CN112630366A - High performance liquid chromatography detection method for content of mecobalamin dispersible tablets - Google Patents
High performance liquid chromatography detection method for content of mecobalamin dispersible tablets Download PDFInfo
- Publication number
- CN112630366A CN112630366A CN202011514856.2A CN202011514856A CN112630366A CN 112630366 A CN112630366 A CN 112630366A CN 202011514856 A CN202011514856 A CN 202011514856A CN 112630366 A CN112630366 A CN 112630366A
- Authority
- CN
- China
- Prior art keywords
- phase
- liquid chromatography
- mecobalamin
- high performance
- performance liquid
- 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
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/89—Inverse chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N2030/042—Standards
- G01N2030/047—Standards external
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The invention discloses a high performance liquid chromatography detection method for the content of mecobalamin dispersible tablets, which comprises the following steps: the determination is carried out by high performance liquid chromatography, a chromatographic column is a reversed-phase chromatographic column which has medium-strength hydrophobicity and takes a hydrogen bonding phase as a chromatographic column filler, and a mobile phase consists of a water phase and an organic solvent; isocratic elution is set according to volume fraction; and carrying out quantitative analysis by adopting an external standard method. The high performance liquid chromatography detection method for the content of the mecobalamin dispersible tablets has the advantages of practicality, reliability, good stability and strong data reproducibility, and can effectively control the quality of the product.
Description
FIELD
The invention relates to the technical field of analytical chemistry, in particular to a high performance liquid chromatography detection method for the content of mecobalamin dispersible tablets.
Background
The mecobalamin dispersible tablet is used for treating peripheral neuropathy, and the active component of the dispersible tablet is mecobalamin with a chemical name of alpha- (5, 6-dimethylbenzimidazolyl) -Co-mecobalamin. The chemical structural formula is C63H91CoN13O14P, the molecular weight is 1344.38 methylcobalamin (Mecobalamin) which is endogenous vitamin B12, the vitamin B is present in blood and medullary fluid, compared with vitamin B12, the vitamin B has good improvement effect on the conduction of neurons, can promote nucleic acid-protein-fat metabolism through methyl conversion reaction, can be used as a coenzyme of methionine synthetase, can convert homocysteine into methionine, participates in the process of synthesizing thymine from deoxynucleoside, promotes nucleic acid and protein synthesis, promotes axonal transport, axonal regeneration and myelination, prevents axonal degeneration, and repairs damaged nerve tissues. The peak blood concentration is reached 3 hours after oral administration, and the absorption is dose-dependent. It can be detected from blood, kidney, adrenal gland, pancreas, liver and stomach tissues in sequence, and has high concentration, while the concentration in muscle, testis, cranial nerve, etc. is low. 8 hours after taking, the excretion amount of the total B12 in urine is 40 to 80 percent of the excretion amount 24 hours after taking the medicine.
In order to ensure the subsequent development and production quality of the mecobalamin dispersible tablets, the quality of the raw material medicaments needs to be controlled. Therefore, the high performance liquid chromatography detection method for researching the content of the mecobalamin dispersible tablets is very urgent for pharmaceutical manufacturing enterprises. However, the existing method for measuring the content of the mecobalamin dispersible tablets is single and is not beneficial to the control of the product quality.
SUMMARY
The high performance liquid chromatography detection method for the content of the mecobalamin dispersible tablets is characterized by comprising the following steps:
the determination is carried out by high performance liquid chromatography, a chromatographic column is a reversed-phase chromatographic column which has medium-strength hydrophobicity and takes a hydrogen bonding phase as a chromatographic column filler, and a mobile phase consists of a water phase and an organic solvent;
isocratic elution is set according to volume fraction; and
quantitative analysis is carried out by adopting an external standard method.
Brief description of the drawings
FIG. 1 shows a high performance liquid chromatography detection analysis chart of a mecobalamin dispersible tablet blank adjuvant according to an embodiment of the present disclosure;
FIG. 2 shows a high performance liquid chromatography assay of a mecobalamin standard according to an embodiment of the disclosure;
figure 3 shows a high performance liquid chromatography assay of a sample of dispersible mecobalamin tablets according to an embodiment of the present disclosure.
Detailed description of the invention
In the following description, certain specific details are included to provide a thorough understanding of various disclosed embodiments. One skilled in the relevant art will recognize, however, that the embodiments can be practiced without one or more of the specific details, or with other methods, components, materials, and so forth.
Unless otherwise required by the disclosure, throughout the specification and the appended claims, the words "comprise", "comprising", and "have" are to be construed in an open, inclusive sense, i.e., "including but not limited to".
Reference throughout the specification to "one embodiment," "an embodiment," "in another embodiment," or "in certain embodiments" means that a particular reference element, structure, or characteristic described in connection with the embodiment is included in at least one embodiment. Thus, the appearances of the phrases "in one embodiment" or "in an embodiment" or "in another embodiment" or "in certain embodiments" in various places throughout this specification are not necessarily all referring to the same embodiment, and furthermore, particular elements, structures, or features may be combined in any suitable manner in one or more embodiments.
Definition of
In the present disclosure, the term "reverse phase liquid chromatography" refers to liquid chromatography in which the mobile phase is polar and the stationary phase is non-polar. The reversed phase chromatography is a liquid chromatography separation mode in which a surface nonpolar carrier is used as a stationary phase and a solvent with stronger polarity than the stationary phase is used as a mobile phase. The reversed phase chromatographic column is a chromatographic column which is filled by taking a carrier bonded with nonpolar groups as a filling agent.
In the present disclosure, the term "external standard method" refers to a method of quantifying by comparing response signals of a control substance and a component to be measured in a sample using a pure product of the component to be measured as the control substance.
In the present disclosure, the term "blank solution" refers to a solution that is measured under conditions completely identical to those used for measuring a sample in order to eliminate interference in a detection analysis method by high performance liquid chromatography. The measurement result is called a "blank value" and should be subtracted from the measurement result of the sample, thereby improving the accuracy of the measurement.
In the present disclosure, the term "test solution" refers to a solution of a sample to be tested in a high performance liquid chromatography assay.
In the present disclosure, the term "control solution" is used to refer to a solution of a standard sample in a high performance liquid chromatography assay.
In the present disclosure, the term "mobile phase" refers to a substance that moves forward carrying a component to be measured during liquid chromatography.
Detailed Description
The high performance liquid chromatography detection method for the content of the mecobalamin dispersible tablets is characterized by comprising the following steps:
the determination is carried out by high performance liquid chromatography, a chromatographic column is a reversed-phase chromatographic column which has medium-strength hydrophobicity and takes a hydrogen bonding phase as a chromatographic column filler, and a mobile phase consists of a water phase and an organic solvent;
isocratic elution is set according to volume fraction; and
quantitative analysis is carried out by adopting an external standard method.
In certain embodiments, further comprising the preparation of a pre-liquid chromatography detection solution comprising:
the blank auxiliary material solution consists of a blank auxiliary material and a mobile phase;
the test solution consists of a mecobalamin dispersible tablet and a mobile phase;
and the control solution consists of a mecobalamin control and a mobile phase.
In certain embodiments, the aqueous phase is 0.01mol/L sodium dihydrogen phosphate buffer salt and the pH of the aqueous phase is 4.0.
In certain embodiments, the volume ratio of aqueous phase to organic phase is (55-65): (45-35).
In certain embodiments, the volume ratio of aqueous phase to organic phase is 60: 40.
in certain embodiments, the organic phase is selected from methanol, ethanol, acetonitrile, or mixtures thereof.
In certain embodiments, the organic phase is selected from methanol.
In certain embodiments, the column temperature of the chromatography column is 30 ℃, the column specification is 150 x 4.6mm,5 μm; the detection wavelength was 264nm and the flow rate was 0.5 mL/min.
In certain embodiments, the test solution is at a concentration of 0.001mg/mL to 1mg/mL and the control solution is at a concentration of 0.001 to 1 mg/mL.
In certain embodiments, the test solution has a concentration of 0.050mg/mL and the control solution has a concentration of 0.050 mg/mL.
Example 1 specificity test
Blank adjuvant solution: taking a proper amount of blank auxiliary materials, adding a solvent to dissolve and dilute the blank auxiliary materials to a scale, shaking up the mixture, and filtering the mixture to obtain the product.
Control solution: taking a proper amount of mecobalamin reference substance, precisely weighing, and preparing into solution containing about 50 μ g of mecobalamin per 1ml as reference substance solution.
Test solution: taking appropriate amount of the product, preparing into solution containing mecobalamin about 50 μ g per 1ml, and filtering to obtain the final product.
Injecting blank adjuvant solution, reference solution, and sample solution 10 μ l each into high performance liquid chromatograph, and recording chromatogram (shown in figures 1-3); the result shows that the test solution and the reference solution peak at the same position, and the blank auxiliary material does not generate any chromatographic peak at the position, which indicates that the blank auxiliary material does not interfere with the determination of mecobalamin in the product.
Example 2 linearity and Range
And (4) avoiding light. Taking appropriate amount of mecobalamin reference substance, adding mobile phase for dissolving and diluting to obtain solution containing about 0.5mg per 1ml as reference substance stock solution; precisely measuring the reference stock solutions 0.5ml, 0.8ml, 1.0ml, 1.2ml and 1.5ml respectively, placing in a 10ml measuring flask, diluting to scale with mobile phase, and shaking up to obtain linear solutions. Precisely measuring 10 mu l, injecting into a liquid chromatograph, recording a chromatogram, and calculating a linear regression equation and a correlation coefficient by taking the concentration as a horizontal coordinate and the peak area as a vertical coordinate. The results are shown in Table 1.
TABLE 1 results of the Linear test
As shown in Table 1, the concentration and the peak area of mecobalamin are in good linear relation within the range of 0.025-0.075 mg/ml, and the correlation coefficients are all larger than 0.999.
EXAMPLE 3 repeatability test
Control solution: and (4) avoiding light. Taking a proper amount of mecobalamin reference substance, precisely weighing, adding mobile phase for dissolving, and diluting to obtain a solution containing 50 μ g of mecobalamin per 1ml as reference substance solution.
Test solution: and (4) avoiding light. Taking appropriate amount of the product, preparing into solution containing mecobalamin about 50 μ g per 1ml, and filtering to obtain the final product.
Sample injection determination: precisely measuring 10 mu l of each solution, respectively injecting the solution into a liquid chromatograph, recording a chromatogram, and calculating the content by adopting an external standard method, wherein the result is shown in the following table 2.
TABLE 2 results of the repeatability tests
As can be seen from Table 2, the RSD content in 6 samples is less than 2.0%, and the method has good repeatability.
Example 4 accuracy test
And determining the difference between the measurement result and the true value of the detection method in the detection concentration range of 80-120%, thereby confirming that the method can obtain an accurate detection result.
Preparation of a reference stock solution: and (4) avoiding light. Taking a proper amount of mecobalamin reference substance, precisely weighing, adding mobile phase for dissolving, and diluting to obtain solution containing about 500 μ g per 1ml, and using as reference substance stock solution.
Preparation of a reference solution: and (4) avoiding light. Taking a proper amount of mecobalamin reference substance, precisely weighing, adding mobile phase for dissolving, and diluting to obtain a solution containing 50 μ g of mecobalamin per 1ml as reference substance solution.
80% concentration recovery sample: taking a proper amount of blank auxiliary materials (about equivalent to the amount of auxiliary materials used for 0.5mg of mecobalamin), placing the blank auxiliary materials into a 10ml measuring flask, precisely adding 0.8ml of reference substance stock solution, adding a mobile phase for dissolving and diluting to a scale, shaking up, and filtering to obtain the product, wherein 3 parts are prepared in parallel;
100% concentration recovery sample: taking a proper amount of blank auxiliary materials (about equivalent to the amount of auxiliary materials used for 0.5mg of mecobalamin), placing the blank auxiliary materials into a 10ml measuring flask, precisely adding 1.0ml of reference substance stock solution, adding a mobile phase for dissolving and diluting to a scale, shaking up, and filtering to obtain the product, wherein 3 parts are prepared in parallel;
120% concentration recovery sample: taking a proper amount of blank auxiliary materials (about equivalent to the amount of auxiliary materials used for 0.5mg of mecobalamin), placing the blank auxiliary materials into a 10ml measuring flask, precisely adding 1.2ml of reference substance stock solution, adding a mobile phase for dissolving and diluting to a scale, shaking up, and filtering to obtain the product, wherein 3 parts are prepared in parallel;
sample injection determination: the control solution and each recovery solution were measured precisely at 10. mu.l each, and injected into a liquid chromatograph, and the chromatogram was recorded, and the recovery was calculated from the measured amount/added amount, with the results shown in Table 3.
TABLE 3 accuracy test results
As can be seen from Table 3: in the test of recovery rate of 80-120% detection concentration, the average recovery rate is between 98.0-101.0%, which shows that the method has better accuracy.
In summary, the following steps: the high performance liquid chromatography detection method for the content of the mecobalamin dispersible tablets has the advantages of practicality, reliability, good stability and strong data reproducibility, and can effectively control the quality of the product.
From the foregoing it will be appreciated that, although specific embodiments of the disclosure have been described herein for purposes of illustration, various modifications or improvements may be made by those skilled in the art without departing from the spirit and scope of the disclosure, and that such modifications or improvements are intended to be within the scope of the appended claims.
Claims (10)
1. The high performance liquid chromatography detection method for the content of the mecobalamin dispersible tablets is characterized by comprising the following steps:
the determination is carried out by high performance liquid chromatography, a chromatographic column is a reversed-phase chromatographic column which has medium-strength hydrophobicity and takes a hydrogen bonding phase as a chromatographic column filler, and a mobile phase consists of a water phase and an organic solvent;
isocratic elution is set according to volume fraction; and
quantitative analysis is carried out by adopting an external standard method.
2. The assay of claim 1, further comprising the preparation of a pre-liquid chromatography detection solution comprising:
the blank auxiliary material solution consists of a blank auxiliary material and a mobile phase;
the test solution consists of a mecobalamin dispersible tablet and a mobile phase;
and the control solution consists of a mecobalamin control and a mobile phase.
3. The analytical method according to claim 1, wherein the aqueous phase is 0.01mol/L sodium dihydrogen phosphate buffer salt and the pH of the aqueous phase is 4.0.
4. An assay method according to claim 3, wherein the volume ratio of aqueous phase to organic phase is (55-65): (45-35).
5. The analytical method of claim 3, wherein the volume ratio of the aqueous phase to the organic phase is 60: 40.
6. the assay of claim 1, wherein the organic phase is selected from methanol, ethanol, acetonitrile, or mixtures thereof.
7. The analytical method of claim 1, wherein the organic phase is selected from methanol.
8. The assay of claim 1, wherein the column temperature of the column is 30 ℃, the column specification is 150 x 4.6mm,5 μ ι η; the detection wavelength was 264nm and the flow rate was 0.5 mL/min.
9. The assay of claim 2, wherein the test solution is at a concentration of 0.001mg/mL to 1mg/mL and the control solution is at a concentration of 0.001 to 1 mg/mL.
10. The assay of claim 2, wherein the test solution concentration is 0.050mg/mL and the control solution concentration is 0.050 mg/mL.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011514856.2A CN112630366A (en) | 2020-12-18 | 2020-12-18 | High performance liquid chromatography detection method for content of mecobalamin dispersible tablets |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011514856.2A CN112630366A (en) | 2020-12-18 | 2020-12-18 | High performance liquid chromatography detection method for content of mecobalamin dispersible tablets |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112630366A true CN112630366A (en) | 2021-04-09 |
Family
ID=75317959
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011514856.2A Pending CN112630366A (en) | 2020-12-18 | 2020-12-18 | High performance liquid chromatography detection method for content of mecobalamin dispersible tablets |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112630366A (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1742748A (en) * | 2005-09-29 | 2006-03-08 | 周卓和 | Methy cobalamine dispersion tablet and preparing method |
| US20130196964A1 (en) * | 2008-11-14 | 2013-08-01 | Bayer Schering Pharma Aktiengesellschaft | Heterocyclically substituted aryl compounds as hif inhibitors |
| US20140116112A1 (en) * | 2012-10-25 | 2014-05-01 | K & D Laboratories, Inc. | Methods for determining the presence or absence of contaminants in a sample |
| CN105218608A (en) * | 2015-10-29 | 2016-01-06 | 无锡福祈制药有限公司 | A kind of preparation method of mecobalamin |
| CN106770726A (en) * | 2016-11-29 | 2017-05-31 | 无锡福祈制药有限公司 | A kind of detection method of residual organic solvent in mecobalamin |
| CN111044635A (en) * | 2019-12-30 | 2020-04-21 | 卓和药业集团有限公司 | Method for analyzing content of mecobalamin particles |
-
2020
- 2020-12-18 CN CN202011514856.2A patent/CN112630366A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1742748A (en) * | 2005-09-29 | 2006-03-08 | 周卓和 | Methy cobalamine dispersion tablet and preparing method |
| US20130196964A1 (en) * | 2008-11-14 | 2013-08-01 | Bayer Schering Pharma Aktiengesellschaft | Heterocyclically substituted aryl compounds as hif inhibitors |
| US20140116112A1 (en) * | 2012-10-25 | 2014-05-01 | K & D Laboratories, Inc. | Methods for determining the presence or absence of contaminants in a sample |
| CN105218608A (en) * | 2015-10-29 | 2016-01-06 | 无锡福祈制药有限公司 | A kind of preparation method of mecobalamin |
| CN106770726A (en) * | 2016-11-29 | 2017-05-31 | 无锡福祈制药有限公司 | A kind of detection method of residual organic solvent in mecobalamin |
| CN111044635A (en) * | 2019-12-30 | 2020-04-21 | 卓和药业集团有限公司 | Method for analyzing content of mecobalamin particles |
Non-Patent Citations (2)
| Title |
|---|
| 崔立勋 等: "注射用甲钴胺含量测定方法研究", 《中国现代药物应用》 * |
| 郭文敏 等: "高效液相色谱法测定不除糖衣之甲钴胺片的含量", 《中国药事》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110398548B (en) | High performance liquid chromatography separation and determination method for related impurities in fosfomycin trometamol raw material medicine and preparation thereof and application | |
| Caudill et al. | Determination of γ-aminobutyric acid by liquid chromatography with electrochemical detection | |
| CN110849980A (en) | Method for detecting content of enantiomer in isopropyl L-alanine | |
| CN114646701B (en) | HPLC test method for related substances in L-prolylamide | |
| CN112630366A (en) | High performance liquid chromatography detection method for content of mecobalamin dispersible tablets | |
| CN117191970A (en) | Method for simultaneously detecting N-bromosuccinimide and N-chlorosuccinimide | |
| CN113533548A (en) | Method for detecting 1-vinyl imidazole in chemical products | |
| CN110954629A (en) | Control method for measuring content of furfuryl amine in furosemide | |
| CN115616133A (en) | Method for detecting cysteine in compound amino acid injection and application thereof | |
| CN108872406B (en) | HPLC analysis detection method for related substances in aspartic acid bulk drug | |
| CN103175930B (en) | A kind of HPLC analytical method measuring sodium sulphite content | |
| CN111983054B (en) | Method for separating and measuring related substances of empagliflozin intermediate by using HPLC (high performance liquid chromatography) | |
| CN115774061A (en) | Method for detecting acetic acid in 1-cyclohexyl piperazine | |
| CN110231416B (en) | Method for measuring 2-iodoxybenzoic acid related substances by using HPLC (high performance liquid chromatography) | |
| CN112630314B (en) | Separation method of L-alanine isopropyl ester hydrochloride and enantiomer thereof | |
| CN114280191A (en) | Method for detecting related substances in bis-cysteine and preparation thereof | |
| CN114280190B (en) | Kit for detecting related substances of double cysteines | |
| CN109374778A (en) | A kind of method of organic impurities in measurement 2-mercaptobenzimidazole | |
| CN112881565B (en) | HPLC detection method of triphenyldiamidine related substances | |
| CN111122742B (en) | Method for detecting residual quantity of dimercaptopolyethylene glycol in sample to be detected | |
| CN117949578A (en) | High performance liquid chromatography analysis method for determining content of adenosylmethionine butanedisulfonate | |
| Wagh et al. | STABILITY INDICATING RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS QUANTIFICATION OF ANTIHISTAMINIC & ASTHMATIC DRUG IN BULK AND TABLET DOSAGE FORM | |
| CN115561367A (en) | High performance liquid chromatography detection method for gout drug related substances | |
| CN115856141A (en) | Method for separating and determining tinib key intermediate and related impurities thereof by HPLC | |
| CN113075331A (en) | Novel method for determining related substances in mecobalamin tablets |
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 | ||
| CB02 | Change of applicant information | ||
| CB02 | Change of applicant information |
Address after: 219 Furong Zhongsi Road, Xishan Economic and Technological Development Zone, Wuxi City, Jiangsu Province, 214000 Applicant after: Zhuohe Pharmaceutical Group Co.,Ltd. Address before: 219 Furong Zhongsi Road, Xishan Economic and Technological Development Zone, Wuxi City, Jiangsu Province, 214000 Applicant before: Zhuohe Pharmaceutical Group Co.,Ltd. |
|
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210409 |