CN107941727A - A kind of method using Erythrosin B as the Spectrophotometric Determination chitosan content of probe - Google Patents
A kind of method using Erythrosin B as the Spectrophotometric Determination chitosan content of probe Download PDFInfo
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Abstract
The present invention relates to a kind of method using Erythrosin B as the Spectrophotometric Determination chitosan content of probe, belong to big Molecular Detection field.The present invention utilizes chitosan under mildly acidic conditions, generation associated matter is reacted with Erythrosin B, measure its absorbance at 530nm with spectrophotometer has obvious reduction compared with Erythrosin B, and the decreasing value of absorbance is directly proportional to the concentration of chitosan, in this, as the quantitative basis of chitosan, the Fading spectrophotometry for measuring chitosan is established, this method measure is influenced by the molecular weight of chitosan, the advantages of it is cheap with reagent, high sensitivity favorable reproducibility, is adapted to promote the use of in practical applications.
Description
Technical field
The present invention relates to a kind of side using Erythrosin B as the Fading spectrophotometry Accurate Determining chitosan content of probe
Method, belongs to big Molecular Detection field.
Background technology
Chitosan (CTS) chemical name is Chitosan (1-4) -2- amino-B-D glucose, also known as deacetylated crust
Element, is made of 2- acetylaminohydroxyphenylarsonic acid 2- deoxidations-β-D-Glucose and 2- amino -2- deoxidations-β-two class monose of D-Glucose, is first
Shell class (shrimp, crab) animal, insect ectoskeleton main component, chitosan is widely distributed in nature, and it is green to belong to macromolecule
Color material.Recent years, with constantly studying, has there is brand-new understanding for chitosan people.Chitosan has control
Cholesterol, inhibit bacteria the functions such as activity, there is many applications in terms of medicine.In addition, chitosan is in field of food
There is outstanding meaning, it is acted on antibacterial, anti-oxidant, fresh-keeping etc..With being understood in depth to chitosan, chitosan also has
There is moisturizing, improve the effect such as skin immunity, therefore also have a wide range of applications in terms of cosmetics.As chitosan is being cured
It is more and more widely used in the every field such as medicine, foods and cosmetics, it is lived with people and health contacts also increasingly
Closely, therefore, it is particularly important that accurate quantitative analysis of chitosan becomes, the method measure chitosan content for establishing simple and effective have emphatically
The meaning wanted, this will be pushed further into application of the chitosan in every field.
Chitosan is that the chitin being widely present by nature is obtained by deacetylation.With safe and non-toxic spy
Property, thus there has been extensive use in food, cosmetics, medicine and other fields.At present, the method for measuring chitosan has spectrophotometric
Method, fluorescence method, resonance rayleigh light scattering method, electrochemical process, high-efficient liquid phase technique etc..In spectrophotometry, high preciousness etc. utilizes madder
The red specific chromogenic with chitosan under the conditions of certain acidity of element reacts, its compound light absorption value and chitosan content are linear
Relation, establishes a kind of spectrophotometry with high selectivity and highly sensitive easy quick measure chitosan, but it has
Have that stability is bad, specificity is poor, setting-out line scope is relatively narrow, Chitosan-phospholipid complex can not be distinguished etc. and lacked
Point.Although fluorescence method has the characteristics that strong antijamming capability, easy to operate, stability is high, fluorescence condition is required more tight
Lattice, otherwise can impact result.Resonance rayleigh light scattering method high sensitivity, but, selective also phase stringent to instrument requirements
To poor, it is difficult to applied in complex system.Since chitosan viscosity is larger in electrochemical process, have a great influence to its current potential, because
And limit the development of this respect.And high-efficient liquid phase technique cost is higher, economically practicality is relatively low.
Based on this, of the invention to provide a kind of easy, quick, new method of sensitivity higher, i.e., one kind is using Erythrosin B as spy
The Spectrophotometric Determination chitosan content of pin.
The content of the invention
It is difficult to quantify for the content of chitosan in finished product in the prior art to overcome, the cumbersome technology of operating technology is not
Foot, the present invention provide a kind of method using Erythrosin B as the Spectrophotometric Determination chitosan content of probe, present invention profit
With chitosan under mildly acidic conditions, combine to form relatively stable Ternary Complex with Erythrosin B and polyvinyl alcohol, make its
Absworption peak gradient in certain wavelength declines, and polyvinyl alcohol is used as stabilizer function wherein.With spectrophotometer at 530nm
Measure its absorbance has obvious reduction compared with Erythrosin B, and the decreasing value of absorbance is directly proportional to the concentration of chitosan, in this, as
The quantitative basis of chitosan, establishes the Fading spectrophotometry of measure chitosan, and this method measures the molecular weight by chitosan
Influence, the advantages of it is cheap with reagent, high sensitivity favorable reproducibility, be adapted to promote the use of in practical applications.
A kind of method using Erythrosin B as the Spectrophotometric Determination chitosan content of probe, it includes following steps
Suddenly：
1) standard curve of the chitosan concentration of △ A and different molecular weight is drawn:
The low molecular chitosan standard solution of 0.5mL a certain concentration gradients is added into 10mL colorimetric cylinders, it is dense to add 0.2mL
The sensitizer for 1%, the BR buffer solutions of 1.0mL, and 4.0mL concentration are spent for 1.0 × 10-4The Erythrosin B solution of mol/L,
Using fully being shaken up after distilled water constant volume, after on ultraviolet specrophotometer when standing 1 is small at room temperature, taken off in the maximum of system
It is reference measurement absorbance that color wavelength 530nm, which sentences water,；Wherein reagent blank is denoted as A0, the solution of chitosan-containing is denoted as A, and counts
Calculate △ A=A0- A, establishes the standard curve C1 of △ A and low molecular chitosan concentration；Replaced using middle-molecular-weihydroxyethyl chitosan solution
Low molecular chitosan solution, establishes the standard curve C2 of △ A and middle molecular chitosan concentration；Use high molecular weight chitosan solution
Low molecular chitosan solution is replaced, establishes the standard curve C3 of △ A and polymer chitosan concentration；
Under optimal experiment condition, measurement result of the system to the chitosan of different molecular weight is investigated.Investigate respectively
The chitosan of low molecular weight, middle-molecular-weihydroxyethyl and high molecular weight.By statistical analysis, the chitosan result of different molecular weight is deposited
In the significance difference opposite sex, this method measure chitosan content is influenced by different molecular weight.
2) preparation of the sample working solution of 10 μ g/mL：The capsule shells of a certain amount of detected sample are weighed, it is molten using glacial acetic acid
Solve and its constant volume is obtained into stock sample solution, filter storing solution with funnel absorbent cotton, filtrate is through 6000r/min, centrifuge
20min, taking supernatant 2.5mL, constant volume, obtains the sample working solution that concentration is 10 μ g/mL in 100mL volumetric flasks；
3) according to chitosan molecule amount selection criteria curve：△ A are drawn using step 1) kind detection method and sample works
The standard curve of liquid, and by it compared with the chitosan standard curve of different molecular weight, according to the △ A and sample working solution of sample
Standard curve regression equation determine chitosan in sample molecular weight size, according to the molecule of corresponding chitosan
The size of amount determines used regression equation；
4) chitosan content determines in sample：Sample working solution 1mL is taken, is measured by step 1) detection method,
Its absorbance A is measured at 530nm, and calculates △ A=A0- A, chitosan in sample is tried to achieve by △ A substitution equations of linear regression
Content, while do mark-on reclaims experiment.
Method using Erythrosin B as the Spectrophotometric Determination chitosan content of probe as described above, it is described
B-R buffer solutions by 0.04mol/L mixed acid [(2.71mL orthophosphoric acid+2.36mL glacial acetic acid+2.47g boric acid)/L] with
0.2mol/LNaOH solution is formulated by different proportion, applicants experimentally found that, system is buffered in the B-R of pH=4.2
Solution medium sensitivity highest, Δ A values are maximum, in addition it is found by the applicant that B-R buffer solutions have preferable Anti-Jamming, can cover
Cover part metals ion, therefore the B-R buffer solutions of optimized buffer solution selected as pH=4.2.
Method using Erythrosin B as the Spectrophotometric Determination chitosan content of probe as described above, it is described
Sensitizer is polyvinyl alcohol, polysorbas20, Tween 80 or OP-10, applicants experimentally found that, polyvinyl alcohol is added as enhanced sensitivity
△ A values during agent are maximum, therefore select polyvinyl alcohol as sensitizer.
Method using Erythrosin B as the Spectrophotometric Determination chitosan content of probe as described above, in step 1
Secondly the addition sequence of solution adds poly-vinyl alcohol solution to be firstly added chitosan standard solution, it is molten to add BR bufferings again
Liquid, is eventually adding Erythrosin B solution.
Method using Erythrosin B as the Spectrophotometric Determination chitosan content of probe as described above, chitosan
The μ g/mL of 0.05 μ g/mL of concentration range~1.50.
Sample pre-treatments：Capsule shells to be detected are taken, weigh a certain amount of capsule shells, sample is obtained using glacial acetic acid dissolving constant volume
Storing solution, storing solution is filtered with funnel absorbent cotton, filtrate through 6000r/min, centrifuge 20min, take supernatant 2.5mL in
100mL volumetric flasks, constant volume, obtains the sample working solution that concentration is 10 μ g/mL；
With sample working solution, empirically method draws sample curves, compared with the chitosan standard curve of different molecular weight,
The size of the molecular weight of the chitosan in sample is determined according to the regression equation of sample, according to the molecular weight of corresponding chitosan
Size determine used in △ A and chitosan concentration standard curve；
Sample working solution 1mL is taken, empirically method is measured, its absorbance A is measured at 500nm, and calculate
△ A=A0- A, the content of chitosan in sample is tried to achieve by △ A substitution equations of linear regression, while does mark-on reclaims
Experiment.
Test example：
The abosrption spectrogram under different CTS concentration of 1.CTS- Erythrosin B systems.
Fig. 1 is the abosrption spectrogram under different CTS concentration of CTS- Erythrosin B systems, and as seen from the figure, which exists
Have absworption peak at 530nm wavelength, when adding chitosan, its absworption peak substantially reduces, at 530nm with zero pipe be reference when
There is maximum absorption band (negative peak), and have obvious graded, from top to bottom chitosan concentration (μ g/mL)：0.4,0.8,
1.2.Light absorption value is with the increase of the concentration of chitosan as can be seen from Figure, and the light absorption value decline of system, there are linear relationship.
2. influence of the stabilization time for system absorbance
Under more excellent experiment condition, the stabilization time of system is investigated, time 2h is investigated, is surveyed at regular intervals in room temperature
Its absorbance, as a result such as Fig. 2, system can reach stabilization in 1h, can keep stable absorbance constant in 2h.
3. influence of the different buffer solutions for system absorbance
3 kinds of B-R buffer solutions, citric acid-sodium citrate buffer and NaAc_HAc buffer solution have been investigated respectively
Influence of the buffer solution to system absorbance.According to experimental method, change the buffer solution for adding different pH value, obtain each slow
Rush the absorbance of solution.
The result is shown in Fig. 3,4, and the system for adding B-R solution is stablized relatively, and Δ A values are larger.According to buffering range, B- is selected
R buffer solutions (pH=2.5,3.5,4.5,5.5) are cooked standard curve for buffer solution, it is known that the Optimal pH of solution should be tied 4.5 or so
Fruit sees Fig. 3.Therefore continue to have done refinement experiment and the screening experiment (Fig. 4) of buffer solution species of pH, body as seen from Figure 4
The B-R buffer solution medium sensitivity highests of pH=4.2 are tied up to, Δ A values are maximum, in addition it is found by the applicant that B-R has preferable resist
Interference effect, energy masked portion metal ion, therefore the B-R buffer solutions of optimized buffer solution selected as pH=4.2.
4. influence of the addition of buffer solution to absorbance
Buffer solution provides suitable acidity combining environmental for experimental system, and formation of its addition to ionic associate has
Certain influence, does not change other experiment conditions, the addition for changing the B-R buffer solutions of pH=4.2 is 0.5,1.0,
1.5,2.0,3.0mL, measure its absorbance.
Fig. 5 is influence of the addition of B-R buffer solutions for absorbance.As a result find out, the addition of B-R buffer solutions
The influence measured to the reaction system is smaller, and in addition in the range of 0.5~3.0mL, the change of light absorption value value is smaller, in 1.0mL
△ A reach maximum during addition.Therefore selection BR buffer solution additions are 1.0mL.5. reaction temperature is for system absorbance
Influence
Under experimental conditions, influence of 20~100 DEG C of different temperatures to system sensitivity has been investigated, the results showed that, such as Fig. 6
There is influence for light absorption value of the temperature displaying function on system.△ A reduce rapidly with the rise of temperature, and under the reaction temperature, weight
Existing property is preferable, therefore temperature of reaction system selection room temperature is the most suitable.
6. the influence of sensitizer species and addition for system absorbance
Do not change other experiment conditions, change sensitizer species (polyvinyl alcohol, polysorbas20, Tween 80, OP-10), measure
Its absorbance.The results are shown in Figure 7, and △ A values when adding polyvinyl alcohol (PVA) as sensitizer are maximum, therefore select PVA conducts
Sensitizer.
Fig. 8 is the screening of PVA additions, it can be seen that with the addition increase of PVA, its △ A value can accordingly reduce, but
If being added without PVA or the less situation of addition, in the reaction system Erythrosin B can autohemagglutination form precipitation, lead to not
Measurement result.Therefore considering various factors (deposited phenomenon occur in 0.1mL and 0mL), the more suitable PVA of selection is added
Measure as 0.2mL.In the system, PVA plays stable effect, and making dyestuff, agglomeration speed declines in acid condition, is easy to
Detection.
7. influence of the Erythrosin B addition for system absorbance
Erythrosin B is combined as probe dye with target substance chitosan, its addition directly affects ionic associate
Formation, do not change other experiment conditions, it is 1.0 × 10 to change concentration-4The addition of the Erythrosin B of mol/L is 1.0,2.0,
3.0,4.0,5.0mL, measure its absorbance.
Fig. 9 is influence of the addition of Erythrosin B for system absorbance.As a result find out, the addition of Erythrosin B is to this
Reaction system has a significant impact, its △ A is maximum when adding 4.0mL Erythrosin Bs as seen from the figure, therefore 1.0 × 10-4The red moss of mol/L
Red B optimal addns are 4.0mL.
8. influence of the reagent addition sequence for system absorbance
Under experimental conditions, dyestuff Erythrosin B has been investigated, BR buffer solutions, 12 kinds of chitosan and polyvinyl alcohol different
Influence of the addition sequence to system sensitivity.Consider all addition sequences as a result, with reference to reappearance, linearly, light absorption value,
The many factors such as stability, stabilization time, have selected the 8th kind " CTS+PVA+BR+ Erythrosin Bs " is optimal addition sequence.
The influence of 1 addition sequence of table
9. influence of the ionic strength for system absorbance
Influence of the ionic strength to the absorbance of system is investigated with NaCl (0.01~3mol/L).The result is shown in Figure 10,
Ionic strength influences system absorbance little during 0.01-0.07mol/L, its relative error is no more than 5%；In 0.07mol/
After L, the absorbance of system is in slow increase tendency with the increase of ionic strength, it may be possible to NaCl and the body in high concentration
Tying, which is combined, raises Δ I.Therefore, experiment is not influenced when the concentration of NaCl is less than 0.007mol/L.
10. influence of the coexisting substances for system absorbance
Interference of 7 kinds of coexisting substances to the system is investigated, the concentration of chitosan is 0.5 μ g/mL in system.In opposite mistake
For difference in the range of ± 5%, the allowance of various coexisting substances is shown in Table 2.Starch, beta cyclodextrin, influence of the glycine to system is very
It is small, it is allowed to measure larger；The allowances such as ascorbic acid, asparagine, lysine, zinc sulfate are relatively small.Can from reaction system
To show that its most important influence for pH value, should strictly control the amount of buffer solution, control pH in the reaction.
The influence of 2 coexisting substances of table
The present invention has following technical advantage compared with prior art：
1) good linearity, detection limit are low：Under optimum experimental condition, according to the chitosan pair of determination of experimental method various concentrations
The △ A answered, draw standard curve, the results showed that, there are good with △ A in the range of 0.05 μ g/mL-1.5 μ g/mL of concentration for chitosan
Good linear relationship, 0.0441 μ g/mL of detection limit.
2) this method measure is influenced by the molecular weight of chitosan, in actual sample measure, need to be considered as close point
The standard items of son amount do quantitative criterion, have the advantages of reagent is cheap, high sensitivity favorable reproducibility.
Brief description of the drawings
The abosrption spectrogram under different CTS concentration of Fig. 1 CTS- Erythrosin B systems.
Influence of Fig. 2 stabilization times for system absorbance.
Influence of Fig. 3 reaction acidity for system absorbance.
The influence of Fig. 4 pH refinements and buffer solution species for system absorbance.
Influence of the addition of Fig. 5 BR buffer solutions for absorbance.
Influence of Fig. 6 reaction temperatures for system absorbance.
Influence of Fig. 7 differences sensitizer for system absorbance.
Influence of Fig. 8 polyvinyl alcohol addition for system absorbance.
Influence of Fig. 9 Erythrosin Bs addition for system absorbance.
Influence of Figure 10 ionic strengths for system absorbance
Embodiment
The present invention is further described below by way of specific embodiment, but the invention does not limit the present invention in any way
The scope of patent protection.
Embodiment
A kind of method using Erythrosin B as the Spectrophotometric Determination chitosan content of probe, it includes following steps
Suddenly：
1) standard curve of the chitosan concentration of △ A and different molecular weight is drawn:
The low molecular chitosan standard solution of 0.5mL a certain concentration gradients is added into 10mL colorimetric cylinders, it is dense to add 0.2mL
The polyvinyl alcohol for 1%, the BR buffer solutions of 1.0mL, and 4.0mL concentration are spent for 1.0 × 10-4The Erythrosin B of mol/L is molten
Liquid, using fully being shaken up after distilled water constant volume, stand at room temperature 1 it is small when after on ultraviolet specrophotometer, in system most
It is reference measurement absorbance that big colour fading wavelength 530nm, which sentences water,；Wherein reagent blank is denoted as A0, the solution of chitosan-containing is denoted as A,
And calculate △ A=A0- A, establishes the standard curve C1 of △ A and low molecular chitosan concentration；Use middle-molecular-weihydroxyethyl chitosan solution
Low molecular chitosan solution is replaced, establishes the standard curve C2 of △ A and middle molecular chitosan concentration；Use high molecular weight chitosan
Solution replaces low molecular chitosan solution, establishes the standard curve C3 of △ A and polymer chitosan concentration；
The chitosan result of 3 different molecular weight of table
2) preparation of the sample working solution of 10 μ g/mL：The capsule shells of a certain amount of detected sample are weighed, it is molten using glacial acetic acid
Solve and its constant volume is obtained into stock sample solution, filter storing solution with funnel absorbent cotton, filtrate is through 6000r/min, centrifuge
20min, taking supernatant 2.5mL, constant volume, obtains the sample working solution that concentration is 10 μ g/mL in 100mL volumetric flasks；
3) according to chitosan molecule amount selection criteria curve：△ A are drawn using step 1) kind detection method and sample works
The standard curve of liquid, and by it compared with the chitosan standard curve of different molecular weight, according to the △ A and sample working solution of sample
Standard curve regression equation determine chitosan in sample molecular weight size, according to the molecule of corresponding chitosan
The size of amount determines used regression equation；
With sample working solution, empirically method draws sample curves, compared with the chitosan standard curve of different molecular weight,
As a result be △ A=-0.6835c-0.0788 for the equation of linear regression of sample, related coefficient 0.9949, as a result with middle-molecular-weihydroxyethyl
Chitosan standard curve it is identical, using the standard curve of middle-molecular-weihydroxyethyl chitosan as quantitation curves.
4) chitosan content determines in sample：Sample working solution 1mL is taken, is measured by step 1) detection method,
Its absorbance A is measured at 530nm, and calculates △ A=A0- A, chitosan in sample is tried to achieve by △ A substitution equations of linear regression
Content, while do mark-on reclaims experiment.
Sample working solution 1mL is taken, empirically method is measured, its absorbance A is measured at 530nm, and calculate △ A
=A0- A, the content of chitosan in sample is tried to achieve by △ A substitution equations of linear regression, while does mark-on reclaims experiment.As a result
Going out, the content for defending chitosan in the difficult to understand capsule that comes unglued is 852mg/g, RSD 1.51%,
Average recovery of standard addition is 101.65%.
4 sample analysis result of table
The low molecular chitosan standard solution of 0.5mL a certain concentration gradients is added into 10mL colorimetric cylinders, it is dense to add 0.2mL
The polyvinyl alcohol for 1%, the BR buffer solutions of 1.0mL, and 4.0mL concentration are spent for 1.0 × 10-4The Erythrosin B of mol/L is molten
Liquid, using fully being shaken up after distilled water constant volume, stand at room temperature 1 it is small when after on U-3010 type ultraviolet specrophotometers,
It is reference measurement absorbance that the maximum colour fading wavelength 530nm of system, which sentences water, in having been surveyed in 2h.Wherein reagent blank is denoted as A0,
The solution of chitosan-containing is denoted as A, and calculates △ A=A0–A.Chitosan in the range of 0.05 μ g/mL-1.5 μ g/mL of concentration with A0Deposit
In good linear relationship, detection is limited to 0.0441 μ g/mL.This method measure is influenced by the molecular weight of chitosan, in reality
In sample measure, the standard items that need to be considered as close molecular weight do quantitative criterion, cheap with reagent, high sensitivity reappearance
The advantages of good.
The range of linearity and detection limit
Under optimum experimental condition, according to the corresponding △ A of the chitosan of determination of experimental method various concentrations, it is bent to draw standard
Line, the results showed that, chitosan in the range of 0.05 μ g/mL-1.5 μ g/mL of concentration with △ A there are good linear relationship, linearly
Regression equation is △ A=-0.6835c-0.0788, related coefficient 0.9949,0.0441 μ g/mL of detection limit.
Claims (8)
1. a kind of method using Erythrosin B as the Spectrophotometric Determination chitosan content of probe, it comprises the following steps：
1) standard curve of the chitosan concentration of △ A and different molecular weight is drawn：
The low molecular chitosan standard solution of 0.5mL a certain concentration gradients is added into 10mL colorimetric cylinders, adding 0.2mL concentration is
1% sensitizer, the BR buffer solutions of 1.0mL, and 4.0mL concentration are 1.0 × 10-4The Erythrosin B solution of mol/L, uses
Fully shaken up after distilled water constant volume, stand at room temperature 1 it is small when after on ultraviolet specrophotometer, in the maximum colour fading ripple of system
It is reference measurement absorbance that long 530nm, which sentences water,；Wherein reagent blank is denoted as A0, the solution of chitosan-containing is denoted as A, and calculates △
A=A0- A, establishes the standard curve C1 of △ A and low molecular chitosan concentration；Low point is replaced using middle-molecular-weihydroxyethyl chitosan solution
Seed chitosan solution, establishes the standard curve C2 of △ A and middle molecular chitosan concentration；Replaced using high molecular weight chitosan solution
Low molecular chitosan solution, establishes the standard curve C3 of △ A and polymer chitosan concentration；
2) preparation of the sample working solution of 10 μ g/mL：The capsule shells of a certain amount of detected sample are weighed, are dissolved using glacial acetic acid,
And its constant volume is obtained into stock sample solution, storing solution is filtered with funnel absorbent cotton, filtrate is through 6000r/min, centrifuge
20min, taking supernatant 2.5mL, constant volume, obtains the sample working solution that concentration is 10 μ g/mL in 100mL volumetric flasks；
3) according to chitosan molecule amount selection criteria curve：△ A and sample working solution are drawn using step 1) kind detection method
Standard curve, and by it compared with the chitosan standard curve of different molecular weight, according to the △ A of sample and the mark of sample working solution
The regression equation of directrix curve determines the size of the molecular weight of the chitosan in sample, according to the molecular weight of corresponding chitosan
Size determines used regression equation；
4) chitosan content determines in sample：Sample working solution 1mL is taken, is measured by step 1) detection method, at 530nm
Its absorbance A is measured, and calculates △ A=A0△ A substitution equations of linear regression, are tried to achieve the content of chitosan in sample by-A,
Do mark-on reclaims experiment at the same time.
2. the method according to claim 1 by Fading spectrophotometry Accurate Determining chitosan content, its feature exists
In the B-R buffer solutions are formulated by 0.04mol/L mixed acid with 0.2mol/LNaOH solution by different proportion.
3. the method according to claim 2 by Fading spectrophotometry Accurate Determining chitosan content, its feature exists
In the mixed acid is made of orthophosphoric acid, glacial acetic acid and boric acid.
4. the method according to claim 1 by Fading spectrophotometry Accurate Determining chitosan content, its feature exists
In the pH of the BR buffer solutions is 4.2.
5. the method according to claim 1 by Fading spectrophotometry Accurate Determining chitosan content, its feature exists
In, the sensitizer be polyvinyl alcohol, polysorbas20, one kind in Tween 80 or OP-10.
6. the method according to claim 5 by Fading spectrophotometry Accurate Determining chitosan content, its feature exists
In the sensitizer is polyvinyl alcohol.
7. the method according to claim 1 by Fading spectrophotometry Accurate Determining chitosan content, its feature exists
In in step 1 secondly the addition sequence of solution adds poly-vinyl alcohol solution to be firstly added chitosan standard solution, adds again
Enter BR buffer solutions, be eventually adding Erythrosin B solution.
8. the method according to claim 1 by Fading spectrophotometry Accurate Determining chitosan content, its feature exists
In the concentration range of chitosan is the μ g/mL of 0.05 μ g/mL~1.50 in step 1) standard curve C1, C2 and C3.
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| CN112525844A (en) * | 2020-11-12 | 2021-03-19 | 德莱福(重庆)医疗器械有限公司 | Method for testing urea concentration in stable dialyzer clearance rate simulation solution |
| CN112525844B (en) * | 2020-11-12 | 2024-06-04 | 德莱福(重庆)医疗器械有限公司 | Stable urea concentration test method in dialyser clearance simulation liquid |
| CN116698776A (en) * | 2023-06-28 | 2023-09-05 | 湖北工程学院 | Color development-free spectrophotometry for quantitatively analyzing chitosan |
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