CN113247882B - Preparation and application of oil-tea meal-based carbon dots - Google Patents

Preparation and application of oil-tea meal-based carbon dots Download PDF

Info

Publication number
CN113247882B
CN113247882B CN202110712113.4A CN202110712113A CN113247882B CN 113247882 B CN113247882 B CN 113247882B CN 202110712113 A CN202110712113 A CN 202110712113A CN 113247882 B CN113247882 B CN 113247882B
Authority
CN
China
Prior art keywords
oil
meal
tea camellia
cds
based carbon
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.)
Active
Application number
CN202110712113.4A
Other languages
Chinese (zh)
Other versions
CN113247882A (en
Inventor
黄启同
林小凤
舒珍珍
闫晨曦
薛晶晶
邰璐
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Gannan Medical University
Original Assignee
Gannan Medical University
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Gannan Medical University filed Critical Gannan Medical University
Priority to CN202110712113.4A priority Critical patent/CN113247882B/en
Publication of CN113247882A publication Critical patent/CN113247882A/en
Application granted granted Critical
Publication of CN113247882B publication Critical patent/CN113247882B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C01—INORGANIC CHEMISTRY
    • C01B—NON-METALLIC ELEMENTS; COMPOUNDS THEREOF; METALLOIDS OR COMPOUNDS THEREOF NOT COVERED BY SUBCLASS C01C
    • C01B32/00—Carbon; Compounds thereof
    • C01B32/15—Nano-sized carbon materials
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G01N21/6456—Spatial resolved fluorescence measurements; Imaging
    • G01N21/6458—Fluorescence microscopy
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/308—Electrodes, e.g. test electrodes; Half-cells at least partially made of carbon

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Electrochemistry (AREA)
  • Materials Engineering (AREA)
  • Nanotechnology (AREA)
  • Inorganic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Optics & Photonics (AREA)
  • Carbon And Carbon Compounds (AREA)

Abstract

The invention discloses preparation and application of oil-tea camellia meal-based carbon dots, which comprises the following steps of crushing and filtering oil-tea camellia meal to obtain oil-tea camellia meal powder; adding the oil-tea camellia meal powder into an ethanol/water solution, placing the mixture into a microwave reactor, and carrying out microwave stirring reaction to obtain a crude product; and finally, centrifuging the crude product, dialyzing the supernatant, and freeze-drying to obtain a solid, namely the oil-tea camellia meal-based carbon dots. The invention firstly takes the camellia oleifera seed meal with high nitrogen content as the carbon source to prepare the camellia oleifera seed meal base carbon dots by the microwave method, the preparation process is simple, economic and environment-friendly, and the camellia oleifera seed meal base carbon dots are prepared by a transmission electron microscope and an X-ray photoelectron spectroscopy pairoThe appearance and the like of CDs are characterized, and the results show that the microwave method can be successfully realizedoPreparation of CDs, the results of the study showing at the same time the results obtainedoCDs have excellent biocompatibility and cell imaging performance, and meanwhile, the conductivity of CDs is found to be excellent through testing the performance of CDs by an electrochemical cyclic voltammetry method, and the CDs are expected to be used as an excellent electrochemical sensor interface material.

Description

Preparation and application of oil-tea meal-based carbon dots
Technical Field
The invention belongs to the technical field of carbon nano materials, and particularly relates to preparation and application of oil-tea camellia meal-based carbon dots.
Background
The appearance of the nanotechnology opens up a new idea for the development and application of nanomaterials in the field of analytical chemistry. The nano particles have the excellent properties of large specific surface area, high surface reaction activity, high catalytic efficiency, strong adsorption capacity and the like, provide a new research approach for biomedical research, and promote the rapid development of chemical and biological sensors. As a novel 'zero-dimensional' Carbon nano material, carbon Dots (CDs), the excellent optical and electrical properties and potential application value thereof become one of the hot spots of the current basic research and application. At present, common methods for synthesizing CDs mainly include an arc discharge method, a laser ablation method, a strong acid oxidation method, a microwave synthesis method, a hydrothermal method, a solvothermal method, an ultrasonic oscillation method, and the like. The arc discharge method and the laser ablation method have higher equipment requirements, more complex operation process and low yield; CDs obtained by a strong acid oxidation method contain a large amount of carboxyl, which is beneficial to further modification, but the particle size of the obtained product is not uniform enough, and the separation step is complicated; currently, hydrothermal method, microwave digestion method, ultrasonic oscillation method, solvothermal method and the like are used as more methods, and the methods are simple and economical in preparation process, but the yield needs to be improved. Therefore, it is important to prepare CDs with high yield by a simple and economical method.
The Jiangnan region has abundant oil-tea camellia, and the oil-tea camellia meal in China each year is not effectively utilized, so that certain resource waste is caused, and the ecological environment is seriously influenced. Therefore, how to change waste into valuable and effectively utilize the waste residues becomes a hotspot and a difficulty of research. The oil tea meal contains rich nitrogen elements, and because the electronegativity (x = 3.04) of nitrogen atoms is higher than that of carbon atoms (x = 2.55), the doping of nitrogen can attract electrons of adjacent carbon atoms, so that the free carrier density is improved, the electrochemical performance of a CDs nano material is further improved, and therefore, the conductivity of CDs can be remarkably improved by doping CDs with nitrogen atoms, and the oil tea meal is expected to be used as an excellent electrochemical sensor interface material.
Disclosure of Invention
The invention aims to provide preparation and application of oil-tea camellia meal-based carbon dots. The invention takes the camellia oleifera seed meal as a carbon source, and prepares the camellia oleifera seed meal base carbon dots (with excellent performance) by a simple and economic methodoCDs) to be widely used in the fields of energy, environment, food analysis, pharmaceutical analysis, etc.
In order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of oil-tea camellia meal-based carbon dots comprises the following steps:
1) Drying oil-tea camellia meal, crushing into powder, and filtering to obtain synthetic oil-tea camellia meal-based carbon dotso-precursors of CDs) -oil tea meal powder;
2) Adding the oil-tea camellia meal powder into an ethanol/water solution, stirring until the mixture is uniformly mixed, placing the mixture into a microwave reactor, carrying out microwave stirring reaction, and cooling to room temperature to obtain a crude product of oil-tea camellia meal-based carbon dots;
3) Centrifuging the crude product of the oil-tea camellia meal-based carbon dots, taking the supernatant, dialyzing for at least 48 h, and finally freeze-drying to obtain orange yellow solid, namely the oil-tea camellia meal-based carbon dots (C: (C))oCDs) which are sealed at 4 ℃ until use.
In the step 1), the aperture of the filter screen is 50-60 meshes.
In the step 2), the dosage ratio of the oil-tea camellia meal powder to the ethanol/water solution is 2 g: 18-22mL, and the volume ratio of ethanol to water in the ethanol/water solution is 1.
In the step 2), the microwave stirring power is 1000-1200W, and the reaction time is 10-15min.
In the step 3), the rotating speed of the centrifugation is 14000-16000 rpm, and the time is 20-25min.
In step 3), dialyzing with dialysis bag with molecular weight cutoff of 500 Da, and changing distilled water every 3.8-4.2 h for 1 time.
The invention firstly takes the camellia oleifera seed meal with high nitrogen content as a carbon source to prepare the camellia oleifera seed meal base carbon dots by a microwave methodoCDs) and the preparation process is simple, economic and environment-friendly. By transmission electron microscopy and X-ray photoelectron spectroscopyoThe appearance and the like of CDs are characterized, and the results show that the microwave method can be successfully realizedoPreparation of CDs, the results of the study showing at the same time the results obtainedoCDs have excellent biocompatibility and cell imaging performance, and meanwhile, the conductivity is found to be excellent through the performance test of electrochemical cyclic voltammetry.
Drawings
FIG. 1 is prepared as in example 1oTransmission electron micrographs of CDs.
FIG. 2 is prepared as in example 1o-X-ray photoelectron spectroscopy of CDs.
FIG. 3 iso-cell viability profile after co-culture of CDs with Hela cells.
FIG. 4 is a drawingoFluorescence mapping after co-culture of NCDs with Hela cells.
FIG. 5 shows different materials at 0.5 mM [ Fe (CN) 6 ] 3-/4- CV diagram (sweep rate of 0.1V S) -1 ）。
Detailed Description
Example 1
A preparation method of oil-tea camellia meal-based carbon dots comprises the following steps:
1) Drying oil-tea camellia meal, crushing the oil-tea camellia meal into powder by a crusher, and filtering the powder by a 60-mesh filter screen to obtain the synthetic oil-tea camellia meal-based carbon dots (C:)o-CDs) -camellia oleifera meal powder.
2) Adding 2.0g of dried oil-tea camellia meal into 20 mL of ethanol/water solution (1, V/V), quickly stirring for 30min by using an electric stirrer, putting the mixture into a microwave reactor, carrying out microwave stirring reaction for 10 min under 1000W of power, and after the reaction is finished, carrying out programmed cooling to room temperature to obtain the oil-tea camellia meal powdero-crude product of CDs.
3) GetoThe crude product of-CDs was centrifuged at 15000 rpm for 20 min. The supernatant was dialyzed (molecular weight cut-off 500 Da) for 48 h, and distilled water was changed 1 time per 4 hours. Finally freeze drying to obtain orange yellow solid, namely the oil-tea camellia meal-based carbon dots (C:)oCDs) which are placed at 4 ℃ and sealed for use.
By Transmission Electron Microscope (TEM) pairsoCDs nanomaterials for topographical mapping, as shown in FIG. 1, preparedoThe CDs are approximately spherical under a transmission electron microscope, are well dispersed, have uniform granularity and are mainly distributed at about 2 nm.
FIG. 2 iso-X-ray photoelectron spectrum of CDs, pair thereofoCharacterization of the surface structure and the elements contained in the CDs, the results showing that the products preparedoCDs contain abundant N elements, and the doping of N can improve the overall conductivity of the material.
Example 2
Oil tea meal based carbon point (o-CDs) and cellular imaging
Biocompatibility: investigation by the MTT methodoToxicity of CDs on cervical cancer cells (Hela cells). At 37 ℃ and 5% CO 2 Under the conditions, PC12 cells were cultured in a 96-well plate at a density of 1X 10 4 One cell/well, cultured for 24 h. Dilution with DMEMoCDs, to achieve the desired final concentration and to add to the cells. Cells without any drug were used as positive control, and medium without cells and drug was used as blank control. After treatment, 20. Mu.L of MTT (5 mg/mL) was added to each well, followed by incubation at 37 ℃ in an incubator for an additional 4 h. The supernatant was then carefully aspirated and the formazan crystals were dissolved with 150 μ L of dimethyl sulfoxide (DMSO). The cell culture plate was gently shaken for 10 min to ensure complete dissolution of the formazan crystals. The absorbance values were measured at 490 nm wavelength with a multifunctional microplate reader.
Cell imaging: heLa cells at 37 o C、CO 2 The culture was carried out in DMEM medium (10% bovine serum, 1% penicillin and 5% streptomycin) at a concentration of 5%. First, the cell concentration was set to 1X 10 4 After overnight culture in a well plate to allow adhesion, 20. Mu.L of 5.0 mg. Multidot.mL was added thereto -1 oCDs, incubated together for 4 hours, and after final fixation of the cells, the imaging effect was tested under a fluorescence confocal microscope.
The experimental results are as follows: the low toxicity and good biocompatibility of nanomaterials are critical to ensure their safe and effective application in biology. This example adopts MTT method to studyo-the effect of CDs on the survival of Hela cells. As shown in figure 3 of the drawings,oCDs are hardly cytotoxic for Hela cells. Meanwhile, in combination with the results of fluorescence imaging of cells shown in FIG. 4, it was revealed thatoCDs have excellent luminescence properties. In conclusion, the invention preparesoCDs have good cell compatibility and cell imaging properties.
Example 3
Oil tea meal based carbon point (oCDs) electrochemical Performance test
Preparing an electrode: charging glassy carbonThe alpha-Al with the thickness of 1.0 mu m, 0.3 mu m and 0.05 mu m is adopted in the order 2 O 3 Polishing the surface of Glassy Carbon Electrode (GCE) with powder on the corresponding chamois, and then polishing with H 2 O、CH 3 CH 2 OH and H 2 And sequentially carrying out ultrasonic cleaning, and then blowing the surface of the product by using nitrogen. Finally, 5.0. Mu.L of 5.0 mg. ML -1 o-CDs nano-material is dropped on the GCE surface and dried in an oven at 35 ℃ for standby. The preparation of other working electrodes is also prepared by adopting the method.
The experimental results are as follows: exploration by Cyclic Voltammetry (CV)oElectrochemical Performance of CDs, FIG. 5 is the addition of CDs prepared with different carbon sources at 0.5 mM [ Fe (CN) on the GCE surface 6 ] 3-/4- Cyclic voltammograms in solution, as is evident from the figureoThe electrochemical signals of NCDs are obviously superior to CDs prepared by taking glucose (CDs 1), banana peel (CDs 2) and Datura flower (CDs 3) as carbon sources, and the possible reason is that the prepared CDs are preparedoThe nitrogen content of CDs is rich, and the doping of N can attract electrons of adjacent carbon atoms, so that the free carrier density is improved, and the conductivity of CDs is further improved. The results illustrate the preparation of the inventionoThe CDs nano material is expected to be an excellent electrochemical sensor interface material.

Claims (6)

1. The preparation method of the oil-tea camellia meal-based carbon dots is characterized by comprising the following steps of:
1) Drying the oil-tea camellia meal, crushing the dried oil-tea camellia meal into powder, and filtering to obtain oil-tea camellia meal powder;
2) Adding oil-tea camellia meal powder into an ethanol/water solution, uniformly stirring, placing the mixture into a microwave reactor, reacting for 10-15min under the microwave stirring of 1000-1200W, and then cooling to room temperature to obtain a crude product of oil-tea camellia meal-based carbon dots;
the dosage ratio of the oil-tea camellia meal powder to the ethanol/water solution is 2 g: 18-22mL, and the volume ratio of ethanol to water in the ethanol/water solution is 1;
3) Centrifuging the crude product of the oil-tea camellia meal-based carbon point, taking supernatant, dialyzing by using a dialysis bag with the molecular weight cutoff of 500 Da, changing distilled water for 1 time every 3.8-4.2 hours, dialyzing for at least 48 hours, and finally freeze-drying to obtain a solid, namely the oil-tea camellia meal-based carbon point.
2. The preparation method of the oil-tea camellia meal-based carbon dots according to claim 1, wherein in the step 1), the aperture of the filter screen for filtering is 50-60 meshes.
3. The preparation method of the oil-tea camellia meal-based carbon dots according to claim 1, wherein in the step 3), the rotation speed of centrifugation is 14000-16000 rpm, and the time is 20-25min.
4. The oil-tea camellia meal-based carbon dots obtained by the preparation method according to any one of claims 1 to 3.
5. The use of the camellia oleifera meal-based carbon dot of claim 4 in cellular imaging.
6. The use of the oil tea meal-based carbon dot of claim 4 in an electrochemical sensor.
CN202110712113.4A 2021-06-25 2021-06-25 Preparation and application of oil-tea meal-based carbon dots Active CN113247882B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202110712113.4A CN113247882B (en) 2021-06-25 2021-06-25 Preparation and application of oil-tea meal-based carbon dots

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202110712113.4A CN113247882B (en) 2021-06-25 2021-06-25 Preparation and application of oil-tea meal-based carbon dots

Publications (2)

Publication Number Publication Date
CN113247882A CN113247882A (en) 2021-08-13
CN113247882B true CN113247882B (en) 2022-11-18

Family

ID=77189720

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202110712113.4A Active CN113247882B (en) 2021-06-25 2021-06-25 Preparation and application of oil-tea meal-based carbon dots

Country Status (1)

Country Link
CN (1) CN113247882B (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115231551B (en) * 2022-09-05 2023-09-01 云南师范大学 Method for preparing carbon quantum dots by using dimethyl phthalate
CN115558496A (en) * 2022-10-20 2023-01-03 中南林业科技大学 Preparation and detection application of biomass carbon dots based on camellia oleifera abel cake

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107418567A (en) * 2017-06-14 2017-12-01 西南交通大学 A kind of biomass-based carbon quantum dot and preparation method thereof
CN109135736A (en) * 2018-08-16 2019-01-04 南昌航空大学 A kind of nitrogen-doped carbon quantum dot and the preparation method and application thereof

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0772083B2 (en) * 1991-02-18 1995-08-02 工業技術院長 How to make plant charcoal
CN103408012A (en) * 2013-08-07 2013-11-27 江汉大学 Method for preparing activated carbon from camellia seed waste residues
CN103864049B (en) * 2014-02-26 2015-10-07 上海大学 Microwave degradation biological material prepares the method for fluorescent carbon nano particle
CN110346336A (en) * 2019-06-19 2019-10-18 黄山学院 The fluorescent carbon point of one step microwave attenuation materials is used for Cu2+Method of selectively determining
CN110862084B (en) * 2019-12-04 2021-06-08 安徽裕民生态农业有限公司 Camellia seed shell activated carbon and preparation process thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107418567A (en) * 2017-06-14 2017-12-01 西南交通大学 A kind of biomass-based carbon quantum dot and preparation method thereof
CN109135736A (en) * 2018-08-16 2019-01-04 南昌航空大学 A kind of nitrogen-doped carbon quantum dot and the preparation method and application thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
Nitrogen doped waste tea residue derived carbon dots for selective quantification of tetracycline in urine and pharmaceutical samples and yeast cell imaging application;Datta B.Gunjal等;《Optical Materials》;20191231;第98卷;第1-7页 *
生物质碳点的制备、表征及在分析检测中的应用;杨子康;《中国优秀硕士学位论文全文数据库（电子期刊）工程科技Ⅰ辑》;20181015(第10期);第B014-283页 *

Also Published As

Publication number Publication date
CN113247882A (en) 2021-08-13

Similar Documents

Publication Publication Date Title
CN113247882B (en) Preparation and application of oil-tea meal-based carbon dots
CN109303922B (en) Rosa roxburghii polysaccharide functionalized nano-selenium compound, preparation method thereof and application thereof in hypoglycemic drugs
Wang et al. Nanostructured cobalt phosphates as excellent biomimetic enzymes to sensitively detect superoxide anions released from living cells
CN107010676B (en) A kind of simple and convenient process for preparing for electrode material for super capacitor cobalt sulfide nickel nano film
CN106784829A (en) A kind of preparation method for loading Graphene and the anode of microbial fuel cell of ferrous disulfide compound
CN111438368B (en) Gold-platinum bimetallic nanoenzyme prepared from silk fibroin solution and application thereof
CN107010624A (en) A kind of nitrogen, boron for electrode of super capacitor adulterates porous charcoal and preparation method thereof
CN112044429B (en) Carbon-doped tungsten oxide hollow microsphere rich in oxygen vacancy as well as preparation and application thereof
Jiang et al. Formation, photoluminescence and in vitro bioimaging of polyethylene glycol-derived carbon dots: The molecular weight effects
CN113046073A (en) Microalgae-based nanocarbon quantum dot solid powder and preparation method and application thereof
CN106876732B (en) Icosahedron shape Pd@PdFe nanocrystal and catalyst preparation, application
Zheng et al. Development of Au–Pd@ UiO-66-on-ZIF-L/CC as a self-supported electrochemical sensor for in situ monitoring of cellular hydrogen peroxide
CN106006763A (en) Method for preparing nickel cobaltate nanocrystalline assembly
Wang et al. Crude fiber and protein rich cottonseed meal derived carbon quantum dots composite porous carbon for supercapacitor
Gang et al. Synthesis and biological evaluation of fluorescent hyaluronic acid modified amorphous calcium phosphate drug carriers for tumor-targeting
CN103316711B (en) The preparation method of one kind Graphene carbonitride catalysis material
CN110387233B (en) Simple mass-producible green fluorescent carbon dot and preparation method and application thereof
CN108410027B (en) Nano graphene gel material and preparation method and application thereof
CN111899982A (en) NiO nano-sheet and preparation method and application thereof
CN110272040B (en) Preparation method of high-purity graphene oxide
CN111524716A (en) Preparation and application of composite electrode material with manila herb as carbon source
CN110591698A (en) Preparation method and application of cyclotriphosphazene doped graphene quantum dots
CN109087816B (en) Electrode composite material and preparation method and application thereof
CN112611792B (en) Multi-walled carbon nanotube/cobaltosic oxide for preparing electrochemical sensor for detecting caffeic acid
CN113666370A (en) Method for preparing capacitance carbon material by secondary microbial fermentation and activation of vinasse

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
GR01 Patent grant
GR01 Patent grant