CN104390925A - Composite stabilizer for aspartate aminotransferase assay kit - Google Patents
Composite stabilizer for aspartate aminotransferase assay kit Download PDFInfo
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- CN104390925A CN104390925A CN201410553001.9A CN201410553001A CN104390925A CN 104390925 A CN104390925 A CN 104390925A CN 201410553001 A CN201410553001 A CN 201410553001A CN 104390925 A CN104390925 A CN 104390925A
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- aspartate aminotransferase
- kit
- compound stabilizer
- reagent
- trishydroxymethylaminomethane
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Abstract
The invention discloses a composite stabilizer for an aspartate aminotransferase assay kit. The composite stabilizer comprises trihydroxymethyl aminomethane, disodium ethylenediaminetetraacetate, trehalose, gelatin, propylene glycol, a surfactant Brij-35 and a liquid biological preservative Proclin300. Through the technical scheme, the composite stabilizer for the aspartate aminotransferase assay kit comprises the components of a buffer solution, a stabilizer, a chelating agent, an organic solvent, a surfactant and a preservative and the like. After the composite stabilizer is added into the aspartate aminotransferase assay kit in a certain volume ratio, not only the accuracy of an assay result can be ensured, but also the stability thereof is higher, and the expiry date after the aspartate aminotransferase assay kit is opened can be effectively prolonged.
Description
Technical field
The present invention relates to in-vitro diagnosis field, particularly relate to a kind of compound stabilizer measuring kit for aspartate aminotransferase.
Background technology
Aspartate aminotransferase (AST), is once called as glutamic-oxalacetic transaminease, is extensively present in cardiac muscle, skeletal muscle, liver, and the highest with the content in cardiac muscle, and liver takes second place.The mensuration of aspartate aminotransferase can reflect hepatocellular injury degree, its rising is common in liver diseases as catarrhal jaundice, myocardial infarction, dermatomyositis, muscle damage etc., very common liver function class clinical biochemical test item, significant to clinical diagnosis.
Detection in current serum mainly adopts continuous monitoring method, and concrete mensuration process is as follows:
。
Aspartic acid and α-ketoglutaric acid generate oxaloacetic acid under aspartate aminotransferase effect, oxaloacetic acid and NADH(reducibility coenzyme I) under malic dehydrogenase (MDH) effect, change into malic acid and NAD
+(oxidisability coenzyme I), and the interference eliminating sample endogenous pyruvic acid under lactic dehydrogenase (LDH) effect.NADH has specificity absorption peak at 340nm place, and its absorbance fall is directly proportional to aspartate aminotransferase content.There are three kinds of enzymes in visible mensuration in kit at aspartate aminotransferase, enzymatic activity stability directly affects stabilization of kit.The activity of enzyme is subject to various factors interference, as pH, temperature, ultraviolet, heavy metallic salt, inhibitor etc.In addition NADH is very easily oxidized, less stable, is unfavorable for preserving.So series of factors causes aspartate aminotransferase mensuration stabilization of kit poor, the effect phase is short, directly affects Clinical practice.
The method of current prolongation stabilization of kit and the term of validity substantially all realizes by adding stabilizing agent and changing storage requirement.Patent CN87100939 provides a kind of NADH stable composition, it comprises propylene glycol, boric acid and alkaline buffer, although said composition can keep the stability of independent NADH, can not be applied in the Detection reagent for aspartate aminotransferase box under neutral environment; Patent CN200510023147.3 provides a kind of stabilizing agent for improvement of rate method detection kit---dithiothreitol (DTT), but for be that the kit that do not break a seal adds the holding time after stabilizing agent, and and effective stationary phase after its Kaifeng undeclared; US Patent No. 5654288 discloses a kind of method utilizing liposome to live in NADH, but liposome manufacturing cost is higher.Visible existing patent is all the stability for independent NADH, and the more aspartate aminotransferase of not mentioned complicated component, enzyme class measures kit, therefore study a kind of prepare simple, with low cost, have the stabilizing agent of good stablizing effect to be very important to this kit.
Summary of the invention
The technical matters that the present invention mainly solves is to provide and a kind ofly measures the compound stabilizer of kit for aspartate aminotransferase, and the preparation of this compound stabilizer is simple, with low cost, have good stablizing effect.
For solving the problems of the technologies described above, the technical scheme that the present invention adopts is: provide a kind of compound stabilizer measuring kit for aspartate aminotransferase, described compound stabilizer comprises trishydroxymethylaminomethane, disodium ethylene diamine tetraacetate, trehalose, gelatin, propylene glycol, surfactant B rij-35, liquid bio antiseptic Proclin 300, the content of each component is: trishydroxymethylaminomethane 2 ~ 10 g/L, disodium ethylene diamine tetraacetate 20 ~ 30 g/L, trehalose 20 ~ 30 g/L, gelatin 15 ~ 25 g/L, propylene glycol 10 ~ 20 g/L, surfactant B rij-35 2 ~ 7 g/L, liquid bio antiseptic Proclin 300 0.5 ~ 2.0 g/L.
In a preferred embodiment of the present invention, the content of each component of described compound stabilizer is: trishydroxymethylaminomethane 6.057 g/L, disodium ethylene diamine tetraacetate 25 g/L, trehalose 25 g/L, gelatin 20 g/L, propylene glycol 15 g/L, Brij-35 5 g/L, Proclin 300 1 g/L.
In a preferred embodiment of the present invention, described compound stabilizer is applied to aspartate aminotransferase and measures in kit, described aspartate aminotransferase measures kit and comprises the first reagent and the second reagent, the component of described first reagent is: trishydroxymethylaminomethane 9.691 g/L, NaOH 0.935 g/L, aspartic acid 23.958 g/L, nicotinamide adenine dinucleotide 0.283 g/L, lactic dehydrogenase 1500 U/L, malic dehydrogenase 840 U/L, the component of described second reagent is: trishydroxymethylaminomethane 9.691 g/L, α-ketoglutaric acid 2.191 g/L.
In a preferred embodiment of the present invention, the volume fraction that the use amount of described compound stabilizer measures in kit at aspartate aminotransferase is 5%.
The invention has the beneficial effects as follows: the compound stabilizer measuring kit for aspartate aminotransferase of the present invention, this compound stabilizer constituent comprises damping fluid, stabilizing agent, sequestrant, organic solvent, surfactant and antiseptic etc.Compound stabilizer is joined according to a certain volume aspartate aminotransferase to measure after in kit, measurement result can not only guarantee accuracy, and stability is more excellent, effectively can extend the uncork effect phase that aspartate aminotransferase measures kit.In addition, this compound stabilizer is with low cost, and preparation is simple, is therefore worth further genralrlization to use.
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 and measure the compound stabilizer of kit for aspartate aminotransferase, the content of each component of described compound stabilizer is: trishydroxymethylaminomethane 6.057 g/L, disodium ethylene diamine tetraacetate 25 g/L, trehalose 25 g/L, gelatin 20 g/L, propylene glycol 15 g/L, Brij-35 5 g/L, Proclin 300 1 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 aspartate aminotransferase and measures kit, and evaluate from stability, accuracy and use amount aspect.
(1) stability and accuracy test
1, for examination stabilizing agent: compound stabilizer described in embodiment 1.
2, for have a try agent be aspartate aminotransferase measure kit, be made up of reagent 1 and reagent 2.Component is as follows:
Reagent 1: trishydroxymethylaminomethane 9.691 g/L, NaOH 0.935 g/L, aspartic acid 23.958 g/L, nicotinamide adenine dinucleotide 0.283 g/L, lactic dehydrogenase 1500 U/L, malic dehydrogenase 840 U/L;
Reagent 2: trishydroxymethylaminomethane 9.691 g/L, α-ketoglutaric acid 2.191 g/L.
3, test method and result
Compound stabilizer embodiment 1 prepared is by 5%(v/v) amount add aspartate aminotransferase and measure (reagent 1 and reagent 2 are all by this amount interpolation) in kit, measure kit with the aspartate aminotransferase not adding compound stabilizer and measure aspartate aminotransferase content in sample respectively.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 60 μ L:1000 μ L:500 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, draw the change of △ A/min(absorbance per minute), according to formula AST(U/L)=△ A/min × F calculates aspartate aminotransferase in sample (AST) content (calculated factor F=4180).In quality controlled serum 2, the target value of AST is 23U/L, and in quality controlled serum 3, the target value of AST is 97 U/L, permissible variation scope ± 10%.
According to above assay method, using with batch with bottled quality controlled serum 2, quality controlled serum 3 as detection sample, respectively with add 5%(v/v) compound stabilizer aspartate aminotransferase measure kit and do not add compound stabilizer aspartate aminotransferase mensuration kit detect, test in every 6 days once, do at every turn three parallel and get its mean value, sense cycle 42 days, obtain the test result for quality controlled serum 2, quality controlled serum 3, calculate standard deviation (SD) and the term of validity number of days of measurement result.Standard deviation this can reflect the stability of measurement result.Test findings sees the following form.
From upper table data, the aspartate aminotransferase not adding compound stabilizer measures kit, its testing result standard deviation is obviously larger, be about 2 times of the kit testing result standard deviation adding 5% compound stabilizer, illustrate that the aspartate aminotransferase not adding compound stabilizer measures kit very unstable, term of validity number of days can only maintain about 18 days, relatively, the aspartate aminotransferase adding 5% compound stabilizer measures kit in whole test period, accuracy is all within the scope of permissible variation, illustrate and still can keep excellent test performance at about 42 days these mensuration kits.As can be seen here, the aspartate aminotransferase that with the addition of 5% compound stabilizer measures kit and shows more excellent in measuring stability and accuracy, compound stabilizer has obvious stablizing effect to it, effectively can extend the pot-life that aspartate aminotransferase measures kit.
(2) use amount contrast test
1, for examination stabilizing agent: compound stabilizer described in embodiment 1.
2, be that aspartate aminotransferase measures kit for agent of having a try, composition is with reference to stability test.
3, test method and result
By compound stabilizer according to 3%(v/v), 10%(v/v) ratio add aspartate aminotransferase respectively and measure (reagent 1 and reagent 2 are all by same amount interpolation) in kit, carry out the mensuration of aspartate aminotransferase content in sample.Measure by the assay method of stability test of the present invention, calculate standard deviation and the term of validity 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, three kinds of different additions of compound stabilizer strengthen all to some extent to the stabilization that aspartate aminotransferase measures kit, effectively can extend the pot-life to 42 day of kit.The kit effect adding 10% compound stabilizer is slightly weak, and stability is not good enough; The kit standard deviation adding 3% compound stabilizer is placed in the middle, illustrates that the compound stabilizer effect of this addition can't reach optimum; The kit standard deviation of 5% compound stabilizer addition is minimum, be respectively 1.06 and 3.51, illustrate that the aspartate aminotransferase adding 5% compound stabilizer measures kit repeatability best, this kit is more stable, within about 42 days after kit uncork, can also keep good test performance.Therefore, 5% can be decided to be the optimum quantum of utilization of compound stabilizer in aspartate aminotransferase mensuration kit.
Can obtain from above data and analysis result, after aspartate aminotransferase mensuration kit adds compound stabilizer of the present invention, can extend the holding time of kit, measurement result stability is better, and the suitableeest volume ratio that adds is approximately 5%.In addition, this compound 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. one kind measures the compound stabilizer of kit for aspartate aminotransferase, it is characterized in that, described compound stabilizer comprises trishydroxymethylaminomethane, disodium ethylene diamine tetraacetate, trehalose, gelatin, propylene glycol, surfactant B rij-35, liquid bio antiseptic Proclin 300, the content of each component is: trishydroxymethylaminomethane 2 ~ 10 g/L, disodium ethylene diamine tetraacetate 20 ~ 30 g/L, trehalose 20 ~ 30 g/L, gelatin 15 ~ 25 g/L, propylene glycol 10 ~ 20 g/L, surfactant B rij-35 2 ~ 7 g/L, liquid bio antiseptic Proclin 300 0.5 ~ 2.0 g/L.
2. compound stabilizer according to claim 1, it is characterized in that, the content of each component of described compound stabilizer is: trishydroxymethylaminomethane 6.057 g/L, disodium ethylene diamine tetraacetate 25 g/L, trehalose 25 g/L, gelatin 20 g/L, propylene glycol 15 g/L, Brij-35 5 g/L, Proclin 300 1 g/L.
3. compound stabilizer according to claim 1, it is characterized in that, described compound stabilizer is applied to aspartate aminotransferase and measures in kit, described aspartate aminotransferase measures kit and comprises the first reagent and the second reagent, the component of described first reagent is: trishydroxymethylaminomethane 9.691 g/L, NaOH 0.935 g/L, aspartic acid 23.958 g/L, nicotinamide adenine dinucleotide 0.283 g/L, lactic dehydrogenase 1500 U/L, malic dehydrogenase 840 U/L, the component of described second reagent is: trishydroxymethylaminomethane 9.691 g/L, α-ketoglutaric acid 2.191 g/L.
4. compound stabilizer according to claim 1, is characterized in that, the volume fraction that the use amount of described compound stabilizer measures in kit at aspartate aminotransferase is 5%.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201410553001.9A CN104390925A (en) | 2014-10-20 | 2014-10-20 | Composite stabilizer for aspartate aminotransferase assay kit |
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| CN201410553001.9A CN104390925A (en) | 2014-10-20 | 2014-10-20 | Composite stabilizer for aspartate aminotransferase assay kit |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105974121A (en) * | 2016-04-18 | 2016-09-28 | 武汉云克隆科技股份有限公司 | Biological product stabilizer containing fish gelatin |
| CN106244671A (en) * | 2016-08-30 | 2016-12-21 | 山东博科生物产业有限公司 | A kind of stable aspartate aminotransferase checkout and diagnosis detection kit |
| CN106282308A (en) * | 2016-08-29 | 2017-01-04 | 湖南海源医疗科技有限公司 | A kind of in-vitro diagnosis biochemical reagents stabilizer |
| CN109724933A (en) * | 2018-12-30 | 2019-05-07 | 山东博科生物产业有限公司 | A kind of reproducible aspartate amino transferase detection kit |
Citations (4)
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| CN87100939A (en) * | 1986-02-24 | 1987-11-04 | 艾博特公司 | Stable dinucleotides based composition and method |
| CN1500884A (en) * | 2002-11-15 | 2004-06-02 | 江西特康科技有限公司 | Nicotinamide agent and preparation method thereof |
| CN102863495A (en) * | 2011-07-06 | 2013-01-09 | 上海执诚生物科技股份有限公司 | Stable composition containing NAD+ or NADH |
| CN103320498A (en) * | 2013-05-24 | 2013-09-25 | 宁波美康生物科技股份有限公司 | Detection reagent for aspartate aminotransferase |
-
2014
- 2014-10-20 CN CN201410553001.9A patent/CN104390925A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN87100939A (en) * | 1986-02-24 | 1987-11-04 | 艾博特公司 | Stable dinucleotides based composition and method |
| CN1500884A (en) * | 2002-11-15 | 2004-06-02 | 江西特康科技有限公司 | Nicotinamide agent and preparation method thereof |
| CN102863495A (en) * | 2011-07-06 | 2013-01-09 | 上海执诚生物科技股份有限公司 | Stable composition containing NAD+ or NADH |
| CN103320498A (en) * | 2013-05-24 | 2013-09-25 | 宁波美康生物科技股份有限公司 | Detection reagent for aspartate aminotransferase |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105974121A (en) * | 2016-04-18 | 2016-09-28 | 武汉云克隆科技股份有限公司 | Biological product stabilizer containing fish gelatin |
| CN105974121B (en) * | 2016-04-18 | 2018-02-06 | 武汉云克隆科技股份有限公司 | A kind of biological products stabilizer containing isinglass |
| CN106282308A (en) * | 2016-08-29 | 2017-01-04 | 湖南海源医疗科技有限公司 | A kind of in-vitro diagnosis biochemical reagents stabilizer |
| CN106244671A (en) * | 2016-08-30 | 2016-12-21 | 山东博科生物产业有限公司 | A kind of stable aspartate aminotransferase checkout and diagnosis detection kit |
| CN109724933A (en) * | 2018-12-30 | 2019-05-07 | 山东博科生物产业有限公司 | A kind of reproducible aspartate amino transferase detection kit |
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Application publication date: 20150304 |