CN107843656B - Detection method of 2, 4-dimethylthiophenol related substances - Google Patents
Detection method of 2, 4-dimethylthiophenol related substances Download PDFInfo
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Abstract
The invention provides a high performance liquid chromatography detection method for 2, 4-dimethylthiophenol related substances. The liquid chromatography measurement conditions used were: the chromatographic column takes octadecylsilane chemically bonded silica as a filler, and the mobile phase is phosphate buffer solution-methanol containing cation pairs. The method has the advantages of simplicity, convenience, good specificity, high sensitivity, good repeatability and the like, and is suitable for qualitative and quantitative detection of related substances in the 2, 4-dimethylthiophenol.
Description
Technical Field
The invention relates to a detection method, in particular to a method for separating and analyzing 2, 4-dimethylthiophenol related substances by using high performance liquid chromatography.
Background
2, 4-dimethylthiophenol, also known as 2, 4-dimethylthiophenol, is a commonly used pharmaceutical intermediate having the following structural formula:
the molecular formula is as follows: C8H10S, molecular weight: 138.23.
2, 4-Dimethylthiophenol is most commonly used for the synthesis of the antidepressant drug Vortioxetine, and according to the route for 2, 4-Dimethylthiophenol, there may be present as impurities 3-Methylthiophenol, 4-Methylthiophenol, 2, 5-Dimethylthiophenol, 3, 4-Dimethylthiophenol, 3, 5-Dimethylthiophenol, 2, 6-Dimethylthiophenol. The presence of these impurities will cause a series of side reactions, which will ultimately affect the purity of the product, and will subsequently require a laborious process for isolation and purification. As in document 1: stable-indicating reversed-phase HPLC method and characteristics of imprints in vortioxetine Analysis LC-MS, IR and NMR (Lei Liu et al, Journal of Pharmaceutical and biological Analysis,117: 325) disclose impurities and by-products involved in 4 routes to synthetic vortioxetine, as follows:
as can be seen from the above routes, routes 1-3 all use 2, 4-dimethylthiophenol as an intermediate for the synthesis of vortioxetine, wherein 2,4 dimethylthiophenol (SMB) of route 3 results in the impurity Imp-E in the product due to the possible presence of the impurity 2,6 dimethylthiophenol (SMB-1). Thus, in reference 1, impurities (a to H) in vortioxetine, which is a product, are measured by high performance liquid chromatography, and impurities and side reactions occurring in the raw material are estimated.
However, document 1 reflects only impurities possibly contained in the raw material, and does not provide a method for directly controlling the impurities in the raw material, and the prior art does not disclose a method suitable for analyzing and detecting substances related to 2, 4-dimethylthiophenol. In order to reduce the occurrence of side reactions, improve the yield and purity of the product and avoid the subsequent complicated separation and purification process, the quality control of the 2, 4-dimethylthiophenol raw material is necessary.
The chromatography method is simple, convenient and rapid, has high sensitivity, can reflect the characteristics of chemical composition, content and the like, and is one of the most important methods with the widest application range. The commonly used chromatography includes High Performance Liquid Chromatography (HPLC) and Gas Chromatography (GC). Considering that 2, 4-dimethylthiophenol has a small molecular weight and a certain volatility, the inventors of the present application also tried to detect 2, 4-dimethylthiophenol-related substances by gas chromatography using OP-WAX and DB-FFAP as chromatographic columns, but the separation effect was not ideal. Therefore, the invention provides a high performance liquid chromatography analysis method suitable for separating and detecting 2, 4-dimethylthiophenol related substances.
Disclosure of Invention
The invention aims to provide a high performance liquid chromatography for separating and analyzing related substances of 2, 4-dimethylthiophenol, thereby realizing the quality control of the 2, 4-dimethylthiophenol.
The inventor finally determines a chromatographic column using octadecylsilane chemically bonded silica as a filler and a mixed solution of an organic phase and an aqueous phase as a mobile phase through a plurality of tests, wherein the organic phase is selected from methanol, the aqueous phase is selected from phosphate buffer solution, and the aqueous phase contains a cation pair reagent so as to effectively separate 2, 4-dimethyl thiophenol and related substances.
The inventors have tried acetonitrile as the organic phase, however, experiments found that the acetonitrile-buffer system could not completely detect each peak. The inventors also tried to perform separation analysis by using methanol-water as a mobile phase and using isocratic elution or gradient elution, however, when using gradient elution, 2, 4-dimethylthiophenol cannot be separated from its adjacent impurity peaks effectively due to the very similar properties of 2, 4-dimethylthiophenol and its related substances. And 2 chromatographic peaks can be separated and detected only within 60min by adopting isocratic elution, and the retention time is too long, so that the detection is not facilitated.
Therefore, the invention finally determines that phosphate buffer is taken as the water phase, and cation pair reagent is added into the water phase to improve the separation effect of the 2,4 dimethyl thiophenol and relevant substances. Wherein the phosphate is selected from one or more of potassium dihydrogen phosphate, sodium dihydrogen phosphate and ammonium dihydrogen phosphate, preferably potassium dihydrogen phosphate. The inventors have tested acetate or formate buffers as the mobile phase, but the baseline noise for chromatographic detection was greater under the low wavelength (230nm) assay conditions selected for the present invention.
The cation pair is selected from one or more of tetrabutylammonium hydroxide, tetrabutylammonium chloride, tetrabutylammonium bromide, tetrabutylammonium iodide, tetramethylammonium hydroxide, tetramethylammonium chloride, tetramethylammonium bromide, tetramethylammonium iodide, tetraethylammonium hydroxide, tetraethylammonium chloride, tetraethylammonium bromide, tetraethylammonium iodide and the like suitable for the cation pair of the meta-acidic compound.
The volume ratio of the methanol to the phosphate buffer solution containing the cation pair is 70-55: 30-45, preferably 60: 40. the inventor finds that when the methanol proportion in the mobile phase is lower than 55%, the retention time of the 2, 4-dimethylthiophenol is too long, and the detection is not facilitated. When the methanol ratio is more than 70%, the separation degree of 4-methylthiophenol is not satisfactory, and 3-methylthiophenol and 4-methylthiophenol cannot be separated effectively.
The concentration of the phosphate buffer solution is 0.05-0.1mol/L, preferably 0.05 mol/L.
The mass concentration of the cation pair in the present invention is 3 to 4.5%, preferably 4%. The inventor finds out through experiments that when the mass concentration of the cation pair in the water phase is less than 3% or more than 4.5%, the separation degree of the 2, 4-dimethylthiophenol from the adjacent impurity peak is not satisfactory and is not suitable for detection.
In the liquid chromatography condition, the column temperature is 20-40 ℃, and preferably 30 ℃.
In the liquid chromatography conditions of the present invention, the detection wavelength is 230 nm. All impurities and 2, 4-dimethylthiophenol in the invention have ultraviolet absorption, and the detection wavelength is 230nm by combining the absorption curve.
In the liquid chromatography condition of the invention, the flow rate is 0.8-1.2ml/min, preferably 1.0 ml/min.
The detection method can be realized by the following steps:
1) taking a proper amount of 2, 4-dimethylthiophenol sample, and diluting with methanol to obtain a solution containing 2mg of 2, 4-dimethylthiophenol in 1 ml;
2) setting the flow rate of the mobile phase at 0.8-1.2ml/min, the detection wavelength at 230nm, and the column temperature of the chromatographic column at 20-40 deg.C;
3) precisely measuring 10-20 μ l of the sample solution in the step 1), and injecting the sample solution into a liquid chromatograph to complete the measurement of related substances.
The method for separating and measuring the related substances of the 2, 4-dimethylthiophenol bulk drug by using the high performance liquid chromatography is simple, convenient and fast, has good specificity, high sensitivity and good repeatability, is suitable for qualitative and quantitative detection of the related substances in the 2, 4-dimethylthiophenol, and is particularly suitable for separating and detecting the contents of the related substances such as 3-methylthiophenol, 4-methylthiophenol, 2, 5-dimethylthiophenol, 3, 4-dimethylthiophenol, 3, 5-dimethylthiophenol, 2, 6-dimethylthiophenol and the like, thereby providing powerful guarantee for controlling the quality of the initial raw material intermediate or the final prepared medicine such as Vortioxetine.
Drawings
FIG. 1 is an HPLC chromatogram of a system suitability solution in comparative example 1;
FIG. 2 is an HPLC chromatogram of a system suitability solution in comparative example 2;
FIG. 3 is an HPLC chromatogram of a system suitability solution in comparative example 3;
FIG. 4 is an HPLC chromatogram of a system suitability solution in comparative example 4;
FIG. 5 is an HPLC chromatogram of a system suitability solution in comparative example 5;
FIG. 6 is an HPLC chromatogram of a system suitability solution in example 1;
FIG. 7 is an HPLC chromatogram of the system suitability solution of test number 1 in example 2;
FIG. 8 is an HPLC chromatogram of the system suitability solution of test number 2 in example 2;
FIG. 9 is an HPLC chromatogram of the system suitability solution of test number 3 in example 2; (ii) a
FIG. 10 is an HPLC chromatogram of the system suitability solution of test number 4 in example 2;
FIG. 11 is an HPLC chromatogram of the system suitability solution of test number 5 in example 2;
FIG. 12 is an HPLC chromatogram of the system suitability solution of test number 6 in example 2;
FIG. 13 is an HPLC chromatogram of the system suitability solution of test number 7 in example 2;
FIG. 14 is an HPLC chromatogram of the system suitability solution of test number 8 in example 2;
FIG. 15 is an HPLC chromatogram of the system suitability solution of test number 9 in example 2;
FIG. 16 is an HPLC chromatogram of the system suitability solution of test number 10 in example 2;
FIG. 17 is an HPLC chromatogram of the system suitability solution of test number 11 in example 2;
FIG. 18 is an HPLC chromatogram of the system suitability solution of test number 12 in example 2;
FIG. 19 is an HPLC chromatogram of sample 1 of example 4;
FIG. 20 is an HPLC chromatogram of sample 2 in example 4.
Detailed Description
The present invention is further illustrated by the following examples, which should not be construed as limiting the scope of the invention.
Comparative examples 1 to 3
1. Drugs and reagents
TABLE 1 list of drugs and reagents
2. Main instrument
TABLE 2 Main Instrument List
3. Detection method
System applicability solution: precisely weighing 10.30mg of 3-methylthiophenol, 11.40mg of 4-methylthiophenol, 10.01mg of 3, 5-dimethylthiophenol, 10.09mg of 3, 4-dimethylthiophenol, 10.25mg of 2, 5-dimethylthiophenol, 10.16mg of 2, 4-dimethylthiophenol and 10.21mg of 2, 6-dimethylthiophenol in the same 50ml measuring flask, diluting the solution to the scale with methanol, precisely sucking 2.5ml of the solution, placing the solution in a 50ml measuring flask, and diluting the solution to the scale with methanol to obtain the compound.
Liquid chromatography detection conditions: octadecylsilane bonded silica gel as filler (Kromasil,100-5C18,5 μm,250 x 4.6 mm); taking water-methanol as a mobile phase; the flow rate was 1.0ml per minute; the detection wavelength is 230 nm; the column temperature was 30 ℃. The gradient elution procedure was as follows:
measuring 20 μ l, injecting into liquid chromatograph, and measuring theoretical plate number not less than 3000 according to 2, 4-dimethylthiophenol, wherein the separation degree of each component is required.
4. Results
The results of the HPLC analysis under the above conditions are shown in FIGS. 1 to 3. As can be seen from the figure, the chromatographic peak of 2, 4-dimethylthiophenol and related substances and the adjacent peaks thereof overlap each other, and the separation effect is poor. It is speculated that 2, 4-dimethylthiophenol is very similar in nature to its related species, making efficient separation of the species difficult under the chromatographic conditions described above.
Comparative example 4
1. Drugs and reagents
Same as comparative example 1
2. Main instrument
Same as comparative example 1
3. Detection method
System applicability solution: as in comparative example 1.
Liquid chromatography detection conditions: octadecylsilane bonded silica gel as filler (Kromasil,100-5C18,5 μm,250 x 4.6 mm); taking water-methanol (40:60) as a mobile phase; the flow rate was 1.0ml per minute; the detection wavelength is 230 nm; the column temperature was 30 ℃. Measuring 20 μ l, injecting into liquid chromatograph, and measuring theoretical plate number not less than 3000 according to 2, 4-dimethylthiophenol, wherein the separation degree of each component is required.
4. Results
The results of the HPLC analysis under the above conditions are shown in FIG. 4. As can be seen from the figure, when isocratic elution is adopted, only 2 chromatographic peaks can be separated and detected within 60min, and the retention time is too long, so that the detection is not facilitated.
Comparative example 5
1. Drugs and reagents
Same as comparative example 1
2. Main instrument
Same as comparative example 1
3. Detection method
System applicability solution: as in comparative example 1.
Liquid chromatography detection conditions: octadecylsilane bonded silica gel as filler (Kromasil,100-5C18,5 μm,250 x 4.6 mm); acetonitrile (50:50) which is 0.05mol/L potassium dihydrogen phosphate solution containing 4 percent tetrabutyl ammonium hydroxide is used as a mobile phase; the flow rate was 1.0ml per minute; the detection wavelength is 230 nm; the column temperature was 30 ℃. Measuring 20 μ l, injecting into liquid chromatograph, and measuring theoretical plate number not less than 3000 according to 2, 4-dimethylthiophenol, wherein the separation degree of each component is required.
4. Results
The results of the HPLC analysis under the above conditions are shown in FIG. 5. As can be seen from the figure, only 4 chromatographic peaks can be detected by the acetonitrile-phosphate buffer salt system, which is also not beneficial to completely separating related substances.
Example 1
1. Drugs and reagents
Same as comparative example 1
2. Main instrument
Same as comparative example 1
3. Detection method
System applicability solution: precisely weighing 10.30mg of 3-methylthiophenol, 11.40mg of 4-methylthiophenol, 10.01mg of 3, 5-dimethylthiophenol, 10.09mg of 3, 4-dimethylthiophenol, 10.25mg of 2, 5-dimethylthiophenol, 10.16mg of 2, 4-dimethylthiophenol and 10.21mg of 2, 6-dimethylthiophenol in the same 50ml measuring flask, diluting the solution to the scale with methanol, precisely sucking 2.5ml of the solution, placing the solution in a 50ml measuring flask, and diluting the solution to the scale with methanol to obtain the compound.
Positioning solution: respectively taking appropriate amounts of 3-methylthiophenol, 4-methylthiophenol, 3, 5-dimethylthiophenol, 3, 4-dimethylthiophenol, 2, 5-dimethylthiophenol, 2, 4-dimethylthiophenol and 2, 6-dimethylthiophenol, respectively placing in 25ml measuring bottles, and diluting with methanol to scale.
Liquid chromatography detection conditions: octadecylsilane bonded silica gel as filler (Kromasil,100-5C18,5 μm,250 x 4.6 mm); 0.05mol/L potassium dihydrogen phosphate buffer solution containing 4% tetrabutyl ammonium hydroxide-methanol (40:60) is used as a mobile phase; the flow rate was 1.0ml per minute; the detection wavelength is 230 nm; the column temperature was 30 ℃. Measuring 20 μ l, injecting into liquid chromatograph, and measuring theoretical plate number not less than 3000 according to 2, 4-dimethylthiophenol, wherein the separation degree of each component is required.
4. Results
The results of the HPLC analysis under the above conditions are shown in Table 3 and FIG. 6. In FIG. 6, the order of appearance of peaks is 3-methylthiophenol, 4-methylthiophenol, 3, 4-dimethylthiophenol (3, 5-dimethylthiophenol), 2, 5-dimethylthiophenol, 2, 4-dimethylthiophenol and 2, 6-dimethylthiophenol. Although the peaks of the 3, 4-dimethylthiophenol chromatogram coincide with those of the 3, 5-dimethylthiophenol chromatogram in FIG. 6, the measurement of substances related to 2, 4-dimethylthiophenol was not affected, and the degrees of separation between the remaining peaks were good.
TABLE 32, 4-Dimethylthiophenol specificity survey data
Example 2
This example examines the durability of the chromatographic method by changing the parameters of the partial chromatographic conditions, the drug and reagent and the main instrument as in comparative example 1, and based on the following chromatographic conditions: octadecylsilane bonded silica gel as filler (Kromasil,100-5C18,5 μm,250 x 4.6 mm); phosphate buffer solution-methanol containing tetrabutylammonium hydroxide is used as a mobile phase; the flow rate was 1.0ml per minute; the detection wavelength is 230 nm; the column temperature was 30 ℃.
This example examines the effect of chromatographic conditions such as concentration of tetrabutylammonium hydroxide, concentration of phosphate buffer, type of phosphate and volume ratio of phosphate buffer to methanol on the detection system's adaptive solution. Mainly considers the influence of changing chromatographic conditions on the separation degree of 2, 4-dimethyl thiophenol, 3-methyl thiophenol, 4-methyl thiophenol, 3, 5-dimethyl thiophenol, 3, 4-dimethyl thiophenol, 2, 5-dimethyl thiophenol and 2, 6-dimethyl thiophenol in the system adaptive solution.
System adaptation solution: precisely weighing 10.30mg of 3-methylthiophenol, 11.40mg of 4-methylthiophenol, 10.01mg of 3, 5-dimethylthiophenol, 10.09mg of 3, 4-dimethylthiophenol, 10.25mg of 2, 5-dimethylthiophenol, 10.16mg of 2, 4-dimethylthiophenol and 10.21mg of 2, 6-dimethylthiophenol in the same 50ml measuring flask, diluting the solution to the scale with methanol, precisely sucking 2.5ml of the solution, placing the solution in a 50ml measuring flask, and diluting the solution to the scale with methanol to obtain the compound.
Under each chromatographic condition, 20. mu.l of the system-adapted solution was taken and injected into a liquid chromatograph for detection, and the detection results are shown in Table 4 and the chromatograms are shown in FIGS. 7-18. The peak sequences are 3-methylthiophenol, 4-methylthiophenol, 3, 4-dimethylthiophenol (3, 5-dimethylthiophenol), 2, 5-dimethylthiophenol, 2, 4-dimethylthiophenol and 2, 6-dimethylthiophenol.
The result shows that under the condition of changing partial chromatographic parameters, for example, the concentration of tetrabutylammonium hydroxide is 3-4.5 wt%, the concentration of phosphate buffer solution is 0.05-0.1mol/L, phosphate is phosphate such as potassium dihydrogen phosphate or sodium dihydrogen phosphate, and the volume ratio of the phosphate buffer solution to methanol is 30-45: 70-55, the separation degree of the system adaptive solution is not greatly influenced, the method is still suitable for detecting 2, 4-dimethylthiophenol related substances under the conditions, and the method has good durability.
TABLE 42 investigation data of 4-dimethylthiophenol under different experimental conditions
Example 3
1. Drugs and reagents
Same as comparative example 1
2. Main instrument
Same as comparative example 1
3. Detection method
Impurity control solution: 11.62mg of 3-methylthiophenol, 14.40mg of 4-methylthiophenol, 13.74mg of 3, 5-dimethylthiophenol, 12.98mg of 2, 5-dimethylthiophenol and 13.57mg of 2, 6-dimethylthiophenol were weighed precisely and placed in a 50ml measuring flask, and diluted to the scale with methanol to prepare an impurity control stock solution. Precisely sucking 2.5ml of the solution, placing the solution into a 50ml measuring flask, and diluting the solution to a scale with methanol to obtain an impurity control solution.
Test solution: 0.1006g of 2, 4-dimethylthiophenol was precisely weighed and placed in a 50ml measuring flask, and diluted to the mark with methanol to be used as a test solution.
Liquid chromatography detection conditions: octadecylsilane bonded silica gel as filler (Kromasil,100-5C18,5 μm,250 x 4.6 mm); 0.05mol/L potassium dihydrogen phosphate buffer solution containing 4% tetrabutyl ammonium hydroxide-methanol (40:60) is used as a mobile phase; the flow rate was 1.0ml per minute; the detection wavelength is 230 nm; the column temperature was 30 ℃. Accurately measuring 20 mul of each of the impurity reference solution and the sample solution, continuously injecting a sample for 6 needles, and recording peak areas.
4. Results
The high performance liquid chromatography detection was performed under the above conditions, and the chromatogram was recorded, and the results are shown in table 5 and below. RSD for calculating the content of 3-methylthiophenol, 4-methylthiophenol, 3, 5-dimethylthiophenol, 2, 6-dimethylthiophenol and 2, 4-dimethylthiophenol B is respectively 1.30%, 1.39%, 1.42%, 1.37%, 0.37% and 0.52%, and all the RSD are less than 2.0%, which shows that the method has good repeatability.
TABLE 5 repeatability test data
Example 4
1. Drugs and reagents
Same as comparative example 1
2. Main instrument
Same as comparative example 1
3. Detection method
Control solution: 11.62mg of 3-methylthiophenol, 14.40mg of 4-methylthiophenol, 13.74mg of 3, 5-dimethylthiophenol, 12.98mg of 2, 5-dimethylthiophenol and 13.57mg of 2, 6-dimethylthiophenol were weighed precisely and placed in a 50ml measuring flask, and diluted to the scale with methanol to prepare an impurity control stock solution. Precisely sucking 2.5ml of the solution, placing the solution into a 50ml measuring flask, and diluting the solution to a scale with methanol to obtain an impurity control solution.
Test solution: 0.1061g of 2, 4-dimethylthiophenol is precisely weighed and placed in a 50ml measuring flask, and the mixture is diluted to the scale with methanol to obtain the compound.
Liquid chromatography detection conditions: octadecylsilane bonded silica gel as filler (Kromasil,100-5C18,5 μm,250 x 4.6 mm); 0.05mol/L potassium dihydrogen phosphate buffer solution containing 4% tetrabutyl ammonium hydroxide-methanol (40:60) is used as a mobile phase; the flow rate was 1.0ml per minute; the detection wavelength is 230 nm; the column temperature was 30 ℃. 20 mul of the solution was measured and injected into a liquid chromatograph, and the measurement was performed according to the method, and the area normalization method was used.
4. Results
The results of the HPLC analysis under the above conditions are shown in Table 6 and FIGS. 19 to 20. As can be seen from Table 6, the contents of 3-methylthiophenol, 3, 5-dimethylthiophenol and 2, 6-dimethylthiophenol were 0.011%, 0.098% and 0.032%, respectively, and the sensitivity of the method was high.
TABLE 6 sample test data
In conclusion, the analysis method provided by the invention has the characteristics of specificity, durability, good reproducibility, high sensitivity and the like in the aspect of measuring 2, 4-dimethylthiophenol related substances.
Claims (14)
1. The method for detecting 2, 4-dimethylthiophenol and related substances thereof is characterized by being carried out by adopting a high performance liquid chromatography, wherein the determination conditions of the liquid chromatography are as follows: the chromatographic column takes octadecylsilane chemically bonded silica as a filler and takes a mixed solvent of an organic phase and a water phase as a mobile phase; the method comprises the following steps of preparing a mobile phase, wherein the organic phase in the mobile phase is methanol, the water phase is a phosphate buffer solution, the water phase contains a cation pair reagent, the cation pair reagent in the water phase is one or more of tetrabutylammonium hydroxide, tetrabutylammonium chloride, tetrabutylammonium bromide, tetrabutylammonium iodide, tetramethylammonium hydroxide, tetramethylammonium chloride, tetramethylammonium bromide, tetramethylammonium iodide, tetraethylammonium hydroxide, tetraethylammonium chloride, tetraethylammonium bromide and tetraethylammonium iodide, and the volume ratio of the organic phase to the water phase is 70-55: 30-45, wherein the related substances are 3-methylthiophenol, 4-methylthiophenol, 2, 5-dimethylthiophenol, 3, 4-dimethylthiophenol, 3, 5-dimethylthiophenol and 2, 6-dimethylthiophenol.
2. The detection method according to claim 1, wherein the volume ratio of the organic phase to the aqueous phase is 60: 40.
3. the detection method according to claim 1, wherein the phosphate is one or more selected from potassium dihydrogen phosphate, sodium dihydrogen phosphate, and ammonium dihydrogen phosphate.
4. The detection method according to claim 3, wherein the phosphate is selected from the group consisting of potassium dihydrogen phosphate.
5. The detection method according to claim 3, wherein the concentration of the phosphate buffer is 0.05 to 0.1 mol/L.
6. The detection method according to claim 5, wherein the concentration of the phosphate buffer is 0.05 mol/L.
7. The detection method according to claim 1, wherein the mass concentration of tetrabutylammonium hydroxide in the aqueous phase is 3-4.5%.
8. The detection method according to claim 7, characterized in that the mass concentration of tetrabutylammonium hydroxide in the aqueous phase is 4%.
9. The detection method according to claim 1, wherein the detection wavelength is 230nm in the liquid chromatography condition.
10. The detection method according to claim 1, wherein the liquid chromatography is performed at a column temperature of 20 to 40 ℃.
11. The detection method according to claim 10, wherein the liquid chromatography is performed under conditions in which a column temperature is 30 ℃.
12. The detection method according to claim 1, wherein the flow rate is 0.8 to 1.2ml/min under the liquid chromatography condition.
13. The detection method according to claim 12, wherein the flow rate in the liquid chromatography is 1.0 ml/min.
14. The detection method according to any one of claims 1 to 13, characterized by comprising the steps of: 1) taking a proper amount of 2, 4-dimethylthiophenol sample, and diluting with methanol to obtain a solution containing 2mg of 2, 4-dimethylthiophenol in 1 ml; 2) setting the flow rate of a mobile phase to be 0.8-1.2ml/min, the detection wavelength to be 230nm, and the column temperature of a chromatographic column to be 20-40 ℃; 3) precisely measuring 10-20 μ l of the sample solution in the step 1), and injecting the sample solution into a liquid chromatograph to complete the measurement of related substances.
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| CN104130212B (en) * | 2014-07-01 | 2016-08-24 | 安徽省逸欣铭医药科技有限公司 | A kind of applicable hydrobromic acid irrigates the synthetic method for western spit of fland industrialized production |
| CN104119298B (en) * | 2014-08-13 | 2016-08-24 | 北京蓝贝望生物医药科技股份有限公司 | Hydrobromic acid Wo Saiting or hydrobromic acid are fertile for Xi Ting |
| WO2016079751A2 (en) * | 2014-11-17 | 2016-05-26 | Megafine Pharma (P) Ltd. | A process for preparation of vortioxetine and polymorphs thereof |
| CN104356092B (en) * | 2014-11-27 | 2017-03-22 | 合肥创新医药技术有限公司 | Preparation method for vortioxetine |
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