CN111505159A - Detection method of related substances in arotinolol hydrochloride - Google Patents
Detection method of related substances in arotinolol hydrochloride Download PDFInfo
- Publication number
- CN111505159A CN111505159A CN202010386440.0A CN202010386440A CN111505159A CN 111505159 A CN111505159 A CN 111505159A CN 202010386440 A CN202010386440 A CN 202010386440A CN 111505159 A CN111505159 A CN 111505159A
- Authority
- CN
- China
- Prior art keywords
- arotinolol hydrochloride
- mobile phase
- related substances
- acetonitrile
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—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
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/36—Control of physical parameters of the fluid carrier in high pressure liquid systems
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
-
- 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
- G01N30/8679—Target compound analysis, i.e. whereby a limited number of peaks is analysed
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)
- Spectroscopy & Molecular Physics (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The invention provides a method for detecting related substances in arotinolol hydrochloride. The invention adopts high performance liquid chromatography for detection, and the chromatographic conditions are as follows: a chromatographic column: octadecyl bonding silica gel column; detection wavelength 315-: acetonitrile-water-phosphoric acid, the mobile phase B is acetonitrile-water, and the elution mode is gradient elution. The detection method provided by the invention can realize effective separation of the main component and known impurities and a plurality of unknown impurities, and the method provided by the invention is researched and verified by methodology such as specificity, sensitivity and the like, and finds that the method provided by the invention is sensitive, accurate and good in reproducibility, can realize the qualitative and quantitative determination of more impurities in the arotinolol hydrochloride raw material by a simpler method, and provides reliable guarantee for improving and better controlling the quality of arotinolol hydrochloride preparation products.
Description
Technical Field
The invention relates to the technical field of drug analysis and detection, in particular to a method for detecting related substances in arotinolol hydrochloride.
Background
Arotinolol hydrochloride with chemical name of 5- [2- [ [ [3- (1, 1-dimethylethyl) amino group]-2-hydroxypropyl radical]Sulfur based radicals]-4-thiazolyl]The hydrochloride salt of (E) -2-thiophenecarboxamide, which was first marketed in Japan in 1985, was a selective β salt1-adrenoceptor antagonists with weak α1The medicine has the effect of antagonizing adrenergic receptors, is mainly used for clinically treating mild to moderate essential hypertension, angina, rapid arrhythmia, essential tremor and the like, does not cause the increase of the physique of patients when being used for treating the obese hypertension, and has good market prospect.
The related substances are starting materials, intermediates, side reaction products, degradation impurities and the like brought in the process of synthesizing the medicine, influence the product quality of the medicine and even cause serious adverse reactions. The development of a detection method of related substances is an important part in the medicine quality research, and the safety, effectiveness and quality controllability of the medicine can be improved. Therefore, an analysis method capable of detecting related substances in the arotinolol hydrochloride is developed, the quality of the raw materials is controlled from the source, and the method has very important significance for improving the quality of arotinolol hydrochloride preparation products and improving the medication safety of patients.
Disclosure of Invention
Aiming at the technical problems, the invention provides a method for detecting related substances in arotinolol hydrochloride.
In order to solve the technical problems, the technical scheme provided by the invention is as follows:
a method for detecting related substances in arotinolol hydrochloride adopts high performance liquid chromatography for detection, and the chromatographic conditions are as follows:
a chromatographic column: octadecylsilane chemically bonded silica gel column;
a UV detector for detecting the wavelength of 315-;
mobile phase A: acetonitrile-water-phosphoric acid, and the mobile phase B is acetonitrile-water;
the elution mode is gradient elution, and the elution procedure is as follows:
0-3min, 100% mobile phase A;
3-45min, 100% → 40% mobile phase a, 0% → 60% mobile phase B;
45-46min, 40% → 100% mobile phase a, 60% → 0% mobile phase B;
46-60min, 100% mobile phase A.
Compared with the prior art, the detection method of related substances in arotinolol hydrochloride provided by the invention adopts an octadecyl bonded silica gel chromatographic column, acetonitrile-water-phosphoric acid is used as a mobile phase A, acetonitrile-water is used as a mobile phase B, the method can realize effective separation of the main component in the arotinolol hydrochloride from known impurities and a plurality of unknown impurities by a specific gradient elution mode through a high performance liquid chromatography, accurately qualitatively and quantitatively detect the impurity condition in the arotinolol hydrochloride, and the research and verification of methodologies such as specificity, sensitivity and the like show that the method is sensitive and accurate, has better reproducibility and low detection limit and quantitative limit, can realize qualitative and quantitative detection of more impurities in the arotinolol hydrochloride by a simpler method, and provides reliable guarantee for improving and better controlling the quality of arotinolol hydrochloride preparation products.
The acetonitrile and phosphoric acid used in the present invention are both commercially available analytical grade acetonitrile and phosphoric acid.
The related substances in the invention refer to impurities introduced or generated by degradation in the synthesis process of arotinolol hydrochloride, wherein the known impurities comprise A LL E-1, SM2 and RS-E, and the structural formula of each known impurity is shown as follows.
Preferably, the flow rate is 0.45-0.60m L/min, and the column temperature is 28-32 ℃.
More preferably, the detection wavelength is 317nm, the flow rate is 0.5m L/min, and the column temperature is 30 ℃.
Preferably, the injection volume is 10 μ L.
The optimal detection condition can ensure that the main component, the unknown impurities and the known impurities in the arotinolol hydrochloride reach higher separation degree, and the effective detection of the unknown impurities can be ensured, thereby achieving the purpose of accurately detecting the content of related substances in the arotinolol hydrochloride.
Preferably, the volume ratio of the acetonitrile to the water in the mobile phase A is 20:80, and the volume percentage of the phosphoric acid is 0.18-0.22% based on the total volume of the acetonitrile and the water as 100%.
More preferably, the phosphoric acid is 0.2% by volume.
Preferably, the volume ratio of acetonitrile to water in mobile phase B is 90: 10.
The preferable mobile phase can better detect impurities in the arotinolol hydrochloride on the premise of not generating baseline interference, is beneficial to detecting related substances, effectively improves the peak shape and ensures that the accuracy and precision of the detection result are higher.
Preferably, the chromatography column has a size of 250 x 4.6mm and a packing diameter of 3 μm.
More preferably, the column is YMC-Triart C18, 4.6 × 250mm, 5 μm.
The optimal chromatographic column specification can ensure that the peak shape, the separation degree and the detection sensitivity of each component are good, and the baseline interference is small, so that the arotinolol hydrochloride and the impurities are effectively separated, the result is accurate and reliable, and the repeatability is good.
Preferably, the concentration of the test solution is 1.0mg/m L.
The preferable concentration of the test sample is favorable for leading the peak shape of the main component and the impurities to be better, the column efficiency to be high and the integral to be more accurate, thereby being favorable for carrying out more accurate calculation on the content of the impurities in the test sample.
Drawings
FIG. 1 is a chromatogram of a mixed contaminant control solution under system suitability in example 2;
FIG. 2 is a chromatogram of a 2.2.1 interference and localization solution of arotinolol hydrochloride in example 2;
FIG. 3 is a chromatogram of a localization solution of A LL E-1 under the 2.2.1 interference and localization test item in example 2;
FIG. 4 is a chromatogram of the SM2 localization solution under the 2.2.1 interference and localization test item in example 2;
FIG. 5 is a chromatogram of the RS-E localization solution under the 2.2.1 interference and localization test item in example 2;
FIG. 6 is a chromatogram of a test solution under the 2.2.1 interference and localization test item in example 2;
FIG. 7 is a chromatogram of an acid-disrupted test solution of example 2 under the 2.2.2 forced degradation test;
FIG. 8 is a chromatogram of a high temperature-destroyed test solution of 2.2.2 forced degradation test item in example 2;
FIG. 9 is a chromatogram of a light-disrupted test solution of example 2 under the 2.2.2 forced degradation test;
FIG. 10 is a chromatogram of a base-disrupted test solution of example 2 under the 2.2.2 forced degradation test;
FIG. 11 is a chromatogram of an oxidatively disrupted test solution of example 2 under the 2.2.2 forced degradation test.
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 embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1
In the following examples A LL E-1, SM2 and RS-E represent related substances having the structural formulae shown below.
The material and the method are as follows:
the instrument comprises the following steps: high performance liquid chromatograph, ultraviolet detector, measuring flask, electronic balance.
The reagent comprises acetonitrile, phosphoric acid, arotinolol hydrochloride reference substances (Shijiazhuang Gerui pharmaceutical Co., Ltd., content of 99.7%), SM2 reference substances (Qianhui pharmaceutical Co., Ltd., content of 99.0%), A LL E-1 reference substances (Shijiazhuang Gerui pharmaceutical Co., Ltd., content of 99.2%) and RS-E reference substances (Shijiazhuang Harui pharmaceutical science and technology Co., Ltd., content of 95.99%).
Blank solvent: acetonitrile-water in a volume ratio of 25: 75;
impurity solvent: a mixed solution of a blank solvent and N, N Dimethylformamide (DMF) in a volume ratio of 1: 1.
1.1 preparation of the solution
A LL E-1 reference stock solution is prepared by weighing 10mg of A LL E-1 reference, precisely weighing, placing in a 100m L volumetric flask, dissolving with impurity solvent and fixing volume, precisely weighing 1m L solution, placing in a 20m L volumetric flask, diluting with impurity solvent and fixing volume, and preparing into A LL E-1 reference stock solution with concentration of 0.005 mg/ml.
And (2) weighing 10mg of reference products of arotinolol hydrochloride, SM2 and RS-E respectively, precisely weighing, placing the reference products in the same 100m L volumetric flask, dissolving the reference products by using an impurity solvent, fixing the volume, precisely weighing the solution 1m L and the A LL E-1 reference product stock solution 1m L, placing the reference products in the same 100m L volumetric flask, diluting the reference products by using a blank solvent, fixing the volume, and preparing the mixed impurity reference product solution with the concentration of A LL E-1 being 0.00005mg/m L, the concentration of arotinolol hydrochloride, the concentration of SM2 and the concentration of RS-E being 0.001mg/m L.
And (3) positioning solution, precisely measuring a proper amount of reference substances of arotinolol hydrochloride, A LL E-1, SM-2 and RS-E respectively, dissolving and diluting with water respectively, and performing constant volume to obtain positioning solution with A LL E-1 of about 0.00005mg/m L and arotinolol hydrochloride, SM2 and RS-E impurity of about 0.001mg/m L respectively.
The sample solution is prepared by weighing 10mg of arotinolol hydrochloride sample, precisely weighing, placing in a 10m L volumetric flask, and preparing into solution with concentration of 1mg/m L with blank solvent.
Conditions of high performance liquid chromatography:
a chromatographic column with 3 mu m of YMC-Triart C18(4.6 × 250mm), a flow rate of 0.5m L/min, a detection wavelength of 317nm, a column temperature of 30 ℃, a sample solution concentration of 1.0mg/ml, a sample injection amount of 10 mu L, a mobile phase A phase of acetonitrile-water with a volume ratio of 20:80, a phosphoric acid volume percentage of 0.18-0.22 percent and a mobile phase B phase of acetonitrile-water with a volume ratio of 90:10 based on 100 percent of the total volume of the acetonitrile and the water.
Elution was performed according to the following gradient program:
the calculation method comprises the following steps: calculating impurity content by reference external standard method
Calculating the formula:
Asample (A): peak area of impurities in the test solution;
Ato pair: peak area of the control solution;
Wsample (A): weighing sample amount of the sample, mg;
Wto pair: weighing the reference substance in mg;
VcontrolControl solution dilution volume, m L;
Vsample (I)The dilution volume of the test solution, m L;
p: content of control.
Example 2
And (3) verification of methodology:
2.1 System applicability
And (3) sampling blank solution and the mixed impurity reference substance solution for detection, and sampling according to the high performance liquid chromatography condition for detection, wherein the sampling amount is 10 mu L, the mixed impurity reference substance solution is continuously sampled for 6 needles, and a chromatogram is recorded, and the results are shown in tables 1-2, and the chromatogram of the mixed impurity reference substance solution is shown in figure 1.
TABLE 1 results of suitability test of the System (retention time, peak area)
TABLE 2 System suitability test results (degree of separation, symmetry factor, theoretical plate number)
The test result shows that the base line is stable and has no interference, and the blank solvent has no interference on the detection of impurities and main components; the mixed impurity reference substance solution is continuously injected into a sample of 6 needles, the retention time RSD values are all less than 2.0%, the peak area RSD values are all less than 2.0%, the resolution is all greater than 2.0, and the symmetry factors are all less than 2.0, so that the method is suitable for checking related substances, and the system applicability of the method is good.
2.2 specificity
2.2.1 interference and location test
And (3) respectively sampling and detecting the blank solution, the positioning solutions, the sample solution and the mixed impurity reference solution, wherein the sampling amount is 10 mu L, recording a chromatogram, the retention time results of the main components of the positioning solutions and the sample solution are shown in table 3, and the chromatograms of the positioning solutions and the sample solution are shown in fig. 2-6.
TABLE 3 results of the positioning test
Test results show that the blank solvent does not interfere with the detection of related substances of the arotinolol hydrochloride, and baseline separation can be achieved between the related substances and the arotinolol hydrochloride, which indicates that the method has good specificity.
2.2.2 forced degradation test
Weighing 10mg of arotinolol hydrochloride test sample, placing the test sample in a volumetric flask with the volume of 10m L, treating according to the conditions in the table 4 (neutralization is needed after acid-base treatment), dissolving the test sample by using a blank solvent, diluting the test sample to a scale, and shaking up the test sample to obtain the test sample solution for the degradation test.
Detecting the normally treated and degraded test solution with a DAD detector, detecting the detection wavelength of 200-400nm, respectively testing the purity angle and the purity threshold of the main peak of arotinolol hydrochloride in the test solution, and calculating the material recovery rate of the solution under each damage condition according to the ratio of the sum of the peak areas of the test solution under each forced degradation condition to the concentration of the test solution. The peak area response values of the unit concentrations were used for comparison, and whether the materials before and after the destruction were in equilibrium was examined with respect to 100% of the undisrupted solution, and the results are shown in Table 5.
10. mu. L of each of the 5 solutions was measured, injected into an HP L C chromatograph, detected under the conditions of the high performance liquid chromatography described above, and a chromatogram was recorded, and the results are shown in FIG. 7-graph, and the content of each impurity was calculated, and the results are shown in Table 6.
TABLE 4 preparation of degradation test samples
| Destructive condition | Preparation of degraded samples | Time of day | Temperature of |
| Acid(s) | 1ml of 1M HCl + about 10mg of starting material | 12h | 60℃ |
| Alkali | 1ml of 1M NaOH + about 10mg of starting Material | 4h | 60℃ |
| Oxidation by oxygen | 1ml of 3% hydrogen peroxide solution + about 10mg of starting material | 5h | At normal temperature |
| Illumination of light | About 30mg of starting material, |
2 days | At normal temperature |
| High temperature of solid | About 10mg of starting material | 12h | 60℃ |
Injection, about 10mg of the sample after the light treatment is weighed to prepare a test solution containing arotinolol hydrochloride with the final concentration of 1.0mg/m L.
TABLE 5 Destruction test chromatographic Peak purities and Material balance test results
Table 6 results of impurity test under various degradation conditions of destructive test
And (4) conclusion: from the above results, it can be seen that the product is unstable under both oxidative and alkaline conditions, resulting in significant degradation products. The retention time of the main degradation product under oxidative degradation conditions was 9.641 min. The retention time of the main degradation product generated under the strong alkaline condition is 23.101 min. Under the chromatographic condition for measuring related substances, after the product is destroyed by conditions of high temperature, oxidation, acid, alkali, light and the like, impurity peaks appearing in a chromatogram can be well separated from main component peaks, the separation degree of the main peak and the impurity peaks adjacent to the main peak is greater than 1.5, the purity of the main peak is good, and the material balance before and after destruction also meets the specification, which indicates that the method has strong specificity and meets the detection requirement.
2.3 precision
2.3.1 repeatability
Taking arotinolol hydrochloride test samples, preparing 6 test sample solutions with the concentration of 1.0mg/m L in parallel, carrying out sample injection detection according to the high performance liquid chromatography, recording a chromatogram, and respectively calculating the content of impurities and RSD value in 6 samples, wherein the results are shown in Table 7.
TABLE 7 results of repeated experiments
| Sample numbering | SM2 | ALLE-1 | RS-E | Maximum single impurity (%) | Total impurities (%) |
| 1 | N/A | N/A | N/A | 0.06 | 0.11 |
| 2 | N/A | N/A | N/A | 0.06 | 0.11 |
| 3 | N/A | N/A | N/A | 0.06 | 0.11 |
| 4 | N/A | N/A | N/A | 0.06 | 0.11 |
| 5 | N/A | N/A | N/A | 0.06 | 0.12 |
| 6 | N/A | N/A | N/A | 0.06 | 0.11 |
| Average (%) | N/A | N/A | N/A | 0.06 | 0.11 |
| RSD(％) | N/A | N/A | N/A | 1.55 | 3.74 |
The conclusion is that SM2, A LL E-1 and RS-E are not detected in 6 samples, the RSD values of the maximum unknown single impurity and total impurity content are all less than 10%, and the repeatability experiment is proved to meet the sample detection requirements.
2.3.2 intermediate precision
Two analysts, on different dates, use different liquid chromatographs and different lot numbers of YMC-TriartC18 chromatographic columns to detect the same batch of arotinolol hydrochloride test samples, each person prepares 6 test sample solutions with the concentration of 1.0mg/m L in parallel, examines the impurity content of the test sample solutions, calculates the average value and RSD value of the maximum unknown single impurity and total impurity in 12 test samples, and the results are shown in tables 8-9.
TABLE 8 intermediate precision experimental data
The conclusion is that 12 samples are not detected SM2, A LL E-1 and RS-E, the RSD values of the maximum unknown single impurity and total impurity content are all less than 10 percent, and the intermediate precision experiment is proved to meet the sample detection requirement.
2.4 detection and quantitation limits
Taking arotinolol hydrochloride reference substance solution and each impurity positioning solution prepared under the specificity item, respectively measuring the quantitative limit (S/N is more than or equal to 10) and the detection limit (S/N is more than or equal to 3) by using a dilution method, wherein the results are shown in tables 9-10, continuously feeding the quantitative limit solution for 6 times, and calculating the RSD value of the peak area, wherein the results are shown in table 11.
TABLE 9 detection Limit test results
TABLE 10 quantitative Limit test results
TABLE 11 results of 6 successive injections of quantitative limiting solutions for each impurity
| Peak area | A LL E control | SM2 | ALLE-1 | RS-E |
| 1 | 1174 | 1536 | 1667 | 1243 |
| 2 | 1131 | 1633 | 1537 | 1213 |
| 3 | 1200 | 1538 | 1594 | 1198 |
| 4 | 1141 | 1537 | 1486 | 1101 |
| 5 | 1137 | 1525 | 1525 | 1182 |
| 6 | 1074 | 1494 | 1454 | 1295 |
| Mean value of | 1143 | 1544 | 1544 | 1205 |
| RSD(％) | 3.74 | 3.03 | 4.98 | 5.38 |
Test results show that the detection limit concentrations of arotinolol hydrochloride, SM2, A LL E-1 and RS-E are 0.0110 mu g/m L, 0.0102 mu g/m L, 0.0100 mu g/m L and 0.0097 mu g/m L respectively, the quantitative limit concentrations are 0.0219 mu g/m L, 0.0204 mu g/m L, 0.0249 mu g/m L and 0.0193 mu g/m L respectively, the detection limit of each impurity is less than 0.002% of the sample concentration, and the quantitative limit is less than 0.005% of the sample concentration, which indicates that the method has high sensitivity.
2.5 Linear Range
Preparing a series of solutions with different levels of concentration according to the limit of quantification of the concentration of the reference solution to 200% of the specified limit, precisely measuring each 10 μ L of the reference solution, injecting into a high performance liquid chromatograph, recording chromatogram, measuring peak area, and performing linear regression with peak area Y as ordinate and concentration X (unit: μ g/ml) as abscissa, the results are shown in tables 12-15.
TABLE 12 Linear Experimental data for arotinolol hydrochloride
TABLE 13 SM2 Linear Experimental data
TABLE 14A LL E-1 Linear Experimental data
TABLE 15 RS-E Linear Experimental data
And (4) conclusion: the known impurities and the alorol hydrochloride have good linear relation when being measured at low concentration, and meet the detection requirement.
2.6 accuracy
The recovery rate is measured by adding three different concentrations of impurities of 80%, 100% and 120% of the index into the test solution, so as to verify that the method has good accuracy; 3 samples were prepared in parallel for each concentration, and the results are shown in tables 16 to 18, where the peak area of each impurity in the chromatogram was recorded, and the average recovery rate and RSD value of the recovery rate were calculated for each concentration of 9 samples.
TABLE 16 SM2 accuracy experimental data
Accuracy test data of Table 17A LL E-1
TABLE 18 accuracy test data of RS-E
The results show that the recovery rate of each impurity at each concentration is in the range of 80-120%, and the RSD value is in the range of 1-10%, which shows that the method has better accuracy.
2.7 solution stability
A test sample solution with the concentration of 1.0mg/m L and a mixed impurity reference solution with the concentration of A LL E-1 of 0.00005mg/m L and the concentration of arotinolol hydrochloride, SM2 and RS-E impurity references of 0.001mg/m L are respectively prepared, the content of impurities is tested at different times according to the high performance liquid chromatography conditions, and the results are shown in tables 19 to 20.
TABLE 19 test article solution stability study
Table 20 control solution stability study of impurities
The results show that: the mixed impurity solution is stable in the standing process of 32 h; the known impurities in the test solution are not detected in 24h detection, the content of the unknown impurities is not changed in the 24h placement process, and the total impurities are basically not changed in the 24h placement process, so that the test solution is stable after being placed for 24 h.
2.8 durability
And (3) investigating the influence on retention time, peak area, separation degree, theoretical plate number and the like of each peak in the solution with system applicability and the influence on the determination of each impurity in the test sample under the conditions of different detection wavelengths, different column temperatures, different flow rates and different phosphoric acid concentrations.
(1) Influence of different wavelengths on the measurement results
TABLE 21 results of investigation of retention time and peak area of each component peak at different wavelengths
TABLE 22 results of the measurement of the degree of separation of peaks of each component at different wavelengths and the theoretical plate number
And (4) conclusion: under the condition that the wavelength slightly changes, 6 needles of system applicability solution are continuously injected, the RSD value of each impurity contrast retention time is less than 2.0%, the RSD value of the peak area is less than 10.0%, the resolution is greater than 2.0, and the result meets the system applicability requirement.
(2) Influence of different column temperatures on the measurement results
TABLE 23 results of investigation of retention time and peak area of each component peak at different column temperatures
TABLE 24 results of the degree of separation of each component peak at different column temperatures and the theoretical plate number
And (4) conclusion: under the condition of slight change of the column temperature, 6 needles of solution with system applicability are continuously injected, the RSD value of each impurity contrast retention time is less than 2.0%, the RSD value of the peak area is less than 10.0%, the resolution is greater than 2.0, and the result meets the requirement of the system applicability.
(3) Effect of different flow rates on the assay results
TABLE 25 results of investigation of retention time and peak area of different flow rates for each component peak
TABLE 26 results of the degree of separation of each component peak and theoretical plate number at different flow rates
And (4) conclusion: under the condition of small change of the flow rate, the system applicability solution is continuously injected into 6 needles, the RSD value of each impurity contrast retention time is less than 2.0%, the RSD value of the peak area is less than 10.0%, the resolution is greater than 2.0, and the result meets the system applicability requirement.
(4) Effect of different phosphoric acid concentrations on the assay results
TABLE 27 results of examination of retention time and peak area of peaks of respective components at different phosphoric acid concentrations
TABLE 28 results of examining the degree of separation of peaks of respective components and the number of theoretical plates under the conditions of different phosphoric acid concentrations
And (4) conclusion: the concentration of phosphoric acid in the mobile phase is slightly changed, the applicability solution of the system is continuously injected into 6 needles, the RSD value of each impurity contrast retention time is less than 2.0%, the RSD value of the peak area is less than 10.0%, the resolution is more than 2.0, and the result meets the requirement of the applicability of the system.
TABLE 29 summary of sample testing
And (4) conclusion: the measuring conditions (wavelength, flow rate, column temperature and phosphoric acid concentration) are slightly changed, the detection results of the samples are basically consistent, and the durability of the method is good.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents or improvements made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (10)
1. A method for detecting related substances in arotinolol hydrochloride is characterized in that high performance liquid chromatography is adopted for detection, and the chromatographic conditions are as follows:
a chromatographic column: octadecylsilane chemically bonded silica gel column;
a UV detector for detecting the wavelength of 315-;
mobile phase A: acetonitrile-water-phosphoric acid, and the mobile phase B is acetonitrile-water;
the elution mode is gradient elution, and the elution procedure is as follows:
0-3min, 100% mobile phase A;
3-45min, 100% → 40% mobile phase a, 0% → 60% mobile phase B;
45-46min, 40% → 100% mobile phase a, 60% → 0% mobile phase B;
46-60min, 100% mobile phase A.
2. The method for detecting related substances in arotinolol hydrochloride according to claim 1, wherein the flow rate is 0.45-0.60m L/min, and the column temperature is 28-32 ℃.
3. The method for detecting related substances in arotinolol hydrochloride according to claim 2 wherein the detection wavelength is 317nm, the flow rate is 0.5m L/min, and the column temperature is 30 ℃.
4. The method for detecting related substances in arotinolol hydrochloride according to claim 1, wherein the injection volume is 10 μ L.
5. The method for detecting related substances in arotinolol hydrochloride according to claim 1, wherein the volume ratio of acetonitrile to water in the mobile phase A is 20:80, and the volume percentage of phosphoric acid is 0.18-0.22% based on 100% of the total volume of acetonitrile and water.
6. The method for detecting related substances in arotinolol hydrochloride according to claim 5, wherein the volume percentage of the phosphoric acid is 0.2%.
7. The method for detecting related substances in arotinolol hydrochloride according to claim 1, wherein the volume ratio of acetonitrile to water in the mobile phase B is 90: 10.
8. The method for detecting related substances in arotinolol hydrochloride according to claim 1, wherein the chromatographic column has a size of 250 x 4.6mm and a packing diameter of 3 μm.
9. The method for detecting related substances in arotinolol hydrochloride according to claim 1 or 8, wherein the chromatographic column is YMC-Triart C18, 4.6mm by 250mm, 3 μm.
10. The method of claim 1, wherein the concentration of the sample solution is 1.0mg/m L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010386440.0A CN111505159A (en) | 2020-05-09 | 2020-05-09 | Detection method of related substances in arotinolol hydrochloride |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010386440.0A CN111505159A (en) | 2020-05-09 | 2020-05-09 | Detection method of related substances in arotinolol hydrochloride |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111505159A true CN111505159A (en) | 2020-08-07 |
Family
ID=71878415
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010386440.0A Pending CN111505159A (en) | 2020-05-09 | 2020-05-09 | Detection method of related substances in arotinolol hydrochloride |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111505159A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112213418A (en) * | 2020-09-17 | 2021-01-12 | 石家庄凯瑞德医药科技发展有限公司 | Method for detecting related substances in sofosbuvir |
| CN113582984A (en) * | 2021-07-08 | 2021-11-02 | 北京百美特生物制药有限公司 | Arotinolol hydrochloride impurity and preparation method and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103626750A (en) * | 2013-11-15 | 2014-03-12 | 北京博爱旺康医药科技有限公司 | High-purity arotinolol hydrochloride and preparation method thereof |
| CN105030718A (en) * | 2015-08-18 | 2015-11-11 | 石家庄格瑞药业有限公司 | Arotinolol hydrochloride preparation and preparation method thereof |
| CN105949184A (en) * | 2016-05-18 | 2016-09-21 | 山东齐都药业有限公司 | Refinement method of arotinolol hydrochloride |
| CN107698576A (en) * | 2016-08-08 | 2018-02-16 | 南京逐陆医药科技有限公司 | A kind of preparation technology of High-purity arotinolol hydrochloride |
| US20190212309A1 (en) * | 2018-01-05 | 2019-07-11 | Taiho Pharmaceutical Co., Ltd. | Method for detecting trifluridine-related substance by high-performance liquid chromatography |
-
2020
- 2020-05-09 CN CN202010386440.0A patent/CN111505159A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103626750A (en) * | 2013-11-15 | 2014-03-12 | 北京博爱旺康医药科技有限公司 | High-purity arotinolol hydrochloride and preparation method thereof |
| CN105030718A (en) * | 2015-08-18 | 2015-11-11 | 石家庄格瑞药业有限公司 | Arotinolol hydrochloride preparation and preparation method thereof |
| CN105949184A (en) * | 2016-05-18 | 2016-09-21 | 山东齐都药业有限公司 | Refinement method of arotinolol hydrochloride |
| CN107698576A (en) * | 2016-08-08 | 2018-02-16 | 南京逐陆医药科技有限公司 | A kind of preparation technology of High-purity arotinolol hydrochloride |
| US20190212309A1 (en) * | 2018-01-05 | 2019-07-11 | Taiho Pharmaceutical Co., Ltd. | Method for detecting trifluridine-related substance by high-performance liquid chromatography |
Non-Patent Citations (4)
| Title |
|---|
| 廖洪娟: "HPLC法测定盐酸阿罗洛尔片的含量", 《海峡药学》 * |
| 张晓晓等: "高效液相色谱法测定盐酸阿罗洛尔有关物质", 《河北师范大学学报(自然科学版)》 * |
| 胡琴等: "盐酸阿罗洛尔含量测定方法研究", 《中国药品标准》 * |
| 苏玉怀等: "用HPLC法测定盐酸阿罗洛尔片的含量", 《药学服务与研究》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112213418A (en) * | 2020-09-17 | 2021-01-12 | 石家庄凯瑞德医药科技发展有限公司 | Method for detecting related substances in sofosbuvir |
| CN112213418B (en) * | 2020-09-17 | 2022-08-23 | 石家庄凯瑞德医药科技发展有限公司 | Method for detecting related substances in sofosbuvir |
| CN113582984A (en) * | 2021-07-08 | 2021-11-02 | 北京百美特生物制药有限公司 | Arotinolol hydrochloride impurity and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111650288A (en) | Method for detecting (R) -3-tert-butyloxycarbonylaminopiperidine enantiomer | |
| CN111505159A (en) | Detection method of related substances in arotinolol hydrochloride | |
| CN118010878A (en) | Determination method and application of impurities in sodium rabeprazole enteric-coated preparation | |
| CN111929372B (en) | HPLC (high performance liquid chromatography) detection method for propranolol hydrochloride genotoxic impurities | |
| CN109324135B (en) | Method for simultaneously detecting 3 emulsifiers in composite emulsifier | |
| CN115480020A (en) | Method for measuring substances related to propodoiodoammonium | |
| CN112213418B (en) | Method for detecting related substances in sofosbuvir | |
| CN113030328B (en) | Method for detecting genotoxic impurities in ivabradine hydrochloride | |
| CN114839287A (en) | Method for detecting sodium tetradecanoate in miboplatin | |
| CN111380978B (en) | Method for simultaneously determining contents of coenzymes NADP and FAD in medicine | |
| CN114689737A (en) | Analysis method of S-o-chlorophenyl glycine methyl ester tartrate related substances | |
| CN110501436B (en) | Detection method of related substances in tinidazole pharmaceutical composition | |
| CN108535400B (en) | Thin-layer chromatography analysis and detection method for impurity 5-methoxytryptamine in melatonin bulk drug | |
| CN108037221B (en) | Method for simultaneously separating and determining methionine sulfoxide and methionine sulfone impurities in compound amino acid injection 18AA by liquid chromatography | |
| CN112595793B (en) | Earthworm injection detection method based on phenol determination | |
| CN111638298A (en) | High performance liquid chromatography determination method for related substances in vitamin C ethyl ether | |
| CN113552238A (en) | Method for measuring methylamine content in temozolomide bulk drug | |
| CN114609268B (en) | Detection method for related substances in sodium rabeprazole bulk drug | |
| CN112305100B (en) | Method for detecting content of genotoxic impurity benzyl bromide in medicine | |
| CN114200050B (en) | HPLC detection method for content of related substances in p-bromoanisole | |
| CN108572223A (en) | A kind of method of activity inducement substance in measurement polypeptide | |
| CN115389681B (en) | Detection method for dithiothreitol residue in thiolated hyaluronic acid derivative | |
| CN116046950A (en) | Method for detecting 5 impurities in abiraterone acetate | |
| CN113109462A (en) | Method for detecting chloroacetic acid in lidocaine | |
| CN112903833A (en) | High performance liquid chromatography analysis method for detecting organic impurities in dibazol |
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 |