CN1201907A - Chromotropic composition and its usage - Google Patents
Chromotropic composition and its usage Download PDFInfo
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- CN1201907A CN1201907A CN 97104461 CN97104461A CN1201907A CN 1201907 A CN1201907 A CN 1201907A CN 97104461 CN97104461 CN 97104461 CN 97104461 A CN97104461 A CN 97104461A CN 1201907 A CN1201907 A CN 1201907A
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Abstract
A composite allochromatic agent contains two kinds of organic indicator solution, one is chosen from the alcohol or diluted alkali solution bromcresol purple, metacresol purple, or paranitrophenol and the other is the alcohol or diluted alkali solution of neutral red. The composition can be effectively used in enzyme-linked immunosorbent diagnosis reagent box for allochromatic indication by adding serum to diluted specimen liquid. It can prevent the operation error of missing or repeating the operation of adding serum.
Description
The present invention relates to contain the chromotropic composition of two kinds of organic acids and base indicator, particularly relate to the chromotropic composition that can be used to finish enzyme-linked immunologic diagnosis kit sample liquid increase serum variable color demonstration.
The color change interval of known general monochromatic acid base indicator is in a 1-2 PH unit, in acid-base titration, select suitable acid base indicator can indicate strong acid titration highly basic, highly basic titration strong acid, strong acid titration weak base is with highly basic titration weak acid, but for the reaction of weak base titration weak acid, because sharpness index is had relatively high expectations, single acid base indicator can not reach actual requirement.Such as in the operation of enzyme-linked immunologic diagnosis kit, when extremely weakly alkaline clinical serum adds utmost point faintly acid sample diluting liquid, adopt single indicator can not obviously indicate change color, because sample liquid was adding clinical serum front and back in should operating, the variation of PH is at 0.05-0.1 unit.
But when the enzyme linked immunological kit clinical practice is operated, because the micro reaction plate plate hole is many, each serum application of sample amount is little, has only 5 microlitres, whether naked eyes can't intuitive judgment to add serum, so the mistake of leaking increase serum or repeating increase serum takes place in the clinical manipulation easily.If after making Trace Blood reset and add sample liquid change color is intuitively arranged, just is not easy during clinical manipulation to make mistakes again.
Purpose of the present invention just provides a kind of colour-changing agent of sensitive variable color, after its adding kit sample liquid, sample liquid becomes a kind of changeable colour sample liquid that can steady in a long-term deposit, the kit user can not be leaked the increase serum sample when operating with this sample liquid, also can not repeat to add blood serum sample.
Have now found that, two kinds of monochromatic acid base indicators are mixed by a certain percentage just can form the colour-changing agent that the small acidity of a kind of sensitive indication changes.
According to the present invention, comprise organic indicator (A) and organic indicator (B) in the above-mentioned chromotropic composition.Component (A) is (i) or (ii).(i) structural formula is:
R1 represents H in the formula, Na, K; R2 represents bromo or H; (ii) structural formula is as follows:
R3 represents H in the formula, Na, K; Component (B) is another kind of organic indicator, and its structural formula is:
Wherein R4 represents I or Cl; These two kinds of components are the common acid neutralization indicator, generally all have commercially available.
Component (A) is prepared with absolute ethyl alcohol or dilute alkaline soln, and it is 40-60% that ethanol is converted water concentration, is preferably 60%, and diluted alkaline concentration is 0.1M-1.0M, preferably uses 0.1M NaOH, and (A) component solid indicator content is 0.1-2%, and optimum content is 1.0%;
Component (B) is prepared with absolute alcohol or dilute alkaline soln, and adopting ethanol to convert water concentration is 40-60%, is preferably 60%, and diluted alkaline concentration is 0.1M-1.0M, is preferably 0.1M NaOH, and (B) component solid indicator content is 0.1-2%, and optimum content is 0.1%.
The present composition is pressed the certain volume preparation by the solution of above-mentioned (A) and solution (B), and VA/VB can be 5-20, and optimal proportion is VA/VB=10.
Directly add sample diluting liquid when colour-changing agent of the present invention uses and use in enzyme-linked immunologic diagnosis kit, matched proportion density is 2.0%-0.05%, and the best is 1%, and promptly per 100 ml sample liquid add 1 milliliter of colour-changing agent.
The present invention has narrated the composition of colour-changing agent, each component structure feature and at the composition proportion, and the compound method of this colour-changing agent and in the using method of enzyme-linked immunologic diagnosis kit.
The composition that the present invention proposes can be applicable to the variable color indication that acidity is changed to a 0.05-0.10 PH unit, is particularly useful in the increase serum variable color demonstration of sample liquid in the enzyme-linked immunologic diagnosis kit.Adopt the sample diluting liquid color matching that colour-changing agent of the present invention is mixed with stable, clinical manipulation adds micro-serum promptly change color directly perceived, and the mistake that anti-leak-stopping increase serum or repeat adds serum takes place.
Embodiment:
The following stated embodiment has illustrated the present invention.But the present invention is not limited only to following embodiment.
Accurately take by weighing bromcresol purple 1.000 gram solids, with 60 milliliters of anhydrous alcohol solutions, it is fixed molten to 100 milliliters to add pure water again, promptly get the indicator solution of (A) formula among the present invention, accurately take by weighing 0.100 gram dimethyl diaminophenazine chloride solid again, with 40 milliliters of anhydrous alcohol solutions, add pure water again and be settled to 100 milliliters, promptly get the indicator that meets (B) formula among the present invention.Get above-mentioned 10 milliliters of bromcresol purple indicator then and add 100 milliliters of above-mentioned dimethyl diaminophenazine chloride ethanol liquid indicator, fully mixing promptly gets color indicator of the present invention.
Prepare the enzyme linked immunological kit sample diluting liquid by following component: (pure water is fixed molten to 100 milliliters)
| Animal blood serum | ????20.0ml |
| Casein reagent | ????0.250g |
| Macrogol 6000 | ????1.5g |
| Tween-20 | ????0.10g |
| Triton×100 | ????0.2g |
| Sodium Mercurothiolate | ????0.05g |
| NaN3 | ????0.01g |
| 0.2M PBS (PH=7.20) buffer solution | ????10ml |
Can adapt to the slightly different sample diluting liquid of practical application acidity in order to observe colour-changing agent of the present invention, adopt several different animals serum to be mixed with five kinds of sample diluting liquids that acidity is slightly variant, be numbered 1., 2., 3., 4., 5., get each 100ml of above-mentioned five kinds of sample diluting liquids and add 1ml color indicator of the present invention, the result all gets one and is the rose translucent liquid, with these five kinds of variable color sample diluting liquid potential of hydrogen of acidity instrumentation, in 96 hole micro reaction plates, respectively add 100 these sample diluting liquids of μ l then, add clinical serum by numbering respectively, each hole adds 5 μ l, observe change color, the result is as follows for record:
| Numbering | ????① | ????② | ????③ | ????④ | ????⑤ |
| PH value before the increase serum | ???7.25 | ???7.28 | ???7.20 | ???7.15 | ???7.25 |
| PH value behind the increase serum | ???7.30 | ???7.32 | ???7.25 | ???7.20 | ???7.30 |
| Add the preceding color of 5 μ l serum | Rose-red | Rose-red | Rose-red | Rose-red | Rose-red |
| Add color behind the 5 μ l serum | Dirty-green | Dirty-green | Dirty-green | Dirty-green | Dirty-green |
| Micro sample adding appliance is directly squeezed into variable color with 5 μ l serum and is taken | 60-90 second | 30-60 second | 30-60 second | 30-90 second | 50-90 second |
| Micro sample adding appliance adds 5 μ l serum and uses micropipet 3 variable color times of pressure-vaccum | 1-3 second | 1-5 second | 0-3 second | 1-3 second | 1-3 second |
| Clap the variable color of shake micro reaction plate behind the adding serum gently and need the time | 1-2 second | 1-2 second | 1-2 second | 1-2 second | 1-2 second |
Claims (10)
1, a kind of chromotropic composition comprises organic indicator (A) and organic indicator (B), and component (A) is (i) or (ii), wherein the general formula of (i) is:
R1 represents H in the formula, K, Na; R2 represents bromo or H, (ii) for structural formula is
R3 represents H in the formula, K, Na; The structural formula of component (B) is:
Wherein R4 represents I or Cl.
2, according to the described composition of claim 1, the content that wherein contains the solid indicator in the component (A) is 0.10-2.0%, and optimum content is 1.0%.
3, according to the described composition of claim 1-2, wherein component (A) is prepared with absolute ethyl alcohol or dilute alkaline soln, and the concentration that absolute ethyl alcohol adds pure water is 40-60%, and the best is 60%, and diluted alkaline concentration is 0.1M-1.0M, and the best is 0.1MNaOH.
4, according to the described composition of claim 1, wherein to contain the content of solid indicator be 0.1-2.0% to component (B), and optimum content is 0.1%.
5, according to the described composition of claim 1～4, wherein component (B) is prepared with absolute ethyl alcohol or dilute alkaline soln, and it is 40-60% that ethanol is converted water concentration, and the best is 60%; Diluted alkaline concentration is 0.1M-1.0M, and the best is 0.1MNaOH.
6, according to the described composition of claim 1, the present invention presses the certain volume preparation by the solution of (A) and solution (B), and VA/VB can be 5～20, and optimal proportion VA/VB is 10.
7, being used for the variable color of elisa diagnostic kit sample diluting liquid increase serum according to the described any composition of claim 1～6 shows.
8, according to the described composition of claim 1～7, be used for enzyme connection diagnostic kit color-changing composition of the present invention and add 1 milliliter of color-changing composition of the present invention by 100～200 ml sample dilutions, wherein the best is that per 100 ml sample dilutions add 1 milliliter of color-changing composition of the present invention.
When 9, being used for sour ELISA diagnostic kit according to the described composition of claim 1～7, the pH value of sample diluting liquid can be 7.00-7.30, and wherein the best is 7.20.
10, according to the described composition of claim 1～8, the present invention is applicable to that any acidity is changed to the variable color requirement of a 0.05-0.10 PH unit.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 97104461 CN1091257C (en) | 1997-06-09 | 1997-06-09 | Chromotropic composition and its usage |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 97104461 CN1091257C (en) | 1997-06-09 | 1997-06-09 | Chromotropic composition and its usage |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1201907A true CN1201907A (en) | 1998-12-16 |
| CN1091257C CN1091257C (en) | 2002-09-18 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 97104461 Expired - Fee Related CN1091257C (en) | 1997-06-09 | 1997-06-09 | Chromotropic composition and its usage |
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| Country | Link |
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| CN (1) | CN1091257C (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107976540A (en) * | 2017-12-20 | 2018-05-01 | 山东省滨州畜牧兽医研究院 | Changeable colour ELISA kit sample diluting liquid |
-
1997
- 1997-06-09 CN CN 97104461 patent/CN1091257C/en not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107976540A (en) * | 2017-12-20 | 2018-05-01 | 山东省滨州畜牧兽医研究院 | Changeable colour ELISA kit sample diluting liquid |
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| Publication number | Publication date |
|---|---|
| CN1091257C (en) | 2002-09-18 |
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