CN108586631A - A kind of extracting method of Dendrobium officinale polysaccharide - Google Patents
A kind of extracting method of Dendrobium officinale polysaccharide Download PDFInfo
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- CN108586631A CN108586631A CN201810664221.7A CN201810664221A CN108586631A CN 108586631 A CN108586631 A CN 108586631A CN 201810664221 A CN201810664221 A CN 201810664221A CN 108586631 A CN108586631 A CN 108586631A
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- dendrobium officinale
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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Abstract
The invention discloses a kind of extracting methods of Dendrobium officinale polysaccharide, are related to active components of plants extractive technique field.The extracting method is：Fresh dendrobium candidum is cleaned, freeze-drying, is crushed, is impregnated in ethyl alcohol, volatilizes solvent, obtain the dregs of a decoction, it is 1 that the dregs of a decoction and water, which are pressed solid-liquid ratio,:Extracting solution is concentrated into the 1/3 of original volume by 20 40 ratio mixing, constant temperature water bath extraction, obtain concentrate, ethyl alcohol is added into concentrate, stands 8 10h, centrifugation, precipitation is taken to be dissolved in distilled water, 0.05 0.1% proteolytic enzyme, water enzyme digestion are added into polysaccharide stoste for much glycogen liquid, enzymolysis liquid is obtained, enzymolysis liquid, chloroform, n-butanol are pressed 25:4:1 mixing shakes, centrifugation, after removing jello, is added activated carbon, filtering, concentration, it is dry to get.The present invention effectively increases the recovery rate and purity of Dendrobium officinale polysaccharide, and enhance bioactivity by optimizing extracting method.
Description
【Technical field】
The present invention relates to active components of plants extractive technique fields, and in particular to a kind of extraction side of Dendrobium officinale polysaccharide
Method.
【Background technology】
Dendrobium candidum is China's tradition rare Chinese herbal medicine, has more than 2000 years medicinal history, main chemical compositions packet
Including alkaloids, polysaccharide, Polyphenols etc., wherein polysaccharide is one of most effective ingredient in dendrobium candidum, pharmacological research surface,
Dendrobium officinale polysaccharide has the pharmacological activity such as stronger anti-aging, anti-oxidant, antitumor and enhancing immunity of organisms.With in recent years
Carry out the enhancing of people's health care consciousness, the demand of Dendrobium officinale polysaccharide is being continuously increased, and market demand is vigorous, and supply falls short of demand,
It need to be further improved Dendrobium officinale polysaccharide extractive technique, increase extracted amount.At present Dendrobium officinale polysaccharide extraction exist containing protein,
The impurity such as pigment are more, and the low problem of purity, excessive impurity can influence its bioactivity, quality is made to reduce.
【Invention content】
In view of the above-mentioned problems, the technical problem to be solved in the present invention is to provide a kind of extracting method of Dendrobium officinale polysaccharide,
This method effectively increases the recovery rate and purity of Dendrobium officinale polysaccharide, and enhance bioactivity by optimizing extracting method.
In order to solve the above technical problems, the technical solution adopted by the present invention is as follows：
A kind of extracting method of Dendrobium officinale polysaccharide, includes the following steps：
(1) fresh dendrobium candidum is cleaned, is freeze-dried, crushed, cross 40-60 mesh sieve, it is 90- to be placed in volume fraction
3-5h is impregnated in 95% ethyl alcohol, volatilizes solvent, obtains the dregs of a decoction, it is spare；
(2) it is 1 by solid-liquid ratio by the above-mentioned dregs of a decoction and water:After the ratio mixing of 20-40, constant temperature water bath extracts 100-
180min repeats extraction 2-3 times, merges extracting solution, extracting solution is concentrated into the 1/3 of original volume, concentrate is obtained, into concentrate
The ethyl alcohol that volumetric concentration is 60-70% is added, stands 8-10h, centrifugation takes precipitation to be dissolved in distilled water, much glycogen liquid, standby
With；
(3) proteolytic enzyme of 0.05-0.1%, the water enzyme digestion at 35 DEG C -40 DEG C are added into above-mentioned polysaccharide stoste
1-2h obtains enzymolysis liquid, and enzymolysis liquid, chloroform, n-butanol are pressed 25:4:1 mixing, shakes 20-30min, and centrifugation removes jello
Afterwards, be added activated carbon, filtering, by filtrate concentration, it is dry to get.
Further, in step (1), the temperature of the freeze-drying is -30 DEG C~-20 DEG C.
Further, in step (2), the bath temperature is 70 DEG C -90 DEG C.
Further, it in step (2), after ethyl alcohol is added into concentrate, is stood at 1 DEG C -5 DEG C.
Further, in step (3), the proteolytic enzyme is papain or pronase.
Further, in step (3), the activated carbon addition is the 0.1-0.5% of filtrate weight after removing jello.
Compared with prior art, the present invention has the advantages that：
The present invention effectively increases the recovery rate and purity of Dendrobium officinale polysaccharide, and enhance by optimizing extracting method
Bioactivity.Through experiment, 31.88%-33.12%, polyoses content 79.2%- are up to using the method for the present invention recovery rate
Absorbance under 81.9%, 2mg/mL concentration is 1.82-1.97.Impurity, enzyme are removed using enzymatic isolation method combination activated carbon adsorption
Protein in the removable polysaccharide of solution, and the impurity after the adsorbable pigment of activated carbon and other enzymolysis, and then improve purity.
【Specific implementation mode】
The specific implementation mode of the present invention is described further with reference to embodiments.
Embodiment 1
A kind of extracting method of Dendrobium officinale polysaccharide of the present embodiment, includes the following steps：
(1) fresh dendrobium candidum is cleaned, is freeze-dried at -30 DEG C, crushed, crossed 40 mesh sieve, be placed in volume fraction
To impregnate 3h in 90% ethyl alcohol, solvent is volatilized, the dregs of a decoction are obtained, it is spare.
(2) it is 1 by solid-liquid ratio by the above-mentioned dregs of a decoction and water:After 20 ratio mixing, constant temperature water bath extracts at 70 DEG C
100min repeats extraction 2 times, merges extracting solution, extracting solution is concentrated into the 1/3 of original volume, obtains concentrate, adds into concentrate
Enter the ethyl alcohol that volumetric concentration is 60%, 8h is stood at 1 DEG C, centrifuges, take precipitation to be dissolved in distilled water, much glycogen liquid, it is spare.
(3) 0.05% papain is added into above-mentioned polysaccharide stoste, the water enzyme digestion 1h at 35 DEG C must be digested
Enzymolysis liquid, chloroform, n-butanol are pressed 25 by liquid:4:1 mixing, shakes 20min, and after removing jello, activated carbon, mistake is added in centrifugation
Filter, by filtrate concentration, it is dry to get；The activated carbon addition is 0.1% of filtrate weight after removing jello.
Embodiment 2
A kind of extracting method of Dendrobium officinale polysaccharide of the present embodiment, includes the following steps：
(1) fresh dendrobium candidum is cleaned, is freeze-dried at -20 DEG C, crushed, crossed 60 mesh sieve, be placed in volume fraction
To impregnate 5h in 95% ethyl alcohol, solvent is volatilized, the dregs of a decoction are obtained, it is spare.
(2) it is 1 by solid-liquid ratio by the above-mentioned dregs of a decoction and water:After the ratio mixing of 20-40, constant temperature water bath extracts at 90 DEG C
180min repeats extraction 3 times, merges extracting solution, extracting solution is concentrated into the 1/3 of original volume, obtains concentrate, adds into concentrate
Enter the ethyl alcohol that volumetric concentration is 70%, 10h is stood at 5 DEG C, centrifuges, take precipitation to be dissolved in distilled water, much glycogen liquid, it is standby
With.
(3) 0.1% pronase is added into above-mentioned polysaccharide stoste, water enzyme digestion 2h, obtains enzymolysis liquid at 40 DEG C,
Enzymolysis liquid, chloroform, n-butanol are pressed 25:4:1 mixing shakes 30min, and centrifugation after removing jello, is added activated carbon, filters,
By filtrate concentration, it is dry to get；The activated carbon addition is 0.5% of filtrate weight after removing jello.
Embodiment 3
A kind of extracting method of Dendrobium officinale polysaccharide of the present embodiment, includes the following steps：
(1) fresh dendrobium candidum is cleaned, is freeze-dried at -25 DEG C, crushed, crossed 50 mesh sieve, be placed in volume fraction
To impregnate 4h in 92% ethyl alcohol, solvent is volatilized, the dregs of a decoction are obtained, it is spare.
(2) it is 1 by solid-liquid ratio by the above-mentioned dregs of a decoction and water:After the ratio mixing of 20-40, constant temperature water bath extracts at 80 DEG C
150min repeats extraction 3 times, merges extracting solution, extracting solution is concentrated into the 1/3 of original volume, obtains concentrate, adds into concentrate
Enter the ethyl alcohol that volumetric concentration is 65%, 9h is stood at 3 DEG C, centrifuges, take precipitation to be dissolved in distilled water, much glycogen liquid, it is spare.
(3) 0.08% papain is added into above-mentioned polysaccharide stoste, the water enzyme digestion 1.5h at 38 DEG C must be digested
Enzymolysis liquid, chloroform, n-butanol are pressed 25 by liquid:4:1 mixing, shakes 25min, and after removing jello, activated carbon, mistake is added in centrifugation
Filter, by filtrate concentration, it is dry to get；The activated carbon addition is 0.2% of filtrate weight after removing jello.
Embodiment 4
A kind of extracting method of Dendrobium officinale polysaccharide of the present embodiment, includes the following steps：
(1) fresh dendrobium candidum is cleaned, is freeze-dried at -23 DEG C, crushed, crossed 50 mesh sieve, be placed in volume fraction
To impregnate 4.5h in 93% ethyl alcohol, solvent is volatilized, the dregs of a decoction are obtained, it is spare.
(2) it is 1 by solid-liquid ratio by the above-mentioned dregs of a decoction and water:After the ratio mixing of 20-40, constant temperature water bath extracts at 85 DEG C
170min repeats extraction 2 times, merges extracting solution, extracting solution is concentrated into the 1/3 of original volume, obtains concentrate, adds into concentrate
Enter the ethyl alcohol that volumetric concentration is 67%, 8.5h is stood at 4 DEG C, centrifuges, take precipitation to be dissolved in distilled water, much glycogen liquid, it is standby
With.
(3) 0.06% papain is added into above-mentioned polysaccharide stoste, the water enzyme digestion 1.5h at 37 DEG C must be digested
Enzymolysis liquid, chloroform, n-butanol are pressed 25 by liquid:4:1 mixing, shakes 28min, and after removing jello, activated carbon, mistake is added in centrifugation
Filter, by filtrate concentration, it is dry to get；The activated carbon addition is 0.4% of filtrate weight after removing jello.
Contrast experiment：
Group 1, group 2, group 3, group 4 use embodiment 1, embodiment 2, embodiment 3, the method for embodiment 4 to extract polysaccharide respectively；
Contrast groups 1：Step (3) unused enzymatic treatment, but be directly added into chloroform, n-butanol shaking, other steps with reality
It is identical to apply example 3；
Contrast groups 2：Step (3) only use enzymatic treatment, be not added chloroform, n-butanol shaking, other steps with 3 phase of embodiment
Together.
Each group polysaccharide extract rate is counted, and using High Performance Gel Permeation Chromatography respectively to the homogeneity of each group dendrobium polysaccharide
It is identified；Reducing power is judged by measurement absorbance to judge the bioactivity of each group.As a result it is：
(1) polysaccharide extract rate is respectively 31.88%, 32.11%, 33.12%, 31.97% in group 1, group 2, group 3, group 4,
1 recovery rate of contrast groups is 21.9%, and 2 recovery rate of contrast groups is 23.1%, and therefore, extracting method of the present invention improves Polyose extraction
Rate.
(2) chromatography result is：Group 1, group 2, group 3,4 polyoses contents of group are respectively 79.2%, 80.2%, 81.9%,
79.8%, 1 polyoses content of contrast groups is 50.1%, and 2 polyoses content of contrast groups is 48.9%, and therefore, extracting method of the present invention is pure
Degree is higher.
(3) absorbance measurement result is：When polysaccharide concentration is 2mg/mL, group 1, group 2, group 3, the absorbance point for organizing 4
Not Wei 1.82,1.88,1.97,1.84, the absorbances of contrast groups 1 is 0.91, and the absorbance of contrast groups 2 is 1.02, therefore, this hair
Polysaccharide bioactivity can be improved in bright extracting method, and (absorbance is higher, and reducing power is stronger, and oxidation resistance is stronger, and bioactivity is got over
By force).
Above description is the detailed description for the present invention preferably possible embodiments, but embodiment is not limited to this hair
Bright patent claim, it is all the present invention suggested by technical spirit under completed same changes or modifications change, should all belong to
In the covered the scope of the claims of the present invention.
Claims (6)
1. a kind of extracting method of Dendrobium officinale polysaccharide, it is characterised in that：Include the following steps：
(1) fresh dendrobium candidum is cleaned, is freeze-dried, crushed, cross 40-60 mesh sieve, it is 90-95%'s to be placed in volume fraction
3-5h is impregnated in ethyl alcohol, volatilizes solvent, obtains the dregs of a decoction, it is spare；
(2) it is 1 by solid-liquid ratio by the above-mentioned dregs of a decoction and water:After the ratio mixing of 20-40, constant temperature water bath extracts 100-180min, weight
It extracts 2-3 times again, merges extracting solution, extracting solution is concentrated into the 1/3 of original volume, concentrate is obtained, volume is added into concentrate
The ethyl alcohol of a concentration of 60-70% stands 8-10h, and centrifugation takes precipitation to be dissolved in distilled water, much glycogen liquid, spare；
(3) proteolytic enzyme of 0.05-0.1%, the water enzyme digestion 1- at 35 DEG C -40 DEG C are added into above-mentioned polysaccharide stoste
2h obtains enzymolysis liquid, and enzymolysis liquid, chloroform, n-butanol are pressed 25:4:1 mixing shakes 20-30min, centrifuges, after removing jello,
Be added activated carbon, filtering, by filtrate concentration, it is dry to get.
2. a kind of extracting method of Dendrobium officinale polysaccharide according to claim 1, it is characterised in that：It is described in step (1)
The temperature of freeze-drying is -30 DEG C~-20 DEG C.
3. a kind of extracting method of Dendrobium officinale polysaccharide according to claim 1, it is characterised in that：It is described in step (2)
Bath temperature is 70 DEG C -90 DEG C.
4. a kind of extracting method of Dendrobium officinale polysaccharide according to claim 1, it is characterised in that：In step (2), to dense
After ethyl alcohol is added in contracting liquid, stood at 1 DEG C -5 DEG C.
5. a kind of extracting method of Dendrobium officinale polysaccharide according to claim 1, it is characterised in that：It is described in step (3)
Proteolytic enzyme is papain or pronase.
6. a kind of extracting method of Dendrobium officinale polysaccharide according to claim 1, it is characterised in that：It is described in step (3)
Activated carbon addition is the 0.1-0.5% of filtrate weight after removing jello.
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Cited By (9)
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| CN109336988A (en) * | 2018-10-19 | 2019-02-15 | 贵州真阳纳米医药科技有限公司 | A kind of extracting method of dendrobium candidum |
| CN109651532A (en) * | 2018-11-21 | 2019-04-19 | 浙江农林大学 | A kind of dendrobium candidum Glucomannan |
| CN110343191A (en) * | 2019-08-27 | 2019-10-18 | 平顶山学院 | The extraction of Funiu Shan Mountain Dendrobium nobile polysaccharide and the method for preparing solid beverage |
| CN111196865A (en) * | 2020-03-09 | 2020-05-26 | 杭州奥得科技有限公司 | Extraction method of active ingredients of dendrobium officinale |
| CN112206203A (en) * | 2020-10-15 | 2021-01-12 | 云南金九地生物科技有限公司 | Preparation method of dendrobium officinale polysaccharide injection |
| CN112940292A (en) * | 2021-02-25 | 2021-06-11 | 上海应用技术大学 | Dendrobium officinale polysaccharide hydrogel as well as preparation method and application thereof |
| CN113480675A (en) * | 2021-08-19 | 2021-10-08 | 遵义医科大学 | Extraction method and application of dendrobium nobile polysaccharide |
| CN113512128A (en) * | 2021-05-13 | 2021-10-19 | 海南云皓生物科技有限公司 | Dendrobium polysaccharide and preparation method thereof |
| CN115919967A (en) * | 2022-12-01 | 2023-04-07 | 遵义医科大学 | Dendrobium nobile extract and application thereof in preparing anti-vascular aging drugs |
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109336988A (en) * | 2018-10-19 | 2019-02-15 | 贵州真阳纳米医药科技有限公司 | A kind of extracting method of dendrobium candidum |
| CN109651532A (en) * | 2018-11-21 | 2019-04-19 | 浙江农林大学 | A kind of dendrobium candidum Glucomannan |
| CN109651532B (en) * | 2018-11-21 | 2021-01-26 | 浙江农林大学 | Dendrobium officinale glucomannan |
| CN110343191A (en) * | 2019-08-27 | 2019-10-18 | 平顶山学院 | The extraction of Funiu Shan Mountain Dendrobium nobile polysaccharide and the method for preparing solid beverage |
| CN111196865A (en) * | 2020-03-09 | 2020-05-26 | 杭州奥得科技有限公司 | Extraction method of active ingredients of dendrobium officinale |
| CN111196865B (en) * | 2020-03-09 | 2022-08-12 | 杭州奥得科技有限公司 | Extraction method of active ingredients of dendrobium officinale |
| CN112206203A (en) * | 2020-10-15 | 2021-01-12 | 云南金九地生物科技有限公司 | Preparation method of dendrobium officinale polysaccharide injection |
| CN112940292A (en) * | 2021-02-25 | 2021-06-11 | 上海应用技术大学 | Dendrobium officinale polysaccharide hydrogel as well as preparation method and application thereof |
| CN113512128A (en) * | 2021-05-13 | 2021-10-19 | 海南云皓生物科技有限公司 | Dendrobium polysaccharide and preparation method thereof |
| CN113480675A (en) * | 2021-08-19 | 2021-10-08 | 遵义医科大学 | Extraction method and application of dendrobium nobile polysaccharide |
| CN115919967A (en) * | 2022-12-01 | 2023-04-07 | 遵义医科大学 | Dendrobium nobile extract and application thereof in preparing anti-vascular aging drugs |
| CN115919967B (en) * | 2022-12-01 | 2024-02-06 | 遵义医科大学 | Dendrobium nobile extract and application thereof in preparation of anti-vascular aging drugs |
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Application publication date: 20180928 |