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Electrophoresis method for detecting effective components of aloe polysaccharide

A technology of aloe polysaccharides and active ingredients, applied in the field of biochemical analysis, can solve the problems of large molecular weight of acetylated mannan, complicated identification and detection process, inability to carry out polysaccharide structure, etc., and achieve high sensitivity, low cost, and shortened experiment time Effect

Pending Publication Date: 2022-03-04
广州蔚析生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Due to the large molecular weight and complex structure of acetylated mannan, the active ingredient of aloe polysaccharide, its identification and detection process is complicated and the cost is high. (or differential detector)
However, polysaccharide analysis in the traditional production field often uses methods such as hydrolysis to measure reducing sugars, phenol-sulfuric acid colorimetry, and anthrone-sulfuric acid colorimetry. However, the polysaccharide content of these methods is only determined by the degraded monosaccharide residues and cannot be determined Polysaccharide structure identification

Method used

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  • Electrophoresis method for detecting effective components of aloe polysaccharide
  • Electrophoresis method for detecting effective components of aloe polysaccharide
  • Electrophoresis method for detecting effective components of aloe polysaccharide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] An electrophoresis method for detecting active ingredients of aloe polysaccharides, comprising the following steps:

[0040] (1) Peel the leaves of Aloe vera whose age is 12 months, June and March, and smash them to obtain sample solutions of fresh aloe polysaccharides A1, A2 and A3; the dry powder of aloe polysaccharides is according to the solid-liquid ratio of 1:60 and 1:40, Distilled water was added to redissolve to obtain aloe polysaccharide dry powder P1 and P2 sample solutions; the aloe product was diluted with distilled water at a ratio of 1:1 to obtain aloe product C1 sample solution.

[0041] (2) Take 5ml of the above sample solution, add 10mg of FITC (fluorescein isothiocyanate) reagent, adjust the pH value to 9.0 with sodium hydroxide, and react on a shaker at 150r / min at 45°C for 2h.

[0042] (3) Take 1ml of the above-mentioned solution that has completed the reaction, add 1ml of medical glycerin, mix evenly to obtain a test solution, and set aside.

[004...

Embodiment 2

[0056] An electrophoresis method for detecting active ingredients of aloe polysaccharides, comprising the following steps:

[0057] (1) Peel the fresh aloe leaves and smash them to obtain the fresh aloe polysaccharide A1 sample solution; the aloe polysaccharide dry powder is redissolved in distilled water according to the solid-liquid ratio of 1:40 to obtain the aloe polysaccharide dry powder P1 sample solution; the aloe product is according to the ratio of 1:40 1 ratio, distilled water was added to dilute to obtain the aloe product C1 sample solution.

[0058] (2) O-acetyl color development: Accurately measure 1ml of the sample and place it in a test tube, add 2ml of freshly prepared alkaline hydroxylamine solution, shake well, let it stand at room temperature for 4 minutes, add 1ml of 4mol / L hydrochloric acid solution, shake well, Add 1ml of 0.37mol / L ferric chloride solution and shake well.

[0059] (3) Take 1ml of the above-mentioned solution that has completed the reacti...

Embodiment 3

[0065] In this embodiment, the gel of the aloe polysaccharide electrophoresis band obtained in Example 1 is taken out, and is passed through high-performance liquid gel permeation chromatography (HPGPC), ion chromatography (IC5000+), gas chromatography-mass spectrometry (GC-MS), nuclear magnetic resonance spectroscopy ( NMR) and other methods were used to analyze the monosaccharide composition, methylation and NMR of the samples.

[0066] Such as image 3 As shown in Table 1, the monosaccharide composition was measured by ion spectrometer, and the result showed that the monosaccharide composition of the polysaccharide in the electrophoresis strip was mainly mannose.

[0067] Table 1 Monosaccharide composition of polysaccharides in the gel

[0068]

[0069]

[0070] As shown in Table 2, the connection mode of the polysaccharide sample was measured by GC-MS after derivatization such as methylation, and the results showed that the connection mode of the polysaccharide in t...

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Abstract

The invention discloses an electrophoresis method for detecting effective components of aloe polysaccharide. The electrophoresis method comprises the following steps: S1, preparing an aloe polysaccharide solution from an aloe sample to be detected; s2, taking the aloe polysaccharide solution, and performing dyeing treatment to obtain a dyed sample; s3, performing electrophoresis treatment on the dyed sample to obtain a polysaccharide electrophoresis band; and S4, recording a polysaccharide electrophoresis band, and determining the content of the effective component of the aloe polysaccharide in the aloe sample to be detected. According to the electrophoresis method for detecting the effective components of the aloe polysaccharide, the effective components of the aloe polysaccharide are marked by adopting the color developing substance, so that the steps of polysaccharide precipitation, dyeing, rinsing and the like in the conventional polysaccharide electrophoresis process are simplified, the experiment time is shortened, and the sensitivity and the accuracy of polysaccharide detection are improved.

Description

technical field [0001] The invention belongs to the field of biochemical analysis, in particular to an electrophoresis method for detecting effective components of aloe polysaccharides. Background technique [0002] Aloe polysaccharide is one of the main components of aloe, accounting for about 18%-30% of the dried aloe juice. The main active ingredient of aloe polysaccharide is acetylated mannan (acemannan) formed by linking linear β-(1,4)-D-mannosyl units. It can not only be used as an energy source for intracellular metabolism, but also as a ligand for regulating immunity, a regulator of intercellular transmission, and an effective inhibitor of pathogens attached to healthy cells, etc., and has significant functions such as immune regulation and anti-tumor . [0003] Due to the large molecular weight and complex structure of acetylated mannan, the active ingredient of aloe polysaccharide, its identification and detection process is complicated and the cost is high. (or...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/447G01N1/28
CPCG01N27/447G01N1/28
Inventor 李杰晶李敏和磊和谐李迪
Owner 广州蔚析生物科技有限公司