative of an estimated concentration of extracellular hemoglobin present in a given separated fraction.
5. The method of claim 4 in which the estimated concentration of extracellular hemoglobin is at least 2 mg/dL.
6. The method of claim 4 in which the estimated concentration of extracellular hemoglobin is at least 20 mg/dL.
7. The method of claim 1 in which said detecting step comprises contacting said fraction with a chromogenic reager.; .
8. The method oi claim 7 in which said fraction contains hemoglobin, and said hemoglobin reacts with said chromogenic reagent to provide a visually detectable color change.
9. The method of claim 8 in which said chromogenic reagent is selected from the group consisting of o-toluidine, diisopropylbenzene dihydroperoxide and tetramethylbenzidine.
10. The method of claim 1 in which said detecting step is performed with the aid of a reflectance meter, said meter providing a reading that is a function of the concentration of extracellular hemoglobin present in said fraction.
11. The method of claim 1 or 10 which further comprises estimating the elevation of an analyte in said sample which is due to the hemolysis of red blood cells.
12. The method of claim 2 or 10 which specifically detects a level of hemolysis sufficient to cause an elevation of the potassium concentration by at least 0.1 mM above the potassium concentration which would otherwise have been present in the plasma fraction had the whole-blood sample been free from hemolysis.
13. The method of Claim 1, wherein said analyte is selected from the group consisting of potassium ion, lactate dehydrogenase, and acid phosphatase.
14. The method of claim 1 which further comprises adjusting the apparent concentration or activity of said analyte to account for the proportion of same which is due to the hemolysis of red blood cells.
15. The method of claim 1 in which the physical characteristics of said dry separation material include dimensions in length of from 5 to 50 mm, in width of from 1 to 10 mm, and in thickness of from 0.1 to 3.0 mm.
16. The method of claim 2 in which said fraction is separated from said intact red blood cells by a wicking action of said dry separation material, wherein said red blood cells are retarded relative to the separated fraction.
17. The method of claim 1 in which said dry separation material comprises a composite medium of glass fibers, cellulose fibers and synthetic textile fibers.
18. A method of determining the elevation in the potassium ion concentration due to hemolysis of intact red blood cells in a whole-blood sample, comprising:
(a) contacting a whole-blood sample comprising intact red blood cells with a dry separation material having a site where said whole blood sample is introduced and a remainder, said material having physical characteristics effective to separate by wicking action a plasma fraction, which contains extracellular hemoglobin that may be present in said sample, from the intact red blood cells;
(b) allowing said sample to remain in contact with said material for a period of time sufficient to effect said separation;
(c) estimating the quantity of extracellular hemoglobin in said plasma fraction at a location in said remainder; and
(d) estimating the elevation of the potassium ion concentration in said sample which is due to hemolysis.
19. The method of claim 1 or 18 which is performed in a single-use whole-blood analysis cartridge which comprises said dry separation material and an ion-selective electrode for measuring the concentration of potassium ion.
20. The method of claim 19 in which said potassium ion- selective is microfabricated on a silicon chip.
21. The method of Claim 15, further comprising comparing said elevation of the potassium ion concentration to a pre¬ selected value of hemolysis, and rejecting as unreliable for clinical purposes an elevation above said pre-selected value.
22. An apparatus for detecting hemolysis in a fluid sample comprising:
(a) a dry separation material having a site to which said fluid sample is applied and a remainder, said material having physical characteristics effective to separate by wicking action a fraction of said sample, which fraction would contain a representative concentration of any extracellular hemoglobin that may be present in said sample; and
(b) a detection means for ascertaining at a location in said remainder the concentration of said extracellular hemoglobin in said fraction.
23. The apparatus of claim 22 in which said fluid sample is whole-blood.
24. The apparatus of claim 22 in which said fluid sample is urine.
25. An apparatus f r detecting hemolysis in a who .-blood sample having intact red blood cells comprising:
(a) a dry separation material having a site where said whole blood sample is applied and a remainder, said material having physical characteristics effective to separate by wicking action a fraction of said sample, which fraction would contain a representative concentration of any extracellular hemoglobin that may be present in said sample, from the intact red blood cells; and
(b) a detection means for ascertaining at a location in said remainder the presence of extracellular hemoglobin in said fraction.
26. The apparatus of claim 22 or 25 in which said detection means comprises a colored chart displaying at least one color hue of an estimated concentration of extracellular hemoglobin present in a given separated fraction.
27. The apparatus of claim 22 or 25 in which said detection means comprises a chromogenic reagent that reacts with said extracellular hemoglobin to provide a color change.
28. The apparatus of claim 22 or 25 in which said detection means is contained in a viewing region that comes into contact with said fraction.
29. The apparatus of claim 27 which further comprises an orifice for introduction of said sample.
30. The apparatus of claim 22 or 25 which is a single- use cartridge.
31. The apparatus of claim 25 in which said sample is separated into a plasma fraction that can be observed through a viewing window, said window substantially excluding from view that portion of the separated sample which contains the intact red blood cells.
32. The apparatus of claim 31 in which a colored chart is positioned within view of said window, said colored chart displaying at least one color hue indicative of an estimated concentration of extracellular hemoglobin present in a given plasma fraction.
33. The apparatus of claim 32 in which said color hue correlates to an elevation in the potassium concentration in said plasma fraction by at least 0.1 millimole per liter.
34. The apparatus of claim 32 in which said color hue correlates to an elevation in the potassium concentration in said plasma fraction by at least 0.5 millimole per liter.
35. The apparatus of Claim 22 or 32, comprising at least two color hues, each of which is indicative of an estimated concentration of extra cellular hemoglobin present in a given plasma fraction.
36. The apparatus of claim 21 in which said detection means comprises a reflectance meter including a light source and a light detector, said light source being positioned to permit an incident light beam from said light source to strike said fraction to provide a reflected light beam and said light detector being positioned to permit detection of said reflected light beam.
37. A single-use cartridge, comprising a plurality of microfabricated biosensors arranged to determine the presence or concentration of one or more physiological analytes in a whole-blood sample, said assembly further comprising a hemolysis detection unit comprising:
(a) a dry separation material having a site where said whole blood sample is applied and a remainder, said material physical characteristics effective to separate by wicking action a fraction of said sample, which fraction would contain a representative concentration of any extracellular hemoglobin that may be present in said sample; and (b) a detection means for ascertaining at a location in said remainder the concentration of said extracellular hemoglobin in said fraction.
38. The single-use cartridge of claim 37 which includes a microfabricated potassium ion selective electrode and reference electrode, used to determine the concentration of potassium ion in said sample.
39. A method of estimating a change in an analyte in a whole-blood sample which is due to the hemolysis of red blood cells, comprising:
(a) contacting a whole-blood sample comprising intact red blood cells with a dry separation material having a site where said whole blood sample is introduced and a remainder, said material having physical characteristics effective to separate by wicking action a fraction, which contains extracellular hemoglobin that may be present in said sample, from the intact red blood cells;
(b) allowing said sample to remain in contact with said material for a period of time sufficient to effect said separation;
(c) detecting at a location in said remainder the presence of extracellular hemoglobin in said fraction; and
(d) estimating the change in said analyte in said sample which is due to the hemolysis of red blood cells.
40. The method of Claim 39, wherein said change is an elevation.
41. The method of Claim 39, wherein said change is in the concentration or activity of said analyte.
42. The method of Claim 18 or 39, further comprising the step of determining the presence, concentration or activity of one or more additional physiological analytes in said whole blood sample.
43. The method of Claim 41, wherein said determining comprises contacting said whole blood sample with a plurality of microfabricated biosensors.
44. The method of Claim 43, wherein said additional physiological analyte is potassium.
45. The method of Claim 1, 18 or 39, wherein said dry separation material has an outermost edge and a dimensional axis defined by said site where said whole blood sample is introduced and said outermost edge, and said location in said remainder where said detecting occurs is between said outermost edge and a point one-third of the distance along said dimensional axis.
46. The method of Claim 1, 18 or 39, wherein said dry separation material does not cause visible hemolysis of said intact red blood cells.
Priority Applications (7)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP94929841A EP0722569B1 (en) | 1993-10-04 | 1994-09-28 | A method and apparatus for detecting hemolysis in a fluid sample |
| KR1019960701796A KR100247327B1 (en) | 1993-10-04 | 1994-09-28 | Method and apparatus for detecting hemolysis in a fluid sample |
| DE69432122T DE69432122T2 (en) | 1993-10-04 | 1994-09-28 | METHOD AND DEVICE FOR DETECTING HEMOLYSIS IN LIQUID SAMPLES |
| AU78758/94A AU696428B2 (en) | 1993-10-04 | 1994-09-28 | A method and apparatus for detecting hemolysis in a fluid sample |
| CA002173461A CA2173461A1 (en) | 1993-10-04 | 1994-09-28 | A method and apparatus for detecting hemolysis in a fluid sample |
| AT94929841T ATE232606T1 (en) | 1993-10-04 | 1994-09-28 | METHOD AND DEVICE FOR DETECTING HEMOLYSIS IN LIQUID SAMPLES |
| JP51082395A JP3527741B2 (en) | 1993-10-04 | 1994-09-28 | Method and apparatus for detecting hemolysis in a body fluid sample |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US08/131,527 US5416026A (en) | 1993-10-04 | 1993-10-04 | Method for detecting the change in an analyte due to hemolysis in a fluid sample |
| US131,527 | 1993-10-04 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| WO1995010044A1 WO1995010044A1 (en) | 1995-04-13 |
| WO1995010044B1 true WO1995010044B1 (en) | 1995-04-27 |
Family
ID=22449838
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/US1994/010588 WO1995010044A1 (en) | 1993-10-04 | 1994-09-28 | A method and apparatus for detecting hemolysis in a fluid sample |
Country Status (10)
| Country | Link |
|---|---|
| US (2) | US5416026A (en) |
| EP (1) | EP0722569B1 (en) |
| JP (1) | JP3527741B2 (en) |
| KR (1) | KR100247327B1 (en) |
| CN (1) | CN1135260A (en) |
| AT (1) | ATE232606T1 (en) |
| AU (1) | AU696428B2 (en) |
| CA (1) | CA2173461A1 (en) |
| DE (1) | DE69432122T2 (en) |
| WO (1) | WO1995010044A1 (en) |
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1993
- 1993-10-04 US US08/131,527 patent/US5416026A/en not_active Expired - Lifetime
-
1994
- 1994-09-28 KR KR1019960701796A patent/KR100247327B1/en not_active IP Right Cessation
- 1994-09-28 JP JP51082395A patent/JP3527741B2/en not_active Expired - Fee Related
- 1994-09-28 EP EP94929841A patent/EP0722569B1/en not_active Expired - Lifetime
- 1994-09-28 DE DE69432122T patent/DE69432122T2/en not_active Expired - Lifetime
- 1994-09-28 CN CN94194141A patent/CN1135260A/en active Pending
- 1994-09-28 CA CA002173461A patent/CA2173461A1/en not_active Abandoned
- 1994-09-28 AT AT94929841T patent/ATE232606T1/en not_active IP Right Cessation
- 1994-09-28 WO PCT/US1994/010588 patent/WO1995010044A1/en active IP Right Grant
- 1994-09-28 AU AU78758/94A patent/AU696428B2/en not_active Ceased
- 1994-12-20 US US08/359,595 patent/US5593638A/en not_active Expired - Lifetime
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