CN109200197A - A kind of dendrobium candidum extract and preparation method thereof - Google Patents
A kind of dendrobium candidum extract and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of dendrobium candidum extracts, the extract includes polysaccharide, stilbene class, flavones and lignanoid, and wherein the percentage composition of polysaccharide is 45 ~ 70%, and the percentage composition of stilbene class is 8 ~ 15%, the percentage composition of flavones is 10 ~ 22%, and the percentage composition of lignanoid is 11 ~ 18%.The extract is used for inhibitory effect of the Inhibition test observation to tumour cell of tumour cell; the result shows that extract has preferable lethal effect to tumour cell; the growth of tumour cell can effectively be inhibited; simultaneously; the preparation method of extract is simple, recovery rate is high, at low cost, safety and environmental protection is pollution-free, can carry out large-scale production.
Description
Technical field
The present invention relates to a kind of medicating active ingredients preparation technical fields, and in particular to a kind of dendrobium candidum effective extract
And preparation method thereof.
Background technique
Dendrobium candidum (scientific name: Dendrobium officinale Kimura et Migo), also known as: ribbed hedyotis herb, Yunnan
Iron sheet.Belong to micro- specific item, the perennial herbaceous plant that grows nonparasitically upon another plant of orchid family, sweet in flavor, cold nature.Dendrobium candidum also there are many effective activity at
Point, it specifically includes that dendrobium polysaccharide, stilbene class, flavones, lignanoid, amino acid etc., there is the effect of reinforcing stomach reg fluid, nourishing Yin and clearing heat.
Shennong's Herbal is classified as top grade, it is described as first of " Chinese nine big mesonas " by Taoism regimen classics " Taoist Scriptures ", civil title
Dendrobium candidum is " help mesona ", " bigflower clinopodium herb ", and as the superfine product of health care, ancient Chinese medicine doctor praises highly letter extremely.
By the way that the study found that dendrobium candidum has multiple efficacies, such as: enhancing immune function, protect liver cholagogue, resists anti-aging
Rheumatism, reduction blood pressure and blood lipoid, inhibition tumour etc., dendrobium candidum is pernicious to lung cancer, oophoroma and promyelocytic leukemia etc.
Certain cells of tumour have killing effect, inhibit the activity of tumour cell, the auxiliary that clinically can be used for malignant tumour is controlled
It treats, improves patient's immunity, improve life quality, extend life span.
Summary of the invention
The object of the present invention is to provide a kind of dendrobium candidum extract, extract includes polysaccharide, stilbene class, flavones and the wooden rouge
Element can be used for the adjuvant treatment of tumour cell as traditional Chinese medicinal components.
Technical solution: including polysaccharide, stilbene class, flavones and lignanoid the invention discloses a kind of dendrobium candidum extract,
The percentage composition of middle polysaccharide is 45 ~ 70%, and the percentage composition of stilbene class is 8 ~ 15%, and the percentage composition of flavones is 10 ~ 22%, lignanoid
Percentage composition be 11 ~ 18%.
It include polysaccharide, stilbene class, flavones and lignanoid, wherein the hundred of polysaccharide in dendrobium candidum extract of the present invention
Dividing content is 63 ~ 65%, and the percentage composition of stilbene class is 10 ~ 12%, and the percentage composition of flavones is 11 ~ 14%, the percentage composition of lignanoid
It is 12 ~ 15%.
The method for preparing dendrobium candidum extract of the present invention, includes the following steps:
(1) degreasing
30 ~ 60 meshes are crossed after dendrobium candidum is crushed, and 10 ~ 15 times of organic solvent is added thereto and impregnates 1 ~ 2h, is then refluxed for 2
~ 4 1.5 ~ 2.5h every time remove organic solvent and obtain the dregs of a decoction after filtering.
(2) Polyose extraction
The distilled water of 10 ~ 20 times of addition in the dregs of a decoction is obtained to step (1), is decocted 2 ~ 4 times, 2 ~ 4h is decocted every time, by decoction liquor mistake
The dregs of a decoction are isolated in filter, and decoction liquor is concentrated 10 times and obtains concentrate, and ethyl alcohol is added into concentrate so that the content of ethyl alcohol be 60 ~
80%, it stirs evenly and stands overnight, isolate ethanol water, obtain alcohol sediment, alcohol hypostasis is filtered with organic solvent
Washing, is dried under reduced pressure, obtains polyoses extract.
(3) stilbene class, flavones and lignanoid extract
95% ethyl alcohol of the filter residue of step (2) separation is extracted 3 ~ 5 times again at room temperature, 10 ~ 15h, merges ethyl alcohol leaching every time
Extract, then merge with the ethanol water isolated in step (2), by Rotary Evaporators, the solution is concentrated, then to
20mL aqueous solution is added in concentrate, is first extracted 3 ~ 5 times with chloroform, isolates chloroform phase, then be extracted with ethyl acetate 3 ~ 5 times,
Ethyl acetate phase is isolated, and retains water phase.
(4) stilbene class, flavones and lignanoid isolate and purify
Step (3) are obtained into chloroform phase and ethyl acetate phase is isolated and purified with silica gel column chromatography column respectively, elution is collected and evaporates
Point, water phase is isolated and purified with aperture resin gel rubber column gel column, eluting fraction is collected, the eluent of three-phase is merged, is obtained
To stilbene class, flavones and the extract of lignanoid.
(5) dry
The extract that step (4) is obtained removes organic solvent by rotary evaporation, is then freeze-dried Liquid Residue, obtains
To stilbene class, flavones and the freeze-dried powder of lignanoid.
The method of the present invention for preparing dendrobium candidum extract, the organic solvent in the step (1) are petroleum
The one or more of ether, ethyl acetate, N,N-dimethylformamide.
The method of the present invention for preparing dendrobium candidum extract, silica gel column chromatography passes through ladder in the step (4)
Eluting fraction is isolated in degree elution, and eluent is the petroleum ether of different volumes proportion and the mixed liquor of ethyl acetate, is eluted suitable
Sequence is that the volume ratio of petroleum ether and ethyl acetate is 1:0,20:1,10:1,5:1,2:1 and 1:1.
The method of the present invention for preparing dendrobium candidum extract, aperture resin gel column layer in the step (4)
Analysis isolates eluting fraction by gradient elution, and eluent is the methanol solution of different percentage compositions, wherein first is used in each elution
The mass concentration of alcoholic solution is followed successively by 0%, 10%, 30%, 50%, 75% and 95%.
Dendrobium candidum extract of the present invention is preparing the application in anti-tumor drug or health care product.
The utility model has the advantages that the present invention provides a kind of dendrobium candidum extract, the extract include polysaccharide, stilbene class, flavones and
The extract is used for inhibitory effect of the Inhibition test observation to tumour cell of tumour cell, the results showed that extract by lignanoid
Object has preferable lethal effect to tumour cell, can effectively inhibit the growth of tumour cell, meanwhile, the preparation of extract
Method is simple, recovery rate is high, at low cost, safety and environmental protection is pollution-free, can carry out large-scale production.
Specific embodiment
(1) measurement of the polysaccharide content
The content that polysaccharide is measured by UV-VIS spectrophotometry, accurately weighs after DEXTROSE ANHYDROUS adds water and is configured to
The solution of 90ug/mL, precision measure glucose control product solution 0mL, 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, respectively
It is placed in 10mL tool plug test tube, respectively plus water is mended to 1.0mL, is added after 5% phenol solution of 1mL shakes up, is added in every pipe
5mL sulfuric acid solution shakes up, and is placed in boiling water bath and heats 20min, takes out cooling 5min in postposition ice bath, measures the suction of 488nm
Light value is that ordinate draws standard curve by abscissa light absorption value of concentration of glucose, and regression equation is Y=0.16X+0.27, R2
=0.9992。
Test sample 0.1g accurately is weighed, is placed in 250mL measuring bottle, is diluted with water to scale, it is accurate after shaking up to measure solution
2mL is placed in 15mL centrifuge tube, and 10mL dehydrated alcohol is added in precision, and 1h is refrigerated after shaking up, and centrifugation removal supernatant, sinks after taking-up
It forms sediment with 80% ethanol washing 2 times, liquid, precipitating plus hot water dissolving is discarded supernatant after centrifugation, is transferred in 25mL measuring bottle after cooling,
Scale is added water to shake up.According to the glucose content of the measuring method measurement sample of the above standard curve, according to the extinction of measurement
Be worth from the amount for reading DEXTROSE ANHYDROUS in test solution on standard curve, calculate to obtain the final product.
(2) measurement of total stilbene compound content
Bibliography [Ni Wangdong, full Rayleigh, Li Zhiming, Lu Hong plum ultraviolet spectrophotometry measure in polygonum cuspidate simultaneously stilbene class with
Anthraquinone analog compound, Chemical Engineer, 2006,125 (2): 35-36,39] method measure the content of total stilbene compound.With
Stilbene compound tree peony alcohol A measures the light absorption value at 338nm using UV-VIS spectrophotometry as standard reference material:
10mg tree peony alcohol A is weighed, PH=8 is adjusted, the stock solution of 100 μ g/mL is configured to using 95% ethyl alcohol, is diluted to 5 μ g/mL, 4
The concentration of μ g/mL, 3 μ g/mL, 2 μ g/mL, 1 μ g/mL, 0.5 μ g/mL measure the UV absorption at 338 nm, with concentration for horizontal seat
Marking ultraviolet absorption value is that ordinate draws standard curve, and regression equation is Y=3.68X+0.56, R2=0.9995.According to same
Method measures the content of stilbene compound in test sample.
(3) measurement of general flavone content
General flavone content is detected referring to GBT20574-2006 method.Prepare various concentration rutin standard items, 4,8,
12,16,20,24 μ g/mL, respectively plus chloroform is to 5mL, and bromocresol green buffer is added and 0.2mol/L NaOH solution is each
1mL stands 30min after shaking 1min, clear chloroform liquid is taken to be measured at 415nm wavelength, is cross with concentration
Coordinate, light absorption value are ordinate, draw standard curve, and regression equation is Y=12.26X+2.68, R2=0.9996.According to standard song
Line measures the content of general flavone in test sample.
(4) total lignan assay
Total lignan content is measured by spectrophotometry, accurately weighs syringaresinol reference substance 1mg, is placed in 10mL brown appearance
In measuring bottle, adds methanol to dissolve and be diluted to scale and shake up.Precision draws 0mL, 0.4mL, 0.6mL, 0.8mL, 1.0mL, sets respectively
In 10mL brown volumetric flask, is mended with methanol solution to 10Ml, measure the ultraviolet absorption value at 272nm wavelength.It is molten with reference substance
The concentration of liquid is abscissa, and using corresponding absorbance as ordinate, drawing standard curve regression equation is Y=8.25X+1.93, R2
=0.9997.The content of total lignin in test sample is measured after the same method.
Embodiment 1
A method of dendrobium candidum extract is prepared, is included the following steps:
(1) degreasing
40 meshes are crossed after 500g dendrobium candidum is crushed, and 12 times of petroleum ether is added thereto and impregnates 1.5h, is then refluxed for 3 times often
Secondary 2h removes organic solvent and obtains the dregs of a decoction after filtering.
(2) Polyose extraction
The distilled water of 15 times of addition in the dregs of a decoction is obtained to step (1), is decocted 3 times, is decocted 3h every time, decoction liquor is filtered to isolate
The dregs of a decoction, decoction liquor are concentrated 10 times and obtain concentrate, and ethyl alcohol is added into concentrate, so that the content of ethyl alcohol is 70%, stir evenly
It stands overnight, isolates ethanol water, obtain alcohol sediment, alcohol hypostasis is subjected to filtering and washing with organic solvent, decompression is dry
It is dry, obtain polyoses extract.
(3) stilbene class, flavones and lignanoid extract
95% ethyl alcohol of the filter residue of step (2) separation is extracted 4 times, each 13h again at room temperature, merges alcohol extract,
Merge again with the ethanol water isolated in step (2), by Rotary Evaporators, the solution is concentrated, then to concentrate
Middle addition 20mL aqueous solution is first extracted 4 times with chloroform, isolates chloroform phase, then be extracted with ethyl acetate 4 times, isolate acetic acid
Ethyl ester phase, and retain water phase.
(4) stilbene class, flavones and lignanoid isolate and purify
Step (3) are obtained into chloroform phase and ethyl acetate phase is isolated and purified with silica gel column chromatography column respectively, eluent is not
The petroleum ether of same volume proportion and the mixed liquor of ethyl acetate, eluting order are that the volume ratio of petroleum ether and ethyl acetate is 1:
0,20:1,10:1,5:1,2:1 and 1:1 collect eluting fraction, water phase are isolated and purified with aperture resin gel rubber column gel column, is washed
De- liquid is the methanol solution of different percentage compositions, wherein the mass concentration of each secondary elution methanol solution is followed successively by 0%, 10%,
30%, 50%, 75% and 95%, eluting fraction is collected, the eluent of three-phase is merged, stilbene class, flavones and lignanoid are obtained
Extract.
(5) dry
The extract that step (4) is obtained removes organic solvent by rotary evaporation, is then freeze-dried Liquid Residue, obtains
To stilbene class, flavones and the freeze-dried powder of lignanoid.
Using polysaccharide, stilbene class, flavones and the lignanoid extract total as dendrobium candidum, survey according to the method described in the present invention
The percentage composition for obtaining polysaccharide is 63%, and the percentage composition of stilbene class is 10%, and the percentage composition of flavones is 13%, and the percentage of lignanoid contains
Amount is 14%.
Embodiment 2
A method of dendrobium candidum extract is prepared, is included the following steps:
(1) degreasing
30 meshes are crossed after 500g dendrobium candidum is crushed, and 10 times of petroleum ether is added thereto and impregnates 1h, is then refluxed for 2 times every time
2h removes organic solvent and obtains the dregs of a decoction after filtering.
(2) Polyose extraction
The distilled water of 10 times of addition in the dregs of a decoction is obtained to step (1), is decocted 2 times, is decocted 2h every time, decoction liquor is filtered to isolate
The dregs of a decoction, decoction liquor are concentrated 10 times and obtain concentrate, and ethyl alcohol is added into concentrate, so that the content of ethyl alcohol is 70%, stir evenly
It stands overnight, isolates ethanol water, obtain alcohol sediment, alcohol hypostasis is subjected to filtering and washing with organic solvent, decompression is dry
It is dry, obtain polyoses extract.
(3) stilbene class, flavones and lignanoid extract
95% ethyl alcohol of the filter residue of step (2) separation is extracted 3 times, each 10h again at room temperature, merges alcohol extract,
Merge again with the ethanol water isolated in step (2), by Rotary Evaporators, the solution is concentrated, then to concentrate
Middle addition 20mL aqueous solution is first extracted 3 times with chloroform, isolates chloroform phase, then be extracted with ethyl acetate 3 times, isolate acetic acid
Ethyl ester phase, and retain water phase.
(4) stilbene class, flavones and lignanoid isolate and purify
Step (3) are obtained into chloroform phase and ethyl acetate phase is isolated and purified with silica gel column chromatography column respectively, eluent is not
The petroleum ether of same volume proportion and the mixed liquor of ethyl acetate, eluting order are that the volume ratio of petroleum ether and ethyl acetate is 1:
0,20:1,10:1,5:1,2:1 and 1:1 collect eluting fraction, water phase are isolated and purified with aperture resin gel rubber column gel column, is washed
De- liquid is the methanol solution of different percentage compositions, wherein the mass concentration of each secondary elution methanol solution is followed successively by 0%, 10%,
30%, 50%, 75% and 95%, eluting fraction is collected, the eluent of three-phase is merged, stilbene class, flavones and lignanoid are obtained
Extract.
(5) dry
The extract that step (4) is obtained removes organic solvent by rotary evaporation, is then freeze-dried Liquid Residue, obtains
To stilbene class, flavones and the freeze-dried powder of lignanoid.
Using polysaccharide, stilbene class, flavones and the lignanoid extract total as dendrobium candidum, survey according to the method described in the present invention
The percentage composition for obtaining polysaccharide is 54%, and the percentage composition of stilbene class is 12%, and the percentage composition of flavones is 16%, and the percentage of lignanoid contains
Amount is 18%.
Embodiment 3
A method of dendrobium candidum extract is prepared, is included the following steps:
(1) degreasing
50 meshes are crossed after 500g dendrobium candidum is crushed, and 15 times of ethyl acetate is added thereto and impregnates 2h, is then refluxed for 3 times often
Secondary 2h removes organic solvent and obtains the dregs of a decoction after filtering.
(2) Polyose extraction
The distilled water of 14 times of addition in the dregs of a decoction is obtained to step (1), is decocted 3 times, is decocted 4h every time, decoction liquor is filtered to isolate
The dregs of a decoction, decoction liquor are concentrated 10 times and obtain concentrate, and ethyl alcohol is added into concentrate, so that the content of ethyl alcohol is 70%, stir evenly
It stands overnight, isolates ethanol water, obtain alcohol sediment, alcohol hypostasis is subjected to filtering and washing with organic solvent, decompression is dry
It is dry, obtain polyoses extract.
(3) stilbene class, flavones and lignanoid extract
95% ethyl alcohol of the filter residue of step (2) separation is extracted 5 times, each 10h again at room temperature, merges alcohol extract,
Merge again with the ethanol water isolated in step (2), by Rotary Evaporators, the solution is concentrated, then to concentrate
Middle addition 20mL aqueous solution is first extracted 5 times with chloroform, isolates chloroform phase, then be extracted with ethyl acetate 5 times, isolate acetic acid
Ethyl ester phase, and retain water phase.
(4) stilbene class, flavones and lignanoid isolate and purify
Step (3) are obtained into chloroform phase and ethyl acetate phase is isolated and purified with silica gel column chromatography column respectively, eluent is not
The petroleum ether of same volume proportion and the mixed liquor of ethyl acetate, eluting order are that the volume ratio of petroleum ether and ethyl acetate is 1:
0,20:1,10:1,5:1,2:1 and 1:1 collect eluting fraction, water phase are isolated and purified with aperture resin gel rubber column gel column, is washed
De- liquid is the methanol solution of different percentage compositions, wherein the mass concentration of each secondary elution methanol solution is followed successively by 0%, 10%,
30%, 50%, 75% and 95%, eluting fraction is collected, the eluent of three-phase is merged, stilbene class, flavones and lignanoid are obtained
Extract.
(5) dry
The extract that step (4) is obtained removes organic solvent by rotary evaporation, is then freeze-dried Liquid Residue, obtains
To stilbene class, flavones and the freeze-dried powder of lignanoid.
Using polysaccharide, stilbene class, flavones and the lignanoid extract total as dendrobium candidum, survey according to the method described in the present invention
The percentage composition for obtaining polysaccharide is 48%, and the percentage composition of stilbene class is 14%, and the percentage composition of flavones is 21%, and the percentage of lignanoid contains
Amount is 17%.
Embodiment 4
A method of dendrobium candidum extract is prepared, is included the following steps:
(1) degreasing
60 meshes are crossed after 500g dendrobium candidum is crushed, and 15 times of n,N-Dimethylformamide is added thereto and impregnates 2h, then
Flow back 4 each 2h, and organic solvent is removed after filtering and obtains the dregs of a decoction.
(2) Polyose extraction
The distilled water of 20 times of addition in the dregs of a decoction is obtained to step (1), is decocted 4 times, is decocted 4h every time, decoction liquor is filtered to isolate
The dregs of a decoction, decoction liquor are concentrated 10 times and obtain concentrate, and ethyl alcohol is added into concentrate, so that the content of ethyl alcohol is 70%, stir evenly
It stands overnight, isolates ethanol water, obtain alcohol sediment, alcohol hypostasis is subjected to filtering and washing with organic solvent, decompression is dry
It is dry, obtain polyoses extract.
(3) stilbene class, flavones and lignanoid extract
95% ethyl alcohol of the filter residue of step (2) separation is extracted 5 times, each 15h again at room temperature, merges alcohol extract,
Merge again with the ethanol water isolated in step (2), by Rotary Evaporators, the solution is concentrated, then to concentrate
Middle addition 20mL aqueous solution is first extracted 3 times with chloroform, isolates chloroform phase, then be extracted with ethyl acetate 3 times, isolate acetic acid
Ethyl ester phase, and retain water phase.
(4) stilbene class, flavones and lignanoid isolate and purify
Step (3) are obtained into chloroform phase and ethyl acetate phase is isolated and purified with silica gel column chromatography column respectively, eluent is not
The petroleum ether of same volume proportion and the mixed liquor of ethyl acetate, eluting order are that the volume ratio of petroleum ether and ethyl acetate is 1:
0,20:1,10:1,5:1,2:1 and 1:1 collect eluting fraction, water phase are isolated and purified with aperture resin gel rubber column gel column, is washed
De- liquid is the methanol solution of different percentage compositions, wherein the mass concentration of each secondary elution methanol solution is followed successively by 0%, 10%,
30%, 50%, 75% and 95%, eluting fraction is collected, the eluent of three-phase is merged, stilbene class, flavones and lignanoid are obtained
Extract.
(5) dry
The extract that step (4) is obtained removes organic solvent by rotary evaporation, is then freeze-dried Liquid Residue, obtains
To stilbene class, flavones and the freeze-dried powder of lignanoid.
Using polysaccharide, stilbene class, flavones and the lignanoid extract total as dendrobium candidum, survey according to the method described in the present invention
The percentage composition for obtaining polysaccharide is 58%, and the percentage composition of stilbene class is 9%, and the percentage composition of flavones is 18%, the percentage composition of lignanoid
It is 15%.
Active testing test:
(1) anti-tumor activity pharmacological research
HELA-60 the and BEL-7402 cell of logarithmic growth phase respectively, with the RPM1640 culture solution for containing 10% fetal calf serum, system
At 5 × 104/mL of single cell suspension, which is added in 96 well culture plates, every hole is added 1 × 104A cell；Culture 24
After h, be added the test drug of above 4 embodiments, the concentration of each embodiment test drug is 0 μ g/mL, 5 μ g/mL, 20 μ g/mL,
40 μ g/mL contain 5% CO by plate at 37 DEG C2It is incubated for 2 days, removes culture solution；The RPM1640 culture solution of MTT serum-free is matched
At 1 mg/mL solution, every hole adds 100 μ L, 37 DEG C of incubation 4h；Supernatant liquor is sucked out, 150 μ L DMSO are added, measures 570 nm
Locate OD value, experimental result is as shown in table 1, obtains from table, is added to Herba Dendrobii extract of the invention, rises to growth of tumour cell
To good inhibiting effect.
Table 1
(2) free-radical scavenging activity
Extract of the present invention is configured to 0mg/mL, 0.2mg/mL, 0.4mg/mL, 0.6mg/mL, 0.8mg/mL, 1.0mg/mL
Concentration, respectively draw 2mL in test tube, thereto be added 2mL 0.2mM DPPH solution, place after mixing
15min is returned to zero with dehydrated alcohol, measures the light absorption value (A sample) of 517nm, then measures 2mL extract and 2mL dehydrated alcohol
Mixed liquor light absorption value (A control), the light absorption value (A blank) of the mixed liquor of 2mL dehydrated alcohol and 2mL DPPH solution, clearly
Except the calculating of rate: DPPH clearance rate/%=(1-) × 100%.The results are shown in Table 2, obtains from table, the present invention
Extract has effects that preferably to remove free radical, and with the increase of extract concentrations, remove the effect of free radical by
It is cumulative strong.
Table 2
Claims (7)
1. a kind of dendrobium candidum extract, which is characterized in that include polysaccharide, stilbene class, flavones and lignanoid, the wherein percentage of polysaccharide
Content is 45 ~ 70%, and the percentage composition of stilbene class is 8 ~ 15%, and the percentage composition of flavones is 10 ~ 22%, and the percentage composition of lignanoid is
11~18%。
2. a kind of dendrobium candidum extract according to claim 1, which is characterized in that include polysaccharide, stilbene class, flavones and wood
Rouge element, wherein the percentage composition of polysaccharide is 63 ~ 65%, and the percentage composition of stilbene class is 10 ~ 12%, the percentage composition of flavones is 11 ~
14%, the percentage composition of lignanoid is 12 ~ 15%.
3. the method for preparing the dendrobium candidum extract as described in claim 1 ~ 2 any one, it is characterised in that including as follows
Step:
(1) degreasing
30 ~ 60 meshes are crossed after dendrobium candidum is crushed, and 10 ~ 15 times of organic solvent is added thereto and impregnates 1 ~ 2h, is then refluxed for 2
~ 4 1.5 ~ 2.5h every time remove organic solvent and obtain the dregs of a decoction after filtering；
(2) Polyose extraction
The distilled water of 10 ~ 20 times of addition in the dregs of a decoction is obtained to step (1), is decocted 2 ~ 4 times, 2 ~ 4h is decocted every time, by decoction liquor mistake
The dregs of a decoction are isolated in filter, and decoction liquor is concentrated 10 times and obtains concentrate, and ethyl alcohol is added into concentrate so that the content of ethyl alcohol be 60 ~
80%, it stirs evenly and stands overnight, isolate ethanol water, obtain alcohol sediment, alcohol hypostasis is filtered with organic solvent
Washing, is dried under reduced pressure, obtains polyoses extract；
(3) stilbene class, flavones and lignanoid extract
95% ethyl alcohol of the filter residue of step (2) separation is extracted 3 ~ 5 times again at room temperature, 10 ~ 15h, merges ethyl alcohol leaching every time
Extract, then merge with the ethanol water isolated in step (2), by Rotary Evaporators, the solution is concentrated, then to
20mL aqueous solution is added in concentrate, is first extracted 3 ~ 5 times with chloroform, isolates chloroform phase, then be extracted with ethyl acetate 3 ~ 5 times,
Ethyl acetate phase is isolated, and retains water phase；
(4) stilbene class, flavones and lignanoid isolate and purify
Step (3) are obtained into chloroform phase and ethyl acetate phase is isolated and purified with silica gel column chromatography column respectively, elution is collected and evaporates
Point, water phase is isolated and purified with aperture resin gel rubber column gel column, eluting fraction is collected, the eluent of three-phase is merged, is obtained
To stilbene class, flavones and the extract of lignanoid；
(5) dry
The extract that step (4) is obtained removes organic solvent by rotary evaporation, is then freeze-dried Liquid Residue, obtains
To stilbene class, flavones and the freeze-dried powder of lignanoid.
4. the method according to claim 3 for preparing dendrobium candidum extract, it is characterised in that in the step (1)
Organic solvent is the one or more of petroleum ether, ethyl acetate, N,N-dimethylformamide.
5. the method according to claim 3 for preparing dendrobium candidum extract, it is characterised in that silicon in the step (4)
Plastic column chromatography isolates eluting fraction by gradient elution, and eluent is the petroleum ether of different volumes proportion and mixing for ethyl acetate
Liquid is closed, eluting order is that the volume ratio of petroleum ether and ethyl acetate is 1:0,20:1,10:1,5:1,2:1 and 1:1.
6. the method according to claim 3 for preparing dendrobium candidum extract, it is characterised in that the step (4) is medium and small
Hole resin gel plastic column chromatography isolates eluting fraction by gradient elution, and eluent is the methanol solution of different percentage compositions,
In each secondary elution methanol solution mass concentration be followed successively by 0%, 10%, 30%, 50%, 75% and 95%.
7. such as the described in any item dendrobium candidum extracts of claim 1 ~ 2, in preparing anti-tumor drug or health care product
Using.
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| CN112826894A (en) * | 2021-02-08 | 2021-05-25 | 上海华源制药安徽广生药业有限公司 | Utilization method of leftovers from processing of traditional Chinese medicine decoction pieces |
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