CN104406835A - Indigo carmine colorant as well as preparation method and applications thereof - Google Patents
Indigo carmine colorant as well as preparation method and applications thereof Download PDFInfo
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- CN104406835A CN104406835A CN201410696749.4A CN201410696749A CN104406835A CN 104406835 A CN104406835 A CN 104406835A CN 201410696749 A CN201410696749 A CN 201410696749A CN 104406835 A CN104406835 A CN 104406835A
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Abstract
The invention discloses an indigo carmine colorant as well as a preparation method and applications thereof. A mixed solution of indigo carmine and antioxidants contains 0.1%-1%(weight/volume percent concentration, g/100ml) of indigo carmine, the antioxidants are selected from one or combinations of ascorbic acid, tea polyphenols and phytic acid, and the amount of addition is 0.05%-0.5%(weight/volume percent concentration, g/100ml). The indigo carmine colorant has advantages as follows: a product is greatly improved in stability, is easy to use and store, and has a good clinical effect; the safety performance is high, and the clinical risk is low; the operation is simple. A prepared indigo carmine solution can be directly used in clinical treatment, dilution and other additional operations are not required, a clinical operation process is greatly simplified, and defects of a colorant obtained with an immediate preparation and use method in the prior art are solved.
Description
Technical field
The present invention relates to a kind of gastrointestinal mucosal coloring agent and compound method thereof, particularly relate to a kind of Indigo Carmine Staining agent that can preserve for a long time and compound method thereof, and establish the detection method of Indigo Carmine Staining agent concentration.
Background technology
Cancer of the stomach is the disease of serious harm human health, and its mortality ratio is only second to the second that lung cancer occupies global cancer mortality, and the whole world every year newly-increased number of patients is about 876000 people, and the number dying from cancer of the stomach is every year about 649000 people.According to the 3rd cause of the death Retrospect spot-check in the whole nation, China's cancer of the stomach population standardized death rate is 16.2 people/100,000 people, account for 18.2% of the malignant tumour cause of the death, China cancer of the stomach world standard mortality ratio male sex is 30.8 people/100,000 people, women is 13.8 people/100,000 people, all higher than developed country and developing country's average level.By stages relevant directly with when diagnosing of the prognosis of cancer of the stomach, early carcinoma of stomach postoperative 5 years survival rate >90%, and postoperative 5 years survival rates of advanced gastric carcinoma are only about 15%.Therefore, the key reducing gastric cancer mortality is early diagnosis, early treatment.At present, no matter the prefered method in the diagnosing gastric cancer link that gastrocopy is still generally acknowledged for the whole world, be early carcinoma of stomach or advanced gastric carcinoma, all can carry out pathological examination through stomach Microscopic observation and biopsy tissue and then clarify a diagnosis.But, a large amount of example show cancer of the stomach especially early carcinoma of stomach in gastrocopy, still there is a large amount of failing to pinpoint a disease in diagnosis, research display, operating gastroscope experience less than 10 years persons can reach 25% to cancer of the stomach rate of missed diagnosis, and within more than 10 years, person still reaches 10%-20%.Therefore, correctly find in time with gastroscope and diagnose early carcinoma of stomach to be try hard to pursue caused by gastroscopist to ask always.
At present, clinically for magnifying endoscope, Narrow-Band Imaging art etc. that the method for scope auxiliary diagnosis has traditional Endoscopy and developed recently to get up, due to the use of these technology, the recall rate of cancer of the stomach and precancerous lesion obtains effective raising.Endoscopic image, also known as microendoscopic, can be amplified tens times of even hundreds of times by magnifying endoscope, thus can be used to observe the early stage minute lesion of mucous membrane.Narrow-Band Imaging scope is the broadband spectral in utilize filter filters to fall red bluish-green light wave that speculum illuminator sends, only leaves narrow-band spectrum, thus clearly illustrates the capillary on mucous membrane top layer and a kind of endoscopy technology of mucous membrane surface Pit pattern.The technical equipment such as magnifying endoscope, Narrow-Band Imaging scope require high, and only concentrate on minority large hospital in China at present, and a large amount of endoscopy work is born by vast basic hospital, workload is very large.Endoscopy is another method for auxiliary diagnosis stomach trouble, there is the simple advantage of application, because Endoscopy does not need specific installation, only need coloring agent (as Indigo Carmine Staining agent) just can play the object of auxiliary diagnosis as supplementary means, therefore, can be carried out this in common Endoscopic medical institutions to check, more practical.
Endoscopy refers on the basis of conventional endoscopic inspection, and auxiliary use pigment preparation, increases the contrast of pathology and normal structure, make the form of focus, scope more clear, thus improve naked eyes recognition capability, and guiding biopsy, improves detection rate of pathological change.Indicarminums etc. are often applied to the detection of early carcinoma of stomach and precancerous lesion alone or in combination.Indigo Carmine Staining agent is contrast stain, be deposited on duplicature, do not absorbed, also mucous membrane of getting along well combines, sharp contrast is formed with normal mucosa color and luster, and the fine irregularities that can demonstrate mucous membrane changes and spatial structure, thus get pathological tissues target target is provided for pincers under scope, improve the recall rate of pathology.
But, the coloring agent that Indigo Carmine Staining agent is commonly used as Endoscopy, due to the instability of itself, can there is following reaction in indicarminum solution in atmosphere:
Indigo Carmine Staining agent originally as blue or dark blue liquid, but under contacting with the oxygen in light and air, will be gone bad in a few days gradually, loses original blueness, loses dyeing function.
Therefore, the method that domestic hospitals generally adopts " now with the current ", namely before use by indicarminum dissolution of solid among the water for injection of prescribed volume, make the Indigo Carmine Staining agent of certain concentration, after having prepared, can clinical detection be directly used in.But there are the following problems for the method: first, because the Indigo Carmine Staining agent of " now with the current " method preparation is not through any sterilizing, product is caused to there is larger clinical risk; Secondly, the concentration of the Indigo Carmine Staining agent of " now with the current " method preparation has uncertainty, because indicarminum solid also can react with oxygen when preserving, causes part indicarminum to lose efficacy, the concentration of indicarminum has uncertainty, even can affect clinical practice; Again, the operation of " now with the current " method is more loaded down with trivial details, is unfavorable for the popularization of Endoscopy technology.
For the shortcoming that above-mentioned " now with the current " method exists, be badly in need of clinically a kind ofly can preserving for a long time, Indigo Carmine Staining agent that stable performance, security are higher.
Summary of the invention
The object of this invention is to provide a kind ofly can to preserve for a long time, Indigo Carmine Staining agent that stable performance, security are higher.
Indigo Carmine Staining agent provided by the present invention, for the mixed solution of indicarminum and antioxidant, wherein containing indicarminum 0.1%-1% (mass/volume percent concentration, g/100ml), the combination of one or more in ascorbic acid, Tea Polyphenols and phytic acid selected by antioxidant, addition is 0.05%-0.5% (mass/volume percent concentration, g/100ml).
Wherein: in mixed solution, antioxidant is ascorbic acid or Tea Polyphenols, concentration is 0.05%-0.5%; Or
In mixed solution, antioxidant is phytic acid, and concentration is 0.1%; Or
In mixed solution, antioxidant is that ascorbic acid and Tea Polyphenols combine by the equivalent of quality; In mixed solution, ascorbic acid concentrations is 0.1%, and Tea Polyphenols concentration is 0.1%; Or
In mixed solution, antioxidant is that ascorbic acid, Tea Polyphenols and phytic acid combine by the equivalent of quality; In mixed solution, ascorbic acid concentrations is 0.1%, and Tea Polyphenols concentration is 0.1%, and phytic acid concentration is 0.1%.
Described Indigo Carmine Staining agent, for being selected from any one of subordinate:
Assembly 1: indicarminum: 0.35g, ascorbic acid: 0.1g, distilled water: 100mL;
Assembly 2: indicarminum: 0.35g, Tea Polyphenols: 0.1g, distilled water: 100mL;
Assembly 3: indicarminum: 0.35g, phytic acid: 0.1g, distilled water: 100mL;
Assembly 4: indicarminum: 0.35g, ascorbic acid: 0.1g, Tea Polyphenols: 0.1g, distilled water: 100mL;
Assembly 5: indicarminum: 0.35g, ascorbic acid: 0.1g, Tea Polyphenols: 0.1g, phytic acid: 0.1g, distilled water: 100mL.
Another object of the present invention is to provide a kind of method can preparing the agent of actual concentrations Indigo Carmine Staining.The method comprises the following steps:
1) calculate also correspondence by concentration values and take indicarminum powder and antioxidant, be dissolved in water for injection (as distilled water), make the combination solution of indicarminum and stabilizing agent;
2) combination solution is carried out moist heat sterilization, sterilising conditions is 121 DEG C, 30 minutes, obtains Indigo Carmine Staining agent.
A present invention also object is to provide the method for the indicarminum solution actual concentrations of a kind of UV-Visible method mensuration said method preparation.The method comprises the following steps:
1) Criterion curve: prepare certain density indicarminum standard solution, reference is made with distilled water, absorbance is measured at 610nm wavelength place, be horizontal ordinate with the concentration of indicarminum standard solution, take absorbance as ordinate drawing standard curve, calculate typical curve regression equation by least square method;
2) indicarminum solution concentration measures: by Indigo Carmine Staining dilution agent, to certain multiple, (multiple is 50-200, extension rate is determined according to the range of ultraviolet light/visible ray scene photometer), reference is made equally with distilled water, measure absorbance at 610nm wavelength place, go out the concentration of indicarminum in solution according to typical curve regression equation calculation.
Indigo Carmine Staining agent provided by the present invention and compound method thereof, have the following advantages:
1, product stability significantly improves, and is easy to use and preserve, and clinical effectiveness is good.Indigo Carmine Staining agent of the present invention not only concentration is accurate, and can preserve for a long time at normal temperatures (storage life can reach 12-24 month), overcomes the shortcoming that indicarminum solution not easily stores.Indigo Carmine Staining agent of the present invention adopts overall preparation due to it, unified filling, and add a certain proportion of stabilizing agent (mainly comprising one or more antioxidants), stabilizing agent can ensure that product is not easily oxidized, thus the stability of Indigo Carmine Staining agent product is significantly improved, the concentration change of indicarminum within the long period (12-24 month) is less, and can meet clinical needs, Clinical practice effect is guaranteed.
2, security performance is high, and clinical risk is little.The compound method of Indigo Carmine Staining agent of the present invention, its all operations all completes hundred grades of aseptic indoor, used tool container is all through 250 DEG C of xeothermic bakings, preparation gained solution is through wet-hot steam sterilizing, ensure to greatest extent final products aseptic, without thermal source, substantially increase the biological safety of Indigo Carmine Staining agent, reduce clinical operation risk.
3, simple to operate.The indicarminum solution prepared can directly use clinically, without the need to operation bidirectionals such as dilutions, enormously simplify clinical operation process.
In sum, according to the Indigo Carmine Staining agent of Indigo Carmine Staining agent prescription provided by the invention and compound method gained, substantially can solve the various defects that " now with the current " method gained coloring agent at present exists, have larger application prospect.
Below in conjunction with specific embodiment, the present invention is described in further details.
Embodiment
Will there is oxidation reaction in Indigo Carmine Staining agent, cause coloring agent to lose efficacy in atmosphere, therefore, clinical in the urgent need to a kind ofly can to preserve for a long time, the higher Indigo Carmine Staining agent product of stable performance, security.The method of usual prolongation indicarminum storage life has: low temperature (temperature is at 0-5 DEG C), filling blanket gas (as used argon replaces to fall air in the exterior and interior packing of indicarminum) and interpolation stabilizing agent (as sodium formaldehyde sulfoxylate etc.).But the method for low tempertaure storage can increase indicarminum undoubtedly and produce, stores and the cost of transport, adds the expenditure of patient; Fill the method for blanket gas and can solve product storage and the high problem of transportation cost, but owing to adding gas shield device, also have higher requirement in packaging, production cost is significantly increased; Add the method for stabilizing agent and be undoubtedly one most economical in these three kinds of methods, having does not need increase equipment, does not have particular/special requirement to storage and transport, to the requirement of the product external packaging also advantage such as lower.Consider and cause the principal element of Indigo Carmine Staining agent inefficacy or sex change to be oxidation reaction, prioritizing selection antioxidant of the present invention is as the stabilizing agent of indicarminum.Existingly in indicarminum solution, add stabilizing agent sodium formaldehyde sulfoxylate, addition manner is interpolation 0.05% sodium formaldehyde sulfoxylate and 0.1% sodium citrate (buffering agent), and regulate pH value to 6-9, its Problems existing is that sodium formaldehyde sulfoxylate toxicity is larger.
The present invention is by the further investigation to oxygenant and indicarminum, indicarminum was lost efficacy or the mechanism of action of sex change factor and all kinds of stabilizing agent, optimize its assembly finally choose ascorbic acid, Tea Polyphenols, phytic acid, sodium pyrosulfite and sodium bisulfite alternatively antioxidant and indicarminum combine, finally determine the Indigo Carmine Staining agent product that stable performance, security are higher.
Oxygenant under study for action, is divided into according to the mechanism of action of antioxidant by inventor: 1) free radical scavenger, and benzene Polyphenols antioxidant (as VE, BHT, BHA, Tea Polyphenols etc.), the anti-sealing compound of sterols etc. belong to this kind of antioxidant; 2) oxygenant hardening agent scavenger, as citric acid, EDTA, phytic acid, polyphosphate, lecithin etc. belong to this kind of antioxidant; 3) oxygen agent is bitten, as Vitamin C acid, sulfurous acids, thio derivative.Based on this, in the selection of antioxidant, inventor considers the following aspects from the angle with indicarminum assembly: 1) field of employment: because indicarminum solution is used for in-vivo diagnostic, the therefore water miscible antioxidant of the better natural class of prioritizing selection security; 2) because system is that (concentration is 0.35% indicarminum solution to acid system, pH value is generally 2-3), product also needs through moist heat sterilization (121 DEG C, 30 minutes), therefore, selected antioxidant will in acid condition can stable existence, and thermal stability is also wanted better; 3) because the distortion of indicarminum solution is the coefficient result of multiple oxidation mechanism, therefore need to choose the different antioxidant of Antioxidation Mechanism; 4) consider to there is synergy between multiple antioxidant, inventor also studied the combination of two or more antioxidant.
The Indigo Carmine Staining agent of the present invention determined is aqueous solution, wherein contain: indicarminum 0.1%-1% (mass/volume percent concentration, g/100ml), antioxidant addition is 0.05%-0.5% (mass/volume percent concentration, g/100ml), antioxidant is be selected from any one in ascorbic acid, Tea Polyphenols and phytic acid, or wherein two or more with the combination of equivalent part (by quality) proportioning.
The invention provides a kind of method can preparing the agent of actual concentrations Indigo Carmine Staining, is that coordinate a certain proportion of antioxidant, compound method mainly comprises the following steps with food-grade indicarminum for principal ingredient:
1) take indicarminum powder and the antioxidant of ormal weight by formula, be dissolved in water for injection (as distilled water), make indicarminum solution;
2) indicarminum solution is carried out moist heat sterilization, sterilising conditions is 121 DEG C, 30 minutes, obtains Indigo Carmine Staining agent.
Present invention also offers the method for the indicarminum solution actual concentrations of a kind of UV-Visible method mensuration said method preparation, concrete grammar comprises the following steps:
1) Criterion curve: prepare certain density indicarminum standard solution, reference is made with distilled water, absorbance is measured at 610nm wavelength place, be horizontal ordinate with the concentration of indicarminum standard solution, take absorbance as ordinate drawing standard curve, calculate typical curve regression equation by least square method;
2) indicarminum solution concentration measures: by Indigo Carmine Staining dilution agent, to certain multiple, (multiple is 50-200, extension rate is determined according to the range of ultraviolet light/visible ray scene photometer), reference is made equally with distilled water, measure absorbance at 610nm wavelength place, go out indicarminum solution concentration according to typical curve regression equation calculation.
Described percent concentration is mass/volume (W/V, unit g100/mL) percent concentration if no special instructions.
Embodiment is implemented under premised on technical solution of the present invention, gives detailed embodiment and concrete operating process, and embodiment will contribute to understanding the present invention, but protection scope of the present invention is not limited to following embodiment.
The present invention's experiment is divided into three groups, and first group is the preferred of antioxidant kind; Second group is the selection of selected antioxidant consumption; Investigate the synergy of polycomponent antioxidant for 3rd group.
By the operating specification of Endoscopy coloring agent, in coloring agent solution, indicarminum can working concentration be 0.1%-1% (mass/volume percent concentration, g/100ml).In the present invention in Indigo Carmine Staining agent during indicarminum concentration change, its stability is unaffected, therefore following all embodiments all select indicarminum concentration to be 0.35% as experimental subjects.
The stability of test 1, detection Indigo Carmine Staining agent of the present invention
Embodiment group primary antibodie oxidant species preferred
The formula of embodiment 1-5 and the agent of reference examples Indigo Carmine Staining is as shown in table 1:
The formula (" % " unit is g/100ml) of table 1 embodiment 1-5 and the agent of reference examples Indigo Carmine Staining
The method of preparation embodiment and the agent of reference examples Indigo Carmine Staining, comprises the following steps:
1) take indicarminum powder and the antioxidant of ormal weight by formula, be dissolved in water for injection (as distilled water), make indicarminum solution;
2) indicarminum solution is carried out moist heat sterilization, sterilising conditions is 121 DEG C, 30 minutes, obtains Indigo Carmine Staining agent.
Above all operations all completes hundred grades of aseptic indoor, and used tool container is all through 250 DEG C of xeothermic bakings.
The stability test of indicarminum solution of the present invention embodies with indicarminum concentration style, requires that the concentration of indicarminum solution must not decline more than 20% in the limited period of product.
In the present invention, Detection of Stability method is: preserved by the Indigo Carmine Staining agent normal temperature of the embodiment of the present invention and reference examples, and measure its residual concentration in section 1 month, 2 months, 3 months, 6 months, 9 months, 12 months and 24 months respectively.
The Detection of Stability result of control group Indigo Carmine Staining agent is as shown in table 2, analyze above-mentioned data known, when not adding any antioxidant, the agent of reference substance Indigo Carmine Staining is difficult to long-time preservation at normal temperatures, almost complete oxidation in one month, product residual concentration levels off to zero.
The stability of table 2 reference examples Indigo Carmine Staining agent and security testing result (" % " unit is g/100ml)
| Cycle | Indicarminum residual concentration (%) |
| 0 month | 0.35 |
| 1 month | 0.08 |
| 2 months | 0 |
| 3 months | 0 |
| 6 months | 0 |
| 9 months | 0 |
| 12 months | 0 |
| 24 months | 0 |
The Detection of Stability result of embodiment 1-5 Indigo Carmine Staining agent is as shown in table 3, analysis data are known, according to the Indigo Carmine Staining agent of filling a prescription obtained shown in embodiment 1-5, in embodiment 1, embodiment 2 and embodiment 3, indicarminum residual concentration amplitude of variation in 24 months is no more than 20%, and antioxidant effect is ideal; In embodiment 4 and embodiment 5, indicarminum residual concentration oxidized degree after 1 month is higher, substantially can not reach stability requirement.
Conclusion: the preparation program of the embodiment of the present invention 1, embodiment 2, the agent of embodiment 3 Indigo Carmine Staining is feasible, and preferred anti-oxidant kind is ascorbic acid, Tea Polyphenols and phytic acid.
The stability of table 3 embodiment 1-5 Indigo Carmine Staining agent and security testing result
The selection of antioxidant consumption selected by embodiment group two
According to embodiment group one, preferred anti-oxidant is ascorbic acid, Tea Polyphenols and phytic acid, is test in 0.05%-0.5% (mass/volume percent concentration, g/100ml) scope at addition.
The formula of embodiment 11-28 and the agent of reference examples Indigo Carmine Staining is as shown in table 4:
Table 4 embodiment 11-28 Indigo Carmine Staining agent antioxidant consumption (" % " unit is g/100ml)
| Antioxidant | Addition (%) | |
| Embodiment 11 | Ascorbic acid | 0.05 |
| Embodiment 12 | Ascorbic acid | 0.1 |
| Embodiment 13 | Ascorbic acid | 0.2 |
| Embodiment 14 | Ascorbic acid | 0.3 |
| Embodiment 15 | Ascorbic acid | 0.4 |
| Embodiment 16 | Ascorbic acid | 0.5 |
| Embodiment 17 | Tea Polyphenols | 0.05 |
| Embodiment 18 | Tea Polyphenols | 0.1 |
| Embodiment 19 | Tea Polyphenols | 0.2 |
| Embodiment 20 | Tea Polyphenols | 0.3 |
| Embodiment 21 | Tea Polyphenols | 0.4 |
| Embodiment 22 | Tea Polyphenols | 0.5 |
| Embodiment 23 | Phytic acid | 0.05 |
| Embodiment 24 | Phytic acid | 0.1 |
| Embodiment 25 | Phytic acid | 0.2 |
| Embodiment 26 | Phytic acid | 0.3 |
| Embodiment 27 | Phytic acid | 0.4 |
| Embodiment 28 | Phytic acid | 0.5 |
Measure its residual concentration in section 1 month, 2 months, 3 months, 6 months, 9 months, 12 months and 24 months respectively, result is as shown in table 5:
Table 5: embodiment 11-28 measurement result
Analysis data are known, and all can reach satisfied antioxidant effect when antioxidants ascorbic acid and Tea Polyphenols addition are at 0.05%-0.5%, the oxidized degree of indicarminum is all not more than 20%; When phytic acid addition is 0.1%, effect is best, and 24 months internal oxidition degree were not more than all the other concentration of 20%, 0.05%-0.5% and substantially can reach satisfied result in 12 months.
Relatively can obtain, the preferred addition of oxygenant is 0.1%.
Embodiment group more than three kind of antioxidant conbined usage
The present embodiment selects optimum antioxidant addition 0.1% as experimental standard.
Embodiment 31-34 and contrast 1-3 proportioning as shown in table 6:
Table 6 embodiment 31-34 and contrast 1-3 proportioning (" % " unit is g/100ml)
Measure its residual concentration in section 1 month, 2 months, 3 months, 6 months, 9 months, 12 months and 24 months respectively, result is as shown in table 7:
Table 7 embodiment 31-34 and contrast 1-3 measurement result
Analysis data are known, and during polycomponent antioxidant conbined usage, to be all used alone effect than it better for antioxidant effect, especially fills a prescription as optimal case described in embodiment 31 and embodiment 34.
Experiment 2: the detection of the product of medical Indigo Carmine Staining agent
Detect according to the product standard of medical Indigo Carmine Staining agent, Indigo Carmine Staining agent product prepared by the present invention is almost identical with the physical and chemical index of matching while using Indigo Carmine Staining agent product on physical and chemical index, but also can overcome matching while using method and cause the uneven first-class shortcoming of product system by operator's operating influence.The indicarminum solution simultaneously adopting this method to prepare then shows higher biological safety on biological property.Because matching while using method all cannot detect most of Biological indicators, be therefore vulnerable to operating environment and the impact of operator's human factor, have bacterium etc. as product may be caused.And adopt the inventive method, the product that quality is homogeneous can be obtained, adopt moist heat sterilization then can ensure the aseptic of product, also be gratifying testing the result that can draw to other biological property of product, as: the bacterial endotoxin of product, cytotoxicity, sensitivity response, acute systemic toxicity and legacy toxicity etc. all meet the requirement (the standby indicarminum solution of matching while using legal system then cannot carry out the detection of these projects) of three class medical device product.This shows that the Indigo Carmine Staining agent adopting the method for embodiment to prepare all meets or exceeds matching while using method on physical and chemical index or in Biological indicators.
Claims (10)
1. an Indigo Carmine Staining agent, for the mixed solution of indicarminum and antioxidant, wherein containing indicarminum 0.1%-1% (mass/volume percent concentration, g/100ml), the combination of one or more in ascorbic acid, Tea Polyphenols and phytic acid selected by antioxidant, addition is 0.05%-0.5% (mass/volume percent concentration, g/100ml).
2. Indigo Carmine Staining agent according to claim 1, is characterized in that: in mixed solution, antioxidant is ascorbic acid or Tea Polyphenols, and concentration is 0.05%-0.5%.
3. Indigo Carmine Staining agent according to claim 1, is characterized in that: in mixed solution, antioxidant is phytic acid, and concentration is 0.1%.
4. Indigo Carmine Staining agent according to claim 1, is characterized in that: in mixed solution, antioxidant is that ascorbic acid and Tea Polyphenols combine by the equivalent of quality.
5. Indigo Carmine Staining agent according to claim 4, is characterized in that: in mixed solution, ascorbic acid concentrations is 0.1%, and Tea Polyphenols concentration is 0.1%.
6. Indigo Carmine Staining agent according to claim 1, is characterized in that: in mixed solution, antioxidant is that ascorbic acid, Tea Polyphenols and phytic acid combine by the equivalent of quality.
7. Indigo Carmine Staining agent according to claim 6, is characterized in that: in mixed solution, ascorbic acid concentrations is 0.1%, and Tea Polyphenols concentration is 0.1%, and phytic acid concentration is 0.1%.
8. Indigo Carmine Staining agent according to claim 1, is characterized in that, for being selected from any one of subordinate:
Assembly 1: indicarminum: 0.35g, ascorbic acid: 0.1g, distilled water: 100mL;
Assembly 2: indicarminum: 0.35g, Tea Polyphenols: 0.1g, distilled water: 100mL;
Assembly 3: indicarminum: 0.35g, phytic acid: 0.1g, distilled water: 100mL;
Assembly 4: indicarminum: 0.35g, ascorbic acid: 0.1g, Tea Polyphenols: 0.1g, distilled water: 100mL;
Assembly 5: indicarminum: 0.35g, ascorbic acid: 0.1g, Tea Polyphenols: 0.1g, phytic acid: 0.1g, distilled water: 100mL.
9. prepare a method for the arbitrary described Indigo Carmine Staining agent of claim 1 to 8, comprise the following steps:
1) calculate also correspondence by concentration values and take indicarminum powder and antioxidant, be dissolved in water for injection (as distilled water), make the combination solution of indicarminum and stabilizing agent;
2) combination solution is carried out moist heat sterilization, sterilising conditions is 121 DEG C, 30 minutes, obtains Indigo Carmine Staining agent.
10. measure a method for indicarminum concentration in Indigo Carmine Staining agent by UV-Visible method, comprise the following steps:
1) Criterion curve: prepare certain density indicarminum standard solution, reference is made with distilled water, absorbance is measured at 610nm wavelength place, be horizontal ordinate with the concentration of indicarminum standard solution, take absorbance as ordinate drawing standard curve, calculate typical curve regression equation by least square method;
2) indicarminum solution concentration measures: by Indigo Carmine Staining dilution agent, to certain multiple, (multiple is 50-200, extension rate is determined according to the range of ultraviolet light/visible ray scene photometer), reference is made equally with distilled water, measure absorbance at 610nm wavelength place, go out the concentration of indicarminum in solution according to typical curve regression equation calculation.
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| CN110567785A (en) * | 2019-08-28 | 2019-12-13 | 迈克生物股份有限公司 | Staining kit and method of use |
| CN111184876A (en) * | 2020-02-20 | 2020-05-22 | 沈伟 | Alimentary canal mucous membrane coloring agent, preparation method and application thereof |
| CN111184876B (en) * | 2020-02-20 | 2022-03-04 | 沈伟 | Alimentary canal mucous membrane coloring agent, preparation method and application thereof |
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