CN101416931B - Plant whitening agent and preparation method thereof - Google Patents
Plant whitening agent and preparation method thereof Download PDFInfo
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- CN101416931B CN101416931B CN2008102393226A CN200810239322A CN101416931B CN 101416931 B CN101416931 B CN 101416931B CN 2008102393226 A CN2008102393226 A CN 2008102393226A CN 200810239322 A CN200810239322 A CN 200810239322A CN 101416931 B CN101416931 B CN 101416931B
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Abstract
The invention discloses a vegetal whitening agent and a preparation method thereof, comprising the following steps: (1) the freeze-dried powder of West Indies cherry and water are mixed, agitated and filtered, the filtrate is collected, and then the extracting solution of cherry powder is obtained; and (2) ion exchange chromatography is carried out to the extracting solution of the cherry powder in Step (1) to carry out desalination and decolorization so as to collect outflow fluid and then obtain the vegetal whitening agent. Organic reagents are not required in the preparation method of the vegetal whitening agent, and the extracting and separating method is simple, therefore, a relatively stable system preventing the natural reductive vitamin C in the extracting solution from being oxidized is formed.
Description
Technical field
The present invention relates to a kind of plant whitening agent and preparation method thereof.
Background technology
The reduced form vitamin C has the desalination mottle, promotes beauty functions such as collagen protein synthesis, antioxidation, defying age and protecting skin aspect beauty treatment.Because vitamin C has reproducibility, the melanin intermediate is played reduction, can restraint of tyrosinase or the activity of dopachrome interconvertible enzyme, thereby hinder the oxidative chain reactions on the each point in the melanin forming process, reduce established pigmentation, as black speck, freckle etc., activation collagen building-up process, disturb the formation of lipid peroxide, thereby play the effect of skin whitening.
At present, the vitamin C major part is synthetic or two-step fermentation production by industry, and " green, nature and environmental protection " become the main flow cry of consumer nondurable goods industries such as global daily use chemicals, food, the various trace element of picked-up needed by human body from daily edible gourd, fruit and vegetable, can defying age, the profit skin, and green natural, economical and practical again.Be rich in vitamin C and sugar, albumen and various mineral in the west Indian cherry lyophilized powder, have good white-skinned face function, but because of wherein containing abundant materials such as Plant fiber, can not be directly as the additive of cosmetics.
Summary of the invention
The purpose of this invention is to provide a kind of plant whitening agent and preparation method thereof.
The preparation method of plant whitening agent provided by the present invention may further comprise the steps:
1) west Indian cherry lyophilized powder and water are mixed, stir, filter, collect filtrate, obtain the cherry powder extracting solution;
2) with above-mentioned steps 1) the cherry powder extracting solution carry out ion-exchange chromatography, with desalination and decolouring, collect effluent, obtain the plant whitening agent.
In the said method, in the described step 1), the mass ratio of described west Indian cherry lyophilized powder and water is 1:(9-12), specifically can be 1:10.
Above-mentioned steps 1) in, described filtration is that the mixed liquor with west Indian cherry lyophilized powder and water filters with kieselguhr.
In the said method, described step 2) in, described ion-exchange chromatography is to use cationic resin T
212With resin anion (R.A.) T
112Carry out.
Said method also is included in the step that adds glycerol in the plant whitening agent, and the mass ratio of described plant whitening agent and described glycerol is 1:0.3.
Said method also comprises the step that the mixed liquor of described plant whitening agent and described glycerol is sterilized, and described sterilising conditions is 90 ℃ of sterilization 20min.
Said method also comprises the mixed liquor cooling after sterilizing, and adds the step of L.G.P antiseptic then in refrigerative mixed liquor; The mass ratio of described L.G.P and described mixed liquor is 1:167.
Adopt the plant whitening agent of method for preparing also to belong to protection scope of the present invention.
The preparation method of plant whitening agent of the present invention does not need organic reagent, and extraction separation method is easy, obtains the metastable system of a kind of natural reduced form vitamin C; Adopt the whitening agent steady quality of the inventive method preparation, white-skinned face function is remarkable.
Description of drawings
Fig. 1 is the technological process of production figure of plant whitening agent
Fig. 2 is the maximum inhibition experimental result of plant whitening agent to the L-tryrosinase
Fig. 3 removes the ultra-oxygen anion free radical experimental result for the plant whitening agent
Fig. 4 removes the hydroxy radical experimental result for the plant whitening agent
Fig. 5 removes DPPH free radical experimental result for the plant whitening agent
The specific embodiment
The production of embodiment 1, plant whitening agent
The technological process of production of plant whitening agent as shown in Figure 1, detailed process is as follows:
1) 100g west Indian cherry lyophilized powder (available from Guangxi Gold Fructus Pyracanthae comprehensive agricultural development company limited) and 1L are boiled and leave standstill the deionized water that is cooled to room temperature and mix with the mass ratio of 1:10, room temperature (20-25 ℃) stirs 1h down, index not occur being precipitated as, obtain the aqueous solution of west Indian cherry lyophilized powder;
2) with centrifugal 10 minutes of 5000 rev/mins of the aqueous solutions of above-mentioned west Indian cherry lyophilized powder, the supernatant that obtains filtered with kieselguhr, collects filtrate, obtains clarifying bright flaxen cherry powder extracting solution;
3) above-mentioned cherry powder extracting solution is crossed cationic resin T respectively
212With resin anion (R.A.) T
112(resin is all available from the extraordinary resin company limited of Dandong jewel) with desalination and its electrical conductivity of reduction, collects effluent;
4) with above-mentioned steps 3) effluent and glycerol mix, the quality percentage composition that makes glycerol in the mixed liquor is 23%;
5) with above-mentioned steps 4) mixed liquor that obtains is in 90 ℃ of sterilizations 20 minutes;
6) with above-mentioned steps 5) mixed liquor be cooled to below 40 ℃, add L.G.P (propylene glycol/pair imidazolidinyl urea/iodo propinyl butyl methylamine acid esters is available from American I SP company), the mass ratio that makes L.G.P and mixed liquor is 1:167, mix homogeneously obtains the plant whitening agent; The opaque container of usefulness of the plant whitening agent that obtains is canned.
Ammonium molybdate can generate the complex of sapphirine after the reductant Vc reduction, by setting up the standard curve equation between Vc and the absorbance, adopt the beam split colorimetric can measure the content of reduced form Vc in the plant whitening agent of above-mentioned acquisition.The result shows that the content of reduced form Vc is 9.3mg/ml in the plant whitening agent of above-mentioned acquisition.
The functional experiment of embodiment 2, plant whitening agent
1, the plant whitening agent is to the maximum inhibition experiment of L-tryrosinase
Tryrosinase is a rate-limiting enzyme main in the melanocyte forming process, and the active size of this enzyme is determining the quantity that melanocyte forms, and the whitening product overwhelming majority on the current cosmetics market is based on tyrosinase inhibitor.Catalytic reaction can take place in L-tryrosinase and its substrate L-tyrosine, after in reaction system, being added with the inhibiting reagent of L-tyrosinase activity, can produce inhibitory action to catalytic reaction, be added with the variation of 475nm place absorbance before and after the inhibiting reagent of L-tyrosinase activity by mensuration, estimating this has the suppression ratio of the inhibiting reagent of L-tyrosinase activity to the L-tyrosinase activity.Concrete experimentation is as follows:
Take by weighing the 17.91g disodium hydrogen phosphate and be dissolved in the 500mL distilled water, obtain solution a; Take by weighing the 7.8g sodium dihydrogen phosphate dihydrate and be dissolved in the 500mL distilled water, obtain solution b; 92.6mL solution a and 107.4mL solution b are mixed, obtain 200mL pH and be 6.8 PBS buffer solution.
Take by weighing 0.05g L-tyrosine,, treat that its dissolving is that 6.8 PBS buffer is settled to 100mL with the pH of above-mentioned preparation afterwards earlier with the dissolving with hydrochloric acid of a small amount of 0.1mol/L.
In the experimentation, the addition of each material is as shown in table 1 in each group.
The agent of table 1 plant whitening suppresses the addition of each material in the experiment to the activity of L-tryrosinase
Annotate: C
1Group and T
1Group all adds the 0.5mL tryrosinase, and enzyme is lived and is 100U/mL.
C
0Organize each solution prepare shake up after, heating in water bath 10min in 37 ℃ of water-baths, wavelength 475nm be zeroing down.C
1After group solution prepared and shakes up, adding 0.5ml enzyme was lived and is the tryrosinase of 100U/mL behind 37 ℃ of water-bath 10min, continues water-bath 10 minutes, measures absorbance.
T
0Organize each solution prepare shake up after, heating in water bath 10min in 37 ℃ of water-baths, wavelength 475nm be zeroing down.T
1After group solution prepared and shakes up, adding 0.5ml enzyme was lived and is the tryrosinase of 100U/mL behind 37 ℃ of water-bath 10min, continues water-bath 10 minutes, measures absorbance.
Calculate the maximum inhibition T (%) of tryrosinase, T (%)=(C
1-T
1)/C
1* 100%.
Three repetitions are established in experiment, and three multiple average results as shown in Figure 2.The result shows, when the concentration of the plant whitening agent of the foregoing description 1 preparation is 2% (content that is Vc is 186ug/ml), when addition is 1ml, the suppression ratio of tyrosinase activity reached 98.4%; When the concentration of the plant whitening agent of the foregoing description 1 preparation is 2.5% (content that is Vc is 232.5ug/ml), when addition is 1ml, the suppression ratio of tyrosinase activity is reached 99.1%.So, when the concentration of the plant whitening agent of the foregoing description 1 preparation is 2% (content that is Vc is 186ug/ml), when addition is 1ml, the activity of restraint of tyrosinase substantially fully, so whitening effect is fine.
2, remove the ultra-oxygen anion free radical experiment
Get the Tris-HCl buffer 4.5ml of 0.05mol/L pH 8.2, place 25 ℃ of water-bath preheating 20min, add the plant whitening agent of the foregoing description 1 preparation of 1ml variable concentrations and the pyrogallol solution of 0.4ml25mmol/L respectively, in 25 ℃ of water-baths, react 5min behind the mixing, the HCl cessation reaction that adds 1.0ml 8mol/L, make reference with the Tris-HCl buffer, measure absorbance at the 299nm place, calculate clearance rate.The blank group replaces sample with the 1ml deionized water, and each is handled and all does three repetitions.
The computing formula of clearance rate: ultra-oxygen anion free radical clearance rate (%)=100 (A
1-A
2)/A
1, A wherein
1Be the mean light absorbency of blank, A
2Mean light absorbency for the plant whitening agent.
The plant whitening agent of the foregoing description 1 preparation to the scavenging action result of ultra-oxygen anion free radical as shown in Figure 3.The result shows, along with the increase of plant whitening agent concentration, the scavenging action of ultra-oxygen anion free radical also strengthened gradually, and when 20 times of plant whitening dilution agents, when promptly wherein Vc content is 465 μ g, be 90.3% to the clearance rate of ultra-oxygen anion free radical; And the identical standard Vc solution of Vc content is 87.8% to the clearance rate of ultra-oxygen anion free radical.When 100 times-25 times of the plant whitening dilution agents of the foregoing description 1 preparation, be that Vc content is when being 93-372 μ g, the plant whitening agent will be apparently higher than the pure Vc solution of the identical standard analysis of Vc content to the removing ability of ultra-oxygen anion free radical, its reason may be that ultra-oxygen anion free radical is removed in the antioxidant and the Vc combined effect in the plant whitening agent that also contain other in the plant whitening agent of the foregoing description 1 preparation.
3, remove hydroxy radical (OH) experiment
Hydroxyl radical free radical is the most active free radical of chemical property in the active oxygen, and it almost can react with any biomacromolecule in the living cells, and response speed is exceedingly fast, and is to the maximum free radical of body harm.Catch the clearance rate that the resulting product of hydroxy radical is determined hydroxy radical by measuring salicylic acid, have the absorbance that the material of removing the hydroxy radical effect can reduce reactant liquor if add in the reaction system.Therefore can the amount of hydroxy radical and the ability that test substance is removed hydroxyl radical free radical be described by spectrophotography.Concrete experimentation is as follows:
Get the plant whitening agent (wherein the concentration of Vc is 186 μ g/ml) of the foregoing description 1 preparation of 50 times of dilutions.Hydroxyl radical free radical is produced by the Fenton reaction, and OH oxidation salicylic acid produces has 2 of characteristic absorption to 510nm light, and the 3-resorcylic acid is caught the clearance rate that the resulting product of OH is determined OH by measuring salicylic acid.In the 25ml color comparison tube, add 3ml 2mmol/L FeSO
4With 3ml 1mmol/L H
2O
2, shake up, then add the 3ml6mmol/L salicylic acid again, shake up, behind 37 ℃ of heating in water bath 15min, take out, measure its absorbance A
0Add concentration then respectively and be plant whitening agent 0.2ml, 0.4ml, 0.6ml, 0.8ml and the 1.0ml of the foregoing description 1 preparation of 2%, and then add distilled water 0.8ml, 0.6ml, 0.4ml, 0.2ml, 0ml respectively, shake up, continue heating in water bath 15min, take out and measure its absorbance A x.Above-mentioned same procedure is adopted in the reduction of the system absorbance that is caused for the common 1.0ml plant whitening agent that adds after eliminating and distilled water, measures its absorbance A behind the constant temperature 15min
00, and then add the 1ml distilled water, survey once its absorbance A xx, A after shaking up again
Reduce=A
00-Axx.
The plant whitening agent to the OH clearance rate is: OH clearance rate (%)=100 (A
0-Ax-A
Reduce)/A
0
The plant whitening agent of the foregoing description 1 preparation to the scavenging action result of hydroxy radical as shown in Figure 4.The result shows, after the plant whitening agent of the foregoing description 1 preparation that adds 50 times of 0.8ml (Vc content is 148.8 μ g) dilutions, the clearance rate of hydroxy radical reached 92.2%; And add after Vc content is the standard Vc solution of 148.8 μ g, only be 63.6% to the clearance rate of hydroxy radical.As can be seen from Figure 4, identical Vc content, the plant whitening agent of the foregoing description 1 preparation to the removing ability of hydroxy radical apparently higher than standard Vc solution.
4, remove DPPH free radical (DPPH) experiment
DPPH be a kind of very stable be the free radical at center with nitrogen, if extract can remove it, then point out extract to have valid density that reduces hydroxy radical, alkane free radical or oxyradical and the effect that interrupts the lipid peroxidation chain reaction.DPPH is highly stable in organic solvent, is purple, and a characteristic absorption peak is arranged at the 517nm place, and when it ran into free radical scavenger, the lone electron of DPPH was matched, and its color is shoaled, and diminishes at the absorbance of maximum absorption wave strong point.Therefore, can estimate its removing effect by the variation of measuring absorbance to DPPH.
Accurately take by weighing 20mg DPPH (sigma company), be settled in the 250ml volumetric flask with dehydrated alcohol, obtaining concentration is the DPPH solution of 20mmol/L, the plant whitening agent of the foregoing description 1 preparation is become the test fluid of variable concentrations respectively with distilled water diluting.Get the test fluid and the 2ml 20mmol/L DPPH of 2ml variable concentrations respectively, mixing afterreaction 30min measures the variation of absorbance under the 517nm wavelength, and control solvent replaces with dehydrated alcohol, is calculated as follows suppression ratio.
Suppression ratio (%)=[1-(A
i-A
j)/Ac] * 100.
In the formula, A
iThe absorbance of the mixed liquor of forming for the test fluid of 2ml DPPH liquid and 2ml variable concentrations; A
jThe absorbance of the mixed liquor of forming for the test fluid of 2ml variable concentrations and 2ml dehydrated alcohol; Ac is the absorbance of the mixed liquor of 2ml DPPH liquid and 2ml dehydrated alcohol composition.
The plant whitening agent of the foregoing description 1 preparation to the scavenging action result of DPPH as shown in Figure 5.The result shows, the concentration of the plant whitening agent of the foregoing description 1 preparation is big more, removing ability to DPPH is strong more, when the Vc content in the plant whitening agent is 18.6 μ g, be 500 times of plant whitening dilution agents, clearance rate to DPPH is 57%, and reaches same DPPH clearance rate, and the content of Vc need be 27.9 μ g in the standard Vc solution.And Vc content is when 37.2 μ g are following, the plant whitening agent will be apparently higher than standard Vc solution to the clearance rate of DPPH, this shows that the plant whitening agent of the foregoing description 1 preparation has the ability of good removing DPPH, also show simultaneously, Vc is not only arranged in action in the plant whitening agent of the foregoing description 1 preparation, also may contain other antioxidant contents, be their coefficient results.By above experiment as can be seen, the plant whitening agent of the foregoing description 1 preparation of low concentration promptly has good senile-resistant efficacy.
Claims (6)
1. the preparation method of a whitening agent may further comprise the steps:
1) west Indian cherry lyophilized powder and water are mixed, stir, filter, collect filtrate, obtain the cherry powder extracting solution; Described filtration is that the mixed liquor with west Indian cherry lyophilized powder and water filters with kieselguhr;
2) with above-mentioned steps 1) the cherry powder extracting solution carry out ion-exchange chromatography, with desalination and decolouring, collect effluent, obtain whitening agent; Described ion-exchange chromatography carries out with cationic resin T212 and resin anion (R.A.) T112.
2. method according to claim 1 is characterized in that: in the described step 1), the mass ratio of described west Indian cherry lyophilized powder and water is 1: (9-12).
3. method according to claim 2 is characterized in that: in the described step 1), the mass ratio of described west Indian cherry lyophilized powder and water is 1: 10.
4. according to arbitrary described method among the claim 1-3, it is characterized in that: described method also is included in and adds the step that glycerol obtains mixed liquor in the whitening agent;
In the described mixed liquor, the mass ratio of described whitening agent and described glycerol is 1: 0.3.
5. method according to claim 4 is characterized in that: described method also comprises the step that described mixed liquor is sterilized, and described sterilising conditions is 90 ℃ of sterilizations 20 minutes.
6. according to the whitening agent of arbitrary described method preparation among the claim 1-5.
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| CN106474005A (en) * | 2015-12-30 | 2017-03-08 | 何瑞林 | A kind of compositionss with whitening function and its external preparation and preparation method |
| CN106880511A (en) * | 2017-03-08 | 2017-06-23 | 上海新肌生物科技有限公司 | The purifying technique and formula of the nanoscale lactalbumin with skin whitening effect |
| CN107164115A (en) * | 2017-04-29 | 2017-09-15 | 安徽阜南县向发工艺品有限公司 | A kind of furniture surface of alternative suds catches oil stain processing clay and preparation method thereof |
| CN107099396A (en) * | 2017-04-29 | 2017-08-29 | 安徽阜南县向发工艺品有限公司 | A kind of furniture surface of alternative suds catches treatment of dirt clay and preparation method thereof |
| CN108403506A (en) * | 2018-04-25 | 2018-08-17 | 姜凤艳 | Crease-resistant reparation eye cream of one kind and preparation method thereof |
| CN111840165A (en) * | 2020-08-14 | 2020-10-30 | 山东花物堂生物科技有限公司 | Preparation method of plant whitening fermentation liquor |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1384713A (en) * | 1999-08-27 | 2002-12-11 | 密执安州大学 | Dietary food supplement containing natural cyclooxygenase inhibitors |
| CN101199471A (en) * | 2007-12-28 | 2008-06-18 | 浙江长生鸟珍珠生物科技有限公司 | Pearl whitening moisturizing astringent and process for preparing same |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1384713A (en) * | 1999-08-27 | 2002-12-11 | 密执安州大学 | Dietary food supplement containing natural cyclooxygenase inhibitors |
| CN101199471A (en) * | 2007-12-28 | 2008-06-18 | 浙江长生鸟珍珠生物科技有限公司 | Pearl whitening moisturizing astringent and process for preparing same |
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