CN104388531B - Compound stabilizer used for lactic dehydrogenase kit - Google Patents
Compound stabilizer used for lactic dehydrogenase kit Download PDFInfo
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- CN104388531B CN104388531B CN201410554433.1A CN201410554433A CN104388531B CN 104388531 B CN104388531 B CN 104388531B CN 201410554433 A CN201410554433 A CN 201410554433A CN 104388531 B CN104388531 B CN 104388531B
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- complex stabilizer
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- kit
- lactic dehydrogenase
- lactic
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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, more particularly to a kind of stable composition for lactic dehydrogenase enzyme reagent kit
Agent.
Background technology
Serum lactate dehydrogenase (SLD) (ldh) is a kind of oxidoreductase, the mutual conversion reaction of catalysis acetone acid and lactic acid.Breast
Acidohydrogenase divides 5 kinds of isozymes according to subunit, has two types: muscularity and myocardium type.The activity of lactic acid dehydrogenase is used for
Evaluate the Injured status of cell or tissue, oxidized form nicotinoyl is converted into based on nicotinamide adenine dinucleotide reduced (nadh)
Amine adenine-dinucleotide (nad+), cause the change of absorbance at 340nm wavelength, thus carrying out quantitative measurements of lactate dehydrogenase
Activity, the content of lactic acid dehydrogenase is directly proportional to the consumption of nadh, and nadh is as a kind of coenzyme, its activity be easily subject to various because
The interference of element, and itself is easily oxidized, directly affects the measurement result of Dehydrogenase Content, causes determination of lactate dehydrogenase
Test kit deficient in stability, the holding time is short, directly influences Clinical practice.
Very limited with regard to the report of determination of lactate dehydrogenase reagent stability at present.Logical in patent cn200980105640
Crossing addition glucose dehydrogenase coenzyme is chemically modified, generating a kind of metastable coenzyme derivant, thus reaching
Stable effect, and do not affect the application of reagent, but this method is relatively costly, is not easy to promote；In patent cn1179792a
Using a kind of enzyme and substrate, already oxidised coenzyme is reduced into reduced coenzyme, is so easy to regenerating coenzyme in storage process,
But this method implementation cost is high, and the enzyme itself adding is also unstable；Patent cn02139298 is passed through to add nad+, utilize
Enzyme-substrate-nad+Or enzyme-substrate-nadp+Oxidized coenzyme is changed into reduced coenzyme by system circulation, when reduced coenzyme amasss
Tire out and can be measured during finite concentration, but the method is complex, and it cannot be guaranteed that in system circulation reduced form auxiliary
The generation speed of enzyme and total amount can meet the requirement of test kit；Patent 200610030784.8 provides a kind of the compound steady of nadh
Determine agent, but can only prove that this complex stabilizer has certain Stabilization to the nadh of individualism, can not illustrate to add
Enter and also can have similar stablizing effect in the complex test kit of composition.Therefore, research and develop one kind and prepare simple, cost
Cheap, the stabilizer having good stablizing effect to determination of lactate dehydrogenase test kit is very important.
Content of the invention
The invention mainly solves the technical problem of providing a kind of complex stabilizer for lactic dehydrogenase enzyme reagent kit, should
Complex stabilizer will be prepared simple, with low cost and have good stablizing effect to determination of lactate dehydrogenase test kit.
For solving above-mentioned technical problem, one aspect of the present invention is: provides one kind to be used for lactic acid dehydrogenase
The complex stabilizer of test kit, described complex stabilizer includes trishydroxymethylaminomethane, disodiumedetate, Sargassum
Sugar, beta-schardinger dextrin-, glycerol, Polyvinylpyrrolidone, liquid bio preservative proclin 300, the content of each component is: three hydroxyls
Aminomethane (tris) 3 ~ 9 g/l, disodiumedetate (edta-2na) 40 ~ 60 g/l, trehalose 20 ~ 30
G/l, beta-schardinger dextrin -15 ~ 25 g/l, glycerol 5 ~ 15 g/l, Polyvinylpyrrolidone 15 ~ 25 g/l, liquid bio preservative
proclin 300 1~10 g/l.
In a preferred embodiment of the present invention, the content of each component of described complex stabilizer is: tris 6.06g/l,
Edta-2na 50 g/l, trehalose 25 g/l, beta-schardinger dextrin -20 g/l, glycerol 10 g/l, Polyvinylpyrrolidone 20
g/l 、proclin300 5 g/l.
In a preferred embodiment of the present invention, described complex stabilizer is applied in lactic dehydrogenase enzyme reagent kit, described
Lactic dehydrogenase enzyme reagent kit includes the first reagent and the second reagent, and the group of described first reagent is divided into: disodium hydrogen phosphate 25.069
G/l, potassium dihydrogen phosphate 2.435 g/l, α -one butanoic acid sodium salt 0.496 g/l, the group of described second reagent is divided into: 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 usage amount of described complex stabilizer is in lactic dehydrogenase enzyme reagent kit
Volume fraction is 5%.
The invention has the beneficial effects as follows: the complex stabilizer for lactic dehydrogenase enzyme reagent kit of the present invention, described compound
Stabilizer is made up of buffer agent, chelating agen, saccharide, surfactant, organic solvent, preservative.The complex stabilizer of the present invention
After being added in determination of lactate dehydrogenase test kit, it is remarkably improved the stability of testing result, effectively extend lactic acid dehydrogenase
Measure the holding time of test kit.In addition, this complex stabilizer is with low cost, prepare simple.
Specific embodiment
The enforcement it is clear that described will be clearly and completely described to the technical scheme in the embodiment of the present invention below
Example is only a part of embodiment of the present invention, rather than whole embodiments.Based on the embodiment in the present invention, this area is common
All other embodiment that technical staff is obtained under the premise of not making creative work, broadly falls into the model of present invention protection
Enclose.
Instrument according to the present invention has: (Beijing LabTech instrument is limited for uv blue star ultraviolet spectrophotometer 1
Company, 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 complex stabilizer for lactic dehydrogenase enzyme reagent kit, the content of each component of described complex stabilizer
For: tris 6.06 g/l, edta-2na 50 g/l, trehalose 25 g/l, beta-schardinger dextrin -20 g/l, glycerol 10 g/l, poly-
Vinylpyrrolidone 20 g/l, proclin300 5 g/l.After above-mentioned each component is fully dissolved, mixes, you can be obtained one
Plant liquid composite stabilizing agent finished product.
Embodiment 1 is applied to determination of lactate dehydrogenase test kit, and carries out in terms of stability, accuracy and usage amount
Evaluate.
(1) stability and accuracy test
1st, for examination stabilizer: complex stabilizer described in embodiment 1.
2nd, for trying test kit for determination of lactate dehydrogenase test kit, it is made up of reagent 1 and reagent 2.Component is as follows:
Reagent 1: disodium hydrogen phosphate 25.069 g/l, potassium dihydrogen phosphate 2.435 g/l, α -one butanoic 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.
3rd, test method and result
The complex stabilizer that embodiment 1 is prepared is by 3%(v/v) amount add (reagent in determination of lactate dehydrogenase test kit
1 and reagent 2 all by this amount add), measure respectively in sample with the determination of lactate dehydrogenase test kit without complex stabilizer
Dehydrogenase Content.This test is with the horizontal 2(Quality Control 2 of Britain's Landau quality controlled serum) and quality controlled serum horizontal 3(Quality Control 3) as sample
This, detected using ultraviolet-uisible spectrophotometer, if skip test, (skip test is to distill for two test samples
Water replaces sample).The ratio of sample and reagent 1 and reagent 2 be 20 μ l:1000 μ l:250 μ l, the dominant wavelength setting as 340nm,
The a length of 405nm of complementary wave, postpones 1min reading after sample and reagent mixing, the reading duration is 1.5min, draws every point of △ a/min(
Clock absorbance change), according to formula ldh(u/l)=△ a/min × f calculate sample ldh content (calculate 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 it is allowed to deviation range ± 10%.
According to above assay method, using same batch with bottled quality controlled serum 2, quality controlled serum 3 as detection sample, difference
With add 3%(v/v) complex stabilizer determination of lactate dehydrogenase test kit and without complex stabilizer lactic acid dehydrogenase survey
Determine test kit to be detected, test in every 5 days once, do every time three parallel and take its meansigma methods, detection cycle 35 days, obtain pin
Test result to quality controlled serum 2, quality controlled serum 3, is calculated standard deviation (sd) and the effect duration natural law of measurement result.Mark
Quasi- deviation this can reflect the stability of measurement result.Result of the test see table.
From upper table data, do not add the determination of lactate dehydrogenase test kit of complex stabilizer, its testing result standard
Deviation is significantly greater, about adds 4 times of test kit testing result standard deviation of 3% complex stabilizer, illustrates not add multiple
The determination of lactate dehydrogenase test kit closing stabilizer is very unstable, and effect duration natural law can only maintain 15 days about, relatively, adds
The determination of lactate dehydrogenase test kit of 3% complex stabilizer in whole test period, accuracy all in the range of tolerance,
Illustrate to remain to keep excellent test performance in 35 days about this mensure test kit.As can be seen here, with the addition of 3% complex stabilizer
Determination of lactate dehydrogenase test kit show more excellent in terms of measuring stability and accuracy, complex stabilizer has bright to it
Aobvious stablizing effect, can effectively extend the pot-life of determination of lactate dehydrogenase test kit.
(2) usage amount contrast test
1st, for examination stabilizer: complex stabilizer described in embodiment 1.
2nd, for agent of having a try for determination of lactate dehydrogenase test kit, composition is with reference to stability and accuracy test.
3rd, test method and result
By complex stabilizer according to 1%(v/v), 5%(v/v) ratio be separately added in determination of lactate dehydrogenase test kit
(reagent 1 and reagent 2 are all added by same amount), carries out the mensure of Dehydrogenase Content in sample.By stability of the present invention examination
The assay method tested measures, and is calculated standard deviation and the effect duration natural law of measurement result, standard deviation this can reflect survey
Determine the stability of result.Result see table.
From upper table data, three kinds of different additions of complex stabilizer are stablized to determination of lactate dehydrogenase test kit
Effect has all strengthened, and all can extend the pot-life of test kit to some extent.The test kit adding 1% complex stabilizer is made
With slightly weak, stability is not good enough；Add the kit standard deviation of 3% complex stabilizer placed in the middle, the stable composition of this addition is described
Agent effect can't reach optimum；The kit standard deviation of 5% complex stabilizer addition is minimum, and respectively 12 and 17, explanation
Add the determination of lactate dehydrogenase test kit repeatability of 5% complex stabilizer most preferably, this test kit is more stable, opens in test kit
Good test performance can also be kept within 35 days about after bottle.Therefore, 5% can be set to complex stabilizer in determination of lactate dehydrogenase examination
Optimum quantum of utilization in agent box.
Can obtain from data above and analysis result, determination of lactate dehydrogenase test kit adds of the present invention compound steady
The holding time of test kit after determining agent, can be extended, measurement result stability is more preferable, and the most suitable addition volume ratio is about 5%.Separately
Outward, this complex stabilizer is with low cost, prepares simple, is therefore worth further genralrlization to use.
The foregoing is only embodiments of the invention, not thereby limit the present invention the scope of the claims, every using this
Equivalent structure or equivalent flow conversion that bright description is made, or directly or indirectly it is used in the technology neck of other correlations
Domain, is included within the scope of the present invention.
Claims (4)
1. a kind of complex stabilizer for lactic dehydrogenase enzyme reagent kit is it is characterised in that described complex stabilizer is by for three hydroxyls
Aminomethane, disodiumedetate, trehalose, beta-schardinger dextrin-, glycerol, Polyvinylpyrrolidone, liquid bio are prevented
Rotten agent proclin 300 forms, and the content of each component is: trishydroxymethylaminomethane (tris) 3 ~ 9 g/l, ethylenediamine tetrem
Acid disodium (edta-2na) 40 ~ 60 g/l, trehalose 20 ~ 30 g/l, beta-schardinger dextrin -15 ~ 25 g/l, glycerol 5 ~ 15 g/l,
Polyvinylpyrrolidone 15 ~ 25 g/l, liquid bio preservative proclin 300 1 ~ 10 g/l.
2. complex stabilizer according to claim 1 is it is characterised in that the content of each component of described complex stabilizer is:
Tris 6.06 g/l, edta-2na 50 g/l, trehalose 25 g/l, beta-schardinger dextrin -20 g/l, glycerol 10 g/l, poly- second
Alkene pyrrolidone 20 g/l, proclin300 5 g/l.
3. complex stabilizer according to claim 1 is it is characterised in that described complex stabilizer is applied to lactic acid dehydrogenase
In test kit, described lactic dehydrogenase enzyme reagent kit includes the first reagent and the second reagent, and the group of described first reagent is divided into: phosphoric acid
Disodium hydrogen 25.069 g/l, potassium dihydrogen phosphate 2.435 g/l, α -one butanoic acid sodium salt 0.496 g/l, the component of described second reagent
For: tris 8.479 g/l, nadh 0.0851 g/l, hydrochloric acid 5.6 ml/l.
4. complex stabilizer according to claim 1 is it is characterised in that the usage amount of described complex stabilizer takes off in lactic acid
Volume fraction in hydrogen enzyme reagent kit 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 |
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| CN104388531A CN104388531A (en) | 2015-03-04 |
| CN104388531B true CN104388531B (en) | 2017-01-18 |
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Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE19508192A1 (en) * | 1995-03-09 | 1996-09-12 | Behringwerke Ag | Stable transglutaminase preparations and process for their preparation |
| AUPN200695A0 (en) * | 1995-03-28 | 1995-04-27 | Trace Scientific Pty Ltd | Transaminase reagent |
| CN100434515C (en) * | 2005-12-23 | 2008-11-19 | 上海复星长征医学科学有限公司 | Enzyme combining stabilizer |
| CN102507915B (en) * | 2011-11-07 | 2014-01-01 | 宁波美康生物科技股份有限公司 | Stable liquid kit for measuring lactic acid |
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