A column chromatography separation method and extraction method of Polygonatum polysaccharide

A Polygonatum polysaccharide and separation method technology, applied in the field of column chromatography separation and extraction of Polygonatum polysaccharide, can solve the problems of single type, low product purity, poor purification effect, etc., achieve utilization rate and purity improvement, high product purity, Good effect of removing impurities

Active Publication Date: 2019-02-26
LONGHUA HOSPITAL SHANGHAI UNIV OF TRADITIONAL CHINESE MEDICINE
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0008] The technical problem to be solved by this invention is in order to overcome the product purity of the extracting technique of Polygonatum polysaccharide and purification process in the prior art is not high, and can only extract the small molecule sugar (or oligosaccharide) in Polygonatum polysaccharide, kind is single, and Polygonatum polysaccharides with a wide range of polysaccharide molecular weights cannot be purified, resulting in low utilization of Polygonatum polysaccharides, or because only water is used for elution, so that the removal of impurities and purification effects are poor, often need to be combined with other separation methods for further follow-up Purification, cumbersome steps, increased cost consumption and other defects, and provide a column chromatography separation method and extraction method of Polygonatum polysaccharide

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  • A column chromatography separation method and extraction method of Polygonatum polysaccharide
  • A column chromatography separation method and extraction method of Polygonatum polysaccharide

Examples

Experimental program
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Effect test

Embodiment 1

[0044] The extraction of embodiment 1 polygonatum polysaccharide

[0045] Mix 1 gram of Rhizoma Polygonatum with 8 grams of water, heat to boil and keep boiling for 60 minutes, separate the Rhizoma Rhizoma Polygonatum from the decoction, then mix the separated Rhizoma Rhizoma Polygonatum with another 8 grams of water, and repeat the above The process of heating and boiling and separating the Rhizoma Rhizoma Rhizoma Polygonatum and the decocted liquid is performed twice, and all the decocted liquids are combined.

[0046] Boil the combined decoction on high heat to concentrate, and when the ratio of medicine to liquid is 1:1, stop the concentration.

[0047] Adding the mass percent of ethanol in the concentrated medicinal solution is 95% ethanol aqueous solution, so that the mass percent of ethanol in the mixed solution after adding is 75%, (slowly and quickly stirring when adding ethanol aqueous solution helps to prevent the occurrence of coprecipitation) , after stirring eve...

Embodiment 3

[0051] The drawing of embodiment 3 standard curve

[0052] 1. References for the determination of the maximum absorption wavelength of Polygonatum polysaccharide [Imbety, Bourne Y, Cambillau C etal, Oligosaccharide Conformation in Protein Carbohydrate Computers, Adv. Biophys, Chem, 1993, 3: 71~117.] or [Yang Li, Wang Zhihua , Tao Jiansheng, etc. Comparison of Astragalus Polysaccharide Content Determination Methods in Radix Astragali [J]. Chinese Journal of Pharmaceutical Industry, 2005, 36 (9): 562-563], and obtained the ultraviolet absorption spectrum of Polygonatum polysaccharide, see figure 1 , where 625nm is the maximum absorption wavelength.

[0053] The determination of sugar content in the following examples in the present invention is all carried out at this wavelength.

[0054] 2. Preparation of glucose standard solution

[0055] Accurately weigh an appropriate amount of glucose standard substance that has been dried to a constant value, and add ultrapure water to d...

Embodiment 4

[0061] The purification of embodiment 4 polygonatum polysaccharide

[0062] NKA-9 macroporous adsorption resin was used as a filler, and the pretreatment in Example 2 was performed first, and then 2.0 g of the pretreated resin was wet-packed with ultrapure water. The column length was 50 cm and the column diameter was 2 cm.

[0063] The Polygonatum polysaccharide crude product obtained through the method of Example 1 is formulated with ultrapure water into a medicinal solution of crude Polygonatum polysaccharide with a concentration of 3.90 mg / mL, and 10 mL of it is taken for loading, and the loading flow rate is 2 mL / min; The resting time is 12 hours; then the eluent is 30% ethanol aqueous solution for elution; the amount of eluent is 120mL; the elution flow rate is 2mL / min. Collect the entire eluate in a weighed clean glass.

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Abstract

The invention discloses a Polygonatum sibiricum polysaccharide column chromatography separation method and extraction method. The Polygonatum sibiricum polysaccharide column chromatography separation method comprises the following step: carrying out column chromatography separation on Polygonatum sibiricum polysaccharides with a macroporous adsorbent resin, wherein the sample loading quantity is 10ml per 100-200ml column volume, the concentration of the Polygonatum sibiricum polysaccharides in the loaded liquor is 3.5-4.5mg / ml; the eluting agent is a 10-50 wt% ethanol water solution, the consumption of the eluting agent is 80-150ml per 100-200ml column volume, and the eluting flow rate is 1-2ml / minute. The Polygonatum sibiricum polysaccharide column chromatography separation method and extraction method are simple to operate, and have the advantages of favorable repetitiveness and high product purity; the separated Polygonatum sibiricum polysaccharides are varied and not limited to oligosaccharides; and thus, the Polygonatum sibiricum polysaccharide column chromatography separation method and extraction method are more suitable for industrial production.

Description

technical field [0001] The present invention specifically relates to a column chromatography separation method and extraction method of polygonatum polysaccharide. Background technique [0002] Polygonatum is one of the commonly used clinical traditional Chinese medicines, and its main active ingredient is Polygonatum polysaccharide. Polygonatum polysaccharides exert a variety of beneficial physiological regulation effects on the human body: its anti-tumor effect can effectively inhibit the occurrence and development of tumors; its anti-aging effect can be used as an effective anti-aging preparation for the human body; its Immunomodulatory effect can effectively enhance the immune function of immune cells; in addition, it also has antibacterial, anti-atherosclerosis, and regulation of blood sugar and blood lipids (He Caitong, Li Wen. Research progress in the pharmacological efficacy of Polygonatum Polysaccharides. New Chinese Medicine, 2014, 46(3):196-199). Therefore, poly...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/00
Inventor 王立芳毛泉明徐振晔张宁高嵩司海龙王鹤潼
Owner LONGHUA HOSPITAL SHANGHAI UNIV OF TRADITIONAL CHINESE MEDICINE
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