CN111116536A - Method for extracting anthocyanin by utilizing mulberry wine dregs - Google Patents
Method for extracting anthocyanin by utilizing mulberry wine dregs Download PDFInfo
- Publication number
- CN111116536A CN111116536A CN202010112702.4A CN202010112702A CN111116536A CN 111116536 A CN111116536 A CN 111116536A CN 202010112702 A CN202010112702 A CN 202010112702A CN 111116536 A CN111116536 A CN 111116536A
- Authority
- CN
- China
- Prior art keywords
- anthocyanin
- mulberry
- extracting
- steps
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/58—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
- C07D311/60—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2
- C07D311/62—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2 with oxygen atoms directly attached in position 3, e.g. anthocyanidins
Abstract
The invention discloses a method for extracting anthocyanin by utilizing mulberry wine lees, which comprises the steps of raw material preparation, crude extraction, separation, purification and the like, wherein the residual mulberry peel lees after wine brewing is used as a raw material for extracting anthocyanin, so that the residual nutritive value of mulberry is fully exerted, and the economic benefit is further improved; the extraction method and the extraction conditions are reasonable, the extraction method has simple steps, and the operation is easy; after vacuum reduced pressure concentration, the content of the extracted mulberry anthocyanin reaches 60-75%, the extraction rate is high, and the extracted mulberry anthocyanin is good in stability and purity and has very good application value.
Description
Technical Field
The invention relates to a method for extracting anthocyanin from mulberry wine dregs.
Technical Field
Mulberry, also known as mulberry, is the fruit of the perennial woody plant mulberry of the genus morus of the family moraceae. Mulberry fruit element has the reputation of folk cherry tomato, contains rich active protein, provitamin A, B1, B2, PP and C, amino acid, malic acid, succinic acid, tartaric acid, carotene, mineral calcium, phosphorus, iron, copper, zinc and other nutritional ingredients, and also contains rutin, anthocyanin, resveratrol and other functional ingredients, has good functions of cancer prevention, aging resistance, ulcer resistance, virus resistance and the like, and is praised as the best health-care fruit product in the twenty-first century by the medical community. At present, the development and utilization modes of mulberries are very many, and the brewing of mulberry wine is a common mode.
Generally, in brewing wine with mulberries, mulberries are squeezed and fermented, and therefore, mulberry peel residues cannot be fully utilized. In fact, the mulberry wine dregs contain rich nutrient substances, and especially, the mulberry anthocyanin mostly exists in the fruit peel of the mulberry and is not utilized, so that huge waste is caused if the mulberry anthocyanin is directly discarded, and the environment is polluted. The mulberry anthocyanin has high color value and strong oxidation resistance, and is an ideal nutrient enhancer and colorant. However, due to the high activity of anthocyanins, factors such as temperature, H, oxygen, ascorbic acid, metal particles, etc. all affect the stability of anthocyanins. Anthocyanins are easily degraded under the influence of external factors, and the color, the clarity and the efficacy of the anthocyanins are influenced. Therefore, the need of finding a suitable method for extracting the anthocyanin component of the mulberry bark residues after the mulberry wine brewing is a key for further utilizing the nutritional value of the mulberry.
Disclosure of Invention
In order to solve the existing problems, the peel residue produced by the brewing processing of the mulberry is fully utilized, the invention provides a method for extracting anthocyanin by utilizing mulberry wine dregs, and the peel residue produced after the juicing and brewing value of the mulberry is used for extracting the anthocyanin, and the stability of the anthocyanin is improved. The specific technical scheme is as follows:
a method for extracting anthocyanin by utilizing mulberry wine dregs comprises the following steps:
s1: preparing raw materials: the method comprises the following steps of (1) squeezing fresh mulberries for wine brewing to obtain mulberry juice for wine brewing, collecting mulberry peel residues obtained by squeezing and filtering the mulberry juice as a raw material I for directly extracting anthocyanin, or taking the mulberry peel residues as a raw material II for extracting anthocyanin after fermenting, brewing and distilling;
s2: crude extraction of anthocyanins: adding the extracted solution into a raw material I obtained after juicing mulberries or a raw material II obtained after fermentation and distillation, stirring and extracting at 50-60 ℃ for 30-45 min, soaking and extracting at 20-25 ℃ for 3-4 h, and then standing, precipitating and layering; collecting the layered upper layer crude extract, centrifuging, and separating supernatant and precipitate; adding the precipitate into the extractive solution again, repeating the above stirring extraction and soaking extraction, and centrifuging again to obtain supernatant; mixing the supernatants obtained by the two centrifugations to obtain anthocyanin crude extract;
s3: and (3) separating the anthocyanin: concentrating the obtained crude anthocyanin extract under vacuum and reduced pressure to obtain an anthocyanin concentrated solution, slowly adding the obtained anthocyanin concentrated solution into a macroporous adsorption resin separation column, eluting with 50% ethanol water solution, and collecting eluent for purification;
s4: and (3) purifying the anthocyanin: and adding the anthocyanin-containing eluent separated by the macroporous resin separation column into a Sephadex LH-20 gel column for purification, wherein the mobile phase is 50% of ethanol water solution, the flow rate is 0.6-0.7 ml/min, collecting the eluent, and performing vacuum concentration again until the content of the mulberry anthocyanin is 60-75%.
As a preferable technical solution, in step S1, the raw material preparation further includes a mulberry processing step, specifically: cleaning the mulberry with clear water before juicing, controlling the water content, soaking the mulberry in a natural plant disinfectant for 10-15 min, cleaning the mulberry with sterile water, air-drying the mulberry, and then placing the mulberry in a refrigeration house for freezing and storing for a period of time under a low-temperature condition to ensure the stability of anthocyanin;
in a further preferable technical scheme, the natural plant disinfectant contains 53.6g/L of lactic acid, 20g/L of citric acid and 7g/L of grapefruit peel essential oil.
In a further preferable technical scheme, the freezing temperature of the mulberry is-20 to-15 ℃, and the freezing time is more than 15 min.
In a preferred embodiment, in step S1, the juicing conditions are as follows: and (4) keeping out of the sun, rotating at 5000-6000 rpm for 5-10 min, and keeping the temperature at 0-4 ℃.
In a preferable technical scheme, in the step S2, the adding proportion of the raw material I and the extracting solution is that 500ml of extracting solution is added to every 100g of raw material I; the addition proportion of the raw material II and the extracting solution is that 300ml of extracting solution is added into every 100g of raw material II.
In a preferred embodiment, in step S2, the extraction solution is an ethanol solution containing 1 to 1.3% citric acid.
In a preferable technical scheme, in the step S2, the infiltration extraction is ultrasonic-assisted extraction, the ultrasonic power is 300-400W, and the duty ratio is 1: 0.2.
In a preferred embodiment, in step S3, the macroporous adsorbent resin is selected from DIOI type or AB-8 type.
In a preferable technical scheme, in the steps S3 and S4, the vacuum reduced pressure concentration process conditions are that the vacuum degree is 0.8MPa, the temperature is 60-80 ℃, and the concentration time is 5-6 hours.
The invention has the beneficial effects that:
the anthocyanin is separated and extracted from the mulberry wine dregs through a reasonable extraction method and extraction conditions, so that the nutritional value of the mulberry is further fully exerted, and the economic benefit is improved. The method provided by the invention is simple in steps and easy to operate, and the extracted mulberry anthocyanin is good in stability, high in extraction rate, good in purity and very good in application value.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be clearly and completely described below with reference to the embodiments.
Example 1
The embodiment is to extract anthocyanin by utilizing mulberry wine dregs, and comprises the following steps:
s1: preparing raw materials: cleaning picked fresh mulberries with clear water, draining, and soaking in a natural plant disinfectant for 10min, wherein the natural plant disinfectant contains 53.6g/L lactic acid, 20g/L citric acid and 7g/L grapefruit peel essential oil; then cleaning with sterile water, air drying, placing in a refrigerator, setting the temperature at-20 deg.C, and freezing for more than 15min to ensure stability of anthocyanin; pulping at 5000rpm for 10min under the conditions of keeping out of the sun and at 2 deg.C; then the obtained mulberry juice is filtered for brewing wine, and the mulberry peel residue is collected as a raw material for extracting anthocyanin.
S2: crude extraction of anthocyanins: adding extracted solution into mulberry peel residues collected after juicing mulberries, wherein the extracted solution is ethanol solution containing 1% citric acid, and the adding proportion is that 500ml of the extracted solution is added into every 100g of the peel residues; then stirring and extracting for 45min at the temperature of 60 ℃, and then performing ultrasonic-assisted infiltration and extraction for 4h at the temperature of 25 ℃, wherein the ultrasonic power is 400W, and the duty ratio is 1: 0.2. Then standing, precipitating and layering, collecting layered upper-layer crude extract, and centrifuging to separate supernatant and precipitate; adding the precipitate into the extractive solution again, repeating the above stirring extraction and soaking extraction, and centrifuging again to obtain supernatant; and combining the supernatants obtained by the two centrifugations to obtain the crude anthocyanin extract.
S3: and (3) separating the anthocyanin: and (3) carrying out vacuum reduced pressure concentration on the obtained crude anthocyanin extract to obtain anthocyanin concentrated solution, wherein the process conditions of the vacuum reduced pressure concentration are as follows: the vacuum degree is 0.8MPa, the temperature is 80 ℃, and the concentration time is 5-6 hours; then, the obtained anthocyanin concentrate is slowly added into a DIOI type macroporous adsorption resin separation column, and is eluted by 50% ethanol water solution, and the eluent is collected for purification.
S4: and (3) purifying the anthocyanin: and adding the eluent containing the anthocyanin and separated by the macroporous resin separation column into a Sephadex LH-20 gel column for purification, wherein the mobile phase is the same 50% ethanol water solution, the flow rate is 0.6ml/min, collecting the eluent, and concentrating the eluent under vacuum and reduced pressure again under the conditions of the vacuum degree of 0.8MPa and the temperature of 60 ℃ until the content of the mulberry anthocyanin is 60-75%.
Example 2
The embodiment also utilizes the mulberry wine dregs to extract the anthocyanin, and comprises the following steps:
s1: preparing raw materials: cleaning picked fresh mulberries with clear water, draining, and soaking in a natural plant disinfectant for 15min, wherein the natural plant disinfectant contains 53.6g/L lactic acid, 20g/L citric acid and 7g/L grapefruit peel essential oil; then cleaning with sterile water, air drying, placing in a refrigerator, setting the temperature at-15 deg.C, and freezing for more than 15min to ensure stability of anthocyanin; pulping at 6000rpm for 5min under dark condition at 4 deg.C; then the mulberry juice obtained by filtering is used for brewing wine, and mulberry peel residue is collected and used as a raw material for extracting anthocyanin after fermentation, brewing and distillation.
S2: crude extraction of anthocyanins: adding 300ml of extracting solution into 100g of peel residues, wherein the extracting solution is an ethanol solution containing 1.3% of citric acid. Then, stirring and extracting for 30min at 50 ℃, and then performing ultrasonic-assisted infiltration and extraction for 3h at 20 ℃, wherein the ultrasonic power is 350W, and the duty ratio is 1: 0.2. Then standing, precipitating and layering, collecting layered upper-layer crude extract, and centrifuging to separate supernatant and precipitate; adding the precipitate into the extractive solution again, repeating the above stirring extraction and soaking extraction, and centrifuging again to obtain supernatant; and combining the supernatants obtained by the two centrifugations to obtain the crude anthocyanin extract.
S3: and (3) separating the anthocyanin: concentrating the obtained crude anthocyanin extract in vacuum under reduced pressure to obtain anthocyanin concentrate, wherein the conditions of vacuum reduced pressure concentration are as follows: the vacuum degree is 0.8MPa, the temperature is 60 ℃, and the concentration time is 5 hours; slowly adding the concentrated anthocyanin solution into an AB-8 type macroporous adsorption resin separation column, eluting with 50% ethanol water solution, and collecting the eluent for purification;
s4: and (3) purifying the anthocyanin: and adding the eluent containing the anthocyanin and separated by the macroporous resin separation column into a Sephadex LH-20 gel column for purification, wherein the mobile phase is the same 50% ethanol water solution, the flow rate is 0.7ml/min, collecting the eluent, and concentrating the eluent under vacuum at the vacuum degree of 0.8MPa and the temperature of 60 ℃ until the content of the mulberry anthocyanin is 60-75%.
While the invention has been described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof without departing from the scope of the invention. However, any simple modification, equivalent change and modification of the above embodiments according to the technical essence of the present invention are within the protection scope of the technical solution of the present invention.
Claims (10)
1. A method for extracting anthocyanin by utilizing mulberry wine dregs is characterized by comprising the following steps: the method comprises the following steps:
s1: preparing raw materials: the method comprises the following steps of (1) squeezing fresh mulberries for wine brewing to obtain mulberry juice for wine brewing, collecting mulberry peel residues obtained by squeezing and filtering the mulberry juice as a raw material I for directly extracting anthocyanin, or taking the mulberry peel residues as a raw material II for extracting anthocyanin after fermenting, brewing and distilling;
s2: crude extraction of anthocyanins: adding the extracted solution into a raw material I obtained after juicing mulberries or a raw material II obtained after fermentation and distillation, stirring and extracting at 50-60 ℃ for 30-45 min, soaking and extracting at 20-25 ℃ for 3-4 h, and then standing, precipitating and layering; collecting the layered upper layer crude extract, centrifuging, and separating supernatant and precipitate; adding the precipitate into the extractive solution again, repeating the above stirring extraction and soaking extraction, and centrifuging again to obtain supernatant; mixing the supernatants obtained by the two centrifugations to obtain anthocyanin crude extract;
s3: and (3) separating the anthocyanin: concentrating the obtained crude anthocyanin extract under vacuum and reduced pressure to obtain an anthocyanin concentrated solution, slowly adding the obtained anthocyanin concentrated solution into a macroporous adsorption resin separation column, eluting with 50% ethanol water solution, and collecting eluent for purification;
s4: and (3) purifying the anthocyanin: and adding the anthocyanin-containing eluent separated by the macroporous resin separation column into a Sephadex LH-20 gel column for purification, wherein the mobile phase is 50% of ethanol water solution, the flow rate is 0.6-0.7 ml/min, collecting the eluent, and performing vacuum concentration again until the content of the mulberry anthocyanin is 60-75%.
2. The method for extracting anthocyanin by utilizing mulberry wine dregs as claimed in claim 1, wherein the method comprises the following steps: in step S1, the preparing raw materials further include a mulberry processing step, specifically: the mulberry juice is washed by clean water before juicing, is soaked in natural plant disinfectant for 10-15 min after moisture is controlled to be dry, is washed by sterile water and is dried in the air, and is then placed in a refrigeration house to be frozen and stored for a period of time under the condition of low temperature, so that the stability of anthocyanin is ensured.
3. The method for extracting anthocyanin by utilizing mulberry wine dregs as claimed in claim 2, wherein the method comprises the following steps: the natural plant disinfectant contains 53.6g/L of lactic acid, 20g/L of citric acid and 7g/L of grapefruit peel essential oil.
4. The method for extracting anthocyanin by utilizing mulberry wine dregs as claimed in claim 2, wherein the method comprises the following steps: the freezing temperature of the mulberry is-20 to-15 ℃, and the freezing time is more than 15 min.
5. The method for extracting anthocyanin by utilizing mulberry wine dregs as claimed in claim 1, wherein the method comprises the following steps: in step S1, the juicing conditions are as follows: and (4) keeping out of the sun, rotating at 5000-6000 rpm for 5-10 min, and keeping the temperature at 0-4 ℃.
6. The method for extracting anthocyanin by utilizing mulberry wine dregs as claimed in claim 1, wherein the method comprises the following steps: in step S2, the raw material I and the extracting solution are added in a proportion that 500ml of extracting solution is added to every 100g of raw material I; the addition proportion of the raw material II and the extracting solution is that 300ml of extracting solution is added into every 100g of raw material II.
7. The method for extracting anthocyanin by utilizing mulberry wine dregs as claimed in claim 1 or 6, wherein the method comprises the following steps: in step S2, the extraction solution is an ethanol solution containing 1 to 1.3% citric acid.
8. The method for extracting anthocyanin by utilizing mulberry wine dregs as claimed in claim 1, wherein the method comprises the following steps: in step S2, the immersion extraction is ultrasonic-assisted extraction, the ultrasonic power is 300-400W, and the duty ratio is 1: 0.2.
9. The method for extracting anthocyanin by utilizing mulberry wine dregs as claimed in claim 1, wherein the method comprises the following steps: in step S3, the macroporous adsorbent resin is selected from DIOI type or AB-8 type.
10. The method for extracting anthocyanin by utilizing mulberry wine dregs as claimed in claim 1, wherein the method comprises the following steps: in the steps S3 and S4, the vacuum decompression concentration process conditions are that the vacuum degree is 0.8MPa, the temperature is 60-80 ℃, and the concentration time is 5-6 hours.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010112702.4A CN111116536A (en) | 2020-02-24 | 2020-02-24 | Method for extracting anthocyanin by utilizing mulberry wine dregs |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010112702.4A CN111116536A (en) | 2020-02-24 | 2020-02-24 | Method for extracting anthocyanin by utilizing mulberry wine dregs |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111116536A true CN111116536A (en) | 2020-05-08 |
Family
ID=70492937
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010112702.4A Pending CN111116536A (en) | 2020-02-24 | 2020-02-24 | Method for extracting anthocyanin by utilizing mulberry wine dregs |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111116536A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111675684A (en) * | 2020-06-29 | 2020-09-18 | 广西壮族自治区农业科学院 | Method for extracting procyanidine from mulberry fruits |
| CN115057899A (en) * | 2022-07-26 | 2022-09-16 | 山东省农业科学院 | Method for extracting anthocyanin from mulberry wine residues |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102659743A (en) * | 2012-05-08 | 2012-09-12 | 李丹 | Extraction rectification method of anthocyanin in mulberry |
| CN105949164A (en) * | 2016-06-08 | 2016-09-21 | 句容市东方紫酒业有限公司 | Mulberry anthocyanin extracting method |
| CN106349205A (en) * | 2016-08-26 | 2017-01-25 | 浙江奇异鸟生物科技有限公司 | Extraction method of blueberry anthocyanins |
| CN106674186A (en) * | 2017-01-09 | 2017-05-17 | 上海师范大学 | Method and reagent for extracting anthocyanin |
| CN109879848A (en) * | 2017-12-06 | 2019-06-14 | 南京科技职业学院 | A kind of new process that mulberries anthocyanidin extracts |
-
2020
- 2020-02-24 CN CN202010112702.4A patent/CN111116536A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102659743A (en) * | 2012-05-08 | 2012-09-12 | 李丹 | Extraction rectification method of anthocyanin in mulberry |
| CN105949164A (en) * | 2016-06-08 | 2016-09-21 | 句容市东方紫酒业有限公司 | Mulberry anthocyanin extracting method |
| CN106349205A (en) * | 2016-08-26 | 2017-01-25 | 浙江奇异鸟生物科技有限公司 | Extraction method of blueberry anthocyanins |
| CN106674186A (en) * | 2017-01-09 | 2017-05-17 | 上海师范大学 | Method and reagent for extracting anthocyanin |
| CN109879848A (en) * | 2017-12-06 | 2019-06-14 | 南京科技职业学院 | A kind of new process that mulberries anthocyanidin extracts |
Non-Patent Citations (2)
| Title |
|---|
| 樊馨怡: "花色苷提取纯化工艺的研究进展", 《农业与技术》 * |
| 胡彦新,王英,刘小莉,董明盛,周剑忠: "响应面法优化桑葚酒渣中花色苷提取工艺研究", 《食品研究与开发》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111675684A (en) * | 2020-06-29 | 2020-09-18 | 广西壮族自治区农业科学院 | Method for extracting procyanidine from mulberry fruits |
| CN115057899A (en) * | 2022-07-26 | 2022-09-16 | 山东省农业科学院 | Method for extracting anthocyanin from mulberry wine residues |
| CN115057899B (en) * | 2022-07-26 | 2023-09-22 | 山东省农业科学院 | Method for extracting anthocyanin from mulberry wine residues |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101912480B (en) | Method for preparing procyanidin extract of lycium ruthenicum murr | |
| CN110643196B (en) | Extraction and separation method of anthocyanin | |
| CN101461514B (en) | Bitter melon extract preparation method | |
| CN103462148A (en) | Preparation method of vaccimium spp. juice with high anthocyanin content | |
| CN101392008A (en) | Extraction technique of high-purity phlorizin | |
| CN111116536A (en) | Method for extracting anthocyanin by utilizing mulberry wine dregs | |
| CN102604424A (en) | Method for extracting anthocyanins from purple sweet potato | |
| AU2020103549A4 (en) | Novel se-enriched tea-derived ace inhibitory peptide and preparation process thereof | |
| CN106722942A (en) | The preparation method of moringa enzyme | |
| CN110437197B (en) | Method for extracting anthocyanin from sphenoidea | |
| CN105053952B (en) | A kind of processing technology of the dried orange peel extracts of no bitter taste | |
| CN105695080A (en) | Preparation method of tea oil | |
| CN105616465A (en) | Method for extracting polyphenol from mango skins | |
| CN108530411A (en) | A method of the extraction and purification anthocyanidin from wild cherry certain kind of berries slag | |
| CN101856427B (en) | Method for extracting solanine from potato peels | |
| CN107235840A (en) | A kind of method of the chlorogenic acid extracting from coffee bean and tannin | |
| CN102613352B (en) | Method for preparing high gamma-amino butyric acid instant tea powder | |
| CN105996036A (en) | Artichoke ferment tablets and preparation method thereof | |
| CN101434972A (en) | Process for preparing Lonicera edulis anthocyanin by solid state biological reaction | |
| CN110483532B (en) | Method for extracting chlorophyll and aromatic oil from Chinese prickly ash leaves | |
| CN113142352A (en) | Preparation method of roxburgh rose and cyclocarya paliurus instant tea | |
| CN110981921B (en) | Continuous method for synchronously extracting multiple effective components from figs | |
| CN111732998A (en) | Method for extracting noni seed oil with high oil yield without solvent | |
| CN107988280B (en) | Method for extracting high-purity isothiocyanate from cruciferous vegetable seeds | |
| CN109021042B (en) | Method for extracting high-purity oleuropein from olive leaves |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200508 |
|
| RJ01 | Rejection of invention patent application after publication |