CN114609268A - Method for detecting related substances in dextral rabeprazole sodium bulk drug - Google Patents
Method for detecting related substances in dextral rabeprazole sodium bulk drug Download PDFInfo
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- CN114609268A CN114609268A CN202210124668.1A CN202210124668A CN114609268A CN 114609268 A CN114609268 A CN 114609268A CN 202210124668 A CN202210124668 A CN 202210124668A CN 114609268 A CN114609268 A CN 114609268A
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- GWXQTTKUYBEZBP-UHFFFAOYSA-N 1h-benzimidazol-1-ium-2-sulfonate Chemical compound C1=CC=C2NC(S(=O)(=O)O)=NC2=C1 GWXQTTKUYBEZBP-UHFFFAOYSA-N 0.000 claims description 3
- XPYNCLYLFSMFQE-UHFFFAOYSA-N 2-(chloromethyl)-4-(3-methoxypropoxy)-3-methylpyridine Chemical compound COCCCOC1=CC=NC(CCl)=C1C XPYNCLYLFSMFQE-UHFFFAOYSA-N 0.000 claims description 3
- RAPCQINSRSZSKF-UHFFFAOYSA-N 2-[(4-chloro-3-methylpyridin-2-yl)methylsulfinyl]-1h-benzimidazole Chemical compound CC1=C(Cl)C=CN=C1CS(=O)C1=NC2=CC=CC=C2N1 RAPCQINSRSZSKF-UHFFFAOYSA-N 0.000 claims description 3
- HZNKDLUKNUEOOX-UHFFFAOYSA-N 2-[(4-methoxy-3-methylpyridin-2-yl)methylsulfanyl]-1h-benzimidazole Chemical compound COC1=CC=NC(CSC=2NC3=CC=CC=C3N=2)=C1C HZNKDLUKNUEOOX-UHFFFAOYSA-N 0.000 claims description 3
- ITARWNDPMUQOCB-UHFFFAOYSA-N 2-[(4-methoxy-3-methylpyridin-2-yl)methylsulfinyl]-1h-benzimidazole Chemical compound COC1=CC=NC(CS(=O)C=2NC3=CC=CC=C3N=2)=C1C ITARWNDPMUQOCB-UHFFFAOYSA-N 0.000 claims description 3
- BSXAHDOWMOSVAP-UHFFFAOYSA-N 2-[[4-(3-methoxypropoxy)-3-methylpyridin-2-yl]methylsulfanyl]-1h-benzimidazole Chemical compound COCCCOC1=CC=NC(CSC=2NC3=CC=CC=C3N=2)=C1C BSXAHDOWMOSVAP-UHFFFAOYSA-N 0.000 claims description 3
- JWYUFVNJZUSCSM-UHFFFAOYSA-N 2-aminobenzimidazole Chemical compound C1=CC=C2NC(N)=NC2=C1 JWYUFVNJZUSCSM-UHFFFAOYSA-N 0.000 claims description 3
- 150000003462 sulfoxides Chemical class 0.000 claims description 3
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- YREYEVIYCVEVJK-RUZDIDTESA-N dexrabeprazole Chemical compound COCCCOC1=CC=NC(C[S@@](=O)C=2NC3=CC=CC=C3N=2)=C1C YREYEVIYCVEVJK-RUZDIDTESA-N 0.000 claims description 2
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- 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
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- 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
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- 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
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- 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
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
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- 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
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
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Abstract
The invention relates to a method for detecting related substances in a dextro-rabeprazole sodium raw material drug, which comprises the steps of selecting a specific chromatographic column, optimizing an elution gradient ratio and a column temperature, monitoring a plurality of types and numbers of impurities, ensuring good separation degree among the impurities and between the impurities and main components, keeping the main components and the impurities and ensuring high response of the components, so that the content of 13 related impurities in the dextro-rabeprazole sodium raw material drug can be presented in one chromatographic action, and related substances in the dextro-rabeprazole sodium raw material drug can be quickly and accurately monitored.
Description
Technical Field
The invention belongs to the technical field of drug analysis, and particularly relates to a detection method of related substances of a dextral rabeprazole sodium raw material drug.
Background
Derabeprazole is a Proton Pump Inhibitor (PPI) that passes through the secretory surface of parietal cells (H)+,K+) The last step of inhibition of gastric acid production by covalent binding at 2 sites of the ATPase system. Rabeprazole sodium is similar to rabeprazole sodium and is a prodrug that is protonated at the parietal cell to form the active sulfenamide. Can be used for short-term treatment of gastric ulcer, duodenal ulcer and gastroesophageal reflux disease (GERD).
The related substances are initial materials, intermediates, side reaction products, degradation impurities and the like brought in the process of synthesizing the medicine, and the quality and the safety of the medicine can be controlled by detecting the related substances. The existing domestic and foreign pharmacopoeia does not contain a detection method of related substances of the dextro-rabeprazole sodium, and the detection method of related substances of the rabeprazole sodium is contained in the Chinese pharmacopoeia, the EP, the USP and the JP pharmacopoeia, wherein the elution conditions of the Chinese pharmacopoeia and the JP pharmacopoeia adopt an isocratic elution mode, and impurities with small polarity are difficult to elute, so that the accuracy of an analysis result is influenced; USP pharmacopoeia mobile phase elution mode is complicated, the mobile phase is double-phase and double-phase mixed, and the number of the research impurities is small (4), so that the detection requirement cannot be met; in the EP pharmacopoeia, the mobile phase is three-phase gradient elution, when the three phases are mixed, bubbles are easily generated to influence the stability of a base line, the effective detection of certain impurities is interfered, meanwhile, the three-phase mixing has higher requirement on the precision of an instrument, and when the elution proportion of the mobile phase is slightly changed, a larger error is easily generated.
Chinese patent CN112834628A discloses a method for determining rabeprazole sodium and its impurities by high performance liquid chromatography, which uses phosphate buffer solution and acetonitrile as mobile phase to perform gradient elution detection to detect that the rabeprazole sodium and 9 common impurities are contained, the types of analysis impurities are few, the sensitivity and accuracy are low, and the requirement of quality monitoring in the production process of d-rabeprazole sodium raw material medicine can not be met.
In order to ensure the safety and effectiveness of the drugs, research, detection and monitoring of related substances in the drugs are required. Related substances are mainly process by-products and degradation products, and the impurity spectrum changes in the process of placing the medicine, so that a proper analysis method needs to be established according to different synthesis routes, production processes and storage conditions, and accurate and effective detection and monitoring of the related substances of the dextro-rabeprazole sodium bulk drug are achieved.
Disclosure of Invention
The invention aims to provide a detection method of related substances of a dextral rabeprazole sodium raw material medicament on the basis of the prior art, which can detect 13 related impurities in one-time chromatographic behavior, is convenient for controlling the product quality in the production process, and has the advantages of simplicity, easy implementation, high accuracy and precision and good reproducibility.
The technical scheme of the invention is as follows:
a detection method for related substances in a dextro-rabeprazole sodium bulk drug adopts high performance liquid chromatography to qualitatively or quantitatively detect dextro-rabeprazole sodium and related substances, and the conditions of the high performance liquid chromatography comprise: the chromatographic column is Shimadzu Wondasil C18-a WR column; gradient elution is carried out by adopting a mobile phase A and a mobile phase B as a mixed mobile phase, wherein the mobile phase A is 15-25 mmol/L potassium dihydrogen phosphate buffer solution, and the pH value of the mixed mobile phase A is adjusted to 6.0-8.0; the mobile phase B is methanol.
The initial ratio of the mobile phase A to the mobile phase B in the gradient elution process is 73-77: 27-23; the volume ratio of the mobile phase A and the mobile phase B is kept unchanged in the initial ratio within 0-8 minutes; the volume ratio of the mobile phase A to the mobile phase B is gradually changed from the initial ratio to 55:45 at a constant speed within 8-18 minutes; the volume ratio of the mobile phase A to the mobile phase B is kept constant at 55:45 within 18-33 minutes; the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 55:45 to 25:75 at a constant speed within 33-75 minutes; the volume ratio of the mobile phase A to the mobile phase B is kept constant at 25:75 within 75-95 minutes; the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 25:75 to the initial ratio at a constant speed within 95-96 minutes; the volume ratio of mobile phase A and mobile phase B was kept constant for 96-106 minutes.
For the present invention, the initial ratio of mobile phase A and mobile phase B is controlled to be 73-77: 27-23, but not limited to 73:27, 74:26, 75:25, 76:24 or 77:23, and in a preferred embodiment, the initial ratio of mobile phase A and mobile phase B is 75:25 during the gradient elution process.
In a preferred embodiment, when the initial ratio of mobile phase a to mobile phase B is 75: 25; the gradient elution comprises the following steps: keeping the volume ratio of the mobile phase A to the mobile phase B constant at 75:25 within 0-8 minutes; the volume ratio of the mobile phase A to the mobile phase B is gradually changed from the initial ratio to 55:45 at a constant speed within 8-18 minutes; the volume ratio of the mobile phase A to the mobile phase B is kept constant at 55:45 within 18-33 minutes; the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 55:45 to 25:75 at a constant speed within 33-75 minutes; the volume ratio of the mobile phase A to the mobile phase B is kept constant at 25:75 within 75-95 minutes; the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 25:75 to 75:25 at a constant speed within 95-96 minutes; the volume ratio of mobile phase A to mobile phase B was kept constant at 75:25 for 96-106 minutes. The specific gradient elution procedure is shown in table 1 below:
TABLE 1 gradient elution procedure
When the high performance liquid chromatography is adopted for detection, the mobile phase A is 15-25 mmol/L potassium dihydrogen phosphate buffer solution, and the pH value of the mobile phase A is adjusted to 6.0-8.0; the mobile phase B is methanol. In a preferable scheme, the mobile phase A is 20mmol/L potassium dihydrogen phosphate buffer solution, and the pH value of the mobile phase A is adjusted to 6.8-7.2. Particularly preferably, the mobile phase A is 20mmol/L potassium dihydrogen phosphate buffer solution, and the pH is adjusted to 7.0.
In chromatography, the choice of the chromatographic column is important and the requirements for the chromatographic column: high column efficiency, good selectivity, high analysis speed and the like. When the high performance liquid chromatography is adopted to qualitatively or quantitatively detect the dextro-rabeprazole sodium and related substances, the chromatographic column is Shimadzu Wondasil C18The WR column has a plurality of impurity types and numbers in the process of chromatographic analysis, has good separation degrees among the impurities and between the impurities and the main component, and can quickly and accurately monitor related substances in the dextro-rabeprazole sodium. Under the condition of not influencing the detection effect, the length of the chromatographic column is preferably 250mm, the diameter of the chromatographic column is preferably 4.6mm, and the particle size of the packing is 5 mu m, namely the chromatographic column adopted by the invention is Shimadzu Wondasil C18-WR columns (250X4.6mm,5 μm). In the experimental process, other similar chromatographic columns, such as Inertsil ODS-3, are found to have poor separation of some impurities from the main component, even failing to achieve baseline separation, during the chromatographic analysis.
For the invention, 13 different impurities including intermediate impurities, process impurities and degradation impurities need to be detected, the types of the impurities are more, the column temperature in the chromatographic analysis process needs to be strictly controlled in the chromatographic analysis process, and when the column temperature selected in the chromatographic analysis is higher or lower, the separation degree of some impurities is lower than 1.5, the separation degree is not good, and baseline separation is not easy to achieve. In the invention, the column temperature is controlled to be 18-22 ℃, and can be but is not limited to 18 ℃, 19 ℃, 20 ℃, 21 ℃ or 22 ℃, and preferably, the column temperature is controlled to be 20 ℃.
Further, the high performance liquid chromatography conditions include: the detection wavelength is 200-400nm, preferably 280 nm.
Further, the flow rate is 0.5-1.5 ml/min; preferably 1.0 ml/min.
Furthermore, the sample amount is 5-20 μ l, but is not limited to 5 μ l, 10 μ l, 15 μ l or 20 μ l; preferably, the sample size is 10. mu.l.
The related substances in the dextro-rabeprazole sodium raw material medicine comprise the following substances: impurity 1: 2-chloromethyl-4- (3-methoxypropoxy) -3-methylpyridine; impurity 2: 2-mercaptobenzimidazole; impurity 3: 2- [ [4- (3-methoxypropoxy) -3-methylpyridin-2-yl ] -methylsulfanyl ] -1H-benzimidazole; impurity 4: 2- [ [ [4- (3-methoxypropoxy) -3-methyl-2-pyridinyl ] methyl ] sulfone ] -1H-benzimidazole sodium salt; impurity 5: 2-aminobenzimidazole; impurity 6: 2- [ [ [4- (3-methoxypropoxy) -3-methyl-2-pyridinyl ] methyl ] sulfoxide ] -1H-benzimidazole nitroxide; impurity 7: 2-hydroxybenzimidazole; impurity 8: 2- [ [ [4- (3-methoxypropoxy) -3-methyl-2-pyridinyl ] methyl ] sulfone ] -1H-benzimidazole nitroxide; impurity 9: benzimidazole-2-sulfonic acid; 10 parts of impurities: 1- (1H-benzoimidazol-2-yl) -1, 4-dihydroxy-3-methyl-4-oxo-2-pyridinesodium formate; 20 parts of impurities: 2- [ [ (4-chloro-3-methyl-2-pyridinyl) methyl ] sulfinyl ] -1H-benzimidazole; impurity 21: 2- [ [ (4-methoxy-3-methyl-2-pyridinyl) methyl ] sulfinyl ] -1H-benzimidazole; impurity 22: 2- [ (4-methoxy-3-methylpyridin-2-yl) methylsulfanyl ] -1H-benzimidazole.
The structural formula of the related substances mentioned above is as follows:
the method adopts the high performance liquid chromatography, respectively screens and optimizes from the aspects of chromatographic conditions and the like, draws up a method of related substances, carries out methodology verification, carries out qualitative research on the impurity 1(SMA), carries out quantitative research on the impurities 2(SMB), 3 (intermediate 1), 4-10 and 20-22 (process impurities and degradation impurities), provides a complete verification scheme, and has simple operation, good stability and reproducibility. Wherein the correction factor for impurity 2 is 0.57, the correction factor for impurity 3 is 0.88, the correction factor for impurity 4 is 1.13, the correction factor for impurity 5 is 0.71, the correction factor for impurity 6 is 0.67, the correction factor for impurity 7 is 0.74, the correction factor for impurity 8 is 0.88, the correction factor for impurity 9 is 0.93, the correction factor for impurity 10 is 0.69, the correction factor for impurity 20 is 0.76, the correction factor for impurity 21 is 0.81, and the correction factor for impurity 22 is 0.81.
In a preferred scheme, the method for detecting related substances in the dexrabeprazole sodium bulk drug provided by the invention comprises the following operation steps:
(1) solution preparation:
taking a proper amount of the dextro-rabeprazole sodium sample, adding a solvent (0.01 mol/L sodium hydroxide-methanol (volume ratio is 40: 60)) to dissolve and quantitatively dilute to prepare a solution containing about 1.0mg of dextro-rabeprazole sodium per 1ml, and taking the solution as a test solution. Precisely measuring 1.0ml of the test solution, placing the test solution in a 100ml measuring flask, adding a solvent (0.01 mol/L sodium hydroxide-methanol (volume ratio of 40: 60)) to dilute to scale, and shaking up to obtain a control solution.
(2) Respectively injecting the test solution and the impurity reference solution into a liquid chromatograph, recording a chromatogram, and calculating by peak area according to an external standard method, wherein the chromatographic conditions are as follows:
a chromatographic column: shimadzu Wondasil C18-WR columns (250 × 4.6mm,5 μm);
the column temperature was: 18-22 ℃, preferably 20 ℃;
detection wavelength: 200-400nm, further, the wavelength is 280 nm;
the flow rate is: 0.5-1.5 ml/min; preferably 1.0 ml/min;
sample introduction amount: 5-20 μ l; preferably 10. mu.l;
a mobile phase A: adjusting the pH of the 20mmol/L potassium dihydrogen phosphate buffer solution to 6.0-8.0; preferably, the pH value is adjusted to 6.8-7.2; more preferably, the pH is adjusted to 7.0;
and (3) mobile phase B: methanol, gradient elution was performed as follows:
by adopting the technical scheme of the invention, the advantages are as follows:
according to the detection method for the related substances in the dextro-rabeprazole sodium bulk drug, a specific chromatographic column is selected, the elution gradient proportion and the column temperature are optimized, the types and the number of the monitored impurities are large, the separation degree among the impurities and the separation degree between the impurities and the main component are good, the retention capacity of the main component and the impurities is high, the response of each component is high, the content of 13 related impurities in the dextro-rabeprazole sodium bulk drug can be presented in one chromatographic behavior, the related substances in the dextro-rabeprazole sodium bulk drug can be rapidly and accurately monitored, and the method is simple and easy to implement, and high in accuracy and precision.
Drawings
FIG. 1 is a chromatogram of a mixed solution of impurities and a sample;
FIG. 2 is a chromatogram of a test solution according to the present invention;
FIG. 3 is a linear graph of impurities 2-10, 20-22, and D-rabeprazole sodium;
FIG. 4 is a chromatogram of a mixed solution of impurities and a sample in comparative example 1;
FIG. 5 is a chromatogram of a mixed solution of impurities and a sample in comparative example 2;
fig. 6 is a chromatogram of a mixed solution of impurities and a sample in comparative example 3.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific examples described herein are intended to be illustrative only and are not intended to be limiting. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.
Example 1: HPLC (high Performance liquid chromatography) detection method for related substances of dextro-rabeprazole sodium
First, experimental materials and instruments
1. Drugs and reagents: dextral rabeprazole sodium (Nanjing Hainan medicine science and technology Co., Ltd.), 2-chloromethyl-4- (3-methoxypropoxy) -3-methylpyridine (impurity 1, Yanchenghua chemical Co., Ltd.), 2-mercaptobenzimidazole (impurity 2, Nanjing Kangman biomedical science and technology Co., Ltd.), 2- [ [4- (3-methoxypropoxy) -3-methylpyridin-2-yl ] methylthio ] -1H-benzimidazole (impurity 3, Nanjing Hainan medicine science and technology Co., Ltd.), 2- [ [ [4- (3-methoxypropoxy) -3-methyl-2-pyridyl ] methyl ] sulfone ] -1H-benzimidazole sodium salt (impurity 4, STD), 2-aminobenzimidazole (impurity 5, STD), 2- [ [ [4- (3-methoxypropoxy) -3-methyl-2-pyridinyl ] methyl ] sulfoxide ] -1H-benzimidazole nitroxide (impurity 6, STD), 2-hydroxybenzimidazole (impurity 7, TRC), 2- [ [ [4- (3-methoxypropoxy) -3-methyl-2-pyridinyl ] methyl ] sulfone ] -1H-benzimidazole nitroxide (impurity 8, STD), benzimidazole-2-sulfonic acid (impurity 9, STD), 1- (1H-benzimidazol-2-yl) -1, 4-dihydroxy-3-methyl-4-oxo-2-pyridinecarboxylic acid sodium (impurity 10, USP), 2- [ [ (4-chloro-3-methyl-2-pyridyl) methyl ] sulfinyl ] -1H-benzimidazole (impurity 20, STD), 2- [ [ (4-methoxy-3-methyl-2-pyridyl) methyl ] sulfinyl ] -1H-benzimidazole (impurity 21, STD), 2- [ (4-methoxy-3-methylpyridin-2-yl) methylthio ] -1H-benzimidazole (impurity 22, STD), methanol (chromatographically pure, Shanghai Kagaku Kogyo Co., Ltd.), potassium dihydrogen phosphate (analytically pure, national drug group chemical Co., Ltd.), sodium hydroxide (analytically pure, Shanghai Tantaceae Co., Ltd.), ultrapure water (self-made, millipore).
2. The instrument comprises the following steps: the names and specifications of the specific instruments are shown in table 2 below.
TABLE 2 name and Specifications of specific instruments
| FA124 one-ten-thousandth balance | SHANGHAI SUNNY HENGPING SCIENTIFIC INSTRUMENT Co.,Ltd. |
| XPE204 one-ten-thousandth balance | Mettler |
| AUW 120D one hundred thousand balance | Shimadzu of Japan |
| sQP one hundred thousand balance | Sadoris scientific instruments Ltd |
| XP6I038 millionth-one-day-flat | Mettler |
| pHS-3C digital acidimeter | SHANGHAI INESA SCIENTIFIC INSTRUMENT Co.,Ltd. |
| Agilent1100 high performance liquid chromatograph | Agilent |
Second, liquid phase chromatographic conditions
The chromatographic column adopts Shimadzu Wondasil C18-WR columns (250 × 4.6mm,5 μm); gradient elution was performed according to the following table 1 using 20mmol/L potassium dihydrogen phosphate buffer (adjusted to pH7.0 with sodium hydroxide) as mobile phase a and methanol as mobile phase B; the flow rate is 1.0 ml/min; the column temperature is 20 ℃; measuring the detection wavelength at 280nm, precisely measuring 10 μ l of the sample solution, injecting into a liquid chromatograph, and recording chromatogram.
Third, the experimental process
1. Detection of related substances in dextro-rabeprazole sodium bulk drug
An appropriate amount of the product (batch number: 180923421) is precisely weighed, dissolved by a solvent [0.01mol/L sodium hydroxide-methanol (40:60) ] and quantitatively diluted to prepare a solution containing about 1mg of dextro-rabeprazole sodium per 1ml, and the solution is used as a test solution. Precisely measuring 1.0ml of the test solution, placing the test solution in a 100ml measuring flask, adding a solvent (0.01 mol/L sodium hydroxide-methanol (volume ratio of 40: 60)) to dilute to scale, and shaking up to obtain a control solution. Measuring 10 mul of the reference solution, injecting into a liquid chromatograph, and adjusting the detection sensitivity according to the liquid chromatographic conditions to ensure that the peak height of the main component chromatographic peak is about 15-20% of the full range; then, 10. mu.l of each of the test solution and the control solution was measured precisely, and the solutions were injected into a liquid chromatograph, respectively, and chromatograms were recorded, as shown in FIG. 1. The known impurities are quantified by using a self-control method added with a correction factor, and the unknown impurities are measured by using a self-control method. The results are shown in Table 3. Samples (lots: 181024421 and 181105421) were tested according to the test methods described above, and the results are shown in Table 3.
TABLE 3 test results of different batches of test articles
2. Methodology validation
2.1 specificity
Taking a proper amount (about 10mg) of a dextro-rabeprazole sodium sample (batch number: 180923421), precisely weighing, placing the sample in a 10ml measuring flask, dissolving and diluting the sample to a scale by using a solvent (0.01 mol/L sodium hydroxide-methanol (volume ratio: 40: 60)), and shaking the sample uniformly to obtain a sample solution. Taking appropriate amount of impurities 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 21 and 22, respectively, dissolving with solvent [0.01mol/L sodium hydroxide-methanol (volume ratio 40:60) ] to obtain solutions each containing 100 μ g per 1ml as impurity mother liquor; weighing about 10mg of a dextro-rabeprazole sodium sample, putting the dextro-rabeprazole sodium sample into a 10ml measuring flask, adding a proper amount of the impurity mother liquor, adding a solvent (0.01 mol/L sodium hydroxide-methanol (volume ratio is 40: 60)) to dilute to scale so that the impurities 2, 3, 5, 6, 7, 8, 9, 10, 20, 21 and 22 are 1 mu g/ml (wherein the impurity 1 is 10 mu g/ml, the impurity 4 is 5 mu g/ml, and the concentration of the main component is 1mg/ml), shaking up to obtain a mixed solution of the impurities and the sample.
Weighing about 10mg of dextro-rabeprazole sodium sample, putting the dextro-rabeprazole sodium sample into a 10ml measuring flask, adding a solvent (0.01 mol/L sodium hydroxide-methanol (volume ratio is 40: 60)) to dissolve and dilute the dextro-rabeprazole sodium sample to a scale mark to obtain a sample solution.
Taking appropriate amount of each impurity mother liquor, diluting with solvent [0.01mol/L sodium hydroxide-methanol (volume ratio 40:60) ] to prepare solution with certain concentration as single impurity positioning solution.
Precisely measuring solvent, impurity and sample mixed solution, test solution, and single impurity positioning solution 10 μ l each, injecting into liquid chromatograph, recording chromatogram, and showing the result in Table 4 and the related chromatogram in figures 1-2.
TABLE 4 results of the specificity test
| Impurity and sample mixed solution | Retention time min | Number of theoretical plates | Degree of separation |
| Impurity 9 | 8.180 | 8149 | / |
| |
9.039 | 6587 | 2.12 |
| |
11.260 | 6452 | 4.41 |
| Impurity 7 | 16.566 | 30797 | 11.29 |
| |
17.593 | 35437 | 2.73 |
| |
31.728 | 26743 | 24.55 |
| |
33.218 | 20960 | 1.76 |
| Impurities 21 | 40.362 | 35624 | 8.05 |
| Impurity 4 | 45.191 | 61183 | 6.08 |
| |
47.479 | 66497 | 3.11 |
| Principal component | 49.966 | 86523 | 3.51 |
| Impurity 1 | 54.760 | 99371 | 6.97 |
| Unknown impurity 1 | 55.774 | 116608 | 1.50 |
| |
58.082 | 153539 | 3.70 |
| Unknown impurity 3 | 59.083 | 167287 | 1.71 |
| Impurities 22 | 60.124 | 136277 | 1.69 |
| Impurity 3 | 65.324 | 192971 | 8.33 |
| Unknown impurity 4 | 67.829 | 212086 | 4.23 |
The results show that: under the chromatographic condition, the base line is stable, the blank solvent has no interference to the measurement of the product, the theoretical plate number of the main peak of the dextral rabeprazole sodium is 88655, and the main peak is well separated from adjacent impurities. In the impurity sample mixed solution, the peak shape of each substance is better, the separation degree of a main peak and adjacent impurities is more than 1.5, and the minimum separation degree among the impurities is 1.50, which shows that the specificity is good.
2.2 destructive testing
In order to examine whether degradation products possibly generated by the D-rabeprazole sodium can be detected under selected chromatographic conditions, the product is destroyed by violent conditions of high temperature, acid, alkali, oxidation, illumination and the like, the destroyed sample is dissolved by a solvent (the volume ratio is 40:60) of 0.01mol/L sodium hydroxide-methanol to prepare a test solution, 10 mu L of each solution is precisely measured, the solution is injected into a liquid chromatograph, and a chromatogram is recorded, wherein the specific method is shown in Table 5. The solvent mentioned in Table 5 is [0.01mol/L sodium hydroxide-methanol (volume ratio 40:60) ].
TABLE 5 destructive test results
The result shows that under the conditions of acid, alkali, high temperature, oxidation and illumination, the separation degree of each degradation product and the main peak is good, and the purity of each main peak of a damaged sample meets the requirement. The material balance is calculated by taking the total peak area/sample weighing of the undamaged sample as 100 percent, and the total peak area/respective sample weighing of other damaged samples and the undamaged ratio of the total peak area/respective sample weighing to the undamaged sample are all between 95 percent and 105 percent, so that the materials are conserved. Therefore, the condition is suitable for detecting related substances of the product.
2.3 determination of limits of quantitation and detection
Taking a right-handed rabeprazole sodium reference substance and appropriate amounts of impurities 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 21 and 22 respectively, preparing samples with certain concentrations, diluting gradually, precisely measuring 10 mu l, injecting into a liquid chromatograph, recording a chromatogram, and measuring by using a signal-to-noise ratio S/N-3 and a signal-to-noise ratio S/N-10, wherein the results are shown in Table 6.
TABLE 6 detection limit and quantitation limit results
The result shows that the dextro-rabeprazole sodium and all known impurities have proper detection sensitivity under the concentration and chromatographic conditions of related substances of the product, and all impurities with the content of 0.05 percent can be accurately quantified.
2.4 sample solution stability test
Taking a proper amount of a dextro-rabeprazole sodium sample, adding a solvent (0.01 mol/L sodium hydroxide-methanol (volume ratio is 40: 60)) to dissolve the dextro-rabeprazole sodium sample into a solution containing about 1mg of dextro-rabeprazole sodium per 1ml of the solution serving as a test solution, respectively standing the solution at room temperature after preparation for 0h, 4h, 8h, 12h, 16h, 20h and 24h for sample injection analysis to examine the content of each impurity and main component in the sample solution, and the result is shown in a table 7 by an area normalization method.
TABLE 7 sample solution stability test (Room temperature 15 deg.C)
The results show that: the sample solution was stable for 24h at room temperature (15 ℃).
2.5 Linear
2.5.1 preparation of impurity stock solution
Taking a right-handed rabeprazole sodium reference substance and a proper amount of impurities 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 21 and 22, adding a solvent [0.01mol/L sodium hydroxide-methanol (volume ratio is 40:60) ] to dissolve and dilute into a solution containing about 2 microgram of impurities 2, 3, 5, 6, 7, 8, 9, 10, 20, 21 and 22, 10 microgram of impurities 4 and 20 microgram of right-handed rabeprazole sodium reference substance per 1ml, and taking the solution as a reference substance stock solution to be stored at the temperature of 2-8 ℃.
2.5.2 Linear
Accurately sucking appropriate amount of control stock solution, and diluting with solvent [0.01mol/L sodium hydroxide-methanol (volume ratio of 40:60) ] to obtain solutions with a series of concentrations. Precisely sucking 10 μ l of each of the above gradient concentration solutions, sequentially injecting sample from quantification limit to high concentration, analyzing, recording chromatogram, performing linear regression with impurity reference solution concentration C (μ g/ml) as abscissa and impurity reference peak area as ordinate, and calculating regression equation, with the result shown in Table 8 and FIG. 3.
Table 8 results of linear examination (n ═ 5)
2.6 sample introduction precision test
Accurately weighing appropriate amount of each impurity and a right-handed rabeprazole sodium reference substance, adding a solvent (0.01 mol/L sodium hydroxide-methanol (volume ratio is 40: 60)) to dilute to prepare a mixed solution containing 10 mu g/ml of each impurity (5 mu g/ml of impurity 4) and 10 mu g/ml of right-handed rabeprazole sodium in each 1ml, continuously feeding and measuring for 6 times, and inspecting the change conditions of peak areas and retention time. The results are shown in Table 9.
TABLE 9 sample introduction precision test results
The result shows that the peak areas RSD of the dextro-rabeprazole sodium and each impurity are less than 2.0%, the retention time RSD is less than 1.0%, and the sample injection precision of the instrument is good.
2.7 stability of Mixed control solution
Taking a right-handed rabeprazole sodium reference substance and a proper amount of each impurity reference substance ( impurities 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 21 and 22), dissolving the right-handed rabeprazole sodium reference substance and each impurity reference substance by using a solvent [0.01mol/L sodium hydroxide-methanol (volume ratio is 40:60) ] to prepare a mixed solution of about 10 mu g/ml of each impurity (impurity 4 is 5 mu g/ml) and 10 mu g/ml of the right-handed rabeprazole sodium, using the mixed solution as a reference substance solution, respectively placing the mixed solution at room temperature for 0h, 4h, 8h, 12h, 16h, 20h and 24h after preparation, and analyzing the results by sample injection, wherein the results are shown in a table 10.
TABLE 10 stability test results of impurity control solution (Room temperature 15 deg.C)
The results show that the impurity control solution is stable after being prepared and placed at room temperature for 24 hours.
2.8 repeatability test
Taking a proper amount of dextro-rabeprazole sodium sample, precisely weighing 6 parts, adding a solvent (0.01 mol/L sodium hydroxide-methanol (volume ratio is 40: 60)) to dissolve, and respectively preparing a solution containing 1mg in each 1ml as a test solution. Precisely measuring 10 μ l of each solution, injecting into a liquid chromatograph, calculating the content of each known impurity in the sample by peak area according to an external standard method, calculating the content of unknown impurities in the sample solution by a main component comparison method, and obtaining the measurement results shown in Table 11.
TABLE 11 results of repeatability tests
Note: "-" indicates no detection, and the RSD value was not calculated at a content of less than 0.02%.
2.9 recovery test
Nine parts of dextro-rabeprazole sodium samples are precisely weighed, each part is about 10mg, impurities with limited amount of impurities of 80%, 100% and 120% are respectively added into a 10ml volumetric flask for comparison, a solvent (0.01 mol/L sodium hydroxide-methanol (volume ratio is 40: 60)) is added for dilution to a scale, 10 mu L of the dextro-rabeprazole sodium samples are precisely weighed respectively, and the dextro-rabeprazole sodium samples are injected into a liquid chromatograph for recovery rate measurement, and the results are shown in tables 12 to 23.
TABLE 12 test results for recovery of impurity 2
TABLE 13 test results for recovery of impurity 3
TABLE 14 test results for recovery of impurity 4
TABLE 15 test results for impurity 5 recovery
TABLE 16 test results on recovery of impurity 6
TABLE 17 test results on recovery of impurity 7
TABLE 18 test results for recovery of impurity 8
TABLE 19 test results for impurity 9 recovery
TABLE 20 test results for impurity 10 recovery
TABLE 21 impurity 20 recovery test results
TABLE 22 test results for recovery of impurity 21
TABLE 23 test results for recovery of impurity 22
2.10 intermediate precision test
Taking 6 parts of a dextro-rabeprazole sodium sample, respectively adding a solvent (0.01 mol/L sodium hydroxide-methanol (volume ratio is 40: 60)) to dissolve and dilute the dextro-rabeprazole sodium sample to prepare a solution containing about 1mg of dextro-rabeprazole sodium per 1ml, and taking the solution as a test solution. Precisely measuring 10 μ l, injecting into liquid chromatograph, and recording chromatogram. The content of each known impurity in the test solution was calculated by peak area according to the external standard method, and the content of unknown impurity in the test solution was calculated according to the principal component control method, and the results are shown in table 24.
TABLE 24 intermediate precision test results
Note: apparatus 1: high performance liquid chromatograph 1100, No.: 100141A; an instrument 2: high performance liquid chromatograph 1100, No.: 100129A.
The test result shows that: the method has good intermediate precision.
2.11 calibration factor determination
On 2 liquid chromatographs, 3 chromatographic columns were used, with P3 2And (3) carrying out permutation and combination for 6 times in total, preparing 6 samples in total within the range of the quantification limit to 200% of the specified limited concentration, carrying out regression on each peak area according to the concentration of each component, and calculating a correction factor of impurities relative to the dextro-rabeprazole sodium, wherein the result is shown in a table 25.
TABLE 25 determination of correction factors
Note: a chromatographic column 1: wondasil C18-WR column (250 mm. times.4.6 mm,5 μm), S/N: 7K 5707-21;
and (3) chromatographic column 2: wondasil C18-WR column (250 mm. times.4.6 mm,5 μm), S/N: 5F 5701-11;
a chromatographic column 3: wondasil C18-WR column (250 mm. times.4.6 mm,5 μm), S/N: 6J 5702-14;
an apparatus A: agilent1100 hplc, instrument number 100137 a;
an apparatus B: agilent1100 hplc, instrument No. 100141 a.
The test result shows that: the correction factor for impurity 2 was 0.57, the correction factor for impurity 3 was 0.88, the correction factor for impurity 4 was 1.13, the correction factor for impurity 5 was 0.71, the correction factor for impurity 6 was 0.67, the correction factor for impurity 7 was 0.74, the correction factor for impurity 8 was 0.88, the correction factor for impurity 9 was 0.93, the correction factor for impurity 10 was 0.69, the correction factor for impurity 20 was 0.76, the correction factor for impurity 21 was 0.81, and the correction factor for impurity 22 was 0.81.
2.12 durability examination
In order to examine the degree of resistance of the method to small changes in conditions, a durability test was conducted. Factors considered include: the initial proportion of the organic phase is changed by 25 +/-2%, the pH of the mobile phase is 7.0 +/-0.2, the column temperature is 20 +/-2 ℃ and different chromatographic column tests are carried out. The examination indexes comprise the retention time of the main component in the test sample, the separation degree (minimum separation degree) of the main peak and the adjacent peak, the number of theoretical plates, the number of detected impurity peaks and the content of impurities (the known impurities are calculated according to the main component contrast and a correction factor method), and the measurement results are shown in tables 26 to 29.
2.12.1 Change in initial organic phase proportion of the Mobile phase
The durability was examined at 23%, 25% and 27% of the initial organic phase (methanol) ratio, respectively, and the results are shown in Table 26.
TABLE 26 durability test (initial organic phase ratio of mobile phase)
The results show that: when the proportion of the initial organic phase (methanol) is changed within the range of 23-27%, the retention time of a main peak, the number of theoretical plates and the separation degree are obviously changed, and other chromatographic behaviors and sample detection results are not obviously changed.
2.12.2 change in pH of mobile phase
The durability of the mobile phase was examined at pH 6.8, 7.0 and 7.2, and the results are shown in Table 27.
TABLE 27 durability examination (mobile phase pH)
The results show that: when the pH value of the mobile phase is changed within the range of 6.8-7.2, the retention time of a main peak, the number of theoretical plates and the separation degree are obviously changed, and other chromatographic behaviors and sample detection results are not obviously changed.
2.12.3 Change in column temperature
In order to examine the influence of the column temperature on the chromatographic behavior, the durability of the column temperature was examined at 18 ℃, 20 ℃ and 22 ℃ respectively, and the measurement results are shown in Table 28.
TABLE 28 durability examination (column temperature)
The results show that: when the column temperature is changed within the range of 18-22 ℃, the retention time of the main peak, the number of theoretical plates and the separation degree are obviously changed, and other chromatographic behaviors and sample detection results are not obviously changed.
2.12.4 variations of the column
The durability of the different columns was examined separately and the results are shown in table 29.
TABLE 29 durability examination (different columns)
Note: column 1 was a Wondasil C18-WR column (250 mm. times.4.6 mm,5 μm), S/N: 6J 5702-14;
Column 3 was a Wondasil C18-WR column (250 mm. times.4.6 mm,5 μm), S/N: 7K 5707-21;
the number of impurities 12 in the columns 1 and 3 was not included in the calculation range because the content of impurity 2 was less than 0.005%.
The results show that: the chromatographic columns of different batches have no influence on chromatographic behavior and sample detection results.
In conclusion, the invention adopts the high performance liquid chromatography, screens and optimizes chromatographic conditions and the like, develops a method for related substances, verifies the methodology, qualitatively studies the impurity 1(SMA), and quantitatively studies the impurities 2(SMB), 3 (intermediate 1) and 4-10, 20-22 (process impurities and degradation impurities). The detection method disclosed by the invention has the advantages of good linear relation between the dextro-rabeprazole sodium and related substances, good accuracy and precision, strong specificity and high stability. The detection method has good reproducibility, can meet the detection requirements of related substances of the dextro-rabeprazole sodium bulk drug, and can be used for quality control of the dextro-rabeprazole sodium bulk drug.
Comparative example 1 Effect of different gradients on the assay results during chromatography
Liquid chromatography conditions: the chromatographic column adopts Shimadzu Wondasil C18-WR pillars (250 × 4.6mm,5 μm); gradient elution was performed with 20mmol/L potassium dihydrogen phosphate buffer (pH adjusted to 7.0 with sodium hydroxide) as mobile phase A and methanol as mobile phase B according to Table 30 below; the flow rate is 1.0 ml/min; the column temperature is 20 ℃; the detection wavelength was 280 nm.
TABLE 30 gradient elution procedure
Taking appropriate amounts of impurities 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 21, 22 and a right-handed rabeprazole sodium sample respectively, putting the appropriate amounts into a 10ml measuring flask, adding a solvent [0.01mol/L sodium hydroxide-methanol (volume ratio is 40:60) ] to dilute to scale so that the impurities 2, 3, 5, 6, 7, 8, 9, 10, 20, 21 and 22 are 1 mu g/ml (wherein the impurity 1 is 10 mu g/ml, the impurity 4 is 5 mu g/ml, and the concentration of the main component is 1mg/ml), shaking up to obtain a mixed solution of the impurities and the sample.
And (4) injecting 10 mu l of the solution into a liquid chromatograph, and recording a chromatogram.
TABLE 31 Effect of different gradient elution conditions on chromatographic analysis results
As can be seen from fig. 4 and table 31, in the gradient elution process, in the mixed solution of the impurities and the sample, the impurities are well separated from the main component, but the impurity 1 and the unknown impurity 1(44.718 min) interfere with each other at 44.425min, the separation degree is 0.78, and the separation degrees of the unknown impurity 2 at 45.993min and the unknown impurity 3 at 46.314min are 0.83, which do not reach the baseline separation.
Comparative example 2 influence of different column temperatures on the test results during chromatographic analysis
Liquid chromatography conditions: the chromatographic column adopts Shimadzu Wondasil C18-WR pillars (250 × 4.6mm,5 μm); gradient elution was performed with 20mmol/L potassium dihydrogen phosphate buffer (pH adjusted to 7.0 with sodium hydroxide) as mobile phase A and methanol as mobile phase B according to Table 30 below; the flow rate is 1.0 ml/min; the column temperature is 25 ℃; the detection wavelength was 280 nm. The specific elution process is shown in Table 1.
Taking appropriate amounts of impurities 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 21, 22 and a right-handed rabeprazole sodium sample respectively, putting the appropriate amounts into a 10ml measuring flask, adding a solvent [0.01mol/L sodium hydroxide-methanol (volume ratio is 40:60) ] to dilute to scale so that the impurities 2, 3, 5, 6, 7, 8, 9, 10, 20, 21 and 22 are 1 mu g/ml (wherein the impurity 1 is 10 mu g/ml, the impurity 4 is 5 mu g/ml, and the concentration of the main component is 1mg/ml), shaking up to obtain a mixed solution of the impurities and the sample.
And (3) injecting 10 mu l of the solution into a liquid chromatograph, and recording a chromatogram.
TABLE 32 influence of different column temperatures on the results of the chromatographic analysis
As is clear from fig. 5 and table 32, under the column temperature condition, the degree of separation between the unknown impurity 3(53.038min) and the unknown impurity 22(53.815min) in the impurity/sample mixed solution was 1.36, and the baseline separation was not achieved.
Comparative example 3 influence of different chromatographic columns on the test results during chromatographic analysis
Liquid chromatography conditions: the chromatographic column adopts an Shimadzu Inertsil ODS-3 column (250x4.6mm,5 μm); gradient elution was performed with 20mmol/L potassium dihydrogen phosphate buffer (pH adjusted to 7.0 with sodium hydroxide) as mobile phase A and methanol as mobile phase B according to Table 30 below; the flow rate is 1.0 ml/min; the column temperature is 20 ℃; the detection wavelength was 280 nm. The specific elution process is shown in Table 1.
Taking appropriate amounts of impurities 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 21, 22 and a right-handed rabeprazole sodium sample respectively, putting the appropriate amounts into a 10ml measuring flask, adding a solvent [0.01mol/L sodium hydroxide-methanol (volume ratio is 40:60) ] to dilute to scale so that the impurities 2, 3, 5, 6, 7, 8, 9, 10, 20, 21 and 22 are 1 mu g/ml (wherein the impurity 1 is 10 mu g/ml, the impurity 4 is 5 mu g/ml, and the concentration of the main component is 1mg/ml), shaking up to obtain a mixed solution of the impurities and the sample.
And (3) injecting 10 mu l of the solution into a liquid chromatograph, and recording a chromatogram.
TABLE 33 influence of different column temperatures on the results of the chromatographic analysis
As is clear from fig. 6 and table 33, under the conditions of the column, the degree of separation between the impurity 20(33.563min) and the main peak (34.076min) in the mixed solution of the impurity and the sample was 1.19, and the separation did not reach the baseline separation.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: modifications of the technical solutions described in the foregoing embodiments are still possible, or some technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.
Claims (10)
1. A detection method for related substances in a dextral rabeprazole sodium bulk drug is characterized in that the detection method adopts high performance liquid chromatography to carry out qualitative or quantitative detection on the dextral rabeprazole sodium and the related substances, and the conditions of the high performance liquid chromatography comprise: the chromatographic column is Shimadzu Wondasil C18-a WR column; gradient elution is carried out by adopting a mobile phase A and a mobile phase B as a mixed mobile phase, wherein the mobile phase A is 15-25 mmol/L potassium dihydrogen phosphate buffer solution, and the pH value of the mixed mobile phase A is adjusted to 6.0-8.0; the mobile phase B is methanol; the initial ratio of the mobile phase A to the mobile phase B in the gradient elution process is 73-77: 27-23; the volume ratio of the mobile phase A and the mobile phase B is kept unchanged in the initial ratio within 0-8 minutes; the volume ratio of the mobile phase A to the mobile phase B is gradually changed from the initial ratio to 55:45 at a constant speed within 8-18 minutes; the volume ratio of the mobile phase A to the mobile phase B is kept constant at 55:45 within 18-33 minutes; the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 55:45 to 25:75 at a constant speed within 33-75 minutes; the volume ratio of the mobile phase A to the mobile phase B is kept constant at 25:75 within 75-95 minutes; the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 25:75 to the initial ratio at a constant speed within 95-96 minutes; the volume ratio of mobile phase A and mobile phase B was kept constant for 96-106 minutes.
2. The method for detecting related substances in the dextro-rabeprazole sodium bulk drug according to claim 1, wherein the initial ratio of the mobile phase A to the mobile phase B in the gradient elution process is 75: 25; keeping the volume ratio of the mobile phase A to the mobile phase B constant at 75:25 within 0-8 minutes; the volume ratio of the mobile phase A to the mobile phase B is gradually changed from the initial ratio to 55:45 at a constant speed within 8-18 minutes; the volume ratio of the mobile phase A to the mobile phase B is kept constant at 55:45 within 18-33 minutes; the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 55:45 to 25:75 at a constant speed within 33-75 minutes; the volume ratio of the mobile phase A to the mobile phase B is kept constant at 25:75 within 75-95 minutes; the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 25:75 to 75:25 at a constant speed within 95-96 minutes; the volume ratio of mobile phase A to mobile phase B was kept constant at 75:25 for 96-106 minutes.
3. The method for detecting related substances in the dextro-rabeprazole sodium bulk drug according to claim 1, wherein the high performance liquid chromatography conditions comprise: the column temperature is 18-22 ℃, and preferably 20 ℃.
4. The method for detecting related substances in a dextral rabeprazole sodium bulk drug according to claim 1, wherein the high performance liquid chromatography conditions comprise: the length of the column was 250mm, the diameter was 4.6mm, and the packing particle size was 5 μm.
5. The method for detecting related substances in the dextro-rabeprazole sodium bulk drug according to claim 1, wherein the mobile phase A is 20mmol/L potassium dihydrogen phosphate buffer solution, and the pH value of the mobile phase A is adjusted to 6.8-7.2; preferably, its pH is adjusted to 7.0.
6. The method for detecting related substances in the dextro-rabeprazole sodium bulk drug according to claim 1, wherein the high performance liquid chromatography conditions comprise: the detection wavelength is 200-400nm, preferably 280 nm.
7. The method for detecting related substances in the dextro-rabeprazole sodium bulk drug according to claim 1, wherein the high performance liquid chromatography conditions comprise: the sample injection amount is 5-20 mu l; preferably 10. mu.l.
8. The method for detecting related substances in the dextro-rabeprazole sodium bulk drug according to claim 1, wherein the high performance liquid chromatography conditions comprise: the flow rate is 0.5-1.5 ml/min; preferably 1.0 ml/min.
9. The method for detecting related substances in the dexrabeprazole sodium bulk drug according to claim 1, wherein the related substances comprise the following substances: impurity 1: 2-chloromethyl-4- (3-methoxypropoxy) -3-methylpyridine; impurity 2: 2-mercaptobenzimidazole; impurity 3: 2- [ [4- (3-methoxypropoxy) -3-methylpyridin-2-yl ] -methylsulfanyl ] -1H-benzimidazole; impurity 4: 2- [ [ [4- (3-methoxypropoxy) -3-methyl-2-pyridinyl ] methyl ] sulfone ] -1H-benzimidazole sodium salt; impurity 5: 2-aminobenzimidazole; impurity 6: 2- [ [ [4- (3-methoxypropoxy) -3-methyl-2-pyridinyl ] methyl ] sulfoxide ] -1H-benzimidazole nitroxide; impurity 7: 2-hydroxybenzimidazole; impurity 8: 2- [ [ [4- (3-methoxypropoxy) -3-methyl-2-pyridinyl ] methyl ] sulfone ] -1H-benzimidazole nitroxide; impurity 9: benzimidazole-2-sulfonic acid; 10 parts of impurities: 1- (1H-benzoimidazol-2-yl) -1, 4-dihydroxy-3-methyl-4-oxo-2-pyridinesodium formate; 20 parts of impurities: 2- [ [ (4-chloro-3-methyl-2-pyridinyl) methyl ] sulfinyl ] -1H-benzimidazole; impurity 21: 2- [ [ (4-methoxy-3-methyl-2-pyridinyl) methyl ] sulfinyl ] -1H-benzimidazole; impurity 22: 2- [ (4-methoxy-3-methylpyridin-2-yl) methylsulfanyl ] -1H-benzimidazole.
10. The method for detecting related substances in a dextro-rabeprazole sodium raw material medicament according to claim 1, wherein the correction factor of impurity 2 is 0.57, the correction factor of impurity 3 is 0.88, the correction factor of impurity 4 is 1.13, the correction factor of impurity 5 is 0.71, the correction factor of impurity 6 is 0.67, the correction factor of impurity 7 is 0.74, the correction factor of impurity 8 is 0.88, the correction factor of impurity 9 is 0.93, the correction factor of impurity 10 is 0.69, the correction factor of impurity 20 is 0.76, the correction factor of impurity 21 is 0.81, and the correction factor of impurity 22 is 0.81.
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