CN113325111B - Method for detecting impurity 9 in imidafenacin tablets - Google Patents
Method for detecting impurity 9 in imidafenacin tablets Download PDFInfo
- Publication number
- CN113325111B CN113325111B CN202110633111.6A CN202110633111A CN113325111B CN 113325111 B CN113325111 B CN 113325111B CN 202110633111 A CN202110633111 A CN 202110633111A CN 113325111 B CN113325111 B CN 113325111B
- Authority
- CN
- China
- Prior art keywords
- mobile phase
- impurity
- imidafenacin
- solution
- tablets
- 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
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N2030/042—Standards
- G01N2030/047—Standards external
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Library & Information Science (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The invention relates to a method for detecting impurity 9 in imidafenacin tablets, which adopts phosphoric acid solution containing sodium octane sulfonate-acetonitrile as a mixed mobile phase, strictly controls the pH value of the mobile phase, optimizes the time of gradient elution and the proportion of the mobile phase in the process of gradient elution, ensures that the impurity 9 has good peak shape, strong retention capacity and higher response, effectively solves the problems of poor peak shape and interference of auxiliary materials on the impurity 9 and a main component (imidafenacin) during the determination of the impurity 9, can quickly and accurately monitor the content of the impurity 9 in the imidafenacin tablets, and has important significance for the quality evaluation of the imidafenacin tablets. The detection method provided by the invention can be used for measuring on a binary pump high performance liquid chromatograph and can also be used for measuring on a quaternary pump high performance liquid chromatograph, is not limited by instruments, and completely realizes the free use of the instruments.
Description
Technical Field
The invention belongs to the technical field of drug analysis, and particularly relates to a method for detecting an impurity 9 in imidafenacin tablets.
Background
Imidafenacin Tablets (Imidafenacin Tablets) are a novel diphenylbutanamide anticholinergic drug developed by apricot oil pharmaceutical co-company and small wild drug industry co-company, have high bladder selectivity, are used for treating overactive bladder with symptoms of urgency, frequency and incontinence, and are marketed in japan in 6 months of 2007.
Overactive bladder is a syndrome characterized by symptoms of urgency, often accompanied by symptoms of frequency and nocturia, with or without urge incontinence. Detrusor instability is one of the important causes of overactive bladder. The imidafenacin contained in the imidafenacin tablet can selectively act on M3 and M1 receptors on detrusor, block the contraction action of choline on the detrusor, relax the detrusor and obviously improve symptoms caused by overactive bladder.
When the imidafenacin tablet is used for treating the symptoms, the dosage is low, usually, an adult takes 0.1mg every time for 2 times a day, the effective dosage is low, the specification of the product is extremely small, the product is 0.1mg, the tablet weight is about 120-138 mg, the proportion of the main drug in the tablet weight is less than one thousandth, and in the pharmaceutical research stage, the comprehensive research on related substances of the imidafenacin tablet becomes an indispensable part for quality control of the product.
The impurity 9 is collected in the Japanese IF file of the imidafenacin tablets, is a very important forced degradation product in the imidafenacin tablets, can be generated under the condition of illumination or oxidative damage, and IF the impurity 9 is detected in a sample stored for a long time, the quality of the product can be directly reflected or whether the product is packaged and stored or not has problems, so the detection of the impurity 9 in the imidafenacin tablets becomes a key step for controlling the quality of the product.
At present, there are reports on the detection method of related substances in the product, or even though there are reports, for example, patent with application number CN201910275617.7 discloses a detection method of related substances in imidafenacin, which includes detection of impurity 9, but the detection target is imidafenacin bulk drug, and cannot avoid the interference and influence of various auxiliary materials in the imidafenacin tablet on the impurity, under the chromatographic condition, the peak of impurity 9 in the imidafenacin tablet is seriously interfered by the auxiliary materials, so that the method is not suitable for detecting impurity 9 in the imidafenacin tablet. The patent with the application number of CN201310384585.7 also discloses a method for detecting related substances of imidafenacin tablets, which is not suitable for detecting related substances in imidafenacin tablets containing povidone accessories on the market in Japan, and has the defects of limited method and narrow application range.
Disclosure of Invention
The invention aims to provide a method for detecting impurity 9 in an imidafenacin tablet on the basis of the prior art, the impurity 9 has good peak shape, strong retention capacity and higher response, the separation degree between the impurity 9 and the main component imidafenacin is high, the problems of detection sensitivity and auxiliary material interference caused by the extremely small specification (0.1mg) of the imidafenacin tablet can be effectively solved, and the content of the impurity 9 in the imidafenacin tablet can be quickly and accurately monitored.
The technical scheme of the invention is as follows:
a detection method for an impurity 9 in an imidafenacin tablet is characterized in that the impurity 9 in the imidafenacin tablet is quantitatively detected by adopting high performance liquid chromatography, and the conditions of the high performance liquid chromatography comprise:
the chromatographic column is Inertsil ODS-3C18, InertSustain C18 or Wondasil C18;
gradient elution is carried out by taking a mobile phase A and a mobile phase B as a mixed mobile phase, wherein the mobile phase A is a phosphoric acid solution containing sodium octane sulfonate, and the pH value is adjusted to 2.4-2.7 by triethylamine; the mobile phase B is acetonitrile; the specific gradient elution procedure was as follows: (1) keeping the volume ratio of the mobile phase A to the mobile phase B unchanged at 77:23 within 0-15 minutes; (2) the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 77:23 to 67:33 at a constant speed within 15-25 minutes; (3) the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 67:33 to 60:40 at a constant speed within 25-35 minutes; (4) the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 60:40 to 20:80 at a constant speed within 35-45 minutes; (5) the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 20:80 to 77:23 at a constant speed within 45-55 minutes; (6) the volume ratio of the mobile phase A to the mobile phase B is kept constant at 77:23 within 55-65 minutes;
dissolving the sample imidafenacin tablet comprises the following steps: adding 5-8 imidafenacin tablets into a solvent, fully shaking up after disintegration, carrying out ultrasonic treatment for 10-30 min, filtering, and taking a subsequent filtrate to obtain a solution containing 0.08-0.16 mg/ml of imidafenacin as a test solution. In dissolving the sample, the selected solvent is a mixed solution composed of a mobile phase A and a mobile phase B, wherein the volume ratio of the mobile phase A to the mobile phase B is 60-70: 40-30, and can be but not limited to 60:40, 62:38, 65:35, 66:34, 67:33, 69:31 or 70:30, in order to better dissolve the imidafenacin tablet and reduce the interference effect of auxiliary materials, in a preferred scheme, when the mixed solution composed of the mobile phase A and the mobile phase B is selected, the volume ratio of the mobile phase A to the mobile phase B is 67: 33.
When the detection method disclosed by the invention is adopted to detect the impurity 9, the structural formula of the impurity 9 is as follows:
the detection method provided by the invention is characterized in that a mobile phase A and a mobile phase B are taken as a mixed mobile phase for gradient elution, during chromatographic analysis, after a chromatographic column is selected, the pH value of the mobile phase needs to be determined, the elution process of the mobile phase is isocratic elution or gradient elution, in the elution process, the proportion of the mobile phase A and the mobile phase B can influence the response of the impurity 9 on the chromatographic column, and the auxiliary material peak can not interfere the analysis of the impurity 9 and the imidafenacin main peak. For the invention, the specification of the imidafenacin tablet is extremely small, the amount of the auxiliary materials is extremely large, the interference factors are extremely many, the pH value of the mobile phase, the time of the gradient elution process and the proportion of the mobile phase are not randomly selected and need to be determined by a large amount of experiments and analysis, otherwise, the impurity 9 has poor peak shape and does not meet the requirement of quantitative analysis, the impurity 9 and the imidafenacin are interfered by the auxiliary material peak, the content of the impurity 9 in the sample can not be accurately determined, and the quality control of the product is influenced.
When the detection method is used for detecting the impurity 9 in the imidafenacin tablet, the proportion of the water phase (mobile phase A) and the organic phase (mobile phase B) in the initial mobile phase needs to be strictly controlled in the gradient elution process, and when the proportion of the organic phase (mobile phase B) in the initial mobile phase is too high or too low, the peak shape of the impurity 9 is not good, the recovery rate does not meet the requirement, and the content of the impurity 9 in the imidafenacin tablet cannot be accurately detected.
For the invention, because the specification of the imidafenacin tablet of the sample is very small (0.1mg) and the quantity of each tablet of auxiliary material is very large (about 130mg), the tablet quantity of the imidafenacin tablet and the volume of the solvent are strictly controlled in the process of preparing the test solution, the ultrasonic treated solution is maintained while the auxiliary material is completely dissolved in the solvent, the concentration of the ultrasonic treated solution is proper and convenient for filtration, and the filtrate is taken. If the dosage of the tablet and the volume of the solvent are not appropriate, the concentration of the sample may be too low to meet the sensitivity requirement of impurity detection, the main component may not be completely dissolved, and the auxiliary materials of the solution after ultrasonic treatment are too many to cause difficulty in filtration, thereby seriously interfering the accuracy of the detection result.
In a preferred embodiment, the dissolution of the sample imidafenacin tablet comprises the following steps: taking 5 imidafenacin tablets, adding the 5 imidafenacin tablets into 5ml of solvent, fully shaking the tablets after disintegration, then carrying out ultrasonic treatment for 10-30 min (for example, 20min), then filtering the solution, and taking the subsequent filtrate to obtain a solution containing 0.1mg/ml of imidafenacin as a test solution, wherein the solvent is a mixed solution consisting of a mobile phase A and a mobile phase B in a volume ratio of 67: 33.
The mobile phase A adopted by the invention is a phosphoric acid solution containing sodium octane sulfonate, in a preferable scheme, the concentration of the sodium octane sulfonate in the phosphoric acid solution containing the sodium octane sulfonate is 0.001-0.01 mol/L, and in order to obtain a better effect, the concentration of the sodium octane sulfonate in the phosphoric acid solution containing the sodium octane sulfonate is preferably 0.003-0.007 mol/L; more preferably 0.005 mol/L.
When the detection method is used for detecting the impurity 9 in the imidafenacin tablet, the pH value of the mobile phase A needs to be strictly controlled, and the pH value of the mobile phase A is too high or too low, so that the accurate determination of the content of the impurity 9 is not facilitated. For the purposes of the present invention, the pH of mobile phase a is 2.4-2.7, but may be, but is not limited to, 2.4, 2.45, 2.5, 2.55, 2.6 or 2.7, and for better results, the pH of mobile phase a is 2.5. When the pH value in the mobile phase a is too low, for example, the pH value is 2.3, the peak shape of the impurity 9 is good, but the retention time variation is large, and the recovery rate is not satisfactory; when the pH value in the mobile phase a is too high, for example, pH 2.8, the peak shape of the impurity 9 collapses or branches, and the content of the impurity 9 in the imidafenacin tablet cannot be accurately measured.
The preparation of the mobile phase A adopted by the invention comprises the following steps: firstly, preparing a phosphoric acid aqueous solution with the volume concentration of 0.05-2.5%, preparing a solution with the concentration of 0.001-0.01 mol/L from sodium octane sulfonate by using the prepared phosphoric acid aqueous solution, and adjusting the pH value to 2.4-2.7 by using triethylamine to obtain the sodium octane sulfonate; preferably, the pH is adjusted to 2.5 with triethylamine.
In a preferred embodiment, the preparation of mobile phase a used in the present invention comprises the following steps: firstly, preparing a phosphoric acid aqueous solution with the volume concentration of 0.1%, preparing sodium octane sulfonate into a solution with the concentration of 0.005mol/L by using the prepared phosphoric acid aqueous solution, and then adjusting the pH value to 2.4-2.7 by using triethylamine to obtain the sodium octane sulfonate; preferably, the pH is adjusted to 2.5 with triethylamine.
The detection method provided by the invention adopts a chromatographic column and octadecylsilane bonded silica gel, such as Inertsil ODS-3C18, InertSustain C18 or Wondasil C18, wherein the effect of Inertsil ODS-3C18 is better. In a preferred embodiment, the column has a length of 250mm, a diameter of 4.6mm and a packing particle size of 5 μm.
Furthermore, the detection wavelength of the detector is 218-222 nm; preferably 220 nm.
Further, the column temperature is 20-30 ℃; preferably 25 deg.c.
Furthermore, the flow rate is 0.5-1.5 ml/min, preferably 1.0 ml/min.
Furthermore, the sample injection amount is 50-150 μ l; preferably 100. mu.l. For example, the sample size may be 50. mu.l, 100. mu.l or 150. mu.l.
In the method for detecting the impurity 9 in the imidafenacin tablet, detailed information of the impurity 9 is shown in table 1.
Table 1 impurity 9 information
In a preferred embodiment, the detection method provided by the present invention may be configured as follows, and when configured as follows, the selected solvent is: and the volume ratio of the mobile phase A to the mobile phase B is 67: 33.
Test solution: precisely adding 5 tablets of the product (imidafenacin tablets) into 5ml of solvent, after disintegration, shaking uniformly, sealing, ultrasonically treating for 20min, shaking uniformly, standing, filtering, and taking subsequent filtrate to obtain a solution containing 0.1mg/ml of imidafenacin.
Control solution: 1% test solution.
Blank adjuvant solution: weighing 5 samples of adjuvants (containing pregelatinized starch, microcrystalline cellulose, polyvidone K30, magnesium stearate and stomach soluble film coating powder), precisely adding 5ml of solvent, shaking, sealing, ultrasonic treating for 20min, shaking, standing, and filtering to obtain filtrate.
Adding an impurity 9 solution into a test sample: taking 5 tablets of the product (imidafenacin tablet), precisely adding 5ml of the impurity 9 reference substance solution, after disintegration, shaking up, sealing, ultrasonically treating for 20min, shaking up, standing, filtering, and taking the subsequent filtrate to obtain the final product.
By adopting the technical scheme of the invention, the advantages are as follows:
(1) the method for detecting the impurity 9 in the imidafenacin tablets adopts phosphoric acid solution containing sodium octane sulfonate-acetonitrile as a mixed mobile phase, controls the pH value of the mobile phase and strictly controls the proportion and time of the mobile phase in the elution process, solves the problem of interference of auxiliary materials on the impurity 9, can be used for measuring on a binary pump high performance liquid chromatograph, can also be used for measuring on a quaternary pump high performance liquid chromatograph, is not limited by instruments, and completely realizes free use of the instruments.
(2) The method for detecting the impurity 9 in the imidafenacin tablet, provided by the invention, needs to strictly control the pH value of the mobile phase, optimizes the gradient elution time and the proportion of the mobile phase in the gradient elution process, so that the impurity 9 has good peak shape, strong retention capacity and higher response, effectively solves the problems that the peak shape is poor and auxiliary materials interfere the impurity 9 and a main component (imidafenacin) when the impurity 9 is measured, can quickly and accurately monitor the content of the impurity 9 in the imidafenacin tablet, and has important significance for quality evaluation of the imidafenacin tablet.
Drawings
FIG. 1 is a high performance liquid chromatogram of a solution of an empty white excipient of example 1;
FIG. 2 is a high performance liquid chromatogram of a control solution of impurity 9 from example 1;
FIG. 3 is a high performance liquid chromatogram of the test solution of example 1;
FIG. 4 is a high performance liquid chromatogram of the sample plus impurity 9 solution of example 1;
FIG. 5 is a high performance liquid chromatogram of a photo-disrupted liquid sample of example 1;
FIG. 6 is a high performance liquid chromatogram of the sample plus mixed impurities solution of comparative example 1;
FIG. 7 is a high performance liquid chromatogram of the sample plus impurity 9 solution of comparative example 1;
FIG. 8 is a high performance liquid chromatogram of a dissolution sample plus impurity 9 solution of comparative example 2;
FIG. 9 is a high performance liquid chromatogram of the sample plus impurity 9 solution of comparative example 3;
FIG. 10 is a high performance liquid chromatogram of the sample plus impurity 9 solution of comparative example 4;
FIG. 11 is a high performance liquid chromatogram of the sample plus impurity 9 solution of comparative example 5;
FIG. 12 is a high performance liquid chromatogram of the sample plus impurity 9 solution of comparative example 6.
Detailed Description
The detection method of the present invention is further illustrated by the following examples in conjunction with the drawings, but the present invention is not limited to these examples.
Example 1
High performance liquid chromatography conditions:
the chromatographic column is an octadecylsilane chemically bonded silica column, the model is Inertsil ODS-3C18(250 x 4.6mm, 5 μm), phosphoric acid solution of sodium octane sulfonate (1.08g of sodium octane sulfonate is dissolved and diluted to 1000ml by adding phosphoric acid solution with volume concentration of 0.1 percent, and the pH value is adjusted to 2.5 by triethylamine) is taken as a mobile phase A, acetonitrile is taken as a mobile phase B, gradient elution is carried out, the flow rate is 1.0ml/min, the detection wavelength is 220nm, the column temperature is 25 ℃, and the sample injection amount is 100 μ l.
The specific gradient elution procedure was as follows: (1) the volume ratio of the mobile phase A to the mobile phase B is kept constant at 77:23 within 0-15 minutes; (2) the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 77:23 to 67:33 at a constant speed within 15-25 minutes; (3) the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 67:33 to 60:40 at a constant speed within 25-35 minutes; (4) the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 60:40 to 20:80 at a constant speed within 35-45 minutes; (5) the volume ratio of the mobile phase A to the mobile phase B is uniformly graded from 20:80 to 77:23 within 45-55 minutes; (6) the volume ratio of mobile phase A to mobile phase B was kept constant at 77:23 for 55-65 minutes.
The solvent is a mixed solution of mobile phase A and mobile phase B in a volume ratio of 67:33, and the solution is prepared as follows:
test solution: placing 5 tablets of the product (imidafenacin tablets) in a volumetric flask, precisely adding 5ml of solvent, after disintegration, shaking up, sealing, ultrasonically treating for 20min, shaking up, standing, filtering, and taking the subsequent filtrate to obtain a solution containing 0.1mg/ml of imidafenacin.
Control solution: a proper amount of the test solution is measured and diluted by a solvent to prepare a solution containing 1 mu g of imidafenacin per 1ml, which is used as a control solution, namely a 1% test solution.
Blank adjuvant solution: weighing 5 samples of adjuvants (containing pregelatinized starch, microcrystalline cellulose, polyvidone K30, magnesium stearate and stomach soluble film coating powder), precisely adding 5ml of solvent, shaking, sealing, ultrasonic treating for 20min, shaking, standing, and filtering to obtain filtrate.
Adding an impurity 9 solution into a test sample: taking 5 tablets of the product (imidafenacin tablets), precisely adding 5ml of the impurity 9 reference substance solution, after disintegration, shaking uniformly, sealing, ultrasonically treating for 20min, shaking uniformly, standing, filtering, and taking the subsequent filtrate to obtain the product.
Taking 100 μ l of each solution, sampling, analyzing, and recording chromatogram, wherein the related chromatogram is shown in figure 1, figure 2, figure 3 and figure 4.
As can be seen from FIGS. 1 to 4, the baseline is stable, the peak shape of the impurity 9 is good, and the blank auxiliary material does not interfere with the determination of the impurity 9; when the sample is added into the impurity 9 solution, the separation degree of the impurity 9 and the main component is good, and the number of theoretical plates is high.
The detection method of related substances is verified as follows:
1. specificity property
The preparation of the blank adjuvant solution, the impurity 9 reference solution, the sample solution and the sample plus impurity 9 solution is as described in example 1 above.
Taking each 100 mu l of blank solvent, blank auxiliary material solution, impurity 9 reference substance solution, sample solution and impurity 9 solution for sample, sampling and analyzing, recording chromatogram, and observing retention time, separation degree and theoretical plate number of impurity 9 and main components, wherein the related chromatogram is shown in figures 1-4, and the result is shown in table 2.
TABLE 2 results of the specificity test
Note: "-" indicates no detection.
And (4) conclusion: under the chromatographic condition, the base line is stable, and the blank solvent and the blank auxiliary materials do not interfere with the determination of the impurity 9; in the impurity 9 reference substance solution, the impurity 9 has good peak shape, higher theoretical plate number and higher purity factor; no impurity 9 is detected in the test solution; when the sample is added into the impurity 9 solution, the separation degree between the impurity 9 and the main component is good, the number of theoretical plates is high, and the specificity is good.
2. Destructive test
In order to examine whether the degradation product impurity 9 generated by the imidafenacin tablet can be detected under the selected chromatographic conditions, the product is destroyed by respectively using acid, alkali, oxidation, high temperature, illumination and other violent conditions, and the impurity 9 is measured by using the high performance liquid chromatography conditions in the example 1. The specific method comprises the following steps:
solvent: and the volume ratio of the mobile phase A to the mobile phase B is 67: 33.
Blank auxiliary materials are not damaged: taking about 1.87g blank adjuvants (containing pregelatinized starch, microcrystalline cellulose, polyvidone K30, magnesium stearate, and gastric soluble film coating powder), precisely adding 15ml solvent, shaking, sealing, ultrasonic processing for 20min, shaking, standing, and filtering to obtain filtrate to obtain undamaged blank adjuvants solution.
Non-destructive test solution: placing 20 tablets of the product into a 50ml measuring flask, precisely adding 20ml of solvent, after disintegration, shaking uniformly, sealing, ultrasonically treating for 20min, filtering, and taking the subsequent filtrate to obtain the undamaged test solution.
Irradiating to destroy solid blank auxiliary materials: taking about 0.625g of 5 tablets of blank auxiliary materials (containing pregelatinized starch, microcrystalline cellulose, polyvidone K30, magnesium stearate and gastric-soluble film coating powder), placing in a transparent volumetric flask, irradiating for 48 hours under strong light (5000lx), precisely adding 5ml of solvent, shaking up after disintegration, sealing, ultrasonically treating for 20min, shaking up, cooling to room temperature, filtering, and taking the subsequent filtrate.
Light damage solid samples: placing 5 tablets of the product in a 25ml transparent volumetric flask, irradiating for 48 hours under strong light (5000lx), precisely adding 5ml of solvent, shaking uniformly after disintegration, sealing, ultrasonically treating for 20min, shaking uniformly, cooling to room temperature, filtering, and taking the subsequent filtrate.
Destroying the liquid blank auxiliary materials by illumination: taking 5 tablets of blank auxiliary materials (containing pregelatinized starch, microcrystalline cellulose, polyvidone K30, magnesium stearate and gastric-soluble film coating powder), placing in a transparent volumetric flask, precisely adding 5ml of solvent, after disintegration, shaking up, sealing, ultrasonic treating for 20min, shaking up, cooling to room temperature, irradiating for 48 hours under strong light (5000lx), filtering, and taking the subsequent filtrate.
Light damage to the liquid sample: placing 5 tablets of the product in a 25ml transparent volumetric flask, precisely adding 5ml of solvent, after disintegration, shaking up, sealing, ultrasonically treating for 20min, shaking up, cooling to room temperature, irradiating under strong light (5000lx) for 48 hours, filtering, and collecting the subsequent filtrate.
Oxidizing and destroying blank auxiliary materials: taking 1ml of the undamaged blank auxiliary material solution, adding 0.25ml of 3% hydrogen peroxide solution, shaking up, sealing with a top hollow cover, placing in an oven at 100 ℃ for 2 hours, taking out, cooling to room temperature, removing the top hollow cover, and taking out supernatant to obtain the product.
Oxidative damage of the samples: taking 2ml of the undamaged sample solution, adding 0.5ml of 3% hydrogen peroxide solution, shaking up, sealing with a top hollow cover, placing in an oven at 100 ℃ for 2 hours, taking out, cooling to room temperature, removing the top hollow cover, and taking out supernatant.
Destroying the blank auxiliary materials by high-temperature liquid: taking 1ml of the undamaged blank auxiliary material solution, adding a top hollow cover for sealing, placing in an oven at 100 ℃ for 2 hours, taking out, cooling to room temperature, removing the top hollow cover, and taking supernatant to obtain the Chinese medicinal composition.
High temperature liquid damage samples: taking 1ml of the undamaged sample solution, adding a top empty cover for sealing, placing in an oven at 100 ℃ for 2 hours, taking out, then placing to room temperature, removing the top empty cover, and taking out supernatant fluid to obtain the finished product.
High temperature solids destroy blank excipients: taking about 0.625g of 5 tablets of blank auxiliary materials (containing pregelatinized starch, microcrystalline cellulose, polyvidone K30, magnesium stearate and gastric-soluble film coating powder) in a 25ml measuring flask, plugging, heating in 100 deg.C water bath for 4 hr, cooling, precisely adding 5ml of solvent, after disintegration, shaking, sealing, ultrasonic treating for 20min, shaking, cooling to room temperature, filtering, and collecting the filtrate.
High temperature solids destruct the sample: placing 5 tablets of the product into a 25ml measuring flask, adding a plug, heating in a water bath at 100 ℃ for 4 hours to destroy, cooling, precisely adding 5ml of solvent, after disintegration, shaking up, sealing, ultrasonically treating for 20min, shaking up, cooling to room temperature, and filtering to obtain a subsequent filtrate.
Acid destruction of blank excipients: taking about 0.625g of 5 tablets of blank auxiliary materials (containing pregelatinized starch, microcrystalline cellulose, polyvidone K30, magnesium stearate and gastric-soluble film coating powder) into a 25ml measuring flask, precisely adding 1ml of 1mol/L hydrochloric acid solution, precisely adding 4ml of solvent, after disintegration, shaking up, sealing, ultrasonically treating for 20min, shaking up, heating in a water bath at 100 ℃ for 4 hours, taking out, cooling, filtering, and taking out the subsequent filtrate.
Acid destruction of the samples: placing 5 tablets of the product into a 25ml measuring flask, precisely adding 1ml of 1mol/L hydrochloric acid solution, precisely adding 4ml of solvent, after disintegration, shaking up, sealing, performing ultrasonic treatment for 20min, shaking up, heating in a water bath at 100 ℃ for 4 hours, taking out, cooling, filtering, and taking out the subsequent filtrate.
Alkali destruction of blank auxiliary materials: taking 1ml of the undamaged blank auxiliary material solution, adding 0.25ml of 1mol/L sodium hydroxide solution, shaking up, adding a top hollow cover for sealing, placing in an oven at 100 ℃ for 2 hours, taking out, cooling to room temperature, removing the top hollow cover, and taking out supernatant fluid to obtain the finished product.
Alkali-destroyed sample: and taking 2ml of the undamaged sample solution, adding 0.5ml of 1mol/L sodium hydroxide solution, shaking up, adding a top hollow cover, sealing, placing in an oven at 100 ℃ for 2 hours, taking out, cooling to room temperature, removing the top hollow cover, and taking out supernatant to obtain the finished product.
And taking 100 mu l of each sample solution under each damage condition, carrying out sample injection analysis, and recording a chromatogram. The results are shown in Table 3.
TABLE 3 destructive test results
And (4) conclusion: the results show that the product can degrade the impurities 9 under the conditions of illumination and oxidative destruction, under the chromatographic condition, the blank auxiliary materials can not interfere the determination of the impurities 9 after being destroyed, the impurities 9 can be accurately and quantitatively researched, the specificity is good, namely, the formulated chromatographic condition is suitable for the detection of the impurities 9 in the product, and the illumination destruction map is shown in figure 5.
3. Limit of detection of quantitative limit
Taking 100 mu l of mixed reference substance solution, carrying out sample injection analysis, recording a chromatogram, and respectively determining the detection limit and the quantification limit of the impurity 9 and the imidafenacin by the signal-to-noise ratios S/N & lt 3 & gt and S/N & lt 10 & gt, wherein the results are shown in tables 4 and 5.
TABLE 4 detection of Limit results
TABLE 5 Limit precision results
And (4) conclusion: from the above results, under the conditions of the concentration of the related substances and the chromatogram, the detection limit of the impurity 9 is about 0.06% (0.06 μ g/ml) of the concentration of the test sample, the detection sensitivity is high, and the impurity 9 can be effectively detected. The quantitative limit is 6 times of sample injection, the retention time RSD of the main component and the impurity 9 is less than 1.0 percent, the peak area RSD is less than 10.0 percent, and the precision is good.
4. Precision of sample introduction
Taking 100 mu l of mixed reference substance solution, carrying out sample injection analysis, recording a chromatogram, and carrying out parallel sample injection for 6 times. The results are shown in Table 6.
TABLE 6 sample introduction precision results
And (4) conclusion: the retention time RSD of the main component and the impurity 9 is less than 1.0 percent, the peak area RSD is less than 2.0 percent, and the sample introduction precision is good.
5. Stability of solution
Test solution: placing 5 tablets of the product (imidafenacin tablets) in a volumetric flask, precisely adding 5ml of solvent, after disintegration, shaking up, sealing, ultrasonically treating for 20min, shaking up, standing, filtering, and taking the subsequent filtrate to obtain a solution containing 0.1mg/ml of imidafenacin.
Mixing the reference solution: accurately weighing an appropriate amount of impurity 9 reference substance, adding acetonitrile with volume concentration of 70% for dissolving, and adding a solvent for diluting to prepare a solution containing 0.1mg per 1ml, wherein the solution is used as a mother solution of the impurity 9; accurately weighing a proper amount of imidafenacin reference, adding a solvent to dissolve and dilute the imidafenacin reference to prepare a solution containing 0.5mg of imidafenacin per 1ml of imidafenacin reference as a mother solution; accurately measuring appropriate amount of the mother liquor, and quantitatively diluting with solvent to obtain mixed reference solution containing impurity 9 and imidafenacin 0.2 μ g and 1 μ g per 1 ml.
Precisely measuring 100 μ l of each of the test solution and the mixed reference solution, respectively, performing sample injection analysis at different times, and recording chromatogram, the results are shown in Table 7.
TABLE 7 solution stability results
And (4) conclusion: mixing the reference solution, standing at room temperature for 29 hr until the peak areas RSD of impurity 9 and main component are less than 2.0%; the test solution was allowed to stand at room temperature for 29 hours, no impurity 9 was detected, and the RSD of the main component peak area was less than 2.0%, indicating that the stability was good when the mixed control solution and test solution were allowed to stand at room temperature for 29 hours under these measurement conditions.
6. Linearity
Accurately weighing an appropriate amount of impurity 9 reference substance, adding acetonitrile with volume concentration of 70% for dissolving, and adding a solvent for diluting to prepare a solution containing 0.1mg per 1ml, wherein the solution is used as a mother solution of the impurity 9; an appropriate amount of the imidafenacin reference substance is accurately weighed, dissolved by a solvent and diluted to prepare a solution containing 0.5mg per 1ml as a mother solution.
Precisely measuring 1ml of the imidafenacin reference mother liquor and 1ml of the impurity 9 reference mother liquor respectively, putting the imidafenacin reference mother liquor and the impurity 9 reference mother liquor into a 50ml measuring flask, diluting the imidafenacin reference mother liquor to a scale by using a solvent, and shaking the imidafenacin reference mother liquor and the impurity 9 reference mother liquor uniformly to obtain a linear stock solution.
Precisely measuring the linear stock solutions 0.5ml, 0.8ml, 1ml, 1.2ml, 1.5ml and 2ml respectively in 10ml measuring bottles, diluting to scale with solvent, and shaking to obtain solutions of 50%, 80%, 100%, 120%, 150% and 200% linear levels.
Precisely sucking 100 mul of the solutions with the series of concentrations, sequentially injecting samples from low concentration to high concentration for analysis, and recording a chromatogram. The concentration C (μ g/ml) of the impurity 9 control solution was taken as the abscissa and the peak area of the impurity 9 control solution was taken as the ordinate, and linear regression was performed and the regression equation was calculated, and the results are shown in Table 8.
TABLE 8 investigation of the Linear relationship (n ═ 7)
And (4) conclusion: imidafenacin and impurity 9 in a certain concentration range, R 2 The Y-axis intercept of the linear equation accounts for 100 percent of the response value and is within 25 percent.
7. Calibration factor determination
Taking an impurity 9 reference substance and an imidafenacin reference substance, preparing a series of solutions with linear horizontal concentrations under the collinearity term, performing linear regression by taking the concentrations as horizontal coordinates and peak areas as vertical coordinates, and calculating a regression equation. The ratio of the slope of the regression equation for imidafenacin to the slope of the regression equation for impurity 9 was used as a correction factor and was determined by different personnel at different times and with different instruments. The results are shown in Table 9.
TABLE 9 calibration factor determination results
Note: a chromatographic column 1: inertsil ODS-3C 18250 mm X4.6 mm 5 μm, No.: 1A7173804
And (3) chromatographic column 2: inertsil ODS-3C 18250 mm X4.6 mm 5 μm, No.: 1A7174339
And (3) chromatographic column: inertsil ODS-3C 18250 mm X4.6 mm 5 μm, No.: 1A7165866
An instrument A: agilent1100-DAD high performance liquid chromatograph; an apparatus B: agilent1100 high performance liquid chromatograph
And (4) conclusion: from the above results, the calibration factor of the impurity 9 is 0.95, which is between 0.9 and 1.1, and no calibration factor calculation is required.
8. Repeatability and accuracy
Because the product does not detect the impurity 9, the repeatability and the recovery rate are measured by adopting a sample-adding recovery mode.
Precisely measuring appropriate amount of the impurity 9 stock solution, quantitatively diluting with solvent to obtain solution containing 0.2 μ g of impurity per 1ml, and shaking to obtain impurity 9 reference solution. (parallel preparation 2 portions)
Precisely measuring appropriate amount of the impurity 9 stock solution, quantitatively diluting with solvent to obtain impurity 9 reference substance solution containing 0.16 μ g, 0.2 μ g and 0.24 μ g per 1ml, and shaking to obtain impurity 9 reference substance solutions with limit levels of 80%, 100% and 120%.
Recovery of solution (80%): taking 5 tablets of the product, precisely adding 5ml of the above 80% limit level mixed impurity reference substance solution, after disintegration, shaking, sealing, ultrasonically treating for 20min, shaking, standing, filtering, and taking the subsequent filtrate. (parallel preparation of 3 portions)
Recovery of the solution (100%): taking 5 tablets of the product, precisely adding 5ml of the above 100% limit level mixed impurity reference substance solution, after disintegration, shaking, sealing, ultrasonically treating for 20min, shaking, standing, filtering, and taking the subsequent filtrate. (parallel preparation 6 portions)
Recovery of the solution (120%): taking 5 tablets of the product, precisely adding 5ml of the 120% limit level mixed impurity reference substance solution, after disintegration, shaking uniformly, sealing, ultrasonically treating for 20min, shaking uniformly, standing, filtering, and taking the subsequent filtrate. (parallel preparation 3 parts)
Control solution: an appropriate amount of the solution with a 100% limit horizontal recovery rate was diluted with a solvent to prepare a solution containing about 1. mu.g per 1ml, which was used as a 1% self-control solution. (6 parts prepared for the solution corresponding to the 100% limit horizontal recovery)
Each 100 μ l of the above solutions was precisely measured, injected into a liquid chromatograph, a chromatogram was recorded, and the recovery rate was calculated from [ (measured amount-background amount)/added amount x 100] of the impurity 9, and the results are shown in tables 10 to 11.
TABLE 10 measurement results of recovery of impurity 9
TABLE 11 repeatability of measurement of impurity 9
And (4) conclusion: the recovery rate of the impurity 9 is between 90 and 108 percent, the average recovery rate is 103.1 percent, the RSD is 2.41 percent and is less than 6.0 percent, and the recovery rate is good; under the repeatability term, the recovery rate is between 90% and 108%, the RSD is 2.82% and less than 6.0%, and the repeatability is good; the method is suitable for measuring the impurity 9 of the product.
The content of impurity 9 was calculated by the external standard method and the 1% self-control method, respectively, and the results of the detection by the two methods were compared, and are shown in table 12.
TABLE 12 comparison of measurement results of impurity 9 content at repeatability (1% self-control method, external standard method)
And (4) conclusion: the results show that in 6 test sample solutions, the results of the calculation of the impurity 9 by adopting an external standard method and a 1% self-contrast method are basically consistent, and the RSD of the results measured by the two methods is less than 6.0%, which shows that the method for detecting the impurity 9 in the product has good repeatability.
10. Intermediate precision
Under the test conditions, the same homogeneous sample was used, and a series of tests were performed by different analysts at different times and with different instruments, and the intermediate precision was examined with the recovery rate of impurity 9, and the results are shown in table 13.
TABLE 13 results of intermediate precision test
And (4) conclusion: the results show that different analysts use different instruments to detect at different times, the recovery rate of the impurity 9 is within the range of 90-108%, and the RSD is less than 6.0%, which shows that the intermediate precision of the method for detecting the impurity 9 in the product is good.
Comparative example 1:
reference is made to the method of example 1 in patent application No. CN201910275617.7
High performance liquid chromatography conditions: the chromatographic column is an octadecylsilane chemically bonded silica column, the model is Inertsil ODS-3C18(250 x 4.6mm, 5 μm), phosphoric acid solution of sodium octane sulfonate (1.08g of sodium octane sulfonate is dissolved and diluted to 1000ml by adding phosphoric acid solution with volume concentration of 0.1 percent, and the pH value is adjusted to 2.8 by triethylamine) is taken as a mobile phase A, acetonitrile is taken as a mobile phase B, gradient elution is carried out, the flow rate is 1.0ml/min, the detection wavelength is 220nm, the column temperature is 25 ℃, and the sample injection amount is 50 μ l. The solvent is a mixed solution of mobile phase A and mobile phase B in a volume ratio of 67: 33.
The gradient elution procedure was: (1) the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 80:20 to 60:40 at a constant speed within 0-40 minutes; (2) the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 60:40 to 20:80 at a constant speed within 40-50 minutes; (3) the volume ratio of the mobile phase A to the mobile phase B is kept to be 20:80 isocratic elution within 50-53 minutes; (4) the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 20:80 to 80:20 at a constant speed within 53-58 minutes; (5) the volume ratio of mobile phase A to mobile phase B was kept constant at 80:20 for 58-70 minutes.
Taking impurity 1, impurity 2, impurity 3, impurity 5, impurity 7, impurity 8, impurity 9 and impurity 10 reported in the patent to prepare a sample and mixed impurity solution. The preparation method comprises the following steps:
accurately weighing appropriate amount of impurity 1, impurity 2, impurity 3 and impurity 5 reference substances respectively, dissolving with solvent, and diluting to obtain solution containing 0.5mg per 1ml as mother liquor of impurity 1, impurity 2, impurity 3 and impurity 5; accurately weighing appropriate amounts of impurities 7, 8, 9 and 10 as reference substances, dissolving with acetonitrile with volume concentration of 70%, adding solvent, diluting to obtain solution containing 0.1mg per 1ml, and using the solution as mother liquor of impurities 7, 8, 9 and 10; precisely measuring appropriate amount of the above mother liquor, and quantitatively diluting with solvent to obtain mixed impurity solution containing 5 μ g of each impurity per 1 ml.
Adding mixed impurity solution into a sample: taking 5 tablets (containing imidafenacin, pregelatinized starch, microcrystalline cellulose, polyvidone K30, magnesium stearate, and gastric soluble film coating powder), precisely adding 5ml of the above mixed impurity solution, after disintegrating, shaking, sealing, ultrasonic treating for 20min, shaking, standing, filtering, and collecting the filtrate.
The solution is taken for sample injection analysis, and chromatogram is recorded, and the results are shown in figures 6-7. As can be seen from FIG. 6, in the detection method provided in the comparative example, the retention time of the impurity 9 is 11.768min, and the peak position is seriously interfered by the auxiliary material; as can be seen from FIG. 7, when the concentration of impurity 9 is 2. mu.g/ml, but less than 5. mu.g/ml, impurity 9 is covered in the minor constituent peak. Because the concentration of the impurity 9 in the sample to be detected is 5 mu g/ml, even if the impurity 9 is interfered by the auxiliary material, the peak shape of the impurity 9 can be seen, if the concentration of the impurity 9 in the imidafenacin tablet is lower than the concentration, the impurity 9 is covered in the auxiliary material peak and cannot be detected, and the quality evaluation of the product is influenced, and in the process of researching and developing the imidafenacin tablet, the concentration of the impurity 9 cannot be higher than 5 mu g/ml, otherwise, the quality of the sample is worrisome, so the detection method of the comparative example 1 is not suitable for measuring the impurity 9 in the imidafenacin tablet.
Comparative example 2:
reference is made to the method of example 1 in patent application No. CN201310384585.7
High performance liquid chromatography conditions: the chromatographic column is an octadecylsilane chemically bonded silica gel column, the model is Inertsil ODS-3C18(250 x 4.6mm, 5 mu m), acetonitrile-water phase (1.08g of sodium octane sulfonate is dissolved in 1000ml of dilute phosphoric acid solution (obtained by diluting 1ml of phosphoric acid with water to 100 ml) is mixed uniformly, saturated sodium hydroxide solution is added to adjust the pH to 2.5) (35:65) to be a mobile phase, the flow rate is 1.0ml/min, the detection wavelength is 220nm, the column temperature is 25 ℃, and the sample injection amount is 200 mu l.
Dissolution sample plus impurity 9 solution: accurately weighing a proper amount of impurity 9 reference substance, adding acetonitrile with the volume concentration of 70% for dissolving, and then adding a mobile phase for diluting to prepare a solution containing 0.1mg of impurity 9 reference substance in each 1ml of solution; taking 1 tablet (containing imidafenacin, pregelatinized starch, microcrystalline cellulose, povidone K30, magnesium stearate and gastric-soluble film coating powder), putting into a dissolution cup preheated to 37 ℃ and filled with 900ml of water medium, taking a dissolution liquid after 20 minutes, filtering, taking 1ml of a subsequent filtrate, precisely adding 0.1ml of the impurity 9 control solution, and uniformly mixing to obtain a solution containing 0.1 mu g/ml of imidafenacin and 10 mu g/ml of impurity 9, wherein the solution is used as a dissolution sample and added with the impurity 9 solution.
The solution was sampled and analyzed, and the chromatogram was recorded, and the results are shown in FIG. 8. As can be seen from FIG. 8, the retention time of imidafenacin is 10.147min, and the auxiliary peak interferes with the main component of imidafenacin, which affects the quantitative accuracy; the peak time of the impurity 9 is about 2 min-4 min, and the interference of the solvent peak is serious.
The results indicate that the method of CN201310384585.7 patent is not suitable for measuring the content of imidafenacin tablets containing povidone as an auxiliary material and the impurity 9 in the imidafenacin tablets marketed in japan.
Comparative example 3:
high performance liquid chromatography conditions: the chromatographic column is an octadecylsilane chemically bonded silica column, the model is Inertsil ODS-3C18(250 x 4.6mm, 5 μm), phosphoric acid solution of sodium octane sulfonate (1.08g of sodium octane sulfonate is dissolved and diluted to 1000ml by adding phosphoric acid solution with volume concentration of 0.1 percent, and the pH value is adjusted to 2.8 by triethylamine) is taken as a mobile phase A, acetonitrile is taken as a mobile phase B, gradient elution is carried out, the flow rate is 1.0ml/min, the detection wavelength is 220nm, the column temperature is 25 ℃, and the sample injection amount is 100 μ l. The solvent is a mixed solution of mobile phase A and mobile phase B in a volume ratio of 67: 33.
The gradient elution procedure was: the same as in example 1.
Preparing a solution:
preparation of sample plus impurity 9 solution: an appropriate amount of impurity 9 as a control was precisely weighed, dissolved in acetonitrile at a volume concentration of 70%, and diluted with a solvent to give a solution containing about 0.2. mu.g of impurity 9 as a control solution. Taking 5 tablets (containing imidafenacin, pregelatinized starch, microcrystalline cellulose, polyvidone K30, magnesium stearate, and gastric soluble film coating powder), adding 5ml of the above reference solution containing impurity 9, disintegrating, shaking, sealing, ultrasonic processing for 20min, shaking, standing, filtering, and collecting the filtrate.
The solution was sampled and analyzed, and the chromatogram was recorded, and the results are shown in FIG. 9. The retention time of the impurity 9 is 13.007min, the peak top is collapsed and is in a shape of steamed bread, the auxiliary material peak is close to the impurity 9 at the front end of the impurity 9, the peak emergence of the impurity 9 is easily influenced, the baseline of the peak emergence position of the impurity 9 is not stable, and the phenomenon of inaccurate quantitative determination is easily caused.
Comparative example 4:
high performance liquid chromatography conditions: the chromatographic column is an octadecylsilane chemically bonded silica column, the model is Inertsil ODS-3C18(250 x 4.6mm, 5 μm), phosphoric acid solution of sodium octane sulfonate (1.08g of sodium octane sulfonate is dissolved and diluted to 1000ml by adding phosphoric acid solution with volume concentration of 0.1 percent, and the pH value is adjusted to 2.3 by triethylamine) is taken as a mobile phase A, acetonitrile is taken as a mobile phase B, gradient elution is carried out, the flow rate is 1.0ml/min, the detection wavelength is 220nm, the column temperature is 25 ℃, and the sample injection amount is 100 μ l. The solvent is a mixed solution of mobile phase A and mobile phase B in a volume ratio of 67: 33.
The gradient elution procedure was: the same as in example 1.
Preparing a solution:
The solution was sampled and analyzed, and the chromatogram was recorded, and the results are shown in FIG. 10. As can be seen from FIG. 10, the impurity 9 has a good peak shape, the retention time is 20.244min, a peak is generated at the front edge of the povidone as the excipient, the recovery rate of the impurity 9 in the solution of the sample and the impurity 9 is calculated to be 87.18%, and the recovery rate does not meet the requirement of 90% -108%.
As can be seen from the mobile phase pH values and experimental results in example 1, comparative example 3, and comparative example 4, when the mobile phase pH value is greater than or equal to 2.8, the impurity 9 peak shape collapses or branches; when the pH value of the mobile phase is 2.3, the peak shape is good, but the retention time of the impurity 9 is changed greatly, the recovery rate does not meet the requirement, and the content of the impurity 9 cannot be accurately measured.
The above comparison results show that the mobile phase pH has a greater effect on impurity 9.
Comparative example 5:
high performance liquid chromatography conditions: the chromatographic column is an octadecylsilane chemically bonded silica column, the model is Inertsil ODS-3C18(250 x 4.6mm, 5 μm), phosphoric acid solution of sodium octane sulfonate (1.08g of sodium octane sulfonate is dissolved and diluted to 1000ml by adding phosphoric acid solution with volume concentration of 0.1 percent, and the pH value is adjusted to 2.5 by triethylamine) is taken as a mobile phase A, acetonitrile is taken as a mobile phase B, gradient elution is carried out, the flow rate is 1.0ml/min, the detection wavelength is 220nm, the column temperature is 25 ℃, and the sample injection amount is 100 μ l. The solvent is a mixed solution of mobile phase A and mobile phase B in a volume ratio of 67: 33.
The gradient elution procedure was: (1) keeping the volume ratio of the mobile phase A to the mobile phase B unchanged at 79:21 within 0-15 minutes; (2) the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 79:21 to 67:33 at a constant speed within 15-25 minutes; (3) the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 67:33 to 60:40 at a constant speed within 25-35 minutes; (4) the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 60:40 to 20:80 at a constant speed within 35-45 minutes; (5) the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 20:80 to 79:21 at a constant speed within 45-55 minutes; (6) the volume ratio of mobile phase A to mobile phase B was kept constant at 79:21 for 55-65 minutes.
Preparing a solution:
The solution was sampled and analyzed, and the chromatogram was recorded, and the results are shown in FIG. 11. As can be seen from fig. 11, in the process of gradient elution, the ratio of the aqueous phase (mobile phase a) to the organic phase (mobile phase B) in the initial mobile phase needs to be strictly controlled, when the ratio of the organic phase (mobile phase B) in the initial mobile phase is reduced by 2%, the retention time of the impurity 9 is 19.778min, the peak shape is broadened, and the peak shape is close to the front end of the auxiliary material peak, and the calculated recovery rate of the impurity 9 in the solution of the sample and the impurity 9 is 89.4%, which does not meet the requirement of the range of 90% to 108%.
Comparative example 6:
high performance liquid chromatography conditions: the chromatographic column is an octadecylsilane chemically bonded silica column, the model is Inertsil ODS-3C18(250 x 4.6mm, 5 μm), phosphoric acid solution of sodium octane sulfonate (1.08g of sodium octane sulfonate is dissolved and diluted to 1000ml by adding phosphoric acid solution with volume concentration of 0.1 percent, and the pH value is adjusted to 2.5 by triethylamine) is taken as a mobile phase A, acetonitrile is taken as a mobile phase B, gradient elution is carried out, the flow rate is 1.0ml/min, the detection wavelength is 220nm, the column temperature is 25 ℃, and the sample injection amount is 100 μ l. The solvent is a mixed solution of mobile phase A and mobile phase B in a volume ratio of 67: 33.
The gradient elution procedure was: (1) keeping the volume ratio of the mobile phase A to the mobile phase B unchanged at 75:25 within 0-15 minutes; (2) the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 75:25 to 67:33 at a constant speed within 15-25 minutes; (3) the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 67:33 to 60:40 at a constant speed within 25-35 minutes; (4) the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 60:40 to 20:80 at a constant speed within 35-45 minutes; (5) the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 20:80 to 75:25 at a constant speed within 45-55 minutes; (6) the volume ratio of mobile phase A to mobile phase B was kept constant at 75:25 for 55-65 minutes.
Preparing a solution:
The solution was sampled and analyzed, and the chromatogram was recorded, and the results are shown in FIG. 12. As can be seen from fig. 12, in the process of gradient elution, the ratio of the aqueous phase (mobile phase a) to the organic phase (mobile phase B) in the initial mobile phase needs to be strictly controlled, when the ratio of the organic phase (mobile phase B) in the initial mobile phase is increased by 2%, the retention time of the impurity 9 is 11.194min, the recovery rate of the impurity 9 in the solution of the sample and the impurity 9 is calculated to be 109.5%, which does not meet the requirement of the range of 90% to 108%, which indicates that the impurity 9 may contain a small auxiliary material peak, and the imidafenacin peaks at the tail of the povidone auxiliary material peak, and the peak area is inaccurate, and when the content of the impurity 9 in the product is measured by using a 1% self-contrast method, the measurement result is extremely inaccurate.
In summary, the mobile phase pH value and gradient elution process selected in this example 1 are suitable for detecting impurity 9 in imidafenacin tablet, the impurity 9 peak shape is good, the peak position baseline is stable, no auxiliary material interference occurs before and after the retention time within one minute, the specificity is good, and the accuracy is high.
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 (3)
1. A detection method of an impurity 9 in a imidafenacin tablet is characterized in that the detection method adopts high performance liquid chromatography to carry out quantitative detection on the impurity 9 in the imidafenacin tablet, and the conditions of the high performance liquid chromatography comprise:
the chromatographic column is Inertsil ODS-3C18, 250 × 4.6mm, 5 μm;
taking phosphoric acid solution of sodium octane sulfonate as a mobile phase A, acetonitrile as a mobile phase B, performing gradient elution at a flow rate of 1.0ml/min, a detection wavelength of 220nm, a column temperature of 25 ℃, and a sample injection amount of 100 mu L; the preparation process of the mobile phase A is as follows: 1.08g of sodium octane sulfonate is taken, dissolved and diluted to 1000ml by adding a phosphoric acid solution with the volume concentration of 0.1 percent, and the pH value is adjusted to 2.5 by triethylamine;
the specific gradient elution procedure was as follows: (1) keeping the volume ratio of the mobile phase A to the mobile phase B unchanged at 77:23 within 0-15 minutes; (2) the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 77:23 to 67:33 at a constant speed within 15-25 minutes; (3) the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 67:33 to 60:40 at a constant speed within 25-35 minutes; (4) the volume ratio of the mobile phase A to the mobile phase B is gradually changed from 60:40 to 20:80 at a constant speed within 35-45 minutes; (5) the volume ratio of the mobile phase A to the mobile phase B is uniformly graded from 20:80 to 77:23 within 45-55 minutes; (6) the volume ratio of the mobile phase A to the mobile phase B is kept constant at 77:23 within 55-65 minutes;
dissolving the sample imidafenacin tablet comprises the following steps: taking 5-8 imidafenacin tablets, adding the imidafenacin tablets into a solvent, fully shaking the imidafenacin tablets after disintegration, carrying out ultrasonic treatment for 10-30 min, filtering the imidafenacin tablets, and taking a subsequent filtrate to obtain a solution containing 0.08-0.16 mg/ml of imidafenacin as a test solution; the solvent is a mixed solution composed of a mobile phase A and a mobile phase B, wherein the volume ratio of the mobile phase A to the mobile phase B is 60-70: 40-30;
the structural formula of the impurity 9 is as follows:
2. the method for detecting the impurity 9 in the imidafenacin tablet as claimed in claim 1, wherein the solvent is a mixed solution of mobile phase a and mobile phase B, wherein the volume ratio of the mobile phase a to the mobile phase B is 67: 33.
3. The method for detecting the impurity 9 in the imidafenacin tablet as claimed in claim 1, wherein the dissolving of the sample imidafenacin tablet comprises the steps of: taking 5 imidafenacin tablets, adding the 5 imidafenacin tablets into 5ml of solvent, fully shaking the tablets after disintegration, carrying out ultrasonic treatment for 10-30 min, filtering the solution, and taking a subsequent filtrate to obtain a solution containing 0.1mg/ml of imidafenacin as a test solution, wherein the solvent is a mixed solution consisting of a mobile phase A and a mobile phase B in a volume ratio of 67: 33.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110633111.6A CN113325111B (en) | 2021-06-07 | 2021-06-07 | Method for detecting impurity 9 in imidafenacin tablets |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110633111.6A CN113325111B (en) | 2021-06-07 | 2021-06-07 | Method for detecting impurity 9 in imidafenacin tablets |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113325111A CN113325111A (en) | 2021-08-31 |
| CN113325111B true CN113325111B (en) | 2022-09-30 |
Family
ID=77419832
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110633111.6A Active CN113325111B (en) | 2021-06-07 | 2021-06-07 | Method for detecting impurity 9 in imidafenacin tablets |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113325111B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115856117A (en) * | 2022-11-21 | 2023-03-28 | 南京联智医药科技有限公司 | Method for detecting related substances in imidafenacin |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104415034A (en) * | 2013-08-29 | 2015-03-18 | 天津药物研究院 | Imidafenacin pharmaceutical composition and preparation method thereof |
| CN107752455A (en) * | 2017-12-08 | 2018-03-06 | 南京科技职业学院 | A kind of shoe rack hung on antitheft door |
| CN110068623A (en) * | 2019-04-08 | 2019-07-30 | 南京海纳医药科技股份有限公司 | Detection method in relation to substance in a kind of imidafenacin |
-
2021
- 2021-06-07 CN CN202110633111.6A patent/CN113325111B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104415034A (en) * | 2013-08-29 | 2015-03-18 | 天津药物研究院 | Imidafenacin pharmaceutical composition and preparation method thereof |
| CN107752455A (en) * | 2017-12-08 | 2018-03-06 | 南京科技职业学院 | A kind of shoe rack hung on antitheft door |
| CN110068623A (en) * | 2019-04-08 | 2019-07-30 | 南京海纳医药科技股份有限公司 | Detection method in relation to substance in a kind of imidafenacin |
Non-Patent Citations (2)
| Title |
|---|
| 咪达那新口腔崩解片的制备及处方优化;李丽然 等;《医药导报》;20121031;第31卷(第10期);全文 * |
| 溶剂分散法制备咪达那新片;杨如意 等;《中国药剂学杂志》;20170731;第15卷(第4期);全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113325111A (en) | 2021-08-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111983113A (en) | Method for detecting content of 6-oxosimvastatin in ezetimibe simvastatin tablets | |
| CN110068623B (en) | Method for detecting related substances in imidafenacin | |
| CN113325111B (en) | Method for detecting impurity 9 in imidafenacin tablets | |
| CN113009029A (en) | Method for determining related substances of rabeprazole sodium enteric-coated preparation | |
| CN111122736B (en) | Method for detecting enantiomer in intermediate of brivaracetam | |
| CN114354789B (en) | Method for simultaneously measuring cabozantinib analogue and related substances thereof | |
| CN113702514A (en) | Method for determining atorvastatin calcium related impurity I | |
| CN114689737B (en) | Analysis method of S-o-chlorophenylglycine methyl tartrate related substances | |
| CN114965754A (en) | Method for detecting related substances and bacteriostatic agent in acetaminophen tablet | |
| CN111380978B (en) | Method for simultaneously determining contents of coenzymes NADP and FAD in medicine | |
| CN106153756B (en) | High performance liquid chromatography for detecting rapamycin in everolimus | |
| CN113109461A (en) | Method for detecting related substances in imidafenacin tablets | |
| CN107884496B (en) | Method for determining content of succinic acid in trelagliptin succinate | |
| CN111579688A (en) | Method for determining content of enantiomer in dapoxetine hydrochloride | |
| CN115078576B (en) | Analytical method for related substances of paracetamol and dihydrocodeine tablet | |
| CN112557558B (en) | Method for detecting SCH59566 impurity content in ezetimibe simvastatin tablets | |
| CN111208214A (en) | Method for detecting Tween content in medicinal preparation by liquid phase method | |
| CN112730637B (en) | HPLC detection method for related substances of L-malic acid | |
| CN116754705B (en) | Method for detecting acetic acid and acetate content | |
| CN112526013B (en) | Method for detecting concentration of related substances in ibuprofen medicament by using ultra-high liquid chromatography | |
| CN114924020B (en) | Quality control method for diastereoisomer in arformoterol tartrate inhalation solution | |
| CN114113402B (en) | Method for measuring pinanediol content in bortezomib by adopting high performance liquid chromatography | |
| CN116973487B (en) | Method for measuring impurity content of saxagliptin and application | |
| CN115876911A (en) | Method for measuring venlafaxine hydrochloride tablet related substances by using liquid chromatography | |
| CN117491539A (en) | Analysis method of diastereoisomeric impurities in flurbiprofen patches |
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 |