CN113433249B - Method for detecting minocycline intermediate related substances by high performance liquid chromatography - Google Patents
Method for detecting minocycline intermediate related substances by high performance liquid chromatography Download PDFInfo
- Publication number
- CN113433249B CN113433249B CN202110825303.7A CN202110825303A CN113433249B CN 113433249 B CN113433249 B CN 113433249B CN 202110825303 A CN202110825303 A CN 202110825303A CN 113433249 B CN113433249 B CN 113433249B
- Authority
- CN
- China
- Prior art keywords
- solution
- impurity
- minocycline
- minocycline intermediate
- following
- 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
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
- G01N30/02—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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- 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
- G01N30/50—Conditioning of the sorbent material or stationary liquid
- G01N30/52—Physical parameters
- G01N30/54—Temperature
-
- 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
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention provides a method for detecting minocycline intermediate related substances by high performance liquid chromatography, which adopts the chromatographic conditions as follows: a chromatographic column: c185.0 μm 4.6 x 250mm; a detector: an ultraviolet detector; column temperature: 30 ℃; wavelength: 355nm; sample introduction amount: 10 mu L of the solution; flow rate: 0.6mL/min. The method adopts high performance liquid chromatography, the detector is an ultraviolet detector or a diode array detector, isocratic elution is adopted, the instrument requirement is low, the preparation of the mobile phase is simple, and the detection is rapid; triethylamine is a tailing inhibitor, so that the separation can be effectively improved, the main peak symmetry is good, and the triethylamine and impurities are effectively separated; the chromatographic condition can effectively separate minocycline intermediate, sancycline, from various related substances, and can be used for quality monitoring of liquid and solid samples in the synthesis process.
Description
Technical Field
The invention relates to the technical field of drug analysis, in particular to a method for detecting minocycline intermediate related substances by high performance liquid chromatography.
Background
Minocycline, also known as minocycline and minocycline, is a semi-synthetic tetracycline antibiotic with broad-spectrum antibacterial effect, has the strongest antibacterial effect in the genus, is characterized by high efficiency, quick acting and long acting, can overcome staphylococcus aureus, streptococcus and escherichia coli of milk tetracycline, and has quick oral administration absorption and no food influence. The sancycline (demethoxytetracycline) is an important intermediate of minocycline, is an important semi-synthetic intermediate raw material, generates byproducts, and is easy to epimerize in the production and storage processes to cause the quality change of products, so that the quality of the sancycline not only influences the overall reaction yield, but also indirectly influences the quality of final products. The analysis and detection of the method have important effects on condition exploration and reaction control of the previous synthesis steps and subsequent reaction yield improvement, so that the establishment of an efficient, rapid, simple, stable and effective analysis and detection method is very necessary for analyzing and detecting the minocycline intermediate.
Disclosure of Invention
The invention aims to solve the defects in the prior art and provides a method for detecting minocycline intermediate related substances by high performance liquid chromatography, which has strong specificity, good linearity and good accuracy.
The technical scheme adopted by the invention is as follows: a method for detecting minocycline intermediate related substances by high performance liquid chromatography, which adopts the chromatographic conditions as follows:
a chromatographic column: c18,5.0 μm 4.6 x 250mm or C18,3.0 μm 4.6 x 150mm;
a detector: an ultraviolet detector or a diode array detector;
column temperature: 30 ℃;
wavelength: 355nm;
sample introduction amount: 10 mu L of the solution;
flow rate: 0.6mL/min;
mobile phase: solution A: methanol: acetonitrile =600:240, 160;
isocratic elution was used.
Further, the preparation method of the solution A comprises the following steps: 0.16M sodium dihydrogen phosphate is adjusted to pH 6 + -0.2 by triethylamine and shaken up for standby.
Further, the method also includes the preparation of a test solution, a control solution and a system suitability solution.
Further, the preparation method of the test solution comprises the following steps: taking a minocycline intermediate sample, dissolving or diluting the minocycline intermediate sample with 0.1M hydrochloric acid, diluting to a constant volume of 1mg/mL, and filtering with a microporous filter membrane before sample injection.
Further, the preparation method of the control solution comprises the following steps: precisely measuring 1mL of sample solution, diluting with 0.1M hydrochloric acid in a 100mL volumetric flask, diluting to constant volume to scale, shaking, and filtering with microporous membrane before sample introduction.
Further, the preparation method of the system applicability solution comprises the following steps: preparing a mixed solution of 7 impurities and a minocycline intermediate control solution to be used as a system applicability solution; the concentration of 7 impurities in the mixed solution is as follows: the concentration of the impurity A, the impurity D and the impurity F is respectively 10ug/mL, the concentration of the impurity B, the impurity C, the impurity E and the impurity G is 2ug/mL, and the concentration of the minocycline intermediate is 1mg/mL.
The beneficial effects obtained by the invention are as follows: the method adopts high performance liquid chromatography, the detector is an ultraviolet detector or a diode array detector, isocratic elution is adopted, the instrument requirement is low, the preparation of the mobile phase is simple, and the detection is rapid; triethylamine is a tailing inhibitor, so that the separation can be effectively improved, the main peak symmetry is good, and the triethylamine and impurities are effectively separated; the chromatographic condition can effectively separate minocycline intermediate, sancycline, from various related substances, and can be used for quality monitoring of liquid and solid samples in the synthesis process; the detection method is accurate and feasible through methodology verification, simple and convenient to operate and good in linearity, and can be used for detecting related substances of the minocycline intermediate.
Drawings
FIG. 1 is a chromatogram of a system of the present invention;
FIG. 2 is a blank solvent chromatogram of the present invention;
FIG. 3 is a chromatogram of crude product of the present invention;
FIG. 4 is a chromatogram of a finished product of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments.
Example 1: experiment of system applicability
Preparing a system applicability solution: the 7 known impurities in the table above and the minocycline intermediate, the sancycline control, are configured into a mixed solution to be used as a system applicability solution, the concentrations of the 7 related impurities in the mixed solution are 10ug/mL respectively of impurity A, impurity D and impurity F, the concentrations of impurity B, impurity C, impurity E and impurity G are 2ug/mL, and the concentration of the minocycline intermediate is 1mg/mL.
And precisely measuring 10uL of the system applicability solution, and injecting the solution into a high performance liquid chromatograph.
The system suitability chromatogram is shown in FIG. 1, and the blank solvent chromatogram is shown in FIG. 2. The peak time and system suitability test results for each impurity are shown in table 1:
TABLE 1 System suitability study
And (4) conclusion: under the wavelength of 355nm, the blank solvent does not generate peaks at the main peak and each impurity peak, the separation degrees among the impurities and between the impurities and the main peak meet the requirements, and the theoretical plate number and the symmetry factor meet the measurement requirements of related substances.
Example 2: linear range test
Precisely weighing appropriate amounts of 7 impurities and minocycline intermediate reference substances, respectively dissolving and diluting with 0.1M hydrochloric acid to obtain a series of linear sample solutions, and shaking up.
And precisely measuring 10uL of the system applicability solution, injecting the solution into a high performance liquid chromatograph, recording peak area, and drawing a linear curve and an equation. The results are shown in Table 2:
TABLE 2 Linear investigation of minocycline intermediate and impurity A, B, C, D, E, F, G
And (4) conclusion: the linear regression equation and R ^2 of the 7 impurities and the minocycline intermediate meet the requirements, and the linear regression is remarkable.
Example 3: recovery test
Preparing an unlabeled sample solution: 20mg of the sample is precisely weighed, dissolved in a 20mL volumetric flask with 0.1M hydrochloric acid, diluted to a constant volume and shaken up.
Impurity control stock solution: taking appropriate amount of reference substances of impurities A, B, D, C, D, E, F and G, dissolving with 0.1M hydrochloric acid to obtain a mixed reference substance solution containing 100ug/mL of impurities A, D and F and 20ug/mL of impurities B, C, E and G.
Preparing a standard solution:
50% spiked solution: precisely weighing 20mg of a test article, dissolving the test article in a 20mL volumetric flask by using 0.1M hydrochloric acid, adding 1mL of impurity reference article stock solution, fixing the volume by using 0.1M hydrochloric acid, and shaking up; triplicate samples were prepared.
50% spiked solution: precisely weighing 20mg of a test article, dissolving the test article in a 20mL volumetric flask by using 0.1M hydrochloric acid, adding 2mL of impurity reference article stock solution, fixing the volume by using 0.1M hydrochloric acid, and shaking up; triplicate samples were prepared.
50% spiked solution: precisely weighing 20mg of a test article, dissolving the test article in a 20mL volumetric flask by using 0.1M hydrochloric acid, adding 3mL of impurity reference article stock solution, performing constant volume by using 0.1M hydrochloric acid, and shaking up; triplicate samples were prepared.
Each of the above samples was diluted 100-fold with 0.1M hydrochloric acid to prepare a corresponding control solution.
10uL of each of the above solutions was injected into a liquid chromatograph, and the content of each impurity was measured.
The results of the sample testing before labeling are shown in Table 3:
TABLE 3 sample test results before labeling
| Name (R) | Content% |
| Impurity A | 0.80 |
| Impurity B | Not detected out |
| Impurity C | Not detected out |
| Impurity D | 0.41 |
| Impurity E | Undetected |
| Impurity F | 0.24 |
| Impurity G | Not detected out |
The recovery of the impurity A, B, C, D, E, F, G was calculated separately and the results are shown in tables 4-10:
TABLE 4 impurity A recovery Rate investigation
TABLE 5 impurity B recovery considerations
TABLE 6 impurity C recovery test
TABLE 7 impurity D recovery considerations
TABLE 8 impurity E recovery test
TABLE 9 impurity F recovery test
TABLE 10 impurity G recovery test
And (4) conclusion: according to the results of the recovery rates of the impurities, the recovery rate of the impurities is between 90% and 110%, the RSD (remote subscriber identification) is not more than 5%, the measurement requirement of the impurities is met, and the method is good in accuracy.
Example 4: actual sample detection
Preparing a test solution:
crude test solution: precisely weighing 25mg of a minocycline intermediate sample, dissolving the minocycline intermediate sample with 0.1M hydrochloric acid, diluting to a constant volume of 25mL, and filtering with a microporous filter membrane before sample injection.
Finished product test solution: precisely weighing 25mg of a minocycline intermediate sample, dissolving the minocycline intermediate sample with 0.1M hydrochloric acid, diluting to a constant volume of 25mL, and filtering with a microporous filter membrane before sample injection.
10uL of the sample was precisely measured, and the results were measured and calculated according to the above chromatographic conditions.
The crude chromatogram is shown in FIG. 3, and the detection results are shown in Table 11:
TABLE 11 crude detection results
The chromatogram of the final product is shown in FIG. 4, and the detection results are shown in Table 12:
TABLE 12 test results of the finished products
And (4) conclusion: the detection result of the actual sample shows that the method can detect 7 impurities in the intermediate and can realize the detection of related substances of the minocycline intermediate sample.
The relevant known impurities in the present invention are, respectively, as shown in table 13:
TABLE 13 structural formula of related impurities
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (3)
1. A method for detecting minocycline intermediate related substances by high performance liquid chromatography is characterized by comprising the following steps: the method adopts the chromatographic conditions as follows:
a chromatographic column: c18,5.0 μm 4.6 x 250mm or C18,3.0 μm 4.6 x 150mm;
a detector: an ultraviolet detector or a diode array detector;
column temperature: 30 ℃;
wavelength: 355nm;
sample introduction amount: 10 mu L of the solution;
flow rate: 0.6mL/min;
mobile phase: solution A: methanol: acetonitrile = 600;
isocratic elution is adopted;
the preparation method of the solution A comprises the following steps: adjusting pH of 0.16M sodium dihydrogen phosphate to 6 + -0.2 with triethylamine, and shaking up for use;
the method also comprises the preparation of a test solution, a control solution and a system applicability solution;
the preparation method of the test solution comprises the following steps: dissolving or diluting minocycline intermediate sample with 0.1M hydrochloric acid, diluting to a constant volume of 1mg/mL, and filtering with microporous membrane before sample injection;
the minocycline intermediate related substances comprise sancycline and related impurities, wherein the related impurities have the following structural formula:
2. the method for detecting minocycline intermediate related substances by high performance liquid chromatography according to claim 1, wherein the method comprises the following steps: the preparation method of the control solution comprises the following steps: precisely measuring 1mL of sample solution, diluting with 0.1M hydrochloric acid in a 100mL volumetric flask, diluting to constant volume to scale, shaking, and filtering with microporous membrane before sample introduction.
3. The method for detecting minocycline intermediate related substances by high performance liquid chromatography according to claim 1, wherein the method comprises the following steps: the preparation method of the system applicability solution comprises the following steps: preparing a mixed solution of 7 impurities and a minocycline intermediate control solution to be used as a system applicability solution; the concentration of 7 impurities in the mixed solution is as follows: the concentration of the impurity A, the impurity D and the impurity F is respectively 10ug/mL, the concentration of the impurity B, the impurity C, the impurity E and the impurity G is 2ug/mL, and the concentration of the minocycline intermediate is 1mg/mL.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110825303.7A CN113433249B (en) | 2021-07-21 | 2021-07-21 | Method for detecting minocycline intermediate related substances by high performance liquid chromatography |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110825303.7A CN113433249B (en) | 2021-07-21 | 2021-07-21 | Method for detecting minocycline intermediate related substances by high performance liquid chromatography |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113433249A CN113433249A (en) | 2021-09-24 |
| CN113433249B true CN113433249B (en) | 2023-03-24 |
Family
ID=77761334
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110825303.7A Active CN113433249B (en) | 2021-07-21 | 2021-07-21 | Method for detecting minocycline intermediate related substances by high performance liquid chromatography |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113433249B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103387512A (en) * | 2012-05-08 | 2013-11-13 | 成都睿智化学研究有限公司 | Preparation method and intermediate of minocycline |
| CN108324722A (en) * | 2008-05-23 | 2018-07-27 | 帕拉特克药品公司 | The salt and polymorph of tetracycline compound |
| WO2020135508A1 (en) * | 2018-12-26 | 2020-07-02 | 浙江昌海制药有限公司 | Minocycline(ii) p-chlorobenzene sulfonate crystal form, preparation method therefor and use thereof |
| CN111855857A (en) * | 2020-07-30 | 2020-10-30 | 武汉药品医疗器械检验所 | High performance liquid chromatography detection method for minocycline hydrochloride and impurities thereof |
-
2021
- 2021-07-21 CN CN202110825303.7A patent/CN113433249B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108324722A (en) * | 2008-05-23 | 2018-07-27 | 帕拉特克药品公司 | The salt and polymorph of tetracycline compound |
| CN103387512A (en) * | 2012-05-08 | 2013-11-13 | 成都睿智化学研究有限公司 | Preparation method and intermediate of minocycline |
| WO2020135508A1 (en) * | 2018-12-26 | 2020-07-02 | 浙江昌海制药有限公司 | Minocycline(ii) p-chlorobenzene sulfonate crystal form, preparation method therefor and use thereof |
| CN111855857A (en) * | 2020-07-30 | 2020-10-30 | 武汉药品医疗器械检验所 | High performance liquid chromatography detection method for minocycline hydrochloride and impurities thereof |
Non-Patent Citations (1)
| Title |
|---|
| 高效液相色谱法测定盐酸米诺环素的有关物质;王珑等;《海峡药学》;20061231;第18卷(第01期);85-86 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113433249A (en) | 2021-09-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113866337B (en) | Mass analysis method for separating and measuring oseltamivir phosphate isomer | |
| CN111579704A (en) | Method for detecting content of N, N' -diacetyl-L-cystine in compound amino acid injection | |
| CN107941970A (en) | The method for separating and detecting of D D-pHPGs and its enantiomter | |
| CN101216468B (en) | 2-methoxymethyl-4-aminophenol and its impurity highly effective liquid phase chromatography analytical method | |
| CN111983055B (en) | Method for separating and measuring rivaroxaban intermediate related substances by using HPLC (high performance liquid chromatography) | |
| CN113433249B (en) | Method for detecting minocycline intermediate related substances by high performance liquid chromatography | |
| CN114778743B (en) | Detection method of trace chiral isomer D-proline in L-proline | |
| CN114646701B (en) | HPLC test method for related substances in L-prolylamide | |
| CN115097023A (en) | High performance liquid chromatography detection method for zolmitamine related substances | |
| CN113049687B (en) | Method for detecting ambroxol hydrochloride raw material and injection related substances | |
| CN109001342B (en) | High performance liquid chromatography method for detecting N-2, 3-dimethoxybenzyl piperonylethylamine and salt content thereof | |
| CN101025407A (en) | Analytical method for determining micro moisture in cyclopropyl amine by gas phase chromatography | |
| CN109239214B (en) | Method for detecting Shakubiqu isomers in Shakubiqu sodium | |
| CN116265937A (en) | Detection method and application of oseltamivir phosphate related impurities | |
| CN108008035B (en) | Method for detecting purity of 3-ethoxy-4-methoxybenzaldehyde | |
| CN110646540B (en) | HPLC-ELSD detection method for Retamolin starting material and intermediate | |
| CN111257441B (en) | Method for detecting impurities in parecoxib sodium synthesis process | |
| CN114324637B (en) | Method for detecting three potential genotoxic impurities in meropenem | |
| CN114200067B (en) | High performance liquid chromatography analysis method for 6-bromo-3-hydroxy pyrazine-2-carboxamide and impurities | |
| CN111521693B (en) | Method for detecting isosorbide mononitrate | |
| CN110873767A (en) | Rapid detection method for dexlansoprazole synthetic intermediate and related substances thereof | |
| CN110850012B (en) | Detection method of 1- (2, 3-dichlorophenyl) piperazine hydrochloride and related substances thereof | |
| CN113552257B (en) | Method for determining isomer impurities in p-fluorobenzothiophenol by HPLC (high performance liquid chromatography) method | |
| CN116223650A (en) | High performance liquid chromatography detection method of imidazole acyl chloride | |
| CN107991404B (en) | Detection method of (2S,5R) -benzyloxyaminopiperidine-2-formamide |
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 |