CN103948670B - The preparation method and purposes of green gram spermoderm active principle - Google Patents
The preparation method and purposes of green gram spermoderm active principle Download PDFInfo
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Abstract
The present invention relates to a kind of preparation method and purposes of green gram spermoderm active principle, belong to traditional Chinese medicine effective ingredient extraction field.Its preparation process comprises the following steps：A, green gram spermoderm is taken, extracting in water, is filtered, concentration, is diluted with water, filters, obtains sample solution；B, on the sample solution after the absorption of HPD450 macroporous absorbent resins, it will first be eluted with water to colourless, and then use ethanol elution again, obtain eluent, filter, and concentrate, produce green gram spermoderm active principle.The beneficial effects of the invention are as follows：This method can improve the purity of product, reduce production cost, and production equipment is simple, efficiency high, be adapted to large-scale production.Gained active principle has anti-oxidant, anti-aging, protection skin, delay skin aging isoreactivity, can be used for the additive of food, health products, cosmetics and medicine or as raw material.
Description
Technical field
The invention belongs to traditional Chinese medicine effective ingredient extraction field, and in particular to a kind of preparation method of green gram spermoderm active principle with
Purposes.
Background technology
Green gram spermoderm is to be soaked after mung bean to rub the kind skin to get off.《Compendium of Materia Medica》Carry：For relieving heat toxin, mesh screen is moved back.Mung bean
Skin is common mung bean products processing, uses as feed, is not developed and utilized sufficiently mostly.
Cheng Shuan etc. carries in " Primary Study of antioxidant in green gram spermoderm "：Flavonoids is extracted from green gram spermoderm using methanol
For material for anti-oxidant, the recovery rate of its flavone compound is 1.32%.
Carried in " Study on extraction of general flavone in green gram spermoderm " text：Utilize green gram spermoderm, extraction time 150min, second
Alcohol volume fraction 50%, 80 DEG C of Extracting temperature, solid-to-liquid ratio 1:10, extraction time 2 times, the extracted amount of general flavone is in green gram spermoderm
3.879mg/g。
Also carried in " extraction and quantitative determination of flavonoid compounds in mung bean hull " text：Inquired into orthogonal experiment
The optimum extraction process of flavonoid compounds in mung bean hull.Result of study shows：With 6 times of mung bean tare weight volumes at 70 DEG C
30% alcohol reflux extracts 2 times, each 2h, and mung bean flavonoid compounds extraction effect is best.Sample Al (NO3)3Colorimetric
Method determines under 510nm, and rutin is criterion calculation content, and the average content for obtaining flavonoid compounds in mung bean hull is
1.459%。
A kind of Chinese invention patent " method for extracting anti-oxidant mung bean polysaccharide " of Application No. 201110203090.0 is public
Open and a kind of mung bean polysaccharide is extracted from green gram spermoderm be used for antioxidation method.
However, the existing composition proposed using green gram spermoderm is used for, oxidation resistant action effect is not notable, and also there is effective
The shortcomings that constituents extraction is insufficient, cause the waste of resource.
The content of the invention
It is an object of the invention to provide a kind of preparation method and purposes of green gram spermoderm active principle.
The green gram spermoderm active principle of the present invention, its preparation process comprise the following steps：
A, green gram spermoderm is taken, extracting in water, is filtered, concentration, is diluted with water, filters, obtains sample solution；
B, on the sample solution after the absorption of HPD450 macroporous absorbent resins, it will first be eluted with water to colourless, and then use second again
Alcohol elutes, and obtains eluent, filters, and concentration, produces green gram spermoderm active principle.
Preferably, the number extracted in the step a is 1~3 time, the time of extraction is 1~3 hour.
Preferably, in the step a during extracting in water, the weight for adding water is 8~20 times of the green gram spermoderm.It is especially excellent
Choosing, in the step a during extracting in water, the weight for adding water is 10 times, 12 times, 14 times, 16 times and 18 times of the green gram spermoderm.
It is concentrated under reduced pressure preferably, concentrating to use in the step a.
Preferably, proportion is concentrated into the step a as 1.05~1.075.
Preferably, the mass percent concentration of ethanol is 70%~90% in the step b.
Preferably, proportion is concentrated into the step b as 1.15~1.175.
Further, the present invention also provides green gram spermoderm active principle made from a kind of preparation method.
Further, the present invention also provides a kind of application of green gram spermoderm active principle in anti-oxidation medicine is prepared.
Further, what the green gram spermoderm active principle that the present invention is also provided described in a kind of utilization made has anti-oxidation function
Cosmetic composition, the cosmetic composition is composed of the following components：In parts by weight, 1,600~800 parts of 3- butanediols,
50~150 parts of green gram spermoderm active principle, 200~400 parts of purified water, 0.005~0.015 part of Phenoxyethanol.It is particularly preferred, should
Cosmetic composition is composed of the following components：In parts by weight, 1,700 parts of 3- butanediols, 100 parts of green gram spermoderm active principle, pure
Change 300 parts of water, 0.01 part of Phenoxyethanol.Wherein, 1,3- butanediols, purified water are added in green gram spermoderm active principle, is contributed to
Said composition is compatible with other supplementary materials, forms stable cosmetics finished product.Add Phenoxyethanol and be used for anti-corrosion.
In the present invention, can with above-mentioned cosmetics or directly based on green gram spermoderm active principle composition, then be equipped with other
Supplementary material, cosmetics finished product is made according to the common process for preparing cosmetics.
The beneficial effects of the present invention are：Using green gram spermoderm as raw material, a kind of system for preparing green gram spermoderm active principle is worked out
Preparation Method.The purity of product is the method increase, reduces production cost, and production equipment is simple, efficiency high, is adapted to extensive
Production.The active principle has anti-oxidant, anti-aging, protection skin, delay skin aging isoreactivity, can be used for food, protects
It is good for product, the additive of cosmetics and medicine or as raw material.
Brief description of the drawings
In order to illustrate more clearly about the embodiment of the present invention or technical scheme of the prior art, embodiment will be described below
In the required accompanying drawing used be briefly described.
Fig. 1 is green gram spermoderm active principle reducing power test chart；
Fig. 2 is the test chart that green gram spermoderm active principle is acted on DPPH radicals scavengings；
Fig. 3 is the test chart of scavenging action of the green gram spermoderm active principle to superoxide anion；
Fig. 4 is test chart of the green gram spermoderm active principle to chelating copper ions ability；
Fig. 5 is the test chart that green gram spermoderm active principle is acted on DPPH radicals scavengings with mung bean seed active principle.
Embodiment
To make those skilled in the art understand the production technology and technique effect of the present invention in detail, below specifically to produce
Example come be further described the present invention application and technique effect.
Embodiment 1：
A, green gram spermoderm is taken, adds the water of 10 times of weight to extract 2 times, the time extracted every time is 2 hours, using with 200 mesh
Filter screen filtered, be concentrated under reduced pressure into proportion as 1.06（Heat determination）, then the unit based on concentrate, adds 4 times of weight
Water dilutes, and is filtered using filter cloth, obtains sample solution；
B, on the sample solution after the absorption of HPD450 macroporous absorbent resins, it will first be eluted with water to colourless, and then use matter again
Measure percent concentration to elute for 75% ethanol solution, flow velocity is 2BV/ hours, obtains eluent, is carried out using the filter screen with 300 mesh
Filtering, concentration proportion are 1.15（Heat determination）, produce green gram spermoderm active principle.
Embodiment 2：
A, green gram spermoderm is taken, adds the water of 20 times of weight to extract 1 time, 1 hour extraction time, is entered using the filter screen with 200 mesh
Row filtering, proportion is concentrated under reduced pressure into as 1.07（Heat determination）, then the unit based on concentrate, adds 5 times of weight water dilutions, adopts
Filtered with filter cloth, obtain sample solution；
B, on the sample solution after the absorption of HPD450 macroporous absorbent resins, it will first be eluted with water to colourless, and then use matter again
Measure percent concentration to elute for 70% ethanol solution, flow velocity is 2BV/ hours, obtains eluent, is carried out using the filter screen with 300 mesh
Filtering, concentration proportion are 1.17（Heat determination）, produce green gram spermoderm active principle.
Embodiment 3：
A, green gram spermoderm is taken, adds the water of 8 times of weight to extract 3 times, the time extracted every time is 3 hours, using with 200 purposes
Filter screen is filtered, and is concentrated under reduced pressure into proportion as 1.06（Heat determination）, then the unit based on concentrate, adds 3 times of weight waters
Dilution, is filtered using filter cloth, obtains sample solution；
B, on the sample solution after the absorption of HPD450 macroporous absorbent resins, it will first be eluted with water to colourless, and then use matter again
Measure percent concentration to elute for 90% ethanol solution, flow velocity is 2BV/ hours, obtains eluent, is carried out using the filter screen with 300 mesh
Filtering, concentration proportion are 1.16（Heat determination）, produce green gram spermoderm active principle.
Embodiment 4：
A, green gram spermoderm is taken, adds the water of 14 times of weight to extract 2 times, the time extracted every time is 3 hours, using with 200 mesh
Filter screen filtered, be concentrated under reduced pressure into proportion as 1.06（Heat determination）, then the unit based on concentrate, adds 3 times of weight
Water dilutes, and is filtered using filter cloth, obtains sample solution；
B, on the sample solution after the absorption of HPD450 macroporous absorbent resins, it will first be eluted with water to colourless, and then use matter again
Measure percent concentration to elute for 90% ethanol solution, flow velocity is 1.5BV/ hours, obtains eluent, using the filter screen with 300 mesh enters
Row filtering, concentration proportion are 1.16（Heat determination）, produce green gram spermoderm active principle.
Comparative example 1：
Green gram spermoderm is taken to be boiled 2 times with 10 times of amount decoctings, the time decocted every time is 2 hours, obtains green gram spermoderm water extract.
Comparative example 2：
The ethanol for taking green gram spermoderm to be 50% with 10 times of amount mass percent concentrations extracts 2 times, and extraction 2 hours, obtain mung bean every time
Skin alcohol extracting thing.
Comparative example 3：
The ethanol for taking green gram spermoderm to be 30% with 10 times of amount mass percent concentrations extracts 2 times, and extraction 2 hours, obtain mung bean every time
Skin alcohol extracting thing.
To verify effect of the present invention, spy makees comparative testing below：
| Product | General flavone content |
| Embodiment 1 | 7.35mg/ml |
| Embodiment 2 | 6.28mg/ml |
| Embodiment 3 | 8.12mg/ml |
| Embodiment 4 | 5.32mg/ml |
| Comparative example 1 | 0.535mg/ml |
| Comparative example 2 | 0.380mg/ml |
| Comparative example 3 | 0.345mg/ml |
As seen from the above table, in the green gram spermoderm active principle as obtained by preparation method of the present invention, general flavone content is higher, separately
Due to by HPD450 resin concentration general flavones, can effectively avoid the dissolution of the impurity such as chlorophyll into green gram spermoderm active principle, from
And it ensure that the purity of green gram spermoderm active principle.
Separately active principle and extract as obtained by embodiment 1-4 and comparative example 1-3 are acted on DPPH radicals scavengings
Research it was found that, the oxidation resistance of green gram spermoderm active principle of gained is 4- extract obtained in comparative example in embodiment
6 times.
Embodiment 5：
Effectively to preserve green gram spermoderm active principle, preferably it is applied in cosmetics.Can be by the mung bean obtained by embodiment 1-4
Skin active principle is equipped with other components and composition needed for cosmetics is made.Said composition is composed of the following components：By weight
Number, takes 0.01 part of 700 parts of 1,3- butanediols, 100 parts of green gram spermoderm active principle, 300 parts of purified water and Phenoxyethanol, mixes, bag
Dress, you can.
Embodiment 6：
Effectively to preserve green gram spermoderm active principle, preferably it is applied in cosmetics.Can be by the mung bean obtained by embodiment 1-4
Skin active principle is equipped with other components and composition needed for cosmetics is made.Said composition is composed of the following components：By weight
Number, takes 0.015 part of 600 parts of 1,3- butanediols, 150 parts of green gram spermoderm active principle, 200 parts of purified water and Phenoxyethanol, mixes, bag
Dress, you can.
Embodiment 7：
Effectively to preserve green gram spermoderm active principle, preferably it is applied in cosmetics.Can be by the mung bean obtained by embodiment 1-4
Skin active principle is equipped with other components and composition needed for cosmetics is made.Said composition is composed of the following components：By weight
Number, takes 0.005 part of 800 parts of 1,3- butanediols, 50 parts of green gram spermoderm active principle, 400 parts of purified water and Phenoxyethanol, mixes, bag
Dress, you can.
For the technique effect of the checking present invention, spy makees tests below：
1st, green gram spermoderm active principle reducing power is tested
Experimental principle：Reducing power is one of index of the potential oxidation resistance of material, and potassium ferricyanide is reduced into yellow blood by sample
Salt, potassium ferrocyanide and Fe3+Effect generation is Prussian blue, and light absorption value is determined at 700nm to detect Prussian blue growing amount, is passed through
Prussian blue growing amount weighs the reducing power of sample.
Experimental method：Green gram spermoderm active principle solution 0.1mL, 0.2mL, 0.5mL, 1mL to be measured are taken respectively, will with distilled water
Volume supplies 1mL；Positive control sample vitamin c solution 0.1mL, 0.2mL, 0.5mL, 1mL are taken, is supplied with distilled water
1mL.1mL phosphate buffers and 1mL potassium ferricyanide solutions are added in each group sample, is mixed, 50 DEG C of water-bath 20min, is taken out,
1mL solution of trichloroacetic acid is added, is mixed.Take 2mL mixed liquor to be placed in 5mL centrifuge tubes, add 2mL distilled water and 0.4mL
Liquor ferri trichloridi is rapid to mix.Determine light absorption value of the solution at 700nm.
Experimental result：See Fig. 1；Green gram spermoderm active principle shows good reducing power, shows green gram spermoderm active principle
With potential oxidation resistance.
2nd, the test that green gram spermoderm active principle is acted on DPPH radicals scavengings
Experimental principle：DPPH methanol solutions are pansy, there is strong light absorption value under 517nm, if being combined with sample, meeting
Light absorption value is reduced, the ability of free radical is thus removed so as to judgement sample.
Experimental method
Experimental group：0.1mL, 0.2mL, 0.5mL are taken respectively, and 1mL green gram spermoderm active principle solution examples are in test tube, with steaming
Distilled water supplies 1mL, adds 4mLDPPH methanol solutions, mixes, and 30min, constantly concussion are placed in dark place, determine the suction at 517nm
Light value.
Blank group：Take the sample solution of same experimental group equal volume respectively in each test tube, supplied with distilled water
1mL, add 4mL methanol solution.30min is placed in dark place after mixing, determines light absorption value at 517nm.
Control group：1mL distilled water adds 4mLDPPH methanol solutions.30min is placed in dark place after mixing, determines at 517nm
Light absorption value.
Experimental result：DPPH inhibiting rates（%）=（A0-A1+A2）/A0*100%
Wherein A0 is the light absorption value of control group, and A1 is the light absorption value of experimental group, and A2 is the light absorption value of corresponding blank group.
As a result Fig. 2 is seen；Sample can effectively remove DPPH free radicals, and reach more than 90% in test scope clearance rate.
3rd, scavenging action test of the green gram spermoderm active principle to superoxide anion
Experimental principle：Pyrogallol autoxidation in weakly alkaline environment, which decomposes, produces superoxide anion, with reaction
Carry out, superoxide anion constantly accumulates in system, causes absorbance of the reaction solution at 420nm to start a period of time in reaction
Interior change that can be over time and linearly increase, therefore determine within this time measured object reaction solution absorbance with the time
Rate of change, and the ability that measured object suppresses superoxide anion can be drawn compared with blank solution.
Experimental method
Experimental group：5.6mLTris HCl buffer solutions are taken, be separately added into 0.1mL, 0.2mL, 0.3mL, 0.4mL treats test sample
Product, 0.4mL is supplied with distilled water, be put into water-bath 10min in 25 DEG C of water-baths, then add the pyrogallol of 25 DEG C of pre-temperatures
（If room temperature can operate at room temperature higher than 25 DEG C）, fully mix, the accurate timing 3min since adding after pyrogallol,
Then add 5% ascorbic acid（Vc）0.05mL terminating reactions.Its absorbance is determined after 10min clocks at 420nm.
Control group：5.6mL Tris HCl buffer solutions are taken, are separately added into the testing sample of same experimental group same volume, are used
Distilled water supplies 0.4mL, adds 12mmol/LHCl, fully mixes, the accurate timing 3min since being added HCl, it is anti-to add 5%
Bad hematic acid terminating reaction.Its absorbance is determined after 10min clocks at 420nm.
Blank group：5.6mL50mmol/L Tris-HCl buffer solutions are taken, add 0.4mL distilled water, hereafter operation is the same as experiment
Group.
Experimental result：E=[(A0-A1+A2)/A0]*100%
A0 is Blank absorbance values, and A1 is experimental group absorbance, and A2 is control group absorbance.
As a result Fig. 3 is seen；Testing sample green gram spermoderm active principle shows the effect of certain removing superoxide anion.
4th, chelating copper ions aptitude tests
Experimental principle：The metal ion such as generation of iron ion, copper ion in lipid oxidation, cellular oxidation and melanin
Play a part of catalyst in journey, the material with metal-chelating effect can also play anti-oxidant and suppression melanin indirectly
The effect of generation, TMM chelating method measure chelating copper ions abilities are used to assess sample chelated mineral ability.
Experimental method
Standard curve：Take 0mL, 0.2mL, 0.4mL respectively, 0.6mL, 0.8mL, 1mL 0.2mM anhydrous slufuric acid copper solutions,
1mL is supplied surely with Hex buffer solutions.4mL Hex buffer solutions and 0.2mLTMM indicator is added, mixes and stands 10min, Hex delays
Fliud flushing returns to zero, and determines the light absorption value at 460nm and at 530nm.Using A460/A530 as ordinate, copper ion concentration is abscissa
Draw standard curve.
Experimental group：0.1mL, 0.2mL, 0.3mL, 0.4mL are taken respectively, and 0.5mL samples are supplied in centrifuge tube with distilled water
To 0.5mL, add 1.5mLHex buffer solutions and 0.5mL is dissolved in the CuSO of Hex buffer solutions4(2mM), 1h, no sample are placed at room temperature
Product blank does same processing, separately takes the sample of series of identical volume to supply 0.5mL in centrifuge tube with distilled water, add
Enter 1mLHex buffer solutions, place 1h at room temperature.
Above-mentioned reaction solution 0.2mL is taken, adds 4.8mLHex buffer solutions and 0.2mLTMM indicator, mixes and stands 10min, with
Hex buffer solutions are cooked blank zeroing, determine the light absorption value at 460nm and 530nm.
Experimental result：E%=[(A0-A1+A3)/A0]*100%
A0 is n.s blank A460/A530 corresponding x values on standard curve, and A1 is experimental group sample A460/A530,
A2 is to compare A460/A530 without copper.
As a result Fig. 4 is seen：Sample green gram spermoderm active principle shows stable chelating copper ions ability.
5th, the research that green gram spermoderm active principle is acted on DPPH radicals scavengings with mung bean seed active principle
Experimental method
Experimental group：Taking green gram spermoderm active principle solution respectively, with green gram spermoderm active principle is prepared prepared by identical preparation method
Mung bean seed active principle solution 1mL, with distilled water constant volume to 25mL, therefrom take 1mL solution respectively in test tube, add
4mLDPPH methanol solutions, mix, 30min, constantly concussion are placed in dark place, determine the light absorption value at 517nm.
Blank group：Take the sample solution of same experimental group equal volume respectively in each test tube, supplied with distilled water
1mL, add 4mL methanol solution.30min is placed in dark place after mixing, determines light absorption value at 517nm.
Control group：1mL distilled water adds 4mLDPPH methanol solutions.30min is placed in dark place after mixing, determines at 517nm
Light absorption value.
Experimental result：DPPH inhibiting rates（%）=（A0-A1+A2）/A0*100%
See Fig. 5, the results showed that the antioxidant effect of green gram spermoderm active principle is better than mung bean seed active principle.
Finally it should be noted that above example is only to illustrative and not limiting technical scheme, although reference
The present invention is described in detail above-described embodiment, it will be appreciated by those skilled in the art that can still be carried out to the present invention
Modification or equivalent substitution, any modification or partial replacement without departing from the spirit and scope of the present invention, it all should cover
In scope of the presently claimed invention.
Claims (8)
1. a kind of green gram spermoderm active principle for being used to prepare antioxidation cosmetic product, it is characterised in that its preparation method includes following
Step：
A, green gram spermoderm is taken, extracting in water, is filtered, concentration, is diluted with water, filters, obtains sample solution；
B, on the sample solution after the absorption of HPD450 macroporous absorbent resins, it will first be eluted with water to colourless, then be washed again with ethanol
It is de-, eluent is obtained, is filtered, concentration, produces green gram spermoderm active principle；
Wherein, the number extracted in the step a is 1~3 time, and the time of extraction is 1~3 hour；Add water extraction in the step a
When taking, the weight for adding water is 8~20 times of the green gram spermoderm；
The mass percent concentration of ethanol is 70%~90% in the step b；Proportion is concentrated into as 1.15~1.175.
2. green gram spermoderm active principle according to claim 1, it is characterised in that：Concentration is dense using depressurizing in the step a
Contracting.
3. green gram spermoderm active principle according to claim 1, it is characterised in that：Being concentrated into proportion in the step a is
1.05~1.075.
4. application of the green gram spermoderm active principle in antioxidation cosmetic product are prepared according to any one of claim 1-3.
A kind of 5. change with anti-oxidation function that green gram spermoderm active principle using any one of claim 1-3 makes
Cosmetic compositions, it is characterised in that：The cosmetic composition is composed of the following components：In parts by weight, 1,3- butanediols 600~
800 parts, 50~150 parts of green gram spermoderm active principle, 200~400 parts of purified water, 0.005~0.015 part of Phenoxyethanol.
6. a kind of preparation method as claimed in claim 1 for being used to prepare the green gram spermoderm active principle of antioxidation cosmetic product, its
It is characterised by, comprises the following steps：
A, green gram spermoderm is taken, extracting in water, is filtered, concentration, is diluted with water, filters, obtains sample solution；
B, on the sample solution after the absorption of HPD450 macroporous absorbent resins, it will first be eluted with water to colourless, then be washed again with ethanol
It is de-, eluent is obtained, is filtered, concentration, produces green gram spermoderm active principle；
Wherein, the number extracted in the step a is 1~3 time, and the time of extraction is 1~3 hour；Add water extraction in the step a
When taking, the weight for adding water is 8~20 times of the green gram spermoderm；
The mass percent concentration of ethanol is 70%~90% in the step b；Proportion is concentrated into as 1.15~1.175.
7. preparation method according to claim 6, it is characterised in that：Concentration is used and is concentrated under reduced pressure in the step a.
8. preparation method according to claim 6, it is characterised in that：Be concentrated into the step a proportion for 1.05~
1.075。
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