CN115452968A - Method for detecting impurity of caraprolol hydrochloride by using HPLC - Google Patents
Method for detecting impurity of caraprolol hydrochloride by using HPLC Download PDFInfo
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- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 title claims abstract description 96
- 239000012535 impurity Substances 0.000 title claims abstract description 66
- 238000004128 high performance liquid chromatography Methods 0.000 title claims abstract description 44
- 238000000034 method Methods 0.000 title claims abstract description 43
- 238000001514 detection method Methods 0.000 claims abstract description 31
- 238000002347 injection Methods 0.000 claims abstract description 24
- 239000007924 injection Substances 0.000 claims abstract description 24
- HUMNYLRZRPPJDN-UHFFFAOYSA-N benzaldehyde Chemical compound O=CC1=CC=CC=C1 HUMNYLRZRPPJDN-UHFFFAOYSA-N 0.000 claims description 102
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- 239000005695 Ammonium acetate Substances 0.000 claims description 10
- 229940043376 ammonium acetate Drugs 0.000 claims description 10
- 235000019257 ammonium acetate Nutrition 0.000 claims description 10
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- 239000012488 sample solution Substances 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 5
- XORXDJBDZJBCOC-UHFFFAOYSA-N azanium;acetonitrile;acetate Chemical group [NH4+].CC#N.CC([O-])=O XORXDJBDZJBCOC-UHFFFAOYSA-N 0.000 claims description 3
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical group CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 3
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- 238000012360 testing method Methods 0.000 description 16
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- 238000002360 preparation method Methods 0.000 description 14
- 239000013558 reference substance Substances 0.000 description 13
- 239000002253 acid Substances 0.000 description 10
- 239000008186 active pharmaceutical agent Substances 0.000 description 10
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- 230000015556 catabolic process Effects 0.000 description 10
- 238000007865 diluting Methods 0.000 description 9
- 239000002994 raw material Substances 0.000 description 9
- 239000012085 test solution Substances 0.000 description 9
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 239000003513 alkali Substances 0.000 description 6
- 238000006731 degradation reaction Methods 0.000 description 6
- 238000005259 measurement Methods 0.000 description 6
- 230000003647 oxidation Effects 0.000 description 6
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- 229940069078 citric acid / sodium citrate Drugs 0.000 description 3
- 239000012362 glacial acetic acid Substances 0.000 description 3
- 238000012417 linear regression Methods 0.000 description 3
- 230000014759 maintenance of location Effects 0.000 description 3
- 230000001590 oxidative effect Effects 0.000 description 3
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
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- 238000001782 photodegradation Methods 0.000 description 2
- 238000003908 quality control method Methods 0.000 description 2
- 239000001509 sodium citrate Substances 0.000 description 2
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 2
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- UCTWMZQNUQWSLP-VIFPVBQESA-N (R)-adrenaline Chemical compound CNC[C@H](O)C1=CC=C(O)C(O)=C1 UCTWMZQNUQWSLP-VIFPVBQESA-N 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- 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
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/062—Preparation extracting sample from raw material
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Abstract
The invention belongs to the technical field of chemical detection, and relates to a method for detecting a caramolol hydrochloride impurity by using HPLC. The HPLC detection method for the related substance of the carralol hydrochloride in the carralol injection provided by the invention is simple to operate, high in accuracy, high in detection efficiency, good in specificity and good in linearity, and the quality of the carralol injection can be further effectively improved by adopting the detection method provided by the invention.
Description
Technical Field
The invention belongs to the technical field of chemical detection, and relates to a method for detecting a caramolol hydrochloride impurity by using HPLC.
Background
The carraolol injection is intramuscular injection comprising carraolol hydrochloride and proper amount of supplementary material. Carragolol is a non-specific beta receptor blocker (beta sympathetic or beta adrenergic receptor antagonist) and is a structural analog of catecholamines (epinephrine and norepinephrine). Thus, it is immobilized by reversible binding to the β receptor and does not cause adrenergic activation. This prevents the catecholamines from being released and acting under pressure. In veterinary medicine, carranolol is injected intramuscularly for the relief of stress conditions in pigs. The preparation is applied to the breeding regulation of pigs and cattle, and can promote parturition, promote placenta discharge and improve fertility level.
The Carranolol hydrochloride is the main component of the Carranolol injection, and the preparation and the raw material medicaments are not included in the current pharmacopoeias of various countries. Quality control of drugs is usually required to ensure their safe use, and related substances are the items that most drugs must be tested for. Because the structure of the Carranolol hydrochloride contains ether bonds and secondary amine groups, the stability of the Carranolol hydrochloride can be influenced by conditions such as temperature, pH, oxidation and the like, a method for effectively detecting related substances of the Carranolol hydrochloride in the Carranolol injection is sought, and the method has important significance for further quality control of the Carranolol injection.
Disclosure of Invention
The invention aims to solve the problems in the prior art and provides a method for detecting the impurities of the caramolol hydrochloride by using HPLC, which is simple to operate, high in accuracy, high in detection efficiency and good in specificity.
The purpose of the invention can be realized by the following technical scheme:
a method for detecting a caramolol hydrochloride impurity by HPLC, the method comprising the steps of:
s1, dissolving benzaldehyde in a mobile phase to obtain a benzaldehyde-containing contrast solution;
s2, dissolving a to-be-detected product containing the caramolol into a mobile phase to obtain a control solution;
and S3, injecting the benzaldehyde-containing control solution, the control solution and the sample containing the caraprolol into a liquid chromatograph respectively, and detecting the content of the caraprolol impurity in the sample solution by using HPLC.
In the method for detecting the impurity of the caramolol hydrochloride by using HPLC, the mobile phase is acetonitrile which is an ammonium acetate solution with pH of 5.5-6.5 and concentration of 1-3 g/L. The ammonium acetate solution has stronger buffer capacity, is beneficial to analyzing compounds with different pH values, and takes the ammonium acetate solution-acetonitrile as a mobile phase, so that the peak shapes of all substances are better.
In the method for detecting the impurity of the caraprolol hydrochloride by using HPLC, the volume ratio of the ammonium acetate solution to the acetonitrile in the ammonium acetate-acetonitrile solution is (9-14): 6.
in the method for detecting the impurity of the caramolol hydrochloride by using HPLC, the flow rate of the flowing phase in the HPLC detection process is 0.9-1.1ml/min. According to the method, impurities of the carraolol can be effectively separated by controlling the flow rate of the mobile phase, so that the impurity content of the carraolol in the test solution can be more accurately calculated.
In the method for detecting the impurity of the caraprolol hydrochloride by using HPLC, the concentration of the benzaldehyde in the benzaldehyde-containing control solution is 15-25 mu g/ml. The Caralol injection contains benzyl alcohol as an auxiliary material, the benzyl alcohol is very easy to oxidize to generate benzaldehyde, and the benzaldehyde is added into a control solution in order to accurately calculate the content of benzaldehyde impurities in a test sample.
In the method for detecting the impurity of the carralol hydrochloride by using HPLC, the volume ratio of the carralol injection to the mobile phase in the control solution is 1: (95-105).
In the method for detecting the impurity of the caramolol hydrochloride by using HPLC, the chromatographic column in the HPLC detection process is a C18 chromatographic column with the particle size of 3-8 mu m, wherein the filler is octadecylsilane chemically bonded silica.
In the method for detecting the impurity of the caramolol hydrochloride by using HPLC, the temperature of a chromatographic column is 25-35 ℃ in the HPLC detection process. The invention can ensure the reproducibility of the retention time of the substance by controlling the column temperature, and simultaneously, the impurities of the caraprolol injection can be effectively separated in the column temperature range, and the chromatographic column can be protected.
In the method for detecting the impurity of the caraprolol hydrochloride by using the HPLC, the detector used in the HPLC detection method is an ultraviolet-visible absorption detector, wherein the detection wavelength is 234-238nm.
Compared with the prior art, the invention has the following beneficial effects: the HPLC detection method for the related substance of the carralol hydrochloride in the carralol injection provided by the invention is simple to operate, high in accuracy, high in detection efficiency, good in specificity and good in linearity, and the quality of the carralol injection can be further effectively improved by adopting the detection method provided by the invention.
Drawings
FIG. 1 is an HPLC chromatogram of a carralol hydrochloride drug without destruction, after acid, base, oxidation, heating and light destruction.
FIG. 2 is a graph of peak purity for undamaged bulk drug of caramolol hydrochloride.
FIG. 3 is a graph of peak purity of acid breakdown of a bulk drug of caramolol hydrochloride.
FIG. 4 is a graph of peak purity of the base breakdown of a bulk drug of caraprolol hydrochloride.
FIG. 5 is a peak purity profile of oxidative destruction of a bulk drug of caraprolol hydrochloride.
FIG. 6 is a peak purity profile of thermal destruction of a bulk drug of caraprolol hydrochloride.
FIG. 7 is a peak purity profile of photodisruption of a bulk drug of caramolol hydrochloride.
FIG. 8 is a graph of the linear relationship of caramolol hydrochloride.
FIG. 9 is a linear relationship of benzaldehyde.
FIG. 10 is a typical map of related substances of Carranolol hydrochloride in the Carranolol injection.
FIG. 11 is a graph comparing results of caraprolol hydrochloride + citric acid at 60 ℃/(4500 Lx. + -. 500 Lx);
FIG. 12 is a graph comparing results of carralol hydrochloride and citric acid/sodium citrate at 60 ℃/(4500 Lx. + -. 500 Lx).
FIG. 13 is a graph comparing results of carralol hydrochloride + benzyl alcohol at 60 ℃/(4500 Lx. + -. 500 Lx).
Detailed Description
The following are specific examples of the present invention and further describe the technical solutions of the present invention, but the present invention is not limited to these examples.
Example 1:
s1, chromatographic condition and system applicability test: welch Ultimate LP-CN 250X 4.6mm,5 μm C18 column, octadecylsilane chemically bonded silica as a packing, 2.0g/L ammonium acetate solution-acetonitrile as a mobile phase, at flow rates: 1ml/min, column temperature 30 ℃;
s2, preparing a benzaldehyde reference substance solution: taking 20mg of benzaldehyde reference substance, accurately weighing, placing in a 50mL measuring flask, adding a mobile phase for dissolving and diluting to a scale, accurately weighing 1mL, diluting to 20mL with the mobile phase, and shaking uniformly to obtain the benzaldehyde reference substance;
preparation of test solution and control solution: taking a sample containing the caramolol to be detected as a sample solution, precisely measuring 1mL of the sample solution, diluting the sample solution to 100mL by using a mobile phase, and shaking up to be used as a reference solution;
s3, measuring: precisely measuring 10 mu L of each of a benzaldehyde reference solution, a reference solution and a test solution, respectively injecting into a liquid chromatograph, measuring according to the chromatographic conditions, recording a chromatogram, wherein if a chromatographic peak with the same retention time as benzaldehyde exists in the chromatogram, the peak area is calculated according to an external standard method; if other impurities exist, comparing with a reference solution according to a proportion;
the ammonium acetate solution: the volume ratio of acetonitrile is 65:35, the 2.0g/L ammonium acetate solution is adjusted to pH 6.0 with glacial acetic acid.
In the chromatographic condition and system applicability test of the step S1, the detection wavelength is 236nm.
Example 2:
at present, there is no similar product in domestic market, and more than thirty years of history animal original research products are marketed in several countries such as the United states and Europe: fatro SPA pharmaceuticals Inc
0.5mg/ml is a hydro-acupuncture preparation, and the preparation has good clinical effect in application. Therefore, n.915326 batch of carraolol injection produced by the company is taken and tested by the HPLC test method of related substances of the carraolol hydrochloride, and the test method comprises the following steps:
the instrument comprises the following steps: waters 2695-VWD 2489 high performance liquid chromatograph; electronic balance METTLER XS205DU; PH meter METTLER PE25S
Comparison products: benzaldehyde reference substance (batch No. 111650-201905, content 99.9%, provided by China institute for food and drug testing);
reagent preparation: acetonitrile (chromatographically pure, batch No. 20210519, supplied by national chemical group, inc.), ammonium acetate (chromatographically pure, batch No. 187057, supplied by Saimer Feishale science, inc.), glacial acetic acid (analytically pure, batch No. C12069452, supplied by Shanghai Michelin Biochemical science, inc.)
Sample preparation: caralol injection (batch No. n.915326, manufactured by Fatro SPA pharmaceutical Co., ltd.).
Chromatographic conditions are as follows: a Welch Ultimate LP-CN 250 × 4.6mm,5 μm C18 chromatography column, packed with octadecylsilane-bonded silica gel, mobile phase with 2.0g/L ammonium acetate solution (pH adjusted to 6.0 with glacial acetic acid) -acetonitrile (65) at flow rates: 1ml/min, column temperature 30 deg.C, detection wavelength 236nm.
Solution preparation: preparation of benzaldehyde control solution: taking 20mg of benzaldehyde as a reference substance, precisely weighing, placing in a 50mL measuring flask, adding a mobile phase for dissolving and diluting to a scale, precisely weighing 1mL, diluting to 20mL with the mobile phase, and shaking uniformly to obtain the benzaldehyde.
Preparation of test solution and control solution:
taking a proper amount of the product as a test solution. 1mL of the test solution was measured precisely, diluted to 100mL with the mobile phase, and shaken up to give a control solution.
The determination method comprises the following steps: precisely measuring 10 μ L of benzaldehyde reference solution, reference solution and sample solution, respectively injecting into liquid chromatograph, measuring according to chromatographic conditions, and recording chromatogram.
The measurement results of the relevant substances are as follows: for batch No. n.915326, fatro SPA pharmaceuticals Inc
Measuring 0.5mg/ml for water injection preparation, calculating benzaldehyde content by external standard method, and calculating maximum single impurity and total impurity content except benzaldehyde by self-control method.
In the formula: a. The For supplying to Is the peak area of benzaldehyde in the test solution;
A to pair The peak area of the main peak of the benzaldehyde reference substance solution;
C for supplying to The concentration of the test solution is the Carranolol concentration, mu g/ml;
C to pair The concentration of the benzaldehyde reference substance solution is shown in mu g/ml;
p is the purity of benzaldehyde reference substance.
In the formula: a. The For supplying to Is the peak area of unknown impurities in the test solution;
A to pair The main peak area of the control solution.
The sample contained 0.42% benzaldehyde, 0.01% maximum single impurity content and 0.44% total impurity content, and the results are shown in Table 1 and the HPLC chromatogram is shown in FIG. 10.
Table 1: summary table of related substance measurement results of Carranolol injection
| Retention time | Content (wt.) |
| 6.09 | 0.01％ |
| 6.99 (benzaldehyde) | 0.42％ |
| 7.96 | 0.01％ |
| General miscellaneous | 0.44％ |
Analysis of related substance measurement results: from the condition of the raw material of the carralol hydrochloride, the carralol hydrochloride is stable, the quantity of degradation products influenced by external conditions is less, and the impurity content is lower. The carralol injection contains a large amount of benzaldehyde impurities, and analysis of citric acid, sodium citrate and benzyl alcohol which are auxiliary materials of the carralol injection infers that the benzyl alcohol can generate benzaldehyde after being oxidized. In order to verify and conclude, the carralol hydrochloride API and the citric acid are mixed according to the pharmaceutical prescription proportion (sample 1), the carralol hydrochloride API and the sodium citrate are mixed according to the pharmaceutical prescription proportion (sample 2), the carralol hydrochloride API and the benzyl alcohol are mixed according to the pharmaceutical prescription proportion (sample 3) to respectively prepare samples, the samples are respectively placed under fluorescent lamps at room temperature and 60 ℃ and with the illumination of 4500Lx +/-500 Lx for 6 days, the HPLC detection method of the carralol hydrochloride related substances is adopted for measurement, the results of the related substances are shown in a table 2, the HPLC map of the sample 1 is shown in a table 11, the HPLC map of the sample 2 is shown in a table 12, the HPLC map of the sample 3 is shown in a table 13, and the benzaldehyde impurities appear in the sample containing the benzyl alcohol and slightly increase, so that the benzaldehyde impurities in the preparation can be brought in by the benzyl alcohol as an auxiliary material. Therefore, a certain amount of benzaldehyde impurities are detected in the preparation. The result also indicates that the production enterprises should pay attention to the quality of the benzyl alcohol, meanwhile, the preparation is prevented from being placed under the condition of high-temperature illumination, and the circulation and use fields should strictly adhere to the storage conditions of the specification.
Table 2: summary table of the measurement results of the related substances of each sample
Specificity test
Acid destruction test: adding 1mol/L hydrochloric acid solution 1.0mL into 11mg of Caralol hydrochloride API, shaking to dissolve, placing in water bath at 60 deg.C, standing for 2 hr, adding 1mol/L sodium hydroxide solution 1.0mL for neutralization, diluting to 20mL with mobile phase, shaking, and filtering. No new impurities are generated, and the content ratio of the impurities brought by the raw materials in 8.3min is slightly increased.
Alkali destruction test: adding 1mol/L sodium hydroxide solution 1.0mL into 11mg of Caralol hydrochloride API, shaking for dissolving, placing in water bath at 60 deg.C for 2 hr, adding 1mol/L hydrochloric acid solution 1.0mL for neutralizing, diluting to 20mL with mobile phase, shaking, and filtering. No new impurity is generated, and the content ratio of the impurity brought by the raw material in 8.3min is not obviously changed.
Oxidative destruction test: adding 1.0mL of 3% hydrogen peroxide into 11mg of Carranolol hydrochloride API, placing in a water bath at 60 ℃ for 2 hours, diluting to 20mL by using a mobile phase, shaking up, and filtering. No new impurities are generated, and the content ratio of the impurities brought by the raw materials in 8.3min is slightly increased.
Heating failure test: taking a proper amount of 11mg of catalol hydrochloride API, placing the catalol hydrochloride API in an oven at 150 ℃ for 1 hour, cooling to room temperature, taking 11mg of the catalol hydrochloride API to dissolve in mobile phase and dilute to 20mL, shaking up, and filtering. New impurities are generated for 6.8min, 7.2min, 7.8min, 8.3min, 9.4min and 10.9min, and the content proportion of the impurities brought in by the raw materials for 8.3min is obviously increased.
Light damage test: proper amount of Caralol hydrochloride API is taken and placed under a fluorescent lamp with the illumination of 4500Lx +/-500 Lx for 24 hours, 11mg of Caralol hydrochloride is taken to be dissolved by mobile phase and diluted to 20mL, shaken up and filtered. New impurities are generated within 6.0min, and the content ratio of the impurities brought by the raw materials within 8.3min is not obviously changed.
The normalized content comparison table of the impurities and the main components generated by the various destruction modes is shown in table 3, wherein a chromatogram of non-destruction, acid, alkali, oxidation, heating and light destruction is shown in fig. 1, a peak purity map of a main peak in a test sample of non-destruction is shown in fig. 2, a peak purity map of a main peak in a test sample of acid destruction is shown in fig. 3, a peak purity map of a main peak in a test sample of alkali destruction is shown in fig. 4, a peak purity map of a main peak in a test sample of oxidation destruction is shown in fig. 5, a peak purity map of a main peak in a test sample of heat destruction is shown in fig. 6, a peak purity map of a main peak in a test sample of light destruction is shown in fig. 7, and the destruction tests of acid, alkali, oxidation, heating, light irradiation and the like are respectively carried out on the caramolol hydrochloride raw material, and the experimental results show that: the content of impurities in the raw material of the caramolol hydrochloride is low, and the quantity of the impurities is small; destructive testing produces a small amount of impurities.
Table 3: table for comparing content of impurities generated by various destruction modes with normalized content of main components
Detection limit and quantification limit of the method:
taking a proper amount of a Carranolol hydrochloride reference substance and a proper amount of a benzaldehyde reference substance, adding a mobile phase for dissolving and diluting into a solution containing 0.01mg of Carranolol hydrochloride and 0.07 mu g of benzaldehyde per 1mL, taking the solution as a benzaldehyde detection limit measuring solution, precisely measuring a proper amount of a solution containing 0.08 mu g of Carranolol hydrochloride diluted by the mobile phase per 1mL, taking the solution as a Carranolol hydrochloride detection limit measuring solution, measuring according to the chromatographic conditions defined in the method, and respectively calculating the detection limit and the quantitative limit according to the S/N of 3 and 10. As a result, the detection limit and the quantitative limit of benzaldehyde were 0.01% and 0.04%, respectively, and the detection limit and the quantitative limit of Carranolol hydrochloride were 0.01% and 0.05%, respectively.
The method is linear:
proper amounts of the Caralol hydrochloride reference substance and the benzaldehyde reference substance were taken, and a mobile phase was added to dissolve and dilute the Caralol hydrochloride reference substance into solutions of various concentrations, as shown in Table 4. The measurements were performed under the chromatographic conditions defined in the method of the invention, the linear relationship was plotted as a function of measured response signal (peak area) versus analyte concentration, and the linear relationship was linear regression using the least squares method, with the linear relationship for caraprolol hydrochloride shown in FIG. 8 and the linear relationship for benzaldehyde shown in FIG. 9. The linear range of the caramolol hydrochloride is between 0.27 mu g/mL and 6.43 mu g/mL, and the linear regression coefficient R in the range 2 Is 1.0000; the linear range of the benzaldehyde is between 0.22 and 52.36 mu g/mL, and the linear regression coefficient R in the range 2 Is 0.9998; the linearity proved to be good.
Table 4: corresponding table of solution concentration and peak area
FIG. 1 is an HPLC chromatogram of a carralol hydrochloride drug without destruction, after acid, base, oxidation, heating and light destruction. As can be seen from the figure, under each destruction condition, the bulk drug of the caraprolol hydrochloride is relatively stable and produces a small amount of impurities.
FIG. 2 is a peak purity profile of a bulk drug of caraprolol hydrochloride without disruption. As can be seen from the figure, the content of impurities in the bulk drug of the caraprolol hydrochloride is low, and the quantity is small.
FIG. 3 is a graph of peak purity of acid breakdown of a bulk drug of caramolol hydrochloride. As can be seen from the figure, under the acid degradation condition, no other peak is contained in the main peak, which indicates that the method is suitable for the determination of the acid degradation impurity of the caraprolol hydrochloride.
FIG. 4 is a graph of peak purity of the base breakdown of a bulk drug of caraprolol hydrochloride. As can be seen from the figure, under the alkali degradation condition, no other peak is contained in the main peak, which indicates that the method is suitable for the determination of the alkali degradation impurity of the caraprolol hydrochloride.
FIG. 5 is a peak purity profile of oxidative destruction of a bulk drug of caraprolol hydrochloride. As can be seen from the figure, under the oxidative degradation conditions, no other peak is contained in the main peak, indicating that the method is suitable for the determination of the oxidative degradation impurity of the caramolol hydrochloride.
FIG. 6 is a peak purity profile of thermal destruction of a bulk drug of caramolol hydrochloride. As can be seen from the figure, under the high temperature degradation condition, no other peak is contained in the main peak, which indicates that the method is applicable to the determination of the high temperature degradation impurity of the caraprolol hydrochloride.
FIG. 7 is a peak purity profile of photodisruption of a bulk drug of caramolol hydrochloride. As can be seen from the figure, under the photodegradation condition, no other peak is contained in the main peak, which indicates that the method is suitable for the determination of the photodegradation impurities of the caraprolol hydrochloride.
FIG. 8 is a graph of the linear relationship of caraprolol hydrochloride. As can be seen from the figure, the linear relationship of the Carragolol hydrochloride is good in the range of 0.27. Mu.g/mL to 6.43. Mu.g/mL.
FIG. 9 is a linear relationship of benzaldehyde. As can be seen from the graph, benzaldehyde has a good linear relationship in the range of 0.22. Mu.g/mL to 52.36. Mu.g/mL.
FIG. 10 is a typical map of related substances of Carranolol hydrochloride in the Carranolol injection. As can be seen from the figure, the injection of the carralol contains a large amount of benzaldehyde impurities compared with the carralol hydrochloride bulk drug.
FIG. 11 is a graph comparing results of carralol hydrochloride + citric acid at 60 ℃/(4500 Lx. + -. 500 Lx). As can be seen from the figure, under the conditions of high temperature and illumination, the caramolol hydrochloride and the citric acid have no benzaldehyde impurity.
FIG. 12 is a graph comparing results of carralol hydrochloride and citric acid/sodium citrate at 60 ℃/(4500 Lx. + -. 500 Lx). As can be seen from the figure, the production of benzaldehyde impurity is avoided when the carralol hydrochloride and the citric acid/sodium citrate are under the conditions of high temperature and illumination.
FIG. 13 is a graph comparing results of carralol hydrochloride + benzyl alcohol at 60 ℃/(4500 Lx. + -. 500 Lx). As can be seen from the figure, in the sample containing benzyl alcohol, benzaldehyde impurity appears, which indicates that the benzaldehyde impurity in the Carranolol injection is brought in by the auxiliary material of benzyl alcohol.
In conclusion, the HPLC detection method for the related substance of the carralol hydrochloride in the carralol injection provided by the invention is simple to operate, high in accuracy, high in detection efficiency, good in specificity and good in linearity, and the quality of the carralol injection can be further effectively improved by adopting the detection method provided by the invention.
The technical scope of the invention claimed by the embodiments of the present application is not exhaustive, and new technical solutions formed by equivalent replacement of single or multiple technical features in the technical solutions of the embodiments are also within the scope of the invention claimed by the present application; in all the embodiments of the present invention, which are listed or not listed, each parameter in the same embodiment only represents an example (i.e., a feasible embodiment) of the technical solution, and there is no strict matching and limiting relationship between the parameters, wherein the parameters may be replaced with each other without departing from the axiom and the requirements of the present invention, unless otherwise specified.
The technical means disclosed by the scheme of the invention are not limited to the technical means disclosed by the technical means, and the technical means also comprises the technical scheme formed by any combination of the technical features. While the foregoing is directed to embodiments of the present invention, it will be appreciated by those skilled in the art that various changes may be made in the embodiments without departing from the principles of the invention, and that such changes and modifications are intended to be included within the scope of the invention.
Claims (9)
1. A method for detecting a caramolol hydrochloride impurity by using HPLC is characterized by comprising the following steps:
s1, dissolving benzaldehyde in a mobile phase to obtain a benzaldehyde-containing contrast solution;
s2, dissolving a sample to be detected containing the caraprolol into the mobile phase to obtain a control solution;
and S3, injecting the benzaldehyde-containing control solution, the control solution and the sample containing the caraprolol into a liquid chromatograph respectively, and detecting the impurity content of the caraprolol in the sample solution by using HPLC.
2. The method for detecting the impurity of the caramolol hydrochloride by using HPLC as claimed in claim 1, wherein the mobile phase is acetonitrile-ammonium acetate solution with pH 5.5-6.5 and concentration of 1-3 g/L.
3. The method for detecting the impurity of the caraprolol hydrochloride by using HPLC as claimed in claim 2, wherein the volume ratio of the ammonium acetate solution to the acetonitrile in the ammonium acetate-acetonitrile solution is (9-14): 6.
4. the method for detecting the impurity of the caraprolol hydrochloride by HPLC as recited in claim 1, wherein the flow rate of the mobile phase during the HPLC detection is 0.9-1.1ml/min.
5. The method for detecting the impurity of caramolol hydrochloride by HPLC as claimed in claim 1, wherein the concentration of benzaldehyde in the control solution containing benzaldehyde is 15-25 μ g/ml.
6. The method for detecting the impurities of the caramolol hydrochloride by using HPLC as claimed in claim 1, wherein the volume ratio of the caramolol injection to the mobile phase in the control solution is 1: (95-105).
7. The method for detecting the impurity of the caraprolol hydrochloride by using HPLC as claimed in claim 1, wherein the chromatographic column in the HPLC detection process is a C18 chromatographic column with a particle size of 3-8 μm, wherein the filler is octadecylsilane chemically bonded silica.
8. The method for detecting the impurity of the caramolol hydrochloride by HPLC as claimed in claim 7, wherein the column temperature of the chromatographic column during the HPLC detection is 25-35 ℃.
9. The method for detecting the caramolol hydrochloride by HPLC as claimed in claim 1, wherein the detector used in the HPLC detection method is an ultraviolet-visible absorption detector, and the detection wavelength is 234-238nm.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109336965A (en) * | 2018-10-30 | 2019-02-15 | 江南大学 | A kind of synthetic method of carazolol artificial antigen |
| CN113092640A (en) * | 2021-03-29 | 2021-07-09 | 河北常山生化药业股份有限公司 | Method for detecting benzyl alcohol and benzaldehyde in heparin sodium injection |
-
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- 2022-07-27 CN CN202210889682.0A patent/CN115452968A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109336965A (en) * | 2018-10-30 | 2019-02-15 | 江南大学 | A kind of synthetic method of carazolol artificial antigen |
| CN113092640A (en) * | 2021-03-29 | 2021-07-09 | 河北常山生化药业股份有限公司 | Method for detecting benzyl alcohol and benzaldehyde in heparin sodium injection |
Non-Patent Citations (9)
| Title |
|---|
| H. J. KEUKENS AND M. M. L. AERTS: "DETERMINATION OF RESIDUES OF CARAZOLOL AND A NUMBER OF TRANQUILLIZERS IN SWINE KIDNEY BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY WITH ULTRAVIOLET AND FLUORESCENCE DETECTION", JOURNAL OF CHROMATOGRAPHY, vol. 464, 31 December 1989 (1989-12-31) * |
| IMRAN ALI • ZEID A. AL-OTHMAN • AFZAL HUSSAIN •: "Chiral Separation of β-Adrenergic Blockers in Human Plasma by SPE-HPLC", CHROMATOGRAPHIA, vol. 73, 11 January 2011 (2011-01-11), XP019881123, DOI: 10.1007/s10337-010-1891-4 * |
| IMRAN ALI，等: "Chiral separation of β -adrenergic blockers on CelluCoat column by HPL C", TALANTA, vol. 78, 25 December 2009 (2009-12-25) * |
| L.A. VAN GINKEL, 等: "LIQUID CHROMATOGRAPHIC METHOD WITH ON-LINE UV SPECTRUM IDENTIFICATION AND OFF-LINE THIN-LAYER CHROMATOGRAPHIC CONFIRMATION FOR THE DETECTION OF TRANQUILLIZERS AND CARAZOLOL IN PIG KIDNEYS", ANALYTLCA CHIMICA ACTA, vol. 225 * |
| 吴春敏等: "HPLC法快速检测苦参素注射液中8个抑菌剂及苯甲醛", 药物分析杂志, no. 07 * |
| 杨龙华;杨娟;陈涛;张磊;于玮洁;丁大中;杨钊;: "HPLC法梯度洗脱同时测定硫辛酸注射液有关物质、苯甲醇和苯甲醛", 中国药师, no. 01 * |
| 王军等: "猪肌肉与肝脏中卡拉洛尔高效液相色谱检测方法的研究", 动物医学进展, vol. 32, no. 4, pages 80 - 81 * |
| 陈雪江等: "长春西汀注射液中的基因毒性杂质研究与含量测定", 药物研究, no. 2, pages 42 - 43 * |
| 魏晋梅等: "高效液相色谱法同时测定羊肉中的 11种镇静剂类药物", 分析检测, vol. 35, no. 10 * |
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