CN101229114B - Free radical scavenging agent - Google Patents
Free radical scavenging agent Download PDFInfo
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- CN101229114B CN101229114B CN2008100575643A CN200810057564A CN101229114B CN 101229114 B CN101229114 B CN 101229114B CN 2008100575643 A CN2008100575643 A CN 2008100575643A CN 200810057564 A CN200810057564 A CN 200810057564A CN 101229114 B CN101229114 B CN 101229114B
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Abstract
The invention discloses a free radical removing agent. The free radical removing agent of the invention is extracted from the materials including flos sophora, olive and pawpaw with an organic solvent. The weight portions of the flos sophora in the material are 5.6 to 51.2; the weight portions of the olive are 2.9 to 38.3; the weight portions of the pawpaw are 10.7 to 43.1. The organic solvent is anhydrous ethanol. The composite free radical network removing agent of the invention can play a great role in the manufacturing of the cosmetics with efficacies of anti-caducity, anti-sunning and repairs after being exposed to the sun.
Description
Technical field
The present invention relates to a kind of free radical scavenger, particularly relate to and from Chinese herbal medicine, screen free radical scavenger.
Background technology
The mechanism of explaining skin aging at present has tens kinds, as hypoendocrinism theory, immunologic function degression theory, cross linkage theory, free radical injury theory etc., wherein the free radical injury theory is present unique theory that can disclose naturally aging of skin and photo-aging.This theoretical main points of view is: 1. free radical mainly contains superoxide anion, hydroxy radical, singlet oxygen etc., because the outermost electron of these molecules is for not arranging in pairs, so its chemism is extremely active.2. being created in the biologic artifact of free radical is a kind of general phenomenon, and on the one hand, organism needs the participation of active oxygen to finish some physiological process; On the other hand, the active oxygen accumulation is superfluous, can produce the damaging action of molecule, cellular level.3. existing with the Hydrogen Peroxide Metabolism in the body is the defence or the control mechanism naturally of core, comprises enzyme mechanism (as superoxide dismutase SOD, catalase, glutathione peroxidase etc.) and chemical mechanism (as antioxidants such as vitamin E, vitamin C, glutathion).
Occurring in nature has several factors can induce the generation of free radical, and as water quality, air pollution, over-drastic exposure in sunshine, and over-exposure is in pollutant and ionizing radiation; And the other factor, as smoking drink, tension pressure, spiritual agitation etc. also can produce free radical.With advancing age, those activity that can resist the scavenger enzyme of free radical invasion and attack reduce gradually, the generation of free radical and removing disequilibrium.Excessive free radical will damage body, causes the oxidative damage of many biomacromolecules in cell, attacks protein, makes it to take place crosslinked, deposition, attacks lipid, makes its peroxidating.These albumen or lipids under fire become lipofuscin, form senile plaque.The crosslinked collagen that makes of connective tissue collagen is stiff, skin is followed the string and gloss, becomes coarse, fold etc. occurs.
The metabolic pathway of free radical is a network system in vivo, between the different free radicals phase co-conversion can take place, and concrete metabolic pathway as shown in Figure 1.
Oxygen is accepted an electronics and is formed ultra-oxygen anion free radical O
2 -, O
2 -Can be under the catalysis of ferrum chelate and H
2O
2Reaction produces hydroxy radical OH, and course of reaction is as follows:
OH is the most active active oxygen species of chemical property.Its reaction characteristics is no specificity, almost with organism in all substances, can both react as sugar, protein, DNA, base, phospholipid and organic acid etc., and reaction rate is fast, can make the non-free radical reactant become free radical.For example, the effect of the biomacromolecule (representing) in OH and cell membrane and the cellular content: OH+RH → H with RH
2O+R.The organic free radical R that generates can continue again to work with oxygen and generate RO
2: R+O
2→ RO
2Like this, free radical can transmit and breed by the way.
Simultaneously, at present the research of free radical scavenger is confined to remove the screening of the active component of a certain free radical, the anti-aging cosmetics of having developed also just has the removing effect at a certain type free base, and as can be seen from Figure 1, have close contact between the metabolism of various free radicals, i.e. the generation of free radical is a network system.Single degree of freedom base scavenger just works to certain metabolic process, only tend to cause the generation of other free radical at the scavenger of single degree of freedom base, (SOD) can remove superoxide anion as superoxide dismutase, if but its metabolite hydrogen peroxide can not in time be removed, can produce hydroxy radical in the presence of ferrous ion, hydroxy radical is at present to the strongest free radical of body biomacromolecule attack.Therefore, if filter out a kind of can be simultaneously to source free radical (superoxide anion), the free radical that attack is the strongest (hydroxy radical) and free radical (R, the RO that can cause chain reaction
2) free radical scavenger, just can efficiently prevent the skin aging that causes owing to free radical injury, can free radical scavenger that remove effect all not arranged to multiple free radical but also prepare at present.
The Flos Sophorae Immaturus is the dry flower of leguminous plant Chinese scholartree (Sophora japonica L.), and the function of cooling blood for hemostasis, clearing away heat-fire is arranged.
Fructus Canarii (Fructus canarii) is the fruit of Burseraceae (Burseraceae) Canarium (Canarium) plant Canariumalbum Raeusch, it is exactly China's conventional Chinese medicine material from ancient times, its nature and flavor are sweet, sour, puckery, warm, flat, nontoxic, return lung stomach two warps, have heat clearing away, sore-throat relieving, eliminate the phlegm, promote the production of body fluid, effects such as spleen invigorating, detoxifcation, be usually used in laryngopharynx swelling and pain, cough, excessive thirst, fish Trionyx sinensis Wiegmann and poison etc.
Fructus Chaenomelis is the fruit of rosaceous plant chaenomeles lagenaria Chaenomeles lagenaria (Loisel.) koidz., has the effects such as anticancer, arthralgia relieving by relaxing muscles and tendons and spleen removing dampness that relieve internal heat.
Flos Caryophylli is the alabastrum of plant clove of myrtaceae Syzygium aromaticum (L.) Merr.et Perry.Xin Wen returns spleen, kidney, stomach warp.Warming middle-JIAO to send down the adverse QI, warming the kidney to activate YANG.
Radix Scutellariae is the root of labiate Radix Scutellariae Scutellaria baicalensis Georgi, has the hot dampness that disappears, eliminating fire and detoxication, hemostasis, antiabortive effect.
Flos Sophorae is the dry flower of leguminous plant Chinese scholartree (Sophora japonica L.), and the function of cooling blood for hemostasis, clearing away heat-fire is arranged.
Summary of the invention
The purpose of this invention is to provide a kind of free radical scavenger.
Free radical scavenger provided by the present invention is to extract from the raw material that comprises the Flos Sophorae Immaturus, Fructus Canarii and Fructus Chaenomelis with organic solvent to obtain;
The parts by weight of the Flos Sophorae Immaturus are in the described raw material: 5.6-51.2 part; The parts by weight of Fructus Canarii are: 2.9-38.3 part; The parts by weight of Fructus Chaenomelis are: 10.7-43.1 part;
Described organic solvent is that methanol, volumn concentration are ethanol water or the dehydrated alcohol more than 70%.
The parts by weight of the described Flos Sophorae Immaturus are specially 51.2 parts; The parts by weight of described Fructus Canarii are specially 21.2 parts; The parts by weight of described Fructus Chaenomelis are specially 10.7 parts.
Described free radical can be at least a in following four kinds of free radicals: total free radical, DPPH free radical, superoxide anion and hydroxy radical.
The raw material of free radical scavenger of the present invention also can comprise Flos Caryophylli or Radix Scutellariae;
Described free radical is total free radical, DPPH free radical and hydroxy radical; The raw material of described free radical scavenger consists of: Flos Sophorae Immaturus 5.6-51.2 weight portion, Fructus Canarii 2.9-38.3 weight portion, Fructus Chaenomelis 10.7-43.1 weight portion, Flos Caryophylli 0.9-33.3 weight portion;
Described free radical is a superoxide anion; The raw material of described free radical scavenger consists of: Flos Sophorae Immaturus 5.6-51.2 weight portion, Fructus Canarii 2.9-38.3 weight portion, Fructus Chaenomelis 10.7-43.1 weight portion, Radix Scutellariae 1.9-17.5 weight portion.
The raw material of free radical scavenger of the present invention also can comprise Flos Sophorae;
Described free radical is total free radical, DPPH free radical and hydroxy radical; The raw material of described free radical scavenger consists of: Flos Sophorae Immaturus 5.6-51.2 weight portion, Fructus Canarii 2.9-38.3 weight portion, Fructus Chaenomelis 10.7-43.1 weight portion, Flos Caryophylli 0.9-33.3 weight portion, Flos Sophorae 3.2-41.4 weight portion;
Described free radical is a superoxide anion; The raw material of described free radical scavenger consists of: Flos Sophorae Immaturus 5.6-51.2 weight portion, Fructus Canarii 2.9-38.3 weight portion, Fructus Chaenomelis 10.7-43.1 weight portion, Radix Scutellariae 1.9-17.5 weight portion, Flos Sophorae 3.2-41.4 weight portion.
The raw material of free radical scavenger of the present invention also can comprise Flos Caryophylli, Radix Scutellariae and Flos Sophorae;
Described free radical is total free radical and DPPH free radical; The raw material of described free radical scavenger consists of: Flos Sophorae Immaturus 10.1-51.2 weight portion, Fructus Canarii 2.9-21.2 weight portion, Fructus Chaenomelis 12.3-16.9 weight portion, Flos Caryophylli 6.7-33.3 weight portion, Flos Sophorae 3.2-41.4 weight portion and Radix Scutellariae 5.2-17.5 weight portion;
Described free radical is a superoxide anion; The raw material of described free radical scavenger consists of: Flos Sophorae Immaturus 5.6-51.2 weight portion, Fructus Canarii 12.6-38.3 weight portion, Fructus Chaenomelis 12.6-43.1 weight portion, Flos Caryophylli 0.9-6.6 weight portion, Flos Sophorae 3.2-23.3 weight portion and Radix Scutellariae 1.9-5.2 weight portion;
Described free radical is a hydroxy radical; The raw material of described free radical scavenger consists of: Flos Sophorae Immaturus 5.6-51.2 weight portion, Fructus Canarii 2.9-38.3 weight portion, Fructus Chaenomelis 12.3-43.1 weight portion, Flos Caryophylli 0.9-18.6 weight portion, Flos Sophorae 3.2-41.4 weight portion and Radix Scutellariae 1.9-14.4 weight portion.
The parts by weight of the described Flos Sophorae Immaturus are specially 51.2 parts; The parts by weight of described Fructus Canarii are specially 21.2 parts; The parts by weight of described Fructus Chaenomelis are specially 12.6 parts; The parts by weight of described Flos Caryophylli are specially 6.7 parts; The parts by weight of described Flos Sophorae are specially 3.2 parts; The parts by weight of described Radix Scutellariae are specially 5.2 parts.
The present invention is based on the characteristics of the secondary metabolites effect complexity of Chinese herbal medicine, is the screening object of reference with the Radix Glycyrrhizae extract, compares by the action effect that existing Chinese herbal medicine is removed various free radicals, obtains removing the free radical scavenger of various free radicals.Free radical scavenger of the present invention is delaying decrepitude of skin effectively, gives new life of skin and vigor, and has the effect of shining the back reparation.Can be used for defying age, sun-proof, shine in the cosmetics such as back reparations, be the green cosmetics functional additive of low stimulation pure natural.
Description of drawings
Fig. 1 is the metabolic pathway of free radical in the organism
Fig. 2 is the standard curve of total radical scavenging activity screening
A represents that the response time is 15 minutes among Fig. 2
B represents that the response time is 45 minutes among Fig. 2
The specific embodiment
1, the preparation of Chinese herbal medicine extract
With pulverizer Radix Glycyrrhizae, the Flos Sophorae Immaturus, Fructus Canarii, Fructus Chaenomelis, Flos Caryophylli, Radix Scutellariae, these 7 kinds of Chinese herbal medicine pulverize separately of Flos Sophorae are also crossed 120 mesh sieves.Each 30g of Chinese herbal medicine that takes by weighing after the pulverizing places 100mL ground triangular flask, adds the 300mL dehydrated alcohol, bathes in the shaking table (55 ℃ of vibrations at full warm air, 150rpm) after 1 hour, use filter paper filtering, collect filtrate, precipitation adds the 300mL dehydrated alcohol again, repeats above step 2 time.After 3 filtrates merging, quantitatively be the sample solution of gained to 900mL with dehydrated alcohol.
2, Chinese herbal medicine extract is removed the mensuration of total free radical ability
Carry out the mensuration that Chinese herbal medicine extract is removed total free radical ability: Fe according to following characteristic
3+-three pyridines three azines (tripyridyl-triazine, TPTZ) Fe in (Sigma-Aldrich company)
3+Can be reduced to Fe by the reducing substances in the testing sample
2+And present blue color, and maximum absorbance is arranged in the 593nm place.Size according to absorbance calculates the power of removing total free radical ability in the testing sample.Absorbance is big more, shows that testing sample has the ability of the strong more total free radical of removing.
Standard curve making: draw the copperas solution that 0.3mL concentration is respectively 25,100,150,200,500,1000 μ mol/L, mix with 0.3mL 10mmol/L TPTZ solution (preparing as solvent) and the acetate buffer of 3mL 0.3mol/L pH3.6, survey its absorbance down in 593nm after placing down 15min or 45min for 37 ℃ with 40mmol/L HCl solution.Concentration with ferrous sulfate is abscissa, and absorbance is the standard curve that vertical coordinate is drawn the differential responses time.Concrete standard curve as shown in Figure 2.As can be seen from Figure 2, the R of the standard curve when keeping 15min (Fig. 2 A) and 45min (Fig. 2 B), drawn of reaction system
2Be respectively 0.9983 and 0.998, show that this reaction system keeps stable in 45min.Carry out the mensuration needs of many group samples for convenience simultaneously, select the response time of 45min as this system, iron concentration and absorbance linear relationship equation are y=1.7777x-0.0392 under this reaction system, R
2=0.998.
The mensuration of sample: the sample solution and 3mL TPTZ working solution (the 0.3mol/LpH3.6 acetate buffer 25mL that draw the 0.4mL step 1 respectively, 10mmol/L TPTZ solution 2.5mL, 20mmol/L ferric chloride solution 2.5mL) mix homogeneously, survey its absorbance down in 593nm behind the placement 45min down for 37 ℃, the gained absorbance is designated as A; Replace sample in contrast with sample solvent (dehydrated alcohol), the gained absorbance is designated as A
0Replace the TPTZ working solution as blank with the 0.3mol/LpH3.6 acetate buffer, with the zeroing of pH3.6 acetate buffer, the gained absorbance is designated as A
1
With A-A
0-A
1The corresponding standard curve of numerical value on corresponding ferrous sulfate concentration (mmol/L) be defined as the FRAP value.Sample solution is removed total free radical ability carry out following regulation: FRAP value=1 is defined as 1 unit of activity (U), and the size that testing sample is removed total free radical ability has what unit of activity (U/ml) expression with every milliliter.
Respectively the total free radical ability of the removing of 7 kinds of Chinese herbal medicine extracts is measured according to the method described above, three repetitions are established in experiment, and three times multiple meansigma methods is as shown in table 1.
The different Chinese herbal medicine extracts of table 1 are removed total free radical ability relatively
| The Chinese herbal medicine ethanol extract | Remove total free radical ability (U/ml) |
| Flos Caryophylli Flos Sophorae Immaturus Flos Sophorae Fructus Canarii Fructus Chaenomelis Radix Scutellariae Radix Glycyrrhizae | 139.75 113.28 66.94 44.44 38.14 28.33 9.44 |
3, Chinese herbal medicine extract is removed the mensuration of hexichol for bitterness acyl group free radical (DPPH) ability
Hexichol is for bitterness acyl group (DPPH, C
18H
12N
5O
6) free radical (Sigma-Aldrich company) be a kind of very stable be the free radical at center with nitrogen, outermost electron is arranged and is single electron on the track, at the 520nm place strong absorption is arranged.Its alcoholic solution is darkviolet, and behind the adding testing sample, it is removed DPPH free radical and causes situation that absorbance reduces to utilize spectrophotometry under 520nm.Can know the ability that testing sample is removed the DPPH free radical by inference according to the absorbance situation of change.If testing sample can be removed the DPPH free radical, show that then this testing sample has the effect of free radicals such as reducing hydroxy radical, alkane free radical or peroxy radical, and have the effect that interrupts the lipid peroxidation chain reaction.
Sample determination: the sample and the 0.9mL distilled water that in centrifuge tube, add the 0.1mL step 1 respectively, mix homogeneously, the DPPH free-atom aqueous solution (is that solvent is made into the dehydrated alcohol) that adds 2mL 0.1777mmol/L again, abundant mixing, survey its absorbance in the 520nm place after placing 45min, the gained absorbance is designated as A; Replace 2mL DPPH free-atom aqueous solution with the 2mL distilled water, with the water zeroing, the gained absorbance is designated as A
1With the 0.1mL sample solution in the 0.1mL distilled water replacement system, the gained absorbance is designated as A
0
Sample calculates as follows to the clearance rate of DPPH free radical:
K(％)＝[A
0-(A-A
1)]/A
0×100％。
Clearance rate to the DPPH free radical that obtains by above-mentioned formula is carried out following regulation: will be defined as 1 unit of activity (U) to 50% of DPPH free radical scavenging activity, testing sample is removed the size of DPPH free radical ability and is represented to have what unit of activity (U/mL) in every milliliter of extracts of Chinese herbal medicine.
According to the method described above, respectively the removing DPPH free radical ability of 7 kinds of Chinese herbal medicine extracts is measured, three repetitions are established in experiment, and three times multiple meansigma methods is as shown in table 2.
The different Chinese herbal medicine extracts of table 2 are removed DPPH free radical ability relatively
| The Chinese herbal medicine ethanol extract | Remove DPPH free radical ability (U/mL) |
| Flos Caryophylli Flos Sophorae Immaturus Flos Sophorae Fructus Canarii Fructus Chaenomelis Radix Scutellariae Radix Glycyrrhizae | 19.22 18.96 18.86 19.05 18.20 19.16 11.81 |
4, Chinese herbal medicine extract is removed the mensuration of superoxide anion ability
Sample determination: in small beaker, add successively 2mL PBS (pH7.8,0.1mol/L), 0.3mL L-methionine (0.026mol/L), 0.3mL nitroblue tetrazolium (NBT) (0.75mmol/L), 0.3mL riboflavin aqueous solution (0.1mmol/L EDTA and 0.02mmol/L riboflavin) and 0.1mL sample solution.Behind the illumination 15min, with the contrast zeroing of placing under the dark, measure its absorbance under 560nm, the gained absorbance is designated as A; (pH7.8 0.1mol/L) replaces sample solution in contrast, and the gained absorbance is designated as A with 0.1mLPBS
0(pH7.8 0.1mol/L) mixes, and (the gained absorbance is designated as A for pH7.8,0.1mol/L) zeroing with 3mL PBS with 0.1mL sample solution and 2.9mL PBS
1
Sample calculates as follows to the clearance rate of superoxide anion:
K(％)＝(A
0-A+A
1)/A
0×100％。
Clearance rate to the superoxide anion that obtains by above-mentioned formula is carried out following regulation: will be defined as 1 unit of activity (U) to 50% of superoxide anion clearance rate, testing sample is removed the size of superoxide anion ability and is represented to have what unit of activity (U/mL) in every milliliter of extracts of Chinese herbal medicine.
According to the method described above, respectively the removing superoxide anion ability of 7 kinds of Chinese herbal medicine extracts is measured, three repetitions are established in experiment, and three times multiple meansigma methods is as shown in table 3.
The different Chinese herbal medicine extracts of table 3 are removed the superoxide anion ability relatively
| The Chinese herbal medicine ethanol extract | Remove superoxide anion ability (U/mL) |
| Flos Caryophylli Flos Sophorae Immaturus Flos Sophorae Fructus Canarii Fructus Chaenomelis Radix Scutellariae Radix Glycyrrhizae | 12.14 7.31 13.46 12.79 13.44 11.58 6.57 |
5, Chinese herbal medicine extract is removed the mensuration of hydroxy radical ability
Sample determination: the PBS solution, the 0.5mL 1mmol/L FeSO that in centrifuge tube, add 0.5mL 0.4mmoL/L crystal violet solution, 1.4mL 0.2mol/LpH7.4 respectively
4Solution and 0.1mL sample solution add 0.5mL6%H at last
2O
2Measure its absorbance under 584nm after placing 1h, the gained absorbance is designated as A; Replace sample in contrast with the 0.1mL distilled water, the gained absorbance is designated as CK
1Replace sample and 0.5mL distilled water to replace H with the 0.1mL distilled water
2O
2, the gained absorbance is designated as CK
2
Sample calculates as follows to the clearance rate of hydroxy radical:
K(％)＝[1-(CK
2-A)/(CK
2-CK
1)]×100％。
Clearance rate to the hydroxy radical that obtains by above-mentioned formula is carried out following regulation: will be defined as 1 unit of activity (U) to 50% of hydroxy radical clearance rate, testing sample is removed the size of hydroxy radical ability and is represented to have what unit of activity (U/mL) in every milliliter of extracts of Chinese herbal medicine.
According to the method described above, respectively the removing hydroxy radical ability of 7 kinds of Chinese herbal medicine extracts is measured, three repetitions are established in experiment, and three times multiple meansigma methods is as shown in table 4.
The different Chinese medicine extract of table 4 are removed the hydroxy radical ability relatively
| The Chinese herbal medicine ethanol extract | Remove hydroxy radical ability (U/mL) |
| Flos Caryophylli Flos Sophorae Immaturus Flos Sophorae Fructus Canarii Fructus Chaenomelis Radix Scutellariae Radix Glycyrrhizae | 7.77 16.03 14.25 9.85 9.40 12.53 6.04 |
Embodiment 2, remove the mensuration of ability with the free radical scavenger of 6 kinds of Chinese herbal medicine extracts preparation
Flos Sophorae, the Flos Sophorae Immaturus, Flos Caryophylli, Fructus Chaenomelis, Fructus Canarii are the Chinese herbal medicine of integration of edible and medicinal herbs, and Radix Scutellariae is conventional Chinese herbal medicine, and the said herbal medicine extract all is not the Chinese herbal medicine extract of listing in the national cosmetic hygienic practice that limits the use of, so safety is also higher.The color and luster of extract is lower simultaneously, can satisfy the needs of cosmetics sense organ.By the DPS software design raw material compound recipe of 12 free radical scavengers.In 12 free radical scavengers, every kind of proportion of raw materials is as shown in table 5.
The raw material of table 5 free radical scavenger is formed (weight: g)
| Numbering | Flos Caryophylli | The Flos Sophorae Immaturus | Flos Sophorae | Fructus Canarii | Fructus Chaenomelis | Radix Scutellariae |
| Free |
17.2 11.7 6.0 0.8 33.3 18.6 11.5 9.0 6.7 4.6 2.6 0.9 | 0.8 3.6 2.8 11.9 10.1 10.4 9.8 3.0 51.2 25.3 13.8 5.6 | 3.8 18.0 52.6 5.1 12.9 41.4 39.3 20.2 3.2 45.8 23.3 10.2 | 12.1 7.9 0.9 31.5 9.3 2.9 2.5 17.9 21.2 0.5 12.6 38.3 | 19.1 24.9 10.7 28.9 16.9 12.3 19.9 31.2 12.6 18.8 41.7 43.1 | 47.0 34.0 26.9 21.8 17.5 14.4 16.9 18.7 5.2 5.0 6.0 1.9 |
Proportioning according to table 5 takes by weighing various Chinese herbal medicine, with pulverizer various Chinese herbal medicine pulverize separately is also crossed 120 mesh sieves.The Chinese herbal medicine powder mixture 30g that gets after the pulverizing places the ground triangular flask, adds the 300mL dehydrated alcohol, bathes in the shaking table (55 ℃ of vibrations at full warm air, 150rpm) after 1 hour, use filter paper filtering, collect filtrate, precipitation adds the 300mL dehydrated alcohol again, repeats above step 2 time.After 3 filtrates merging, quantitatively to 900mL, be free radical scavenger 1-12 with dehydrated alcohol.
Measure total free radical ability, removing DPPH free radical ability, the removing superoxide anion ability of free radical scavenger 1-12 and remove the hydroxy radical ability according to the method among the embodiment 1.Wherein, remove the size of total free radical ability, removing DPPH free radical ability, removing superoxide anion ability and removing hydroxy radical ability and all represent that to have what unit of activity (U/mL) in every milliliter of free radical scavenger the definition of unit of activity is with embodiment 1.The result is as shown in table 6.Table 6 shows among the free radical scavenger 1-12, remove total free radical ability three free radical scavengers the strongest and be respectively free radical scavenger 5, free radical scavenger 6 and free radical scavenger 9, be respectively specific to each component effective range of free radical scavenger raw material: Flos Caryophylli 6.7-33.3g, Flos Sophorae Immaturus 10.1-51.2g, Flos Sophorae 3.2-41.4g, Fructus Canarii 2.9-21.2g, Fructus Chaenomelis 12.3-16.9g, Radix Scutellariae 5.2-17.5g with removing DPPH free radical ability; Remove three the strongest free radical scavengers of superoxide anion ability and be respectively free radical scavenger 9, free radical scavenger 11 and free radical scavenger 12, be respectively specific to each component effective range of free radical scavenger raw material: Flos Caryophylli 0.9-6.6g, Flos Sophorae Immaturus 5.6-51.2g, Flos Sophorae 3.2-23.3g, Fructus Canarii 12.6-38.3g, Fructus Chaenomelis 12.6-43.1g, Radix Scutellariae 1.9-5.2g; Remove three the strongest free radical scavengers of hydroxy radical ability and be respectively free radical scavenger 6, free radical scavenger 9 and free radical scavenger 12, be respectively specific to each component effective range of free radical scavenger raw material: Flos Caryophylli 0.9-18.6g, Flos Sophorae Immaturus 5.6-51.2g, Flos Sophorae 3.2-41.4g, Fructus Canarii 2.9-38.3g, Fructus Chaenomelis 12.3-43.1g, Radix Scutellariae 1.9-14.4g.Therefore, the effective range of the various herbal components of compound free radical network scavenger raw material is: Flos Caryophylli 0.9-33.3g, Flos Sophorae Immaturus 5.6-51.2g, Flos Sophorae 3.2-41.4g, Fructus Canarii 2.9-38.3g, Fructus Chaenomelis 12.3-43.1g, Radix Scutellariae 1.9-17.5g.
Table 6 free radical scavenger is removed energy unit of force: the U/mL of various free radicals
| Numbering | Total radical scavenging activity | The DPPH radical scavenging activity | Superoxide anion is removed ability | Hydroxy radical is removed ability |
| Free |
87.72 58.72 69.78 38.32 126.94 106.75 68.44 58.46 172.42 104.88 87.98 71.68 | 69.96 58.40 63.96 46.12 77.72 75.09 57.20 55.38 89.00 71.16 69.64 60.59 | 73.25 68.42 72.21 16.69 71.65 71.19 68.96 71.41 73.70 68.24 75.14 74.23 | 11.17 12.11 15.62 11.31 15.36 17.38 12.21 15.36 18.74 13.45 16.61 17.71 |
By as can be seen comprehensive, free radical scavenger 9 is to remove total free radical ability, remove DPPH free radical ability, remove the superoxide anion ability and remove the free radical scavenger that the hydroxy radical ability all comes front three, the best free radical scavenger that screens for the present invention.The raw material of free radical scavenger 9 consists of: Flos Caryophylli 6.7g, Flos Sophorae Immaturus 51.2g, Flos Sophorae 3.2g, Fructus Canarii 21.2g, Fructus Chaenomelis 12.6g, Radix Scutellariae 5.2g.The total free radical ability of the removing of free radical scavenger 9, remove DPPH free radical ability, remove the superoxide anion ability and remove the hydroxy radical ability and be respectively 172.42,89.00,73.70 and 18.74 active unit/ml samples, be respectively Radix Glycyrrhizae extract the total free radical ability of removing, remove DPPH free radical ability, remove the superoxide anion ability and remove 18.27 times, 7.54 times, 11.22 times and 3.10 times of hydroxy radical ability.
Embodiment 3, remove the mensuration of ability with the free radical scavenger of 3 kinds of Chinese herbal medicine extracts preparation
With the Flos Sophorae Immaturus, Fructus Chaenomelis and Fructus Canarii is raw material, has designed the raw material compound recipe of 4 free radical scavengers.In 4 free radical scavengers, every kind of proportion of raw materials is as shown in table 7.
The raw material of table 7 free radical scavenger is formed (weight: g)
| Numbering | The Flos Sophorae Immaturus | Fructus Canarii | Fructus Chaenomelis |
| Free radical scavenger 13 free radical scavengers 14 free radical scavengers 15 free radical scavengers 16 | 5.6 5.6 51.2 51.2 | 2.9 21.2 2.9 21.2 | 12.6 10.7 12.6 10.7 |
Proportioning according to table 7 takes by weighing various Chinese herbal medicine, with pulverizer various Chinese herbal medicine pulverize separately is also crossed 120 mesh sieves.The Chinese herbal medicine powder mixture 30g that gets after the pulverizing places the ground triangular flask, adds the 300mL dehydrated alcohol, bathes in the shaking table (55 ℃ of vibrations at full warm air, 150rpm) after 1 hour, use filter paper filtering, collect filtrate, precipitation adds the 300mL dehydrated alcohol again, repeats above step 2 time.After 3 filtrates merging, quantitatively to 900mL, be free radical scavenger 13-16 with dehydrated alcohol.
Measure total free radical ability, removing DPPH free radical ability, the removing superoxide anion ability of free radical scavenger 13-16 and remove the hydroxy radical ability according to the method among the embodiment 1.Wherein, remove the size of total free radical ability, removing DPPH free radical ability, removing superoxide anion ability and removing hydroxy radical ability and all represent that to have what unit of activity (U/mL) in every milliliter of free radical scavenger the definition of unit of activity is with embodiment 1.The result is as shown in table 8.Table 8 shows among the free radical scavenger 13-16 that removing total free radical ability, removing DPPH free radical ability, remove the superoxide anion ability and remove the strongest free radical scavenger of hydroxy radical ability is free radical scavenger 16.Each component of this free radical scavenger raw material is respectively: Flos Sophorae Immaturus 51.2g, Fructus Canarii 21.2g, Fructus Chaenomelis 10.7g.The total free radical ability of the removing of this free radical scavenger 16, remove DPPH free radical ability, remove the superoxide anion ability and remove the hydroxy radical ability and be respectively 147.57,66.84,64.70 and 13.47 active unit/ml samples, be respectively Radix Glycyrrhizae extract the total free radical ability of removing, remove DPPH free radical ability, remove the superoxide anion ability and remove 15.63 times, 5.66 times, 9.85 times and 2.23 times of hydroxy radical ability.
Table 8 free radical scavenger is removed energy unit of force: the U/mL of various free radicals
| Numbering | Total radical scavenging activity | The DPPH radical scavenging activity | Superoxide anion is removed ability | Hydroxy radical is removed ability |
| Free radical scavenger 13 free radical scavengers 14 free radical scavengers 15 free radical scavengers 16 | 37.97 49.13 146.40 147.57 | 15.83 24.24 64.53 66.87 | 63.12 61.69 61.91 64.70 | 2.31 2.54 13.20 13.47 |
Embodiment 4, remove the mensuration of ability with the free radical scavenger of 4 kinds of Chinese herbal medicine extracts preparation
On the basis of the free radical scavenger 16 for preparing with the Flos Sophorae Immaturus, Fructus Canarii and three kinds of Chinese herbal medicine of Fructus Chaenomelis that embodiment 3 filters out, investigate any one or two kinds of in Flos Caryophylli, Radix Scutellariae, the Flos Sophorae and mixed its extract of back with free radical scavenger 16 as auxiliary material and remove the ability of various free radicals, designed the raw material compound recipe of 4 free radical scavengers.In 4 free radical scavengers, every kind of proportion of raw materials is as shown in table 9.
The raw material of table 9 free radical scavenger is formed (weight: g)
| Numbering | The Flos Sophorae Immaturus | Fructus Canarii | Fructus Chaenomelis | Flos Caryophylli | Radix Scutellariae | Flos Sophorae |
| Free radical scavenger 17 free radical scavengers 18 free radical scavengers 19 free radical scavengers 20 | 51.2 51.2 51.2 51.2 | 21.2 21.2 21.2 21.2 | 10.7 10.7 10.7 10.7 | 6.7 0 0 6.7 | 0 5.2 5.2 0 | 0 3.2 0 3.2 |
Proportioning according to table 9 takes by weighing various Chinese herbal medicine, with pulverizer various Chinese herbal medicine pulverize separately is also crossed 120 mesh sieves.The Chinese herbal medicine powder mixture 30g that gets after the pulverizing places the ground triangular flask, adds the 300mL dehydrated alcohol, bathes in the shaking table (55 ℃ of vibrations at full warm air, 150rpm) after 1 hour, use filter paper filtering, collect filtrate, precipitation adds the 300mL dehydrated alcohol again, repeats above step 2 time.After 3 filtrates merging, quantitatively to 900mL, be free radical scavenger 17-20 with dehydrated alcohol.
Measure total free radical ability, removing DPPH free radical ability, the removing superoxide anion ability of free radical scavenger 17-20 and remove the hydroxy radical ability according to the method among the embodiment 1.Wherein, remove the size of total free radical ability, removing DPPH free radical ability, removing superoxide anion ability and removing hydroxy radical ability and all represent that to have what unit of activity (U/mL) in every milliliter of free radical scavenger the definition of unit of activity is with embodiment 1.The result is as shown in table 10.Table 10 shows on the basis of free radical scavenger 16, independent increase Flos Caryophylli is respectively 152.52,86.02,70.00 and 17.26 active unit/ml samples as the total free radical ability of the removing of the free radical scavenger 17 of auxiliary material, removing DPPH free radical ability, removing superoxide anion ability and removing hydroxy radical ability, has improved 3.35%, 28.64%, 8.19% and 28.14% respectively than the total free radical ability of the removing of free radical scavenger 16, removing DPPH free radical ability, removing superoxide anion ability and removing hydroxy radical ability.On the basis of free radical scavenger 17, increase Flos Sophorae again and be respectively 154.72,81.99,67.87 and 18.42 active unit/ml samples as the total free radical ability of the removing of the free radical scavenger 20 of auxiliary material, removing DPPH free radical ability, removing superoxide anion ability and removing hydroxy radical ability, improved 1.44% and 6.72% respectively than total free radical ability of the removing of free radical scavenger 17 and removing hydroxy radical ability, then descended 4.92% and 3.14% respectively with removing superoxide anion ability but remove DPPH free radical ability.
Use the Flos Sophorae Immaturus what embodiment 3 filtered out, on the basis of the free radical scavenger 16 of three kinds of Chinese herbal medicine preparations of Fructus Canarii and Fructus Chaenomelis, increase Radix Scutellariae separately as the total free radical ability of the removing of the free radical scavenger 19 of auxiliary material, remove DPPH free radical ability, remove the superoxide anion ability and remove the hydroxy radical ability and be respectively 151.18,76.31,71.31 and 12.78 active unit/ml samples, than the total free radical ability of the removing of free radical scavenger 16, remove DPPH free radical ability and improved 2.45% respectively with removing superoxide anion ability, 14.12% and 10.22%, then descended 5.12% but remove the hydroxy radical ability.On the basis of free radical scavenger 19, increase Flos Sophorae again and be respectively 142.06,76.54,73.23 and 13.28 active unit/ml samples as the total free radical ability of the removing of the free radical scavenger 18 of auxiliary material, removing DPPH free radical ability, removing superoxide anion ability and removing hydroxy radical ability, than the total free radical ability drop of the removing of free radical scavenger 19 6.03%, remove DPPH free radical ability, remove the superoxide anion ability and remove the hydroxy radical ability and then improved 0.30%, 2.69% and 3.91%.
Table 10 free radical scavenger is removed energy unit of force: the U/mL of various free radicals
| Numbering | Total radical scavenging activity | The DPPH radical scavenging activity | Superoxide anion is removed ability | Hydroxy radical is removed ability |
| Free radical scavenger 17 free radical scavengers 18 free radical scavengers 19 free radical scavengers 20 | 152.52 142.06 151.18 154.72 | 86.02 76.54 76.31 81.99 | 70.00 73.23 71.31 67.87 | 17.26 13.28 12.78 18.42 |
Claims (14)
1. a free radical scavenger that comes from Chinese herbal medicine is to extract from the raw material that comprises the Flos Sophorae Immaturus, Fructus Canarii and Fructus Chaenomelis with organic solvent to obtain;
The parts by weight of the Flos Sophorae Immaturus are 5.6-51.2 part in the described raw material; The parts by weight of Fructus Canarii are 2.9-38.3 part; The parts by weight of Fructus Chaenomelis are 10.7-43.1 part;
Described organic solvent is that methanol, volumn concentration are ethanol water or the dehydrated alcohol more than 70%.
2. free radical scavenger according to claim 1 is characterized in that: described organic solvent is a dehydrated alcohol.
3. free radical scavenger according to claim 1 is characterized in that: described free radical is at least a in following four kinds of free radicals: total free radical, DPPH free radical, superoxide anion and hydroxy radical.
4. according to claim 1 or 2 or 3 described free radical scavengers, it is characterized in that: described raw material is made up of the Flos Sophorae Immaturus, Fructus Canarii and Fructus Chaenomelis; The parts by weight of the described Flos Sophorae Immaturus are 51.2 parts; The parts by weight of described Fructus Canarii are 21.2 parts; The parts by weight of described Fructus Chaenomelis are 10.7 parts.
5. according to claim 1 or 2 or 3 described free radical scavengers, it is characterized in that: also comprise Flos Caryophylli in the described raw material; The raw material of described free radical scavenger consists of: Flos Sophorae Immaturus 5.6-51.2 weight portion, Fructus Canarii 2.9-38.3 weight portion, Fructus Chaenomelis 10.7-43.1 weight portion, Flos Caryophylli 0.9-33.3 weight portion;
Described free radical is total free radical, DPPH free radical and hydroxy radical.
6. free radical scavenger according to claim 5 is characterized in that: also comprise Flos Sophorae in the described raw material; The raw material of described free radical scavenger consists of: Flos Sophorae Immaturus 5.6-51.2 weight portion, Fructus Canarii 2.9-38.3 weight portion, Fructus Chaenomelis 10.7-43.1 weight portion, Flos Caryophylli 0.9-33.3 weight portion, Flos Sophorae 3.2-41.4 weight portion;
Described free radical is total free radical, DPPH free radical and hydroxy radical.
7. according to claim 1 or 2 or 3 described free radical scavengers, it is characterized in that: also comprise Radix Scutellariae in the described raw material; The raw material of described free radical scavenger consists of: Flos Sophorae Immaturus 5.6-51.2 weight portion, Fructus Canarii 2.9-38.3 weight portion, Fructus Chaenomelis 10.7-43.1 weight portion, Radix Scutellariae 1.9-17.5 weight portion;
Described free radical is a superoxide anion.
8. free radical scavenger according to claim 7 is characterized in that: also comprise Flos Sophorae in the described raw material; The raw material of described free radical scavenger consists of: Flos Sophorae Immaturus 5.6-51.2 weight portion, Fructus Canarii 2.9-38.3 weight portion, Fructus Chaenomelis 10.7-43.1 weight portion, Radix Scutellariae 1.9-17.5 weight portion, Flos Sophorae 3.2-41.4 weight portion;
Described free radical is a superoxide anion.
9. according to claim 1 or 2 or 3 described free radical scavengers, it is characterized in that: also comprise Flos Caryophylli, Radix Scutellariae and Flos Sophorae in the described raw material; The raw material of described free radical scavenger consists of: Flos Sophorae Immaturus 10.1-51.2 weight portion, Fructus Canarii 2.9-21.2 weight portion, Fructus Chaenomelis 12.3-16.9 weight portion, Flos Caryophylli 6.7-33.3 weight portion, Flos Sophorae 3.2-41.4 weight portion and Radix Scutellariae 5.2-17.5 weight portion;
Described free radical is total free radical and DPPH free radical.
10. according to claim 1 or 2 or 3 described free radical scavengers, it is characterized in that: also comprise Flos Caryophylli, Radix Scutellariae and Flos Sophorae in the described raw material; The raw material of described free radical scavenger consists of: Flos Sophorae Immaturus 5.6-51.2 weight portion, Fructus Canarii 12.6-38.3 weight portion, Fructus Chaenomelis 12.6-43.1 weight portion, Flos Caryophylli 0.9-6.6 weight portion, Flos Sophorae 3.2-23.3 weight portion and Radix Scutellariae 1.9-5.2 weight portion;
Described free radical is a superoxide anion.
11. according to claim 1 or 2 or 3 described free radical scavengers, it is characterized in that: also comprise Flos Caryophylli, Radix Scutellariae and Flos Sophorae in the described raw material; The raw material of described free radical scavenger consists of: Flos Sophorae Immaturus 5.6-51.2 weight portion, Fructus Canarii 2.9-38.3 weight portion, Fructus Chaenomelis 12.3-43.1 weight portion, Flos Caryophylli 0.9-18.6 weight portion, Flos Sophorae 3.2-41.4 weight portion and Radix Scutellariae 1.9-14.4 weight portion;
Described free radical is a hydroxy radical.
12. according to claim 1 or 2 or 3 described free radical scavengers, it is characterized in that: also comprise Flos Caryophylli, Radix Scutellariae and Flos Sophorae in the described raw material; The raw material of described free radical scavenger consists of: the Flos Sophorae Immaturus 51.2 weight portions; Fructus Canarii 21.2 weight portions; Fructus Chaenomelis 12.6 weight portions; Flos Caryophylli 6.7 weight portions; Flos Sophorae 3.2 weight portions; Radix Scutellariae 5.2 weight portions.
13. the application of arbitrary described free radical scavenger in the preparation cosmetics among the claim 1-12.
14. one kind is the cosmetics of active component preparation with arbitrary described free radical scavenger among the claim 1-12.
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| CN101978996B (en) * | 2010-10-25 | 2012-01-25 | 中国人民解放军第三军医大学第一附属医院 | Preparation method of traditional Chinese medicinal composition for preventing and treating sunburn and cream, and cream thereof |
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