CN108383921A - A kind of Ginkgo biloba polysaccharide extracting method, and the inoxidizability assay method with it - Google Patents
A kind of Ginkgo biloba polysaccharide extracting method, and the inoxidizability assay method with it Download PDFInfo
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- CN108383921A CN108383921A CN201810304353.9A CN201810304353A CN108383921A CN 108383921 A CN108383921 A CN 108383921A CN 201810304353 A CN201810304353 A CN 201810304353A CN 108383921 A CN108383921 A CN 108383921A
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- ginkgo biloba
- ginkgo
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- 235000008100 Ginkgo biloba Nutrition 0.000 title claims abstract description 137
- 150000004676 glycans Chemical class 0.000 title claims abstract description 88
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 88
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 88
- 244000194101 Ginkgo biloba Species 0.000 title claims abstract description 76
- 238000000034 method Methods 0.000 title claims abstract description 30
- 238000003556 assay Methods 0.000 title claims description 9
- 241000218628 Ginkgo Species 0.000 claims abstract description 61
- 235000011201 Ginkgo Nutrition 0.000 claims abstract description 61
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 45
- 235000009508 confectionery Nutrition 0.000 claims abstract description 24
- 239000000843 powder Substances 0.000 claims abstract description 23
- 239000012141 concentrate Substances 0.000 claims abstract description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 15
- 238000001556 precipitation Methods 0.000 claims abstract description 14
- 230000003078 antioxidant effect Effects 0.000 claims abstract description 6
- 238000003809 water extraction Methods 0.000 claims abstract description 6
- 235000019441 ethanol Nutrition 0.000 claims description 26
- 239000000243 solution Substances 0.000 claims description 26
- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 claims description 18
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 claims description 15
- 239000013641 positive control Substances 0.000 claims description 12
- 150000003254 radicals Chemical class 0.000 claims description 12
- 239000006228 supernatant Substances 0.000 claims description 12
- 239000012224 working solution Substances 0.000 claims description 12
- 239000007788 liquid Substances 0.000 claims description 11
- 238000002835 absorbance Methods 0.000 claims description 9
- 238000012545 processing Methods 0.000 claims description 9
- 238000001035 drying Methods 0.000 claims description 8
- 239000012153 distilled water Substances 0.000 claims description 6
- 238000012869 ethanol precipitation Methods 0.000 claims description 6
- 238000007710 freezing Methods 0.000 claims description 6
- 230000008014 freezing Effects 0.000 claims description 6
- 102000004169 proteins and genes Human genes 0.000 claims description 6
- 108090000623 proteins and genes Proteins 0.000 claims description 6
- 238000009423 ventilation Methods 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 6
- 239000003153 chemical reaction reagent Substances 0.000 claims description 3
- 238000002474 experimental method Methods 0.000 claims description 3
- 239000011888 foil Substances 0.000 claims description 3
- 230000007760 free radical scavenging Effects 0.000 claims description 3
- 230000000873 masking effect Effects 0.000 claims description 3
- 238000011160 research Methods 0.000 abstract description 7
- 238000005119 centrifugation Methods 0.000 abstract description 6
- 241000196324 Embryophyta Species 0.000 abstract description 4
- 235000013361 beverage Nutrition 0.000 abstract description 4
- 230000033228 biological regulation Effects 0.000 abstract description 4
- 230000001900 immune effect Effects 0.000 abstract description 4
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 230000003110 anti-inflammatory effect Effects 0.000 abstract description 3
- 239000002994 raw material Substances 0.000 abstract description 3
- 230000003712 anti-aging effect Effects 0.000 abstract description 2
- 238000010298 pulverizing process Methods 0.000 abstract 1
- 238000000605 extraction Methods 0.000 description 9
- 230000000694 effects Effects 0.000 description 5
- 239000000284 extract Substances 0.000 description 4
- 230000002000 scavenging effect Effects 0.000 description 3
- 230000008859 change Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- -1 ABTS Radicals Chemical class 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 150000003797 alkaloid derivatives Chemical class 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000005352 clarification Methods 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000004880 explosion Methods 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 235000015110 jellies Nutrition 0.000 description 1
- 239000008274 jelly Substances 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 239000002932 luster Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 239000002893 slag Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 150000003505 terpenes Chemical class 0.000 description 1
- 235000007586 terpenes Nutrition 0.000 description 1
- 238000009777 vacuum freeze-drying Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N2021/755—Comparing readings with/without reagents, or before/after reaction
Abstract
The present invention provides a kind of Ginkgo biloba polysaccharide extracting method includes the following steps：By ginkgo leaf dries pulverizing, ginkgo leaf Thick many candies concentrate is made in water extraction and alcohol precipitation method, is freeze-dried into powder；The present invention is using ginkgo leaf as raw material, using the method for water extract-alcohol precipitation, is concentrated in vacuo, centrifugation, obtain having after refrigerator frozen dried immunological regulation, anti-aging, anti-inflammatory activity Ginkgo biloba polysaccharide powder；Its production method not only has higher yield compared to currently used plant polyose extracting method, but also its technique is relatively easy.And the antioxidant activity on this basis, having studied Ginkgo biloba polysaccharide, the researches and exploitation for polysaccharides from ginkgo biloba beverage provide theoretical foundation.
Description
Technical field
The present invention relates to effective ingredients in plant to extract field, and in particular to a kind of Ginkgo biloba polysaccharide extracting method, and have
Its inoxidizability assay method.
Background technology
A kind of peculiar veteran kind of the ginkgo as China, is widely distributed in the multiple areas in China.Include a variety of objects in its leaf
Matter, such as flavonoids, terpene, alkaloid and water-soluble polysaccharide.It has been investigated that these substances are lived with various biological
Property, it is of great significance to being normally carried out for vital movement, therefore the sections such as medicine, food are increasingly becoming to the research of ginkgo leaf
One of the research hotspot in field.
Antitumor since ginkgo leaf has anti-aging, immunological regulation value, the research and development of ginkgo leaf are unanimously by people
Concern.But mostly concentrate the development and utilization of ginkgo fruit.It is also not rarely seen to the research report of Ginkgo biloba polysaccharide.Such as Shen
Please number be 201010569011.3 Chinese patent, disclose a kind of process for extracting ginkgo leaf polysaccharide with aid of ultrasonic, ultrasonic wave
Process for extracting ginkgo leaf polysaccharide with aid, extraction process is as follows, ginkgo leaf (ginkgo leaf after autumn) → dry → pulverize and sieve (40
Mesh) → (distilled water is solvent, and temperature is controlled at 80 DEG C) → centrifugation (removing slag) → vacuum concentration is ultrasonically treated (to concentration preceding 1/
10) → alcohol precipitation → centrifugation (taking precipitation) → vacuum freeze drying → polysaccharide crude.It is generated using ul-trasonic irradiation in liquid empty
Change acts on, and leads to bubble formation, increases, and explosion compression increases to make solid sample disperse between sample and extractant
Contact area improves the delivery rate that object is transferred to liquid phase from solid phase, shortens extraction time, but also has energy consumption
Height, the high disadvantage of extraction cost, there is an urgent need to be improved.
Invention content
To solve the above problems, the present invention provides a kind of Ginkgo biloba polysaccharide extracting method, and the inoxidizability with it
Assay method.The present invention using the method for water extract-alcohol precipitation, is concentrated in vacuo, centrifugation, refrigerator frozen dried using ginkgo leaf as raw material
Obtain that there is immunological regulation, anti-inflammatory, activity of fighting against senium Ginkgo biloba polysaccharide powder afterwards；Its production method is not only normal compared at present
Plant polyose extracting method has higher yield, and its technique is relatively easy.And on this basis, have studied ginkgo
The antioxidant activity of leaf polyose, the researches and exploitation for polysaccharides from ginkgo biloba beverage provide theoretical foundation.
To realize the technical purpose, the technical scheme is that：A kind of Ginkgo biloba polysaccharide extracting method, including it is following
Step：
S1：By ginkgo leaf, drying to constant weight, is crushed to and at least crosses 60 mesh sieve；
S2：Ginkgo leaf after crushing is utilized into water extraction and alcohol precipitation method, ginkgo leaf Thick many candies concentrate is made；
S3：The ginkgo leaf Thick many candies concentrate is redissolved, and Sevag reagents removal foreign protein is added；
S4：It is removed into step S3 in the ginkgo leaf Thick many candies after foreign protein, precipitation, washing drying after ethyl alcohol is added, leads to
After wind volatilizees to ethyl alcohol, it is freeze-dried into powder, Ginkgo biloba polysaccharide powder is made.
Further, the preferred ginkgo leaf fallen leaves of ginkgo leaf in the step S1.
Further, in the step S2 water extraction and alcohol precipitation method be made ginkgo leaf Thick many candies the step of include：
T1：1 part of ginkgo leaf after being crushed in step S1 is taken, at least 30 parts of water is added, it is small that at least 5 are extracted at 100 DEG C
When；
T2：After 4000r/min centrifuges 10min, supernatant is taken, and supernatant is placed in Rotary Evaporators and is concentrated in vacuo,
Obtain initial concentration liquid；
T3:By initial concentration liquid obtained by step T2, centrifuged with 4000r/min after standing for 24 hours with 70% ethanol precipitation
10min obtains ginkgo leaf Thick many candies concentrate.
As a preference of the present invention, it is based on the above, unlike, in the step S4, the concentration of ethyl alcohol is added extremely
It is less 70%, the volume of ethyl alcohol is added at least by the three times volume of the ginkgo leaf Thick many candies concentrate.
Further, in the step S4, after ethanol precipitation, washing drying is added, at least placed in ventilation makes ethyl alcohol for 24 hours
Volatilization, and freezing processing is carried out at least -20 DEG C, it is put into freeze drier that at least 48h is lyophilized into powder, Ginkgo biloba polysaccharide is made
Powder.
A kind of Ginkgo biloba polysaccharide inoxidizability assay method, the ginkgo of various concentration is made including the use of Ginkgo biloba polysaccharide powder
Leaf polyose solution prepares the Vc of corresponding concentration as positive controls, and prepare distilled water as blank control as experimental group
Group；
One kind of dropwise addition DPPH working solutions or ABTS working solutions into the polysaccharide solution of various concentration, and observation experiment group,
Positive controls, the absorbance of blank control group, calculate different Ginkgo biloba polysaccharide concentration to DPPH free radicals and ABTS from
By base clearance rate, the antioxidative intensity of Ginkgo biloba polysaccharide is further shown as.
Further, using the Ginkgo biloba polysaccharide solution of 0.5mL various concentration gradients, a concentration of 6.5 × 10 are sequentially added- 5After standing 30min after mol/L DPPH solution 2.5mL, each concentration is measured at 517nm wavelength using ultraviolet specrophotometer
The absorbance of polysaccharides from ginkgo biloba solution compares the blank control group and positive controls, obtains the Ginkgo biloba polysaccharide of various concentration
To the clearance rate of DPPH free radicals.
Further, using the Ginkgo biloba polysaccharide solution of 0.3mL various concentrations, ABTS working solution 4.7mL are sequentially added, are used
ABTS working solutions are as a control group；After being protected from light processing 30min to the experimental group, positive controls, control group masking foil, utilize
Ultraviolet specrophotometer measures the absorbance of each concentration polysaccharides from ginkgo biloba solution at 734nm wavelength, compares the experimental group, sun
Property control group, control group, obtain the Ginkgo biloba polysaccharide of various concentration to ABTS free radical scavenging activities.
The beneficial effects of the present invention are：
The present invention using the method for water extract-alcohol precipitation, is concentrated in vacuo, centrifugation, refrigerator frozen dried using ginkgo leaf as raw material
Obtain that there is immunological regulation, anti-inflammatory, activity of fighting against senium Ginkgo biloba polysaccharide powder afterwards；Its production method is not only normal compared at present
Plant polyose extracting method has higher yield, and its technique is relatively easy.And on this basis, have studied ginkgo
The antioxidant activity of leaf polyose, the researches and exploitation for polysaccharides from ginkgo biloba beverage provide theoretical foundation.
Description of the drawings
Fig. 1 is the influence curve figure that different feed liquid compares polysaccharides from ginkgo biloba yield；
Fig. 2 is influence curve figure of the different extraction times to polysaccharides from ginkgo biloba yield；
Fig. 3 is influence curve figure of the different extraction temperatures to polysaccharides from ginkgo biloba yield；
Fig. 4 is scavenging effect of the polysaccharides from ginkgo biloba solution of various concentration to DPPH free radicals；
Fig. 5 is scavenging effect of the polysaccharides from ginkgo biloba solution of various concentration to ABTS free radicals.
Specific implementation mode
Technical scheme of the present invention will be clearly and completely described below.
As shown in Figure 1, a kind of Ginkgo biloba polysaccharide extracting method, includes the following steps：
S1：By ginkgo leaf, drying to constant weight, is crushed at least 60 mesh sizes and is sieved；
S2：Ginkgo leaf after crushing is utilized into water extraction and alcohol precipitation method, ginkgo leaf Thick many candies concentrate is made；
S3：The ginkgo leaf Thick many candies concentrate is redissolved, and Sevag reagents removal foreign protein is added；
S4：It is gone into step S3 in the ginkgo leaf Thick many candies after removing protein, precipitation, washing drying after ethyl alcohol, ventilation is added
After volatilizing to ethyl alcohol, it is freeze-dried into powder, Ginkgo biloba polysaccharide powder is made.And the ginkgo leaf extracted according to the above extracting method
Polysaccharide under dissolved state color and luster clarification free from admixture, be solid-state when be in brown color.
Further, the preferred ginkgo leaf fallen leaves of ginkgo leaf in the step S1.The cost of production technology is further reduced,
State-owned resource is made full use of, is turned waste into wealth.Further, the step of ginkgo leaf Thick many candies are made in water extraction and alcohol precipitation method in the step S2
Including：
T1：It is the influence curve figure that different feed liquid compares polysaccharides from ginkgo biloba yield as shown in Figure 1；Fig. 2 is different extraction times
To the influence curve figure of polysaccharides from ginkgo biloba yield；Fig. 3 is influence curve figure of the different extraction temperatures to polysaccharides from ginkgo biloba yield；
It is therefore preferable that taking 1 part of ginkgo leaf after being crushed in step S1, at least 30 parts of water is added, with solid-liquid ratio for 1:30
Ratio, at least 5h is extracted at 100 DEG C；It is the influence curve figure that different feed liquid compares polysaccharides from ginkgo biloba yield as shown in Figure 1；Figure
2 be influence curve figure of the different extraction times to polysaccharides from ginkgo biloba yield；Fig. 3 is shadow of the different extraction temperatures to polysaccharides from ginkgo biloba yield
Ring curve graph；
T2：After 4000r/min centrifuges 10min, supernatant is taken, and supernatant is placed in Rotary Evaporators and is concentrated in vacuo,
Obtain initial concentration liquid；Wherein gained polysaccharide solution is concentrated using Rotary Evaporators, reduction polysaccharide loss can be reached
Advantageous effect.
T3:After standing for 24 hours, 10 are centrifuged with 4000r/min with 70% ethanol precipitation for initial concentration liquid obtained by step T2
Minute, obtain ginkgo leaf Thick many candies concentrate.
As a preference of the present invention, it is based on the above, unlike, in the step S4, the concentration of ethyl alcohol is added extremely
It is less 70%, the volume of ethyl alcohol is added at least by the three times volume of the ginkgo leaf Thick many candies concentrate.
Further, in the step S4, after ethanol precipitation, washing drying is added, at least placed in ventilation makes ethyl alcohol for 24 hours
Volatilization, and freezing processing is carried out at least -20 DEG C, it is put into freeze drier that at least 48h is lyophilized into powder, Ginkgo biloba polysaccharide is made
Powder.
Embodiment 1:
The, which falls leaves ginkgo, crushes, and takes 5g ginkgo powders to place in conical flask, distilled water, material-water ratio 1 is added：40, it will bore
Shape bottle, which is put into water-bath, extracts 5h, and bath temperature is 100 DEG C.Then 4000r/min centrifuges 10min and takes supernatant, by ginkgo
Supernatant is concentrated in vacuo in Rotary Evaporators, until concentrate probably arrives 65mL or so.After again with 70% ethyl alcohol to concentration
Ginkgo supernatant is precipitated.After placing for 24 hours, the ginkgo Thick many candies for the bottom being precipitated out are centrifuged using 4000r/min
The mode of 10min is centrifuged out Thick many candies, and Thick many candies are cold in -20 DEG C of progress after ventilation placement a period of time makes ethyl alcohol volatilize
Sample after freezing processing, is finally put into freeze drier and is lyophilized into powder by jelly processing, and polysaccharide extract rate is up to 13.72%.
Embodiment 2:
Ginkgo fallen leaves be crushed into 60 mesh sieve, take 5g ginkgo powders to place in conical flask, distilled water, material-water ratio 1 is added：
30, conical flask is put into water-bath and extracts 5h, bath temperature is 100 DEG C.Then 4000r/min centrifuges 10min and takes supernatant,
Ginkgo supernatant is concentrated in vacuo in Rotary Evaporators, until concentrate probably arrives 65mL or so.Again with 70% ethyl alcohol to dense
Ginkgo supernatant after contracting is precipitated.After placing for 24 hours, the ginkgo Thick many candies for the bottom being precipitated out are used into 4000r/min
Centrifugation 10min mode be centrifuged out Thick many candies, by Thick many candies ventilation place a period of time make ethyl alcohol volatilize after -20 DEG C into
Sample after freezing processing is finally put into freeze drier and is lyophilized into powder by row freezing processing, and polysaccharide extract rate reaches
18.12%.
A kind of Ginkgo biloba polysaccharide inoxidizability assay method, the ginkgo of various concentration is made including the use of Ginkgo biloba polysaccharide powder
Leaf polyose solution prepares the Vc of corresponding concentration as positive controls, and prepare distilled water as blank control as experimental group
Group；
One kind of dropwise addition DPPH working solutions or ABTS working solutions into the polysaccharide solution of various concentration, and observation experiment group,
Positive controls, the absorbance of blank control group, calculate different Ginkgo biloba polysaccharide concentration to DPPH free radicals and ABTS from
By base clearance rate, the antioxidative intensity of Ginkgo biloba polysaccharide is further shown as.The present invention has studied the antioxygen of Ginkgo biloba polysaccharide
Change activity, the researches and exploitation for polysaccharides from ginkgo biloba beverage provides theoretical foundation.
Further, using the Ginkgo biloba polysaccharide solution of 0.5mL various concentration gradients, a concentration of 6.5 × 10 are sequentially added- 5After standing 30min after mol/L DPPH solution 2.5mL, each concentration is measured at 517nm wavelength using ultraviolet specrophotometer
The absorbance of polysaccharides from ginkgo biloba solution compares the blank control group and positive controls, obtains the Ginkgo biloba polysaccharide of various concentration
To the clearance rate of DPPH free radicals.As shown in figure 4, be the various concentration measured by the present invention polysaccharides from ginkgo biloba solution to DPPH from
By the clearance rate curve graph of base.It can be shown from the curve graph in Fig. 4 and Fig. 5, polysaccharides from ginkgo biloba is to DPPH free radicals and ABTS
Radicals scavenging function well, especially ABTS free radicals understand significant effect, clearance rate when a concentration of 0.5mg/mL
54.31% is had reached, there is good inoxidizability.
Further, using the Ginkgo biloba polysaccharide solution of 0.3mL various concentrations, ABTS working solution 4.7mL are sequentially added, are used
ABTS working solutions are as a control group；After being protected from light processing 30min to the experimental group, positive controls, control group masking foil, utilize
Ultraviolet specrophotometer measures the absorbance of each concentration polysaccharides from ginkgo biloba solution at 734nm wavelength, compares the experimental group, sun
Property control group, control group, obtain the Ginkgo biloba polysaccharide of various concentration to ABTS free radical scavenging activities.As shown in figure 5, being the present invention
Clearance rate curve graph of the polysaccharides from ginkgo biloba solution of measured various concentration to ABTS free radicals.
For those of ordinary skill in the art, without departing from the concept of the premise of the invention, it can also do
Go out several modifications and improvements, these are all within the scope of protection of the present invention.
Claims (8)
1. a kind of Ginkgo biloba polysaccharide extracting method, which is characterized in that include the following steps：
S1：By ginkgo leaf, drying to constant weight, is crushed to and at least crosses 60 mesh sizes sieve；
S2：Ginkgo leaf after crushing is utilized into water extraction and alcohol precipitation method, ginkgo leaf Thick many candies concentrate is made；
S3：The ginkgo leaf Thick many candies concentrate is redissolved, and Sevag reagents removal foreign protein is added；
S4：It is removed into step S3 in the Ginkgo biloba polysaccharide after foreign protein, precipitation, washing drying after ethyl alcohol is added, is vented to second
After alcohol volatilization, it is freeze-dried into powder, Ginkgo biloba polysaccharide powder is made.
2. a kind of Ginkgo biloba polysaccharide extracting method according to claim 1, which is characterized in that the ginkgo in the step S1
The preferred ginkgo leaf fallen leaves of leaf.
3. a kind of Ginkgo biloba polysaccharide extracting method according to claim 1, which is characterized in that water extracting alcohol in the step S2
Heavy legal system obtains the step of ginkgo leaf Thick many candies and includes：
T1：1 part of ginkgo leaf after being crushed in step S1 is taken, at least 30 parts of water is added, is extracted at 100 DEG C at least 5 hours；
T2：After 4000r/min centrifuges 10min, supernatant is taken, and supernatant is placed in Rotary Evaporators and is concentrated in vacuo, obtained
Initial concentration liquid；
T3:After standing for 24 hours, 10min is centrifuged into 4000r/min with 70% ethanol precipitation for initial concentration liquid obtained by step T2,
Obtain ginkgo leaf Thick many candies concentrate.
4. a kind of Ginkgo biloba polysaccharide extracting method according to claim 1, which is characterized in that in the step S4, be added
The concentration of ethyl alcohol is at least 70%, and the volume of ethyl alcohol is added at least by the three times volume of the ginkgo leaf Thick many candies concentrate.
5. a kind of Ginkgo biloba polysaccharide extracting method according to claim 1, which is characterized in that in the step S4, be added
After ethanol precipitation, washing are dry, at least being placed 24 hours in ventilation makes ethyl alcohol volatilize, and is carried out at freezing at least -20 DEG C
Reason, is put into freeze drier at least 48 hours and is lyophilized into powder, Ginkgo biloba polysaccharide powder is made.
6. a kind of Ginkgo biloba polysaccharide inoxidizability assay method, which is characterized in that difference is made including the use of Ginkgo biloba polysaccharide powder
The Ginkgo biloba polysaccharide solution of concentration prepares the Vc of corresponding concentration as positive controls as experimental group, and prepares distilled water work
For blank control group；
One kind of DPPH solution or ABTS working solutions is added dropwise into the polysaccharide solution of various concentration, and observation experiment group, the positive are right
According to group, the absorbance of blank control group, it is clear to DPPH free radicals and ABTS free radicals to calculate different Ginkgo biloba polysaccharide concentration
Except rate, the antioxidative intensity of Ginkgo biloba polysaccharide is further shown as.
7. a kind of Ginkgo biloba polysaccharide inoxidizability assay method according to claim 6, which is characterized in that use 0.5mL
The Ginkgo biloba polysaccharide solution of various concentration gradient, sequentially adds a concentration of 6.5 × 10-5It is quiet after mol/L DPPH working solutions 2.5mL
30min is set, measures the absorbance of each concentration polysaccharides from ginkgo biloba solution at 517nm wavelength using ultraviolet specrophotometer, is compared
The blank control group and positive controls obtain clearance rate of the Ginkgo biloba polysaccharide to DPPH free radicals of various concentration.
8. a kind of Ginkgo biloba polysaccharide inoxidizability assay method according to claim 6, which is characterized in that use 0.3mL
The Ginkgo biloba polysaccharide solution of various concentration sequentially adds ABTS working solution 4.7mL, as a control group using ABTS working solutions；It is right
After the experimental group, positive controls, control group masking foil are protected from light processing 30min, using ultraviolet specrophotometer in 734nm waves
Strong point measures the absorbance of each concentration polysaccharides from ginkgo biloba solution, compares the experimental group, positive controls, control group, obtains not
Ginkgo biloba polysaccharide with concentration is to ABTS free radical scavenging activities.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810304353.9A CN108383921A (en) | 2018-04-08 | 2018-04-08 | A kind of Ginkgo biloba polysaccharide extracting method, and the inoxidizability assay method with it |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810304353.9A CN108383921A (en) | 2018-04-08 | 2018-04-08 | A kind of Ginkgo biloba polysaccharide extracting method, and the inoxidizability assay method with it |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108383921A true CN108383921A (en) | 2018-08-10 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110577657A (en) * | 2019-08-15 | 2019-12-17 | 株洲千金药业股份有限公司 | Method for improving inoxidizability of noni polysaccharide |
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| CN103446195A (en) * | 2013-08-22 | 2013-12-18 | 陕西洋县志建药业科技有限公司 | Preparation method of ginkgo biloba extract |
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| CN102002112A (en) * | 2010-12-02 | 2011-04-06 | 蒋熠霞 | Process for extracting ginkgo leaf polysaccharide with aid of ultrasonic |
| CN103446195A (en) * | 2013-08-22 | 2013-12-18 | 陕西洋县志建药业科技有限公司 | Preparation method of ginkgo biloba extract |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110577657A (en) * | 2019-08-15 | 2019-12-17 | 株洲千金药业股份有限公司 | Method for improving inoxidizability of noni polysaccharide |
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