CN104388531A - Compound stabilizer used for lactic dehydrogenase kit - Google Patents
Compound stabilizer used for lactic dehydrogenase kit Download PDFInfo
- Publication number
- CN104388531A CN104388531A CN201410554433.1A CN201410554433A CN104388531A CN 104388531 A CN104388531 A CN 104388531A CN 201410554433 A CN201410554433 A CN 201410554433A CN 104388531 A CN104388531 A CN 104388531A
- Authority
- CN
- China
- Prior art keywords
- package stabilizer
- lactic dehydrogenase
- kit
- reagent
- compound stabilizer
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a compound stabilizer used for a lactic dehydrogenase kit. The compound stabilizer comprises tris hydroxy methyl aminomethan, edetic acid disodium, mycose, beta-cyclodextrin, glycerol, polyvinylpyrrolidone and liquid biological preservatives Proclin300. By means of the mode, the compound stabilizer used for the lactic dehydrogenase kit comprises buffer agents, chelating agents, saccharides, surface active agents, the organic solvents and corrosion removers. After the compound stabilizer is added into the lactic dehydrogenase measurement kit, the stability of testing results can be obviously improved, and the retention time of the lactic dehydrogenase measurement kit can be effectively prolonged.
Description
Technical field
The present invention relates to in-vitro diagnosis field, particularly relate to a kind of one package stabilizer for lactic dehydrogenase enzyme reagent kit.
Background technology
Serum lactic dehydrogenase (SLDH) (LDH) is a kind of oxydo-reductase, the mutual conversion reaction of catalysis pyruvic acid and lactic acid.Serum lactic dehydrogenase divides 5 kinds of isozymes according to subunit, has two types: muscularity and myocardium type.The activity of serum lactic dehydrogenase is used for evaluating the Injured status of cell or tissue, is converted into NAD (NAD based on nicotinamide adenine dinucleotide reduced (NADH)
+), cause the change of 340nm wavelength place absorbancy, thus carrying out the activity of quantitative measurements of lactate desaturase, the content of serum lactic dehydrogenase is directly proportional to the consumption of NADH, and NADH is as a kind of coenzyme, its activity is subject to the interference of various factors, and self is very easily oxidized, directly affect the measurement result of Dehydrogenase Content, cause determination of lactate dehydrogenase test kit deficient in stability, shelf time is short, directly has influence on Clinical practice.
Very limited about the report of determination of lactate dehydrogenase reagent stability at present.By adding glucose dehydrogenase, chemically modified being carried out to coenzyme in patent CN200980105640, generate a kind of metastable coenzyme derivative, thus make it reach stable effect, and do not affect the application of reagent, but this method cost is higher, be not easy to promote; Utilize a kind of enzyme and substrate that already oxidised coenzyme is reduced into reduced coenzyme in patent CN1179792A, be convenient to regenerating coenzyme in storage process like this, but this method implementation cost is high, and the enzyme added self is also unstable; Patent CN02139298 is by adding NAD
+, utilize enzyme-substrate-NAD
+or enzyme-substrate-NADP
+oxidized coenzyme is changed into reduced coenzyme by systemic circulation, can measure when reduced coenzyme runs up to finite concentration, but the method is comparatively complicated, and can not ensure that the generation speed of reduced coenzyme in systemic circulation and total amount can meet the requirement of test kit; Patent 200610030784.8 provides the one package stabilizer of a kind of NADH, but can only prove that the NADH of this one package stabilizer to Individual existence has certain stabilization, can not illustrate to join in the comparatively complicated test kit of composition also can have similar stabilising effect.Therefore, a kind of preparation of research and development is simple, with low cost, has the stablizer of good stabilising effect to be very important to determination of lactate dehydrogenase test kit.
Summary of the invention
The technical problem that the present invention mainly solves is to provide a kind of one package stabilizer for lactic dehydrogenase enzyme reagent kit, and this one package stabilizer will be prepared simple, with low cost and have satisfactory stability effect to determination of lactate dehydrogenase test kit.
For solving the problems of the technologies described above, the technical scheme that the present invention adopts is: provide a kind of one package stabilizer for lactic dehydrogenase enzyme reagent kit, described one package stabilizer comprises Tutofusin tris, disodium ethylene diamine tetraacetate, trehalose, beta-cyclodextrin, glycerine, polyvinylpyrrolidone, liquid bio sanitas Proclin 300, the content of each component is: Tutofusin tris (Tris) 3 ~ 9 g/L, disodium ethylene diamine tetraacetate (EDTA-2Na) 40 ~ 60 g/L, trehalose 20 ~ 30 g/L, beta-cyclodextrin 15 ~ 25 g/L, glycerine 5 ~ 15 g/L, polyvinylpyrrolidone 15 ~ 25 g/L, liquid bio sanitas Proclin 300 1 ~ 10 g/L.
In a preferred embodiment of the present invention, the content of each component of described one package stabilizer is: Tris 6.06g/L, EDTA-2Na 50 g/L, trehalose 25 g/L, beta-cyclodextrin 20 g/L, glycerine 10 g/L, polyvinylpyrrolidone 20 g/L, Proclin300 5 g/L.
In a preferred embodiment of the present invention, described one package stabilizer is applied in lactic dehydrogenase enzyme reagent kit, described lactic dehydrogenase enzyme reagent kit comprises the first reagent and the second reagent, the component of described first reagent is: Sodium phosphate dibasic 25.069 g/L, potassium primary phosphate 2.435 g/L, α-one butyric acid sodium salt 0.496 g/L, and the component of described second reagent is: Tris 8.479 g/L, NADH 0.0851 g/L, hydrochloric acid 5.6 mL/L.
In a preferred embodiment of the present invention, the volume fraction of usage quantity in lactic dehydrogenase enzyme reagent kit of described one package stabilizer is 5%.
The invention has the beneficial effects as follows: the one package stabilizer for lactic dehydrogenase enzyme reagent kit of the present invention, described one package stabilizer is made up of buffer reagent, sequestrant, carbohydrate, tensio-active agent, organic solvent, sanitas.One package stabilizer of the present invention joins after in determination of lactate dehydrogenase test kit, can significantly improve the stability of detected result, effectively extends the shelf time of determination of lactate dehydrogenase test kit.In addition, this one package stabilizer is with low cost, and preparation is simple.
Embodiment
Be clearly and completely described to the technical scheme in the embodiment of the present invention below, obviously, described embodiment is only a part of embodiment of the present invention, instead of whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art, not making other embodiments all obtained under creative work prerequisite, belong to the scope of protection of the invention.
The instrument that the present invention relates to has: UV Blue Star ultraviolet spectrophotometer 1 (Beijing Labtech Instruments Co., Ltd, also or other brand), constant temperature digital display water-bath 1 (Shanghai Mei Xiang Instrument Ltd., also or other brand).
Embodiment 1
There is provided a kind of one package stabilizer for lactic dehydrogenase enzyme reagent kit, the content of each component of described one package stabilizer is: Tris 6.06 g/L, EDTA-2Na 50 g/L, trehalose 25 g/L, beta-cyclodextrin 20 g/L, glycerine 10 g/L, polyvinylpyrrolidone 20 g/L, Proclin300 5 g/L.After above-mentioned each component fully being dissolved, mixing, a kind of liquid composite stabilizing agent finished product can be obtained.
Embodiment 1 is applied to determination of lactate dehydrogenase test kit, and evaluates from stability, accuracy and usage quantity aspect.
(1) stability and accuracy test
1, for examination stablizer: one package stabilizer described in embodiment 1.
2, supply examination test kit to be determination of lactate dehydrogenase test kit, be made up of reagent 1 and reagent 2.Component is as follows:
Reagent 1: Sodium phosphate dibasic 25.069 g/L, potassium primary phosphate 2.435 g/L, α-one butyric acid sodium salt 0.496 g/L;
Reagent 2:Tris 8.479 g/L, NADH 0.0851 g/L, hydrochloric acid 5.6 mL/L.
3, test method and result
One package stabilizer embodiment 1 prepared is by 3%(v/v) amount to add in determination of lactate dehydrogenase test kit (reagent 1 and reagent 2 all add by this amount), measure Dehydrogenase Content in sample respectively with the determination of lactate dehydrogenase test kit not adding one package stabilizer.This test is with Britain's Landau quality controlled serum horizontal 2(Quality Control 2) and the horizontal 3(Quality Control 3 of quality controlled serum) for sample, adopt ultraviolet-visible pectrophotometer to detect, if a skip test, two test samples (skip test replaces sample with distilled water).The ratio of sample and reagent 1 and reagent 2 is 20 μ L:1000 μ L:250 μ L, the predominant wavelength of setting is 340nm, commplementary wave length is 405nm, 1min reading is postponed after sample and reagent mix, reading duration is 1.5min, show that △ A/min(per minute absorbancy changes), according to formula LDH(U/L)=△ A/min × F calculates sample LDH content (calculated factor F=12500).In Quality Control 2, the target value of LDH is 402U/L, and in Quality Control 3, the target value of LDH is 754U/L, permissible variation scope ± 10%.
According to above measuring method, using with batch with bottled quality controlled serum 2, quality controlled serum 3 as detection sample, respectively with adding 3%(v/v) the determination of lactate dehydrogenase test kit of one package stabilizer and the determination of lactate dehydrogenase test kit that do not add one package stabilizer detect, test in every 5 days once, do at every turn three parallel and get its mean value, sense cycle 35 days, obtain the test result for quality controlled serum 2, quality controlled serum 3, calculate standard deviation (SD) and the validity period number of days of measurement result.Standard deviation this can reflect the stability of measurement result.Test-results sees the following form.
From upper table data, do not add the determination of lactate dehydrogenase test kit of one package stabilizer, its detected result standard deviation is obviously larger, be about 4 times of the test kit detected result standard deviation adding 3% one package stabilizer, illustrate that the determination of lactate dehydrogenase test kit not adding one package stabilizer is very unstable, validity period number of days can only maintain about 15 days, relatively, add the determination of lactate dehydrogenase test kit of 3% one package stabilizer in whole test period, accuracy is all within the scope of permissible variation, illustrate and still can keep excellent test performance at about 35 days these mensuration test kits.As can be seen here, the determination of lactate dehydrogenase test kit that with the addition of 3% one package stabilizer shows more excellent in measuring stability and accuracy, and one package stabilizer has obvious stabilising effect to it, effectively can extend the storage life of determination of lactate dehydrogenase test kit.
(2) usage quantity simultaneous test
1, for examination stablizer: one package stabilizer described in embodiment 1.
2, be determination of lactate dehydrogenase test kit for agent of having a try, composition is with reference to stability and accuracy test.
3, test method and result
By one package stabilizer according to 1%(v/v), 5%(v/v) ratio to add respectively in determination of lactate dehydrogenase test kit (reagent 1 and reagent 2 all add by same amount), carry out the mensuration of Dehydrogenase Content in sample.Measure by the measuring method of stability test of the present invention, calculate standard deviation and the validity period number of days of measurement result, standard deviation this can reflect the stability of measurement result.The results are shown in following table.
From upper table data, the stabilization of three kinds of different add-ons to determination of lactate dehydrogenase test kit of one package stabilizer strengthens all to some extent, all can extend the storage life of test kit to some extent.The test kit effect adding 1% one package stabilizer is slightly weak, and stability is not good enough; The kit standard deviation adding 3% one package stabilizer is placed in the middle, illustrates that the one package stabilizer effect of this add-on can't reach optimum; The kit standard deviation of 5% one package stabilizer addition is minimum, be respectively 12 and 17, illustrate that the determination of lactate dehydrogenase test kit repeatability adding 5% one package stabilizer is best, this test kit is more stable, within about 35 days after test kit uncork, can also keep good test performance.Therefore, 5% can be decided to be the optimum quantum of utilization of one package stabilizer in determination of lactate dehydrogenase test kit.
Can obtain from above data and analytical results, after determination of lactate dehydrogenase test kit adds one package stabilizer of the present invention, can extend the shelf time of test kit, measurement result stability is better, and the suitableeest volume ratio that adds is approximately 5%.In addition, this one package stabilizer is with low cost, and preparation is simple, is therefore worth further genralrlization to use.
The foregoing is only embodiments of the invention; not thereby the scope of the claims of the present invention is limited; every utilize description of the present invention to do equivalent structure or equivalent flow process conversion; or be directly or indirectly used in other relevant technical field, be all in like manner included in scope of patent protection of the present invention.
Claims (4)
1. the one package stabilizer for lactic dehydrogenase enzyme reagent kit, it is characterized in that, described one package stabilizer comprises Tutofusin tris, disodium ethylene diamine tetraacetate, trehalose, beta-cyclodextrin, glycerine, polyvinylpyrrolidone, liquid bio sanitas Proclin 300, the content of each component is: Tutofusin tris (Tris) 3 ~ 9 g/L, disodium ethylene diamine tetraacetate (EDTA-2Na) 40 ~ 60 g/L, trehalose 20 ~ 30 g/L, beta-cyclodextrin 15 ~ 25 g/L, glycerine 5 ~ 15 g/L, polyvinylpyrrolidone 15 ~ 25 g/L, liquid bio sanitas Proclin 300 1 ~ 10 g/L.
2. one package stabilizer according to claim 1, it is characterized in that, the content of each component of described one package stabilizer is: Tris 6.06 g/L, EDTA-2Na 50 g/L, trehalose 25 g/L, beta-cyclodextrin 20 g/L, glycerine 10 g/L, polyvinylpyrrolidone 20 g/L, Proclin300 5 g/L.
3. one package stabilizer according to claim 1, it is characterized in that, described one package stabilizer is applied in lactic dehydrogenase enzyme reagent kit, described lactic dehydrogenase enzyme reagent kit comprises the first reagent and the second reagent, the component of described first reagent is: Sodium phosphate dibasic 25.069 g/L, potassium primary phosphate 2.435 g/L, α-one butyric acid sodium salt 0.496 g/L, and the component of described second reagent is: Tris 8.479 g/L, NADH 0.0851 g/L, hydrochloric acid 5.6 mL/L.
4. one package stabilizer according to claim 1, is characterized in that, the volume fraction of usage quantity in lactic dehydrogenase enzyme reagent kit of described one package stabilizer is 5%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410554433.1A CN104388531B (en) | 2014-10-20 | 2014-10-20 | Compound stabilizer used for lactic dehydrogenase kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410554433.1A CN104388531B (en) | 2014-10-20 | 2014-10-20 | Compound stabilizer used for lactic dehydrogenase kit |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104388531A true CN104388531A (en) | 2015-03-04 |
| CN104388531B CN104388531B (en) | 2017-01-18 |
Family
ID=52606488
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410554433.1A Active CN104388531B (en) | 2014-10-20 | 2014-10-20 | Compound stabilizer used for lactic dehydrogenase kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104388531B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1137564A (en) * | 1995-03-09 | 1996-12-11 | 柏林魏克股份公司 | Stable transglutaminase preparations and processes for producing them |
| CN1179792A (en) * | 1995-03-28 | 1998-04-22 | 微量科学有限公司 | Reagent stabilized using coenzyme reduction system |
| CN1986785A (en) * | 2005-12-23 | 2007-06-27 | 上海复星长征医学科学有限公司 | Enzyme combining stabilizer |
| CN102507915A (en) * | 2011-11-07 | 2012-06-20 | 宁波美康生物科技股份有限公司 | Stable liquid kit for measuring lactic acid |
-
2014
- 2014-10-20 CN CN201410554433.1A patent/CN104388531B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1137564A (en) * | 1995-03-09 | 1996-12-11 | 柏林魏克股份公司 | Stable transglutaminase preparations and processes for producing them |
| CN1179792A (en) * | 1995-03-28 | 1998-04-22 | 微量科学有限公司 | Reagent stabilized using coenzyme reduction system |
| CN1986785A (en) * | 2005-12-23 | 2007-06-27 | 上海复星长征医学科学有限公司 | Enzyme combining stabilizer |
| CN102507915A (en) * | 2011-11-07 | 2012-06-20 | 宁波美康生物科技股份有限公司 | Stable liquid kit for measuring lactic acid |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104388531B (en) | 2017-01-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101717814B (en) | Liquid double reagent diagnostic reagent kit for determining content of potassium ions in serum and blood plasma | |
| CN101750327A (en) | Reagent (kit) for diagnosing/determining amino acid and method for determining concentration of amino acid | |
| CN105203472A (en) | Stable NEFA (non-esterified fatty acid) measuring kit | |
| CN104195222A (en) | Compound stabilizer for total cholesterol measurement kits | |
| CN104198422A (en) | Composite stabilizer for alpha-hydroxybutyric dehydrogenase assay kit | |
| CN105002263B (en) | A kind of reduced coenzyme complex reagent and its application | |
| CN101498662A (en) | Reagent kit for monoamine oxidase MAO single-reagent measurement | |
| CN103558370A (en) | Carbon dioxide measurement kit | |
| CN106644976A (en) | Aspartate amino transferase detection kit | |
| CN104390925A (en) | Composite stabilizer for aspartate aminotransferase assay kit | |
| CN106290202A (en) | A kind of preparation method of reagent thereof using continuous monitoring method quantitative determination AST | |
| CN102564979A (en) | Method for determining alcohol concentration by using enzyme cycling method and alcohol determination kit | |
| CN107991252B (en) | Stabilizer for alpha-hydroxybutyrate dehydrogenase determination kit and preparation method thereof | |
| CN104388531A (en) | Compound stabilizer used for lactic dehydrogenase kit | |
| CN106244672A (en) | A kind of stabilizer for uric acid detectable | |
| CN104195221A (en) | Composite stabilizer for glucose assay kit | |
| CN104049091A (en) | Method and kit for detecting ethanol dehydrogenase | |
| KR20210122258A (en) | Stabilization of NADPH or NADH in Ammonia Detection Assay | |
| CN101750337A (en) | Reagent (kit) for diagnosing/determining amino acid and method for determining concentration of amino acid | |
| CN101750315A (en) | Reagent (kit) for diagnosing/determining amino acid and method for determining concentration of amino acid | |
| CN101082569A (en) | Method for measuring amino acid concentration and amino acid diagnose reagent kit | |
| CN101750322A (en) | Reagent (kit) for diagnosing/determining amino acid and method for determining concentration of amino acid | |
| CN101750291A (en) | Reagent (kit) for diagnosing/determining amino acid and method for determining concentration of amino acid | |
| CN101750292A (en) | Reagent (kit) for diagnosing/determining amino acid and method for determining concentration of amino acid | |
| CN101750296A (en) | Reagent (kit) for diagnosing/determining amino acid and method for determining concentration of amino acid |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |