CN114441684B - Qualitative and quantitative method for analyzing high molecular saccharide compound - Google Patents
Qualitative and quantitative method for analyzing high molecular saccharide compound Download PDFInfo
- Publication number
- CN114441684B CN114441684B CN202210108229.1A CN202210108229A CN114441684B CN 114441684 B CN114441684 B CN 114441684B CN 202210108229 A CN202210108229 A CN 202210108229A CN 114441684 B CN114441684 B CN 114441684B
- Authority
- CN
- China
- Prior art keywords
- sample
- molecular weight
- standard
- concentration
- curve
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- -1 saccharide compound Chemical class 0.000 title claims abstract description 23
- 238000004445 quantitative analysis Methods 0.000 title claims abstract description 22
- 238000000034 method Methods 0.000 claims abstract description 35
- 239000000126 substance Substances 0.000 claims abstract description 27
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 23
- 230000014759 maintenance of location Effects 0.000 claims abstract description 19
- 230000004044 response Effects 0.000 claims abstract description 19
- 238000004451 qualitative analysis Methods 0.000 claims abstract description 13
- 229920001218 Pullulan Polymers 0.000 claims abstract description 7
- 239000004373 Pullulan Substances 0.000 claims abstract description 7
- 238000001514 detection method Methods 0.000 claims abstract description 7
- 235000019423 pullulan Nutrition 0.000 claims abstract description 7
- 238000001914 filtration Methods 0.000 claims abstract description 3
- 238000000611 regression analysis Methods 0.000 claims abstract description 3
- 239000000523 sample Substances 0.000 claims description 37
- 229920001282 polysaccharide Polymers 0.000 claims description 30
- 239000005017 polysaccharide Substances 0.000 claims description 30
- 150000004676 glycans Chemical class 0.000 claims description 29
- 241001506047 Tremella Species 0.000 claims description 9
- 241000196252 Ulva Species 0.000 claims description 8
- 238000005227 gel permeation chromatography Methods 0.000 claims description 7
- 238000000926 separation method Methods 0.000 claims description 6
- 239000000243 solution Substances 0.000 claims description 6
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 6
- 239000012498 ultrapure water Substances 0.000 claims description 6
- 238000011002 quantification Methods 0.000 claims description 5
- 238000002347 injection Methods 0.000 claims description 4
- 239000007924 injection Substances 0.000 claims description 4
- 239000012488 sample solution Substances 0.000 claims description 4
- 239000012528 membrane Substances 0.000 claims description 2
- 239000012086 standard solution Substances 0.000 claims description 2
- 150000001720 carbohydrates Chemical class 0.000 abstract description 4
- 239000012982 microporous membrane Substances 0.000 abstract 1
- 229920000642 polymer Polymers 0.000 abstract 1
- 150000001875 compounds Chemical class 0.000 description 4
- 235000014633 carbohydrates Nutrition 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- CTYRPMDGLDAWRQ-UHFFFAOYSA-N phenyl hydrogen sulfate Chemical compound OS(=O)(=O)OC1=CC=CC=C1 CTYRPMDGLDAWRQ-UHFFFAOYSA-N 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 229920002307 Dextran Polymers 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 238000004164 analytical calibration Methods 0.000 description 1
- RJGDLRCDCYRQOQ-UHFFFAOYSA-N anthrone Chemical compound C1=CC=C2C(=O)C3=CC=CC=C3CC2=C1 RJGDLRCDCYRQOQ-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000013365 molecular weight analysis method Methods 0.000 description 1
- 238000010606 normalization Methods 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N30/14—Preparation by elimination of some components
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N30/14—Preparation by elimination of some components
- G01N2030/146—Preparation by elimination of some components using membranes
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The application relates to a method for analyzing the nature and the quantity of high molecular saccharide compounds, which comprises the following steps: 1) Selecting pullulan as a standard substance; 2) Filtering with microporous membrane, and detecting with high performance liquid chromatography and differential refraction detector; 3) Detecting standard substances with different molecular weights to obtain chromatograms and retention times of the standard substances, and carrying out regression analysis by taking log Mw as an ordinate and the retention time of chromatographic peaks as an abscissa, wherein the molecular weight standard curves are obtained; 4) Performing chromatographic detection on standard substances with different concentrations to obtain a fitting curve; 5) And (3) detecting the sample to be detected by using the method of the step (2), obtaining a response value and retention time of the chromatogram, and calculating the weight-average relative molecular mass and concentration of the sample to be detected by using the standard curve of the step (3) and the fitting curve of the step (4) to obtain an HPLC quantitative curve, so that the qualitative and quantitative analysis of the high polymer carbohydrate is facilitated.
Description
Technical Field
The application belongs to the technical field of analysis of saccharide compounds, and particularly relates to a qualitative and quantitative method for analyzing high-molecular saccharide compounds.
Background
The disclosure of this background section is only intended to increase the understanding of the general background of the application and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art already known to those of ordinary skill in the art.
The pharmacological activity of the polysaccharide is closely related to the relative molecular weight (Mw) and the distribution thereof, and the high molecular saccharide compound has large molecular weight, complex structure and lack of easy-to-detect luminescent groups, so that the related qualitative and quantitative methods are not completely established, further research and application of people are limited, and how to rapidly and accurately identify and qualitatively and quantitatively analyze the saccharide compound becomes a key problem in the saccharide industrialization process.
At present, a phenol sulfate method, an anthrone sulfate method and the like are often used as a method for measuring polysaccharide, and although the measurement can be performed relatively rapidly, the method can only measure the total sugar content of a sample, and cannot accurately perform quantitative analysis on high-molecular-weight saccharide compounds with specific molecular weights. High Performance Liquid Chromatography (HPLC) is a widely used analytical method for measuring and quantifying the molecular weight of polysaccharides, and can separate, qualify and quantify polysaccharide compounds with high and low molecules. However, the method can only establish a quantitative curve under the corresponding polysaccharide standard substance, so that only target compounds with limited molecular weight determined or close to the standard substance can be quantitatively analyzed, and other sugar compounds with unknown or higher molecular weight cannot be quantitatively analyzed. In the existing method, in order to accurately determine the content of polysaccharide and the relative molecular mass distribution in a sample, dextran with the molecular weight similar to that of the sample is selected as a standard substance, and the content of each component is calculated by adopting an area normalization method. Although this method can achieve quantification of similar molecular weight samples to some extent, it is extremely inaccurate for quantitative analysis when the molecular weight difference between the sample and the standard is too large.
Disclosure of Invention
In view of the above problems in the prior art, an object of the present application is to provide a method for analyzing qualitative and quantitative properties of a high molecular saccharide compound.
In order to solve the technical problems, the technical scheme of the application is as follows:
a method for the qualitative and quantitative analysis of high molecular carbohydrate compounds, the method comprising:
1) Dissolving Pullulan (Pullulan KIT) into a standard substance solution by using a mobile phase, and dissolving a sample to be detected into a sample solution by using the mobile phase;
2) Filtering the sample solution to be detected and the standard substance solution in the step 1) through a microporous filter membrane, and detecting by using a high performance liquid chromatography and a differential refraction detector;
3) Detecting standard substance solutions with different molecular weights by using the method of the step 2) to obtain a chromatogram and Retention Time (RT) of the standard substance, and carrying out regression analysis by taking logarithmic log Mw of the molecular weight of the standard substance as an ordinate and the retention time of a chromatographic peak as an abscissa to obtain a regression equation and a standard curve for calculating the molecular weight;
4) Performing chromatographic detection on standard substances with different molecular weights to obtain chromatograms and response values (Mv) of the standard substances, and fitting according to the standard curve of the step 3) and the concentrations of different standard substances to obtain fitting curves under different concentrations;
5) And (3) detecting the sample to be detected by using the method of the step (2), obtaining a response value (Mv) and Retention Time (RT) of the chromatogram, and calculating the weight-average relative molecular mass and concentration of the sample to be detected by using the standard curve of the step (3) and the fitting curve of the step (4).
The response value is Mv of the chromatogram, and the retention time can be obtained by the abscissa corresponding to the peak of the chromatogram. The retention time of the peaks appearing is obtained by means of a chromatogram, the molecular weight log Mw is obtained by means of the molecular weight standard curve of step 3), and the molecular weight is obtained. Further obtaining an HPLC quantitative curve through the fitting curve of the step 4), and obtaining the corresponding concentration through the molecular weight and the response value. Therefore, the high molecular saccharide compound can be qualitatively and quantitatively determined.
Compared with other polysaccharides, the pullulan polysaccharide has the advantages of higher solubility, higher plasticity and viscosity, easy dissolution in water, no color and smell, and the like, and is very suitable for the calibration of instruments.
In some embodiments of the application, the detection conditions in step 2) are that the separation columns are Shodex SB-804 and 806 in series, and the mobile phase is ultrapure water or 0.1M NaNO 3 The flow rate is 0.5-1ml/min, the column temperature is 35-40 ℃, and the sample injection amount is 10-20 mu L. The separation column is selected to be Shodex SB-804 and Shodex SB-806 in series, and the exclusion limit of Shodex SB-806 is selectedIs 2 x 10 7 The molecular weight analysis of higher molecular weight saccharide compounds or polysaccharides can be satisfied. Using 0.1M NaNO 3 Can meet the separation of polysaccharide with similar molecular weight.
In some embodiments of the application, the pullulan in step 2) has a molecular weight Mw of 180Da, 504Da, 991Da, 6600Da, 9900Da, 23000Da, 50600Da, 115000Da, 202000Da, 343000Da, 805000Da, 1330000Da.
In some embodiments of the application, the concentration of the standard solution obtained in step 1) is 1.0mg/ml, 0.8mg/ml, 0.5mg/ml, 0.2mg/ml, 0.1mg/ml.
In some embodiments of the application, the chromatogram is a chromatogram with time on the abscissa and voltage on the ordinate.
In some embodiments of the present application, the method of fitting the curve in step 4) is: obtaining a retention time of the chromatogram corresponding to the molecular weight of the standard substance through the molecular weight standard curve in the step 3), and then fitting the molecular weight concentration with the response value of the chromatogram through the correspondence of the molecular weight and the concentration of the standard substance to obtain a fitting curve. Further, an HPLC quantitative curve at an arbitrary molecular weight can be obtained.
In some embodiments of the present application, the regression equation of the fitted curve obtained in step 4) is y=ae -Bx ，R 2 = 0.9236-0.9698. Further, the concentration was 1.0mg/ml, a=363.07, b=0.349; concentration was 0.8mg/ml, a=295.18, b=0.361; concentration was 0.5mg/ml, a=143.32, b=0.334; concentration was 0.2mg/ml, a=65.347, b=0.372; the concentration was 0.1mg/ml, a=54.458, b=0.485.
In some embodiments of the application, the specific steps of step 5) are: obtaining retention time and response value of a chromatogram by detecting a sample to be detected, calculating molecular weight of the sample by using the molecular weight standard curve obtained in the step 3) to obtain logarithmic value log Mw of the molecular weight, obtaining corresponding HPLC quantitative curve under the molecular weight by using the fitting curve of different concentrations in the step 4), and obtaining the concentration of the sample to be detected by substituting the response value.
In some embodiments of the application, retention Times (RT) of the chromatograms in steps 3) and 5) and response values etc. are calculated by GPC gel chromatography software.
In some embodiments of the present application, the sample to be detected may be a high molecular polysaccharide such as tremella polysaccharide, enteromorpha polysaccharide, etc.
In some embodiments of the application, the HPLC quantification curve has the equation y=ax-B, R 2 = 0.9958-0.9959, x is the concentration, y is the response value; wherein the values of A and B are determined according to the molecular weights of the different saccharide compounds.
One or more of the technical schemes of the application has the following beneficial effects:
(1) The application establishes a novel qualitative and quantitative analysis method for the high molecular carbohydrate by using the high-efficiency gel chromatography, gets rid of the limitation of the current high molecular carbohydrate HPLC quantification by a standard substance, has the advantages of precision, accuracy, good reproducibility, high efficiency, rapidness and the like, and realizes qualitative and quantitative analysis in any molecular weight range.
(2) Compared with the traditional classical methods such as a phenol sulfate method and the like, the method disclosed by the application is used for evaluating the molecular weight difference of different methods for preparing the sugar compounds and accurately calculating the concentration of a sample with a certain molecular weight or a molecular weight range, solves the problem of inaccurate quantitative analysis when the molecular weight difference between the sample and a standard substance is too large, makes up the defects of the polysaccharide detection method in the traditional national standard, and promotes the research progress of the high-molecular sugar compounds in the fields of chemistry, biology, medicine and the like.
Drawings
The accompanying drawings, which are included to provide a further understanding of the application and are incorporated in and constitute a part of this specification, illustrate embodiments of the application and together with the description serve to explain the application.
FIG. 1 is a graph of fit at various concentrations;
FIG. 2 is a GPC chromatogram of tremella polysaccharide;
FIG. 3 is a quantitative HPLC plot of tremella polysaccharide sample peak 1;
FIG. 4 is a quantitative HPLC plot of tremella polysaccharide sample peak 2;
FIG. 5 is a GPC chromatogram of Enteromorpha polysaccharide;
fig. 6 is a HPLC quantification curve of enteromorpha polysaccharide sample peak 1.
Detailed Description
It should be noted that the following detailed description is illustrative and is intended to provide further explanation of the application. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the present application. As used herein, the singular is also intended to include the plural unless the context clearly indicates otherwise, and furthermore, it is to be understood that the terms "comprises" and/or "comprising" when used in this specification are taken to specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof. The application will be further illustrated by the following examples
Example 1
This example details the establishment of HPLC fitted curves.
The chromatographic results of 12 standards with different molecular weights at different concentrations are processed, the logMw is obtained by means of a molecular weight standard curve, and then special fitting is carried out on the results and response values (Mv), so that fitting curves with the concentration of 1.0mg/ml, 0.8mg/ml, 0.5mg/ml, 0.2mg/ml and 0.1mg/ml at 5 different concentrations are constructed, and the curves are used for establishing quantitative curves at different molecular weights (see figure 1).
The regression equation of the resulting fitted curve is y=ae -Bx ，R 2 = 0.9236-0.9698. Further, the concentration was 1.0mg/ml, a=363.07, b=0.349; concentration was 0.8mg/ml, a=295.18, b=0.361; concentration was 0.5mg/ml, a=143.32, b=0.334; concentration was 0.2mg/ml, a=65.347, b=0.372; the concentration was 0.1mg/ml, a=54.458, b=0.485.
Example 2
In this example, the quantitative analysis of the sample was performed by constructing a quantitative HPLC curve using tremella polysaccharide as an example.
Dissolving tremella polysaccharide sample with ultrapure water of mobile phase, measuring molecular weight distribution and specific concentration of tremella polysaccharide sample under each molecular weight, detecting by HPLC-RID, wherein separation columns are Shodex SB-804 and 806 connected in series, the mobile phase is ultrapure water with flow rate of 1ml/min, column temperature is 35 ℃, and sample injection amount is 10 mu L; the time was 30min.
The GPC chromatogram of the tremella polysaccharide is shown in FIG. 2, wherein peak 1 and peak 2 are calculated, the retention time RT of peak 1 is 8.768min, the retention time RT of peak 2 is 13.439min, the log Mw of peak 1 is 7.63, the Mw is 42657951Da, the log Mw of peak 2 is 6.15, and the Mw is 1412537Da; HPLC quantitative curves for the corresponding molecular weights of peak 1 and peak 2 were constructed by fitting curves for the different concentrations in example 1 (see fig. 3 and 4), respectively, and the HPLC concentration of peak 1 was calculated to be 0.393mg/ml and the HPLC concentration of peak 2 was calculated to be 0.133mg/ml.
Example 3
In the embodiment, the enteromorpha polysaccharide sample is used as an example, and the quantitative analysis of the sample is performed by constructing an HPLC quantitative curve.
Dissolving enteromorpha polysaccharide sample with ultrapure water in a mobile phase, measuring molecular weight distribution and specific concentration of the enteromorpha polysaccharide sample under each molecular weight, detecting by adopting HPLC-RID, wherein separation columns are Shodex SB-804 and 806 which are connected in series, the mobile phase is ultrapure water with a flow rate of 1ml/min, the column temperature is 35 ℃, and the sample injection amount is 10 mu L; the time was 50min.
The GPC chromatogram of Enteromorpha polysaccharide is shown in FIG. 5, and by taking peak 1 as an example, the retention time RT of peak 1 is 9.1min, and by using molecular weight standard curve, the log Mw of peak 1 is 7.51, and Mw is 32359365Da; the HPLC quantitative curve for peak 1 (see FIG. 6) was constructed by fitting curves for different concentrations in example 1, and the HPLC concentration of peak 1 was calculated to be 0.267mg/ml.
The above description is only of the preferred embodiments of the present application and is not intended to limit the present application, but various modifications and variations can be made to the present application by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the protection scope of the present application.
Claims (9)
1. A method for qualitative and quantitative analysis of high molecular saccharide compounds, characterized in that: the method comprises the following specific steps: the method comprises the following steps:
1) Dissolving pullulan with a mobile phase to obtain a standard substance solution, and dissolving a sample to be detected with the mobile phase to obtain a sample solution;
2) Filtering the sample solution to be detected and the standard substance solution in the step 1) through a microporous filter membrane, and detecting by using a high performance liquid chromatography and a differential refraction detector;
3) Detecting standard substance solutions with different molecular weights by using the method of the step 2) to obtain a chromatogram and retention time of the standard substance, and carrying out regression analysis by taking the log Mw of the molecular weight of the standard substance as an ordinate and the retention time of a chromatographic peak as an abscissa to obtain a regression equation and a standard curve for calculating the molecular weight;
4) Performing chromatographic detection on standard substances with different molecular weights at different concentrations to obtain a chromatogram and a response value of the standard substances, and fitting by taking the log Mw of the molecular weights of the standard substances as an abscissa and the response value (Mv) as an ordinate to obtain a fitting curve y=ae at different concentrations -Bx The method comprises the steps of carrying out a first treatment on the surface of the Wherein A and B are constants, which vary according to different concentrations of the standard;
5) Detecting a sample to be detected by using the method of the step 2), obtaining a response value and retention time of a chromatogram, calculating the molecular weight of the sample to be detected by using the standard curve of the step 3), and further obtaining an HPLC quantitative curve Y=A1X+B1 by using the fitting curve of the step 4) under different concentrations, wherein Y is a response value (MV), X is a concentration, A1 and B1 are constants, and calculating the concentration of the sample to be detected under the molecular weight according to the change of the molecular weight of the sample to be detected;
the detection conditions in the step 2) are that the mobile phase is ultrapure water or 0.1M NaNO 3 。
2. The method for qualitative and quantitative analysis of a high molecular saccharide compound according to claim 1, wherein: the detection condition in the step 2) is that the separation columns are Shodex SB-804 and Shodex 806 which are connected in series, the flow rate is 0.5-1ml/min, the column temperature is 35-40 ℃, and the sample injection amount is 10-20 mu L.
3. The method for qualitative and quantitative analysis of a high molecular saccharide compound according to claim 1, wherein: the molecular weight Mw of the pullulan in step 2) is 180Da, 504Da, 991Da, 6600Da, 9900Da, 23000Da, 50600Da, 115000Da, 202000Da, 343000Da, 805000Da, 1330000Da.
4. The method for qualitative and quantitative analysis of a high molecular saccharide compound according to claim 1, wherein: the concentration of the standard solution obtained in step 1) was 1.0mg/ml, 0.8mg/ml, 0.5mg/ml, 0.2mg/ml, 0.1mg/ml.
5. The method for qualitative and quantitative analysis of a high molecular saccharide compound according to claim 1, wherein: the regression equation of the fitted curve obtained in step 4) is y=ae -Bx ，R 2 = 0.9236-0.9698; further, the concentration was 1.0mg/ml, a=363.07, b=0.349; concentration was 0.8mg/ml, a=295.18, b=0.361; concentration was 0.5mg/ml, a=143.32, b=0.334; concentration was 0.2mg/ml, a=65.347, b=0.372; the concentration was 0.1mg/ml, a=54.458, b=0.485.
6. The method for qualitative and quantitative analysis of a high molecular saccharide compound according to claim 1, wherein: the specific steps of the step 5) are as follows: obtaining retention time and response value of a chromatogram by detecting a sample to be detected, calculating molecular weight of the sample by using the molecular weight standard curve obtained in the step 3) to obtain logarithmic value log Mw of the molecular weight, and obtaining a quantitative curve by substituting the response value into the fitting curve with different concentrations in the step 4), thereby calculating the concentration of the sample to be detected.
7. The method for qualitative and quantitative analysis of a high molecular saccharide compound according to claim 1, wherein: the retention times and response values of the chromatograms in steps 3) and 5) were calculated by GPC gel chromatography software.
8. The method for qualitative and quantitative analysis of a high molecular saccharide compound according to claim 1, wherein: the sample to be detected can be tremella polysaccharide and enteromorpha polysaccharide high molecular polysaccharide.
9. The method for qualitative and quantitative analysis of a high molecular saccharide compound according to claim 1, wherein: the equation of the HPLC quantification curve is y=a1x+b1, R 2 = 0.9958-0.9959, x is concentration, y is response value, A1 and B1 are constants, and vary according to the molecular weight of the sample to be measured.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210108229.1A CN114441684B (en) | 2022-01-28 | 2022-01-28 | Qualitative and quantitative method for analyzing high molecular saccharide compound |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210108229.1A CN114441684B (en) | 2022-01-28 | 2022-01-28 | Qualitative and quantitative method for analyzing high molecular saccharide compound |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114441684A CN114441684A (en) | 2022-05-06 |
| CN114441684B true CN114441684B (en) | 2023-09-26 |
Family
ID=81371565
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210108229.1A Active CN114441684B (en) | 2022-01-28 | 2022-01-28 | Qualitative and quantitative method for analyzing high molecular saccharide compound |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114441684B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116124928A (en) * | 2022-12-24 | 2023-05-16 | 中国检验检疫科学研究院 | Method for determining molecular weight of low molecular weight glycosaminoglycan by gel permeation chromatography |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008266458A (en) * | 2007-04-20 | 2008-11-06 | Sanai Seiyaku Kk | Chitosan mixture, chitosan, and chitin mixture |
| CN110274972A (en) * | 2019-06-21 | 2019-09-24 | 海南大学 | A method of series connection gel chromatography Dendrobium nobile polysaccharide molecular weight distribution |
| WO2020055207A1 (en) * | 2018-09-12 | 2020-03-19 | 주식회사 엘지화학 | Method for analyzing molecular weight of each component of polymer compound, and analysis system used therefor |
| CN113281443A (en) * | 2021-06-07 | 2021-08-20 | 广州广电计量检测股份有限公司 | Polysaccharide detection method |
-
2022
- 2022-01-28 CN CN202210108229.1A patent/CN114441684B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008266458A (en) * | 2007-04-20 | 2008-11-06 | Sanai Seiyaku Kk | Chitosan mixture, chitosan, and chitin mixture |
| WO2020055207A1 (en) * | 2018-09-12 | 2020-03-19 | 주식회사 엘지화학 | Method for analyzing molecular weight of each component of polymer compound, and analysis system used therefor |
| CN110274972A (en) * | 2019-06-21 | 2019-09-24 | 海南大学 | A method of series connection gel chromatography Dendrobium nobile polysaccharide molecular weight distribution |
| CN113281443A (en) * | 2021-06-07 | 2021-08-20 | 广州广电计量检测股份有限公司 | Polysaccharide detection method |
Non-Patent Citations (1)
| Title |
|---|
| 基于高效凝胶渗透色谱法的银耳多糖质量控制研究;刘芹;宁嘉玲;丁侃;;中草药(09);第1-3节，图1，表1-2 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114441684A (en) | 2022-05-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Liu et al. | Electrochemical microfluidic chip based on molecular imprinting technique applied for therapeutic drug monitoring | |
| CN114441684B (en) | Qualitative and quantitative method for analyzing high molecular saccharide compound | |
| WO2022067533A1 (en) | Method for simultaneously testing phylloquinone and menaquinone-4 in trace blood | |
| Kirthi et al. | A review on bioanalytical method development and validation by RP-HPLC | |
| CN115308319A (en) | Quantitative method for non-targeted screening of perfluoro and polyfluoroalkyl compounds | |
| CN112903885A (en) | Application of combined metabolic marker for screening diabetes and kit thereof | |
| Pelit et al. | Preparation and application of a polythiophene solid‐phase microextraction fiber for the determination of endocrine‐disruptor pesticides in well waters | |
| CN111443151A (en) | Method for detecting content of trace cysteine in compound amino acid injection | |
| CN111521695A (en) | Method for rapidly detecting free toluene diisocyanate in polyurethane coating | |
| Mehta | The validation criteria for analytical methods used in pharmacy practice research | |
| CN105548423A (en) | Quantitative detection method for residual crylic acid in polycarboxylate superplasticizer synthetic process | |
| Yang et al. | Analysis of neutral sugars of Asparagus officinalis Linn. polysaccharide by CZE with amperometric detection | |
| CN117517231B (en) | Analysis method, system and readable medium of total nitrogen water quality online analyzer | |
| CN110174417A (en) | The quantitative detecting method of glyceryl triacetate in a kind of filter stick | |
| CN111141809B (en) | Soil nutrient ion content detection method based on non-contact type conductivity signal | |
| CN117191974A (en) | Method for determining concentration of ibuprofen in blood plasma by eliminating interference of matrix effect | |
| Adamowicz et al. | Simple approach for evaluation of matrix effect in the mass spectrometry of synthetic cannabinoids | |
| CN101071104A (en) | Method for determining chloride content in tobacco | |
| CN115112441B (en) | Analysis method for simultaneously measuring ozone-depleting substances and fluorine-containing greenhouse gases in ambient air | |
| Pourjavid et al. | Separation and direct detection of heavy lanthanides using new ion-exchange chromatography: fast Fourier transform continuous cyclic voltammetry system | |
| CN103018192B (en) | Method of determining polyether polyol content in drilling fluid in field | |
| CN110927282A (en) | Method for quickly quantifying various monosaccharide components in xylose mother liquor | |
| CN108717088B (en) | Method for evaluating volatile components of radix curcumae medicinal material by one test and multiple tests | |
| CN110632186B (en) | Method for measuring vitamin D2 content in vitamin D2 injection by UPCC method | |
| CN112964808A (en) | Biological body fluid total homocysteine detection kit and detection method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20240524 Address after: 437000, 2nd Floor, Building 9, Development New Space, Zhonghuopu Town, Chibi City, Xianning City, Hubei Province, China Patentee after: Hubei Zaozhilian Biotechnology Co.,Ltd. Country or region after: China Address before: 266109 No. 700, the Great Wall Road, Chengyang District, Shandong, Qingdao Patentee before: Qingdao Agricultural University Country or region before: China |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20240710 Address after: Building 9, Development New Space, Zhonghuo Guanggu Industrial Park, Zhonghuopu Town, Chibi City, Xianning City, Hubei Province, China 437300 Patentee after: Hubei Zaolan Biotechnology Co.,Ltd. Country or region after: China Address before: 437000, 2nd Floor, Building 9, Development New Space, Zhonghuopu Town, Chibi City, Xianning City, Hubei Province, China Patentee before: Hubei Zaozhilian Biotechnology Co.,Ltd. Country or region before: China |