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A method for measuring chitosan oligosaccharide content

A chitosan oligosaccharide and content technology, applied in the field of macromolecular detection, can solve the problems of low content measurement range, large measurement error, cumbersome operation process, etc., and achieve the effect of simple and fast measurement method, accurate and reliable results, and wide detection range

Active Publication Date: 2020-05-01
GUANGDONG PHARMA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The HPLC method uses differential detector detection or ultraviolet detector detection. This method has high sensitivity, accuracy and reliability, but the operation process is cumbersome, and the sample processing and analysis time is long.
The TLC method has poor stability, large measurement errors, and low accuracy, which cannot guarantee the quality of the measured products.
Spectrophotometry is a relatively traditional method for the detection of chitosan oligosaccharides, which has been improved many times, but there are still problems such as incomplete hydrolysis and low and narrow content measurement range, which increases the error of the measurement results
In addition, as China Patent Application No. 201711493051.2 discloses a UV-visible spectrophotometric method using water-soluble aniline blue as a probe to determine chitosan oligosaccharides, hydrolyzed chitosan oligosaccharides can also be accurately measured by UV spectrophotometry. However, in this method, the aniline blue-chitooligosaccharide system is affected by temperature and pH value, and the operation steps are many and the stability is poor.

Method used

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  • A method for measuring chitosan oligosaccharide content
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  • A method for measuring chitosan oligosaccharide content

Examples

Experimental program
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Effect test

Embodiment 1

[0020] A method for measuring oligochitosan content, measures the content of oligochitosan in the oligochitosan tablet, it comprises the steps:

[0021] 1) Draw the standard curve of chitosan oligosaccharides (COS): Add a certain amount of 4.00 mg / ml chitosan oligosaccharide standard solution fully dissolved in distilled water into a 10ml colorimetric tube, and then use a pipette to draw 100 μl at a concentration of 5 % phenol aqueous solution was added to the colorimetric tube, and distilled water was fixed to the scale line, and shaken to prepare COS standard solutions with concentration gradients of 0, 0.40, 0.80, 1.20, 1.60, 2.00, and 2.40 mg / ml respectively. Measure the fluorescence intensity F value on the spectrophotometer, the detection condition is λex / λem=285 / 300nm, slit 5nm, and establish the standard curve lnF=9.022-2.272X between lnF and chitosan oligosaccharide concentration X, r=0.997.

[0022] 2) Preparation of 4.00mg / ml oligochitosan tablet working solution: t...

Embodiment 2

[0027] A method for measuring oligochitosan content, measuring the content of oligochitosan in oligochitosan granule, it comprises the steps:

[0028] 1) Draw the standard curve of chitosan oligosaccharides (COS): Add a certain amount of 4.00 mg / ml chitosan oligosaccharide standard solution fully dissolved in distilled water into a 10ml colorimetric tube, and then use a pipette to draw 100 μl at a concentration of 5 % phenol aqueous solution was added to the colorimetric tube, and distilled water was fixed to the scale line, and shaken to prepare COS standard solutions with concentration gradients of 0, 0.40, 0.80, 1.20, 1.60, 2.00, and 2.40 mg / ml respectively. Measure the fluorescence intensity F value on the spectrophotometer, the detection condition is λex / λem=285 / 300nm, slit 5nm, and establish the standard curve lnF=9.008-2.248X between lnF and chitosan oligosaccharide concentration X, r=0.997.

[0029] 2) Preparation of 4.00mg / ml oligochitosan granules working solution: W...

Embodiment 3

[0034] A method for measuring the content of oligochitosan, measuring the content of oligochitosan in the oligochitosan dripping pill, it comprises the steps:

[0035] 1) Draw the standard curve of chitosan oligosaccharides (COS): Add a certain amount of 4.00 mg / ml chitosan oligosaccharide standard solution fully dissolved in distilled water into a 10ml colorimetric tube, and then use a pipette to draw 100 μl at a concentration of 5 % phenol aqueous solution was added to the colorimetric tube, and distilled water was fixed to the scale line, and shaken to prepare COS standard solutions with concentration gradients of 0, 0.40, 0.80, 1.20, 1.60, 2.00, and 2.40 mg / ml respectively. Measure the fluorescence intensity F value on the spectrophotometer, the detection condition is λex / λem=285 / 300nm, slit 5nm, and establish the standard curve lnF=8.998-2.269X between lnF and chitosan oligosaccharide concentration X, r=0.9996.

[0036] 2) Preparation of 16.00mg / ml chitosan oligosaccharid...

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Abstract

The present invention relates to the technical field of macromolecule measurement, and relates to a method for measuring the content of chitosan oligosaccharide. According to the present invention, a phenol-fluorescence spectrophotometry is used, after chitosan oligosaccharide is mixed with a phenol solution, the amino group of chitosan oligosaccharide is bound with the hydroxyl group of phenol, so that the pH value of the mixed aqueous solution is changed, and fluorescence quenching occurs to the phenol, resulting in exponential fall in fluorescence intensity, so that the concentration of the chitosan oligosaccharide is in a linear relationship with a fluorescence log value within a certain range, so as to measure the content of the chitosan oligosaccharide. The method has low costs, high sensibility, high stability and good reproducibility, the concentration measurement range is 0 to 2.4 mg / ml, the method is widely used, can be used for measuring the contents of chitosan oligosaccharide in different dosage forms, and solves the problems in the prior art of complex operations, high costs, low accuracy, and a small measurement range.

Description

technical field [0001] The invention belongs to the field of macromolecule detection and relates to a method for measuring chitosan oligosaccharide content. Background technique [0002] Oligochitosan (also known as oligomeric glucosamine, chitooligosaccharide, Chitosan Oligosaccharide, Chito-oligosaccharide, Oligochitosan) is obtained from the degradation of chitosan, a product of chitin (chitin) deacetylation, and is obtained by 2-10 It is an oligosaccharide formed by connecting glucosamine through β-1,4 glycosidic bonds. It is also the only basic aminopolysaccharide that exists in large quantities in natural sugars. It has good water solubility and is easily absorbed by objects. Oligochitosan raw materials have a wide range of sources, are safe and non-toxic, and have physiological activities such as anti-inflammatory, enhancing immunity, anti-tumor, antibacterial, scavenging free radicals, and anti-oxidation, which have attracted widespread attention from domestic and fo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
CPCG01N21/643G01N2021/6432
Inventor 苏政权曹华
Owner GUANGDONG PHARMA UNIV