CN1281981A - Serum low-density LP determination reagent - Google Patents
Serum low-density LP determination reagent Download PDFInfo
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- CN1281981A CN1281981A CN 99116564 CN99116564A CN1281981A CN 1281981 A CN1281981 A CN 1281981A CN 99116564 CN99116564 CN 99116564 CN 99116564 A CN99116564 A CN 99116564A CN 1281981 A CN1281981 A CN 1281981A
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- quinoline
- damping fluid
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Abstract
The serum low-density lipoprotein assay reagent includes reagent A formed from alpha-sulfonated cyclodextrin, dextran sulfate, MgCl2, dichlorophenol, N-ethyl-N(3-tolyl)-N'-succinylvinyldiamine and 3-[N-morpholine]-2-hydroxypropyl sulfoacid buffer solution and reagent B formed from cholesterol esterase, cholesterol oxidase, peroxidase, 4-ampyrone, polyoxyethylene-polyoxypropylene polyether and 3-[N-morpholine]-2-hydroxypropyl sulfoacid buffer solution. It prossesses the advantages of that it has no need of precipitation of sample, separating operation is simple, convenient and quick, cost is low and accuracy is high and good, etc..
Description
The present invention relates to biochemical technology and biochemical test technology, the particularly diagnosis of clinical medicine in-vitro diagnosis and clinical angiocardiopathy.
LDL is the atherogenicity factor, and LDL increases explanation and has the effect that promotes that atherosclerotic forms.LDL is changed by VLDL in the blood plasma, its function be by liver with cholesterol transport to each histiocytic principal mode of whole body, so and the formation of hypercholesterolemia closely related.Measure the clinical normal employing Friedewald formula of LDL and calculate, the polyvinyl sulfuric acid precipitation method, the electrophoresis technique determining serum low-density LP (Low densitylipoprotein, LDL), the former is that indirect calculation goes out the LDL level, and influence factor is many, and error is big; Although the polyvinyl sulfuric acid precipitation method are common method in the routine work, trouble, error is big; Electrophoresis is not suitable for carrying out in routine work.The method that needs a kind of direct mensuration LDL level in the clinical examination.
The purpose of this invention is to provide a kind of detectable of direct mensuration serum LDL and the detection method of this reagent of use, it is had does not need sample is made precipitation, lock out operation, easy, quick, inexpensive, analysis ability is strong, more convenient than the precipitation method, easier than adopting equation to calculate, and advantage such as accuracy height, degree of accuracy be good, be more suitable for the needs of clinical medical inspection.In addition, reagent of the present invention also is suitable for operating on self-reacting device, can improve the specificity of analytical approach, eliminates the chaff interference influence, improves accuracy.
Detectable of the present invention and detection side's ratio juris are: polyoxyethylene-polyoxypropylene polyethers (POE-POP copolyether) can stop HDL in the serum, very low density lipoprotein (VLDL) (VLDL), chylomicron (CM) to contact (catalysis) with cholesterol oxidase (COD), cholesterol esterase (CEH); And LDL energy freedom and COD, CEH contacts and catalysis.LDL generates 4-courage steroid-3-ketenes and H under cholesterol esterase, cholesterol oxidase effect
2O
2, H
2O
2Under the peroxidase effect, replace than quinoline (4-APP), chlorophenesic acid (DCP) reaction, generate red quinones, utilize the colorimetric analysis principle can obtain LDL content in the sample with the amino peace of 4-.
Serum low-density LP determination reagent provided by the invention comprises by α-sulfonation cyclodextrine, dextran sulfate, MgCl
2, N-ethyl-N-(3-tolyl)-N '-succinyl ethylene diamine, 3-[N-morphine quinoline]-reagent I that 2-hydroxypropyl sulfonic acid damping fluid is formed and by cholesterol esterase, cholesterol oxidase, peroxidase, the amino peace of 4-for than quinoline, polyoxyethylene-polyoxypropylene polyethers, 3-[N-morphine quinoline]-reagent II that 2-hydroxypropyl sulfonic acid damping fluid is formed.Each component concentration is respectively in reagent I, the reagent II:
Reagent I (each component concentration in every liter of reagent):
α-sulfonation cyclodextrine 0.1-2.0mmol,
Dextran sulfate 0.1-1.0g,
MgCl
21-3mmol
Chlorophenesic acid 40-60mmol
N-ethyl-N-(3-tolyl)-N '-succinyl ethylene diamine 1-4mmol
3-[N-morphine quinoline]-2-hydroxypropyl sulfonic acid damping fluid 30-70mmol
Reagent II (each component concentration in every liter of reagent):
Cholesterol esterase 0.5-5.0ku,
Cholesterol oxidase 1-10ku,
Peroxidase 10-100ku,
4-amino-antipyrine 1-5.0mmol
Polyoxyethylene-polyoxypropylene polyethers 1-10g
3-[N-morphine quinoline]-2-hydroxypropyl sulfonic acid damping fluid 30-70mmol
It is the main lipid hazards that development takes place atherosclerotic that LDL increases.Past only surveys TC and estimates the LDL level, but the TC level also is subjected to the influence of HDL level, so present opinion preferably adopts LDL to replace the hazards index of TC as atherosclerosis.
Compare with art methods, reagent provided by the invention and detection method are easy to operate, need not separate each component of lipoprotein, directly measure LDL content in the sample, accuracy, precision, singularity and practicality are good, use detectable of the present invention and detect, and its recovery can reach 98-102%, degree of accuracy reaches: in a few days cv is 0.5-1.0%, and cv is 1.2-2.0% in the daytime; And the recovery of the existing precipitation method only is 90-110%, and degree of accuracy is 0.8-1.5% for cv in a few days then.In the daytime cv is 2-4%; The present invention also is applicable to automatic biochemistry analyzer.Reagent of the present invention can be made into dried frozen aquatic products, is suitable for storage and transport.
Accompanying drawing 1 is an embodiment of the invention typical curve.
The preparation of embodiment 1. reagent is got: α-sulfonation cyclodextrine 0.5mmol, dextran sulfate 0.5g, MgCl
22.0mmol chlorophenesic acid 35mmolN-ethyl-N-(3-tolyl)-N '-succinyl ethylene diamine 1.4mmol3-[N-morphine quinoline]-2-hydroxypropyl sulfonic acid damping fluid 50mmol is mixed with 1 liter of solution generate a reagent I; Get: cholesterol esterase 1ku, cholesterol oxidase 3ku, peroxidase 30ku, the amino peace of 4-is for than quinoline 2.5mmol polyoxyethylene-polyoxypropylene polyethers 4g3-[N-morphine quinoline]-2-hydroxypropyl sulfonic acid damping fluid 50mmol is mixed with 1 liter of solution generate a reagent II.2. detecting operation
| Sample blank sample standard zero standard |
| Serum 200ul 200ul |
| Standard 200ul 200ul |
| Reagent I 1000ul 1000ul 1000ul 1000ul |
| 37 ℃ of insulations of mixing 5 minutes add |
| Reagent II 250ul 250ul |
| Distilled water 250ul 250ul |
The preparation of cholesterol standard items: cholesterol standard items (100mg/dL) are purchased the company in Sigma, required cholesterol is dissolved in the 10mmol/L Nacl solution, add 1 ‰ Sodium azide used as stabilizers, to add the above-mentioned standard items of 2.0ml in the per ampoule, make and freeze thousand product, add the 2.0ml pure water before the use and redissolve, place after 20 minutes and use
3. calculate and the normal value serum low-density LP
Normal value＜3.36mmol/l
4. typical curve
Embodiment of the invention typical curve is stated from Figure of description, is that mensuration is done in French Palcor automatic clinical chemistry analyzer.
According to laboratory condition, optional mensuration by hand or automatic clinical chemistry analyzer revised experiment parameter and made the reagent I: reagent II: sample=20: 4: 5; . when serum VLDL was very high, the result understood on the low side sometimes.So when serum is serious muddy, measure after serum can being diluted one times.
Claims (4)
1. a serum low-density LP determination reagent comprises by α-sulfonation cyclodextrine, dextran sulfate, chlorophenesic acid, MgCl
2, N-ethyl-N-(3-tolyl)-N '-succinyl ethylene diamine, 3-[N-morphine quinoline]-reagent I that 2-hydroxypropyl sulfonic acid damping fluid is formed and by cholesterol esterase, cholesterol oxidase, peroxidase, the amino peace of 4-for than quinoline, polyoxyethylene-polyoxypropylene polyethers, 3-[N-morphine quinoline]-reagent II that 2-hydroxypropyl sulfonic acid damping fluid is formed.
2. serum low-density LP determination reagent according to claim 1 is characterized in that each components contents is in described reagent I, reagent II:
(1) each component concentration is in every liter of reagent I:
α-magnetization cyclodextrine 0.1-2.0mmol,
Dextran sulfate 0.1-1.0g,
MgCl
2?1-3mmol
Chlorophenesic acid 40-60mmol
N-ethyl-N-(3-tolyl)-N '-succinyl ethylene diamine 1-4mmol
3-[N-morphine quinoline]-2-hydroxypropyl sulfonic acid damping fluid 30-70mmol
(2) each component concentration in every liter of reagent II:
Cholesterol esterase 0.5-5.0ku,
Cholesterol oxidase 1-10ku,
Peroxidase 10-100ku,
The amino peace of 4-is replaced than quinoline 1-5.0mmol
Polyoxyethylene-polyoxypropylene polyethers 1-10g
3-[N-morphine quinoline]-2-hydroxypropyl sulfonic acid damping fluid 30-70mmol
3. according to claim 1,2 described serum low-density LP determination reagents, it is characterized in that each components contents is in said reagent I, reagent II:
(1) each component concentration in every liter of reagent I:
α-sulfonation cyclodextrine 0.5mmol,
Dextran sulfate 0.5g,
MgCl
2?2．0mmol
Chlorophenesic acid 35mmol
N-ethyl-N-(3-tolyl)-N '-succinyl ethylene diamine 1.4mmol
3-[N-morphine quinoline]-2-hydroxypropyl sulfonic acid damping fluid 50mmol
(2) each component concentration in every liter of reagent II:
Cholesterol esterase 1ku,
Cholesterol oxidase 3ku,
Peroxidase 30ku,
The amino peace of 4-is replaced than quinoline 2.5mmol
Polyoxyethylene-polyoxypropylene polyethers 4g
3-[N-morphine quinoline]-2-hydroxypropyl sulfonic acid damping fluid 50mmol
4. a serum low-density LP determination method is characterized in that having used each described serum low-density LP determination reagent in the claim 1 to 3.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 99116564 CN1281981A (en) | 1999-07-26 | 1999-07-26 | Serum low-density LP determination reagent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 99116564 CN1281981A (en) | 1999-07-26 | 1999-07-26 | Serum low-density LP determination reagent |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1281981A true CN1281981A (en) | 2001-01-31 |
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|---|---|---|---|
| CN 99116564 Pending CN1281981A (en) | 1999-07-26 | 1999-07-26 | Serum low-density LP determination reagent |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100365409C (en) * | 2005-06-01 | 2008-01-30 | 王贤理 | Reagent for measuring low-density lipoproteincholesterol and preparation method |
| CN1867828B (en) * | 2003-10-15 | 2010-09-29 | 第一化学药品株式会社 | Method for Separation and Determination of Adiponectin Multimer |
| CN103122372A (en) * | 2004-03-31 | 2013-05-29 | 电化生研株式会社 | Method of multiquantification for cholesterol of low-density lipoprotein |
| CN108690869A (en) * | 2017-04-05 | 2018-10-23 | 广州市伊川生物科技有限公司 | A kind of high-density lipoprotein cholesterol detection kit and its detection method |
| CN109097436A (en) * | 2018-08-10 | 2018-12-28 | 山东博科生物产业有限公司 | A kind of low density lipoprotein cholesterol detection reagent of the single agents of precise and high efficiency |
| CN109916890A (en) * | 2019-04-12 | 2019-06-21 | 吉林省汇酉生物技术股份有限公司 | A kind of dry chemistry reagent piece and preparation method thereof quantitative determining total cholesterol concentration |
-
1999
- 1999-07-26 CN CN 99116564 patent/CN1281981A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1867828B (en) * | 2003-10-15 | 2010-09-29 | 第一化学药品株式会社 | Method for Separation and Determination of Adiponectin Multimer |
| CN103122372A (en) * | 2004-03-31 | 2013-05-29 | 电化生研株式会社 | Method of multiquantification for cholesterol of low-density lipoprotein |
| CN103122372B (en) * | 2004-03-31 | 2016-01-13 | 电化生研株式会社 | The method of multiple quantification of cholesterol in low-density lipoproteins |
| CN100365409C (en) * | 2005-06-01 | 2008-01-30 | 王贤理 | Reagent for measuring low-density lipoproteincholesterol and preparation method |
| CN108690869A (en) * | 2017-04-05 | 2018-10-23 | 广州市伊川生物科技有限公司 | A kind of high-density lipoprotein cholesterol detection kit and its detection method |
| CN109097436A (en) * | 2018-08-10 | 2018-12-28 | 山东博科生物产业有限公司 | A kind of low density lipoprotein cholesterol detection reagent of the single agents of precise and high efficiency |
| CN109916890A (en) * | 2019-04-12 | 2019-06-21 | 吉林省汇酉生物技术股份有限公司 | A kind of dry chemistry reagent piece and preparation method thereof quantitative determining total cholesterol concentration |
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