CN106950095B - Method for reducing detection interference of lipemia sample - Google Patents
Method for reducing detection interference of lipemia sample Download PDFInfo
- Publication number
- CN106950095B CN106950095B CN201710211978.6A CN201710211978A CN106950095B CN 106950095 B CN106950095 B CN 106950095B CN 201710211978 A CN201710211978 A CN 201710211978A CN 106950095 B CN106950095 B CN 106950095B
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- lipase
- sample
- buffer solution
- detection
- lipemia
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- 238000001514 detection method Methods 0.000 title claims abstract description 29
- 238000000034 method Methods 0.000 title claims abstract description 16
- 239000004367 Lipase Substances 0.000 claims abstract description 26
- 102000004882 Lipase Human genes 0.000 claims abstract description 22
- 108090001060 Lipase Proteins 0.000 claims abstract description 22
- 235000019421 lipase Nutrition 0.000 claims abstract description 22
- 239000008055 phosphate buffer solution Substances 0.000 claims abstract description 9
- SPOMEWBVWWDQBC-UHFFFAOYSA-K tripotassium;dihydrogen phosphate;hydrogen phosphate Chemical compound [K+].[K+].[K+].OP(O)([O-])=O.OP([O-])([O-])=O SPOMEWBVWWDQBC-UHFFFAOYSA-K 0.000 claims abstract description 9
- 239000000243 solution Substances 0.000 claims abstract description 8
- 239000007853 buffer solution Substances 0.000 claims abstract description 7
- 230000001000 lipidemic effect Effects 0.000 claims abstract description 5
- 239000011521 glass Substances 0.000 claims abstract description 4
- 235000019626 lipase activity Nutrition 0.000 claims abstract description 4
- 239000000872 buffer Substances 0.000 claims description 3
- 210000004369 blood Anatomy 0.000 description 13
- 239000008280 blood Substances 0.000 description 13
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 12
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 8
- 235000014113 dietary fatty acids Nutrition 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 229930195729 fatty acid Natural products 0.000 description 6
- 239000000194 fatty acid Substances 0.000 description 6
- 150000004665 fatty acids Chemical class 0.000 description 6
- 150000002632 lipids Chemical class 0.000 description 6
- 210000002966 serum Anatomy 0.000 description 4
- 239000000126 substance Substances 0.000 description 3
- 238000005199 ultracentrifugation Methods 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 2
- 238000003113 dilution method Methods 0.000 description 2
- 238000002798 spectrophotometry method Methods 0.000 description 2
- 102000004895 Lipoproteins Human genes 0.000 description 1
- 108090001030 Lipoproteins Proteins 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000002834 transmittance Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/34—Purifying; Cleaning
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to a method for reducing the interference of the detection of a lipemic sample, which comprises the following steps of preparing a lipase buffer solution: taking 1-2.5 g of lipase, 1-2 g of co-lipase and 4-10ml of dipotassium hydrogen phosphate-potassium dihydrogen phosphate buffer solution, placing the three into a glass container, uniformly mixing and dissolving to obtain a prepared solution, wherein the lipase activity in the prepared solution is higher than 500 u/L; adding 5-10% of the prepared lipase buffer solution into a lipemia sample, and incubating for 10-20 min; and (4) centrifuging by using a centrifuge, wherein the rotating speed of the centrifuge is controlled to be 1800-3000 rpm. The method does not need to adopt an ultracentrifuge with high requirement and high price, and has simple detection process and small detection result error.
Description
Technical Field
The invention relates to a method for reducing the detection interference of a lipemia sample.
Background
The existing methods for detecting blood samples by using a biochemical analyzer are all spectrophotometry, and the lipemia sample in the blood sample has obvious scattering effect on light with various wavelengths, so that the results of the biochemical analyzer are generally greatly influenced, and the detection results of the biochemical analyzer are inaccurate. Currently, for the detection of a lipemia sample, an ultracentrifugation method or a dilution method is usually used for processing the lipemia sample before the detection so as to reduce the influence of the lipemia on the detection result. However, both of these detection methods have drawbacks: the ultracentrifugation method is characterized in that the lipid and serum of a lipid blood sample are layered by an ultracentrifuge and then are respectively detected, and the ultracentrifuge is expensive and can not be prepared in a common laboratory, and other substances in the blood are layered after ultracentrifugation, namely, not only two layers of the lipid and the serum are formed, so that the detection trouble is caused; the dilution method is to dilute the lipemia sample to a thinner concentration in order to ensure the light transmittance of the spectrophotometry, and the detection result of the lipemia sample with the thinner concentration has a great error directly. Therefore, the invention provides a method for reducing the detection interference of the lipemia sample, which becomes a research problem of the lipemia sample detection in a laboratory.
Disclosure of Invention
The invention aims to provide a method for reducing the detection interference of a lipemia sample, which does not need to adopt an ultracentrifuge with high requirement and high price, and has simple detection process and small detection result error.
In order to achieve the above purpose, the invention adopts the technical scheme that: a method of reducing interference in the detection of a lipemic sample comprising the steps of:
(1) preparing a lipase buffer: taking 1-2.5 g of lipase, 1-2 g of co-lipase and 4-10ml of dipotassium hydrogen phosphate-potassium dihydrogen phosphate buffer solution, placing the three into a glass container, uniformly mixing and dissolving to obtain a prepared solution, wherein the lipase activity in the prepared solution is higher than 500 u/L;
(2) adding 5-10% of the lipase buffer solution prepared in the step (1) into a lipemia sample, and incubating for 10-20 min;
(3) and (4) centrifuging by using a centrifuge, wherein the rotating speed of the centrifuge is controlled to be 1800-3000 rpm.
Further, the pH of the dipotassium hydrogen phosphate-potassium dihydrogen phosphate buffer solution is 7.5.
The invention has the technical effects that: because the thick chylemia is caused by chylemon in blood, and the main component of the chylemon is triglyceride, the triglyceride is decomposed by adding lipase, the glycerol and the fatty acid which are decomposed by the triglyceride are micromolecule substances, and the glycerol and the fatty acid are components of blood fat, and then the triglyceride and the fatty acid are centrifugally treated by a common centrifugal machine (the rotating speed reaches 1800 revolutions per minute), only a blood fat layer and a blood serum layer are separated after centrifugation, so that the scattering of the chylemon to light is eliminated, the detection interference of a lipemia sample is reduced, an ultracentrifugal machine with high requirement and high price is not needed, the detection process is simple, and the detection result error is small. The auxiliary lipase has the function of increasing the activity of lipase, and the dipotassium hydrogen phosphate-potassium dihydrogen phosphate buffer solution has the function of ensuring the activity of the lipase, so that the effective use of the lipase buffer solution is ensured.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
A method of reducing interference in the detection of a lipemic sample comprising the steps of:
(1) preparing a lipase buffer: taking 1-2.5 g of lipase, 1-2 g of co-lipase and 4-10ml of dipotassium hydrogen phosphate-potassium dihydrogen phosphate buffer solution, placing the three into a glass container, uniformly mixing and dissolving to obtain a prepared solution, wherein the lipase activity in the prepared solution is higher than 500 u/L;
(2) adding 5-10% of the lipase buffer solution prepared in the step (1) into a lipemia sample, and incubating for 10-20 min;
(3) and (4) centrifuging by using a centrifuge, wherein the rotating speed of the centrifuge is controlled to be 1800-3000 rpm.
Further, the pH of the dipotassium hydrogen phosphate-potassium dihydrogen phosphate buffer solution is 7.5.
Because the thick chylemia is caused by chylemon in blood, and the main component of the chylemon is triglyceride, the triglyceride is decomposed by adding lipase, the glycerol and the fatty acid which are decomposed by the triglyceride are micromolecule substances, and the glycerol and the fatty acid are components of blood fat, and then the triglyceride and the fatty acid are centrifugally treated by a common centrifugal machine (the rotating speed reaches 1800 revolutions per minute), only a blood fat layer and a blood serum layer are separated after centrifugation, so that the scattering of the chylemon to light is eliminated, the detection interference of a lipemia sample is reduced, an ultracentrifugal machine with high requirement and high price is not needed, the detection process is simple, and the detection result error is small. In addition, after the step (2), a slight flocculent precipitate, which is a lipoprotein component remaining after the decomposition of the blood lipid particles and is also a blood lipid component, may appear in the severe blood lipid sample, and thus the detection result is not affected, and then the step (3) is similarly started. The auxiliary lipase has the function of increasing the activity of lipase, and the dipotassium hydrogen phosphate-potassium dihydrogen phosphate buffer solution has the function of ensuring the activity of the lipase, so that the effective use of the lipase buffer solution is ensured.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof.
Claims (2)
1. A method for reducing interference in the detection of a lipemic sample, comprising the steps of:
(1) preparing a lipase buffer: 1-2.5 g of lipase, 1-2 g of co-lipase and 4-10ml of dipotassium hydrogen phosphate-potassium dihydrogen phosphate buffer solution are taken, the three are placed in a glass container and uniformly mixed and dissolved, and the lipase activity in the obtained prepared solution is higher than 500 u/L;
(2) adding 5-10% of the lipase buffer solution prepared in the step (1) into a lipemia sample, and incubating for 10-20 min;
(3) and (4) centrifuging by using a centrifuge, wherein the rotating speed of the centrifuge is controlled to be 1800-3000 rpm.
2. The method of claim 1, wherein the step of reducing interference in the detection of the lipemic sample comprises: the pH value of the dipotassium hydrogen phosphate-potassium dihydrogen phosphate buffer solution is 7.5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710211978.6A CN106950095B (en) | 2017-04-01 | 2017-04-01 | Method for reducing detection interference of lipemia sample |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710211978.6A CN106950095B (en) | 2017-04-01 | 2017-04-01 | Method for reducing detection interference of lipemia sample |
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| Publication Number | Publication Date |
|---|---|
| CN106950095A CN106950095A (en) | 2017-07-14 |
| CN106950095B true CN106950095B (en) | 2020-01-10 |
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|---|---|---|---|
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1888862A (en) * | 2005-06-29 | 2007-01-03 | 中生北控生物科技股份有限公司 | Low-density lipoprotein cholesterol quantitative determining method reagent and reagent kit |
| CN1914333A (en) * | 2004-01-29 | 2007-02-14 | 协和梅迪克斯株式会社 | Method, reagent, and kit for determining cholesterol in very-low-density lipoprotein remnant (vldl remnant) |
| CN101065495A (en) * | 2004-11-29 | 2007-10-31 | 株式会社Jimro | Method of measuring cholesterol in remnant-like lipoproteins |
| CN102539731A (en) * | 2012-01-09 | 2012-07-04 | 宁波天康生物科技有限公司 | Reagent and kit for quantitatively determining low-density lipoprotein cholesterol (LDL-C) in serum |
| CN104583419A (en) * | 2012-04-20 | 2015-04-29 | 电化生研株式会社 | Method for removal of triglycerides in lipoproteins other than low-density lipoproteins |
| CN104995310A (en) * | 2012-12-10 | 2015-10-21 | 赛拉诺斯股份有限公司 | Rapid, low-sample-volume cholesterol and triglyceride assays |
-
2017
- 2017-04-01 CN CN201710211978.6A patent/CN106950095B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1914333A (en) * | 2004-01-29 | 2007-02-14 | 协和梅迪克斯株式会社 | Method, reagent, and kit for determining cholesterol in very-low-density lipoprotein remnant (vldl remnant) |
| CN101065495A (en) * | 2004-11-29 | 2007-10-31 | 株式会社Jimro | Method of measuring cholesterol in remnant-like lipoproteins |
| CN1888862A (en) * | 2005-06-29 | 2007-01-03 | 中生北控生物科技股份有限公司 | Low-density lipoprotein cholesterol quantitative determining method reagent and reagent kit |
| CN102539731A (en) * | 2012-01-09 | 2012-07-04 | 宁波天康生物科技有限公司 | Reagent and kit for quantitatively determining low-density lipoprotein cholesterol (LDL-C) in serum |
| CN104583419A (en) * | 2012-04-20 | 2015-04-29 | 电化生研株式会社 | Method for removal of triglycerides in lipoproteins other than low-density lipoproteins |
| CN104995310A (en) * | 2012-12-10 | 2015-10-21 | 赛拉诺斯股份有限公司 | Rapid, low-sample-volume cholesterol and triglyceride assays |
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| CN106950095A (en) | 2017-07-14 |
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Address after: 230000, building 9, Xing Lu tech Industrial Park, Luyang Industrial Zone, Hefei, Anhui Patentee after: Hefei Dean Medical Laboratory Co., Ltd. Address before: 230000, building 9, Xing Lu tech Industrial Park, Luyang Industrial Zone, Hefei, Anhui Patentee before: HEFEI DIAN MEDICAL LABORATORY Co.,Ltd. |
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Effective date of registration: 20201217 Address after: 242000 buildings 9 and 10, 15 Huancheng North Road, Xuancheng City, Anhui Province Patentee after: Xuancheng Dean medical laboratory Co.,Ltd. Address before: 230000 Building 9, Xinglu science and Technology Industrial Park, Luyang Industrial Park, Hefei City, Anhui Province Patentee before: Hefei Dean Medical Laboratory Co.,Ltd. |
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Effective date of registration: 20211018 Address after: 230000 Building 9, Xinglu science and Technology Industrial Park, Luyang Industrial Park, Hefei City, Anhui Province Patentee after: Hefei Dean Medical Laboratory Co.,Ltd. Address before: 242000 buildings 9 and 10, 15 Huancheng North Road, Xuancheng City, Anhui Province Patentee before: Xuancheng Dean medical laboratory Co.,Ltd. |