CN111208236A - Method for measuring related substances of lidocaine hydrochloride by high performance liquid chromatography - Google Patents
Method for measuring related substances of lidocaine hydrochloride by high performance liquid chromatography Download PDFInfo
- Publication number
- CN111208236A CN111208236A CN202010148004.XA CN202010148004A CN111208236A CN 111208236 A CN111208236 A CN 111208236A CN 202010148004 A CN202010148004 A CN 202010148004A CN 111208236 A CN111208236 A CN 111208236A
- Authority
- CN
- China
- Prior art keywords
- impurity
- solution
- lidocaine hydrochloride
- high performance
- performance liquid
- 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
- 229960004393 lidocaine hydrochloride Drugs 0.000 title claims abstract description 30
- YECIFGHRMFEPJK-UHFFFAOYSA-N lidocaine hydrochloride monohydrate Chemical compound O.[Cl-].CC[NH+](CC)CC(=O)NC1=C(C)C=CC=C1C YECIFGHRMFEPJK-UHFFFAOYSA-N 0.000 title claims abstract description 30
- 238000000034 method Methods 0.000 title claims abstract description 28
- 239000000126 substance Substances 0.000 title claims abstract description 17
- 238000004128 high performance liquid chromatography Methods 0.000 title claims abstract description 11
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical group O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 14
- 238000001514 detection method Methods 0.000 claims abstract description 14
- 239000007788 liquid Substances 0.000 claims abstract description 10
- 239000000945 filler Substances 0.000 claims abstract description 7
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims abstract description 7
- 239000000377 silicon dioxide Substances 0.000 claims abstract description 7
- 230000005526 G1 to G0 transition Effects 0.000 claims abstract description 6
- 239000002245 particle Substances 0.000 claims abstract description 6
- 239000003960 organic solvent Substances 0.000 claims abstract description 5
- 239000008055 phosphate buffer solution Substances 0.000 claims abstract description 5
- 239000007853 buffer solution Substances 0.000 claims abstract description 4
- 239000000243 solution Substances 0.000 claims description 58
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 12
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 12
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 8
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 8
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 8
- 239000003643 water by type Substances 0.000 claims description 2
- 239000012535 impurity Substances 0.000 abstract description 74
- 239000003814 drug Substances 0.000 abstract description 4
- 229940079593 drug Drugs 0.000 abstract 1
- 238000012795 verification Methods 0.000 abstract 1
- 239000011550 stock solution Substances 0.000 description 20
- 238000011084 recovery Methods 0.000 description 10
- XUKUURHRXDUEBC-SXOMAYOGSA-N (3s,5r)-7-[2-(4-fluorophenyl)-3-phenyl-4-(phenylcarbamoyl)-5-propan-2-ylpyrrol-1-yl]-3,5-dihydroxyheptanoic acid Chemical compound C=1C=CC=CC=1C1=C(C=2C=CC(F)=CC=2)N(CC[C@@H](O)C[C@H](O)CC(O)=O)C(C(C)C)=C1C(=O)NC1=CC=CC=C1 XUKUURHRXDUEBC-SXOMAYOGSA-N 0.000 description 6
- 239000012085 test solution Substances 0.000 description 6
- 239000000463 material Substances 0.000 description 5
- 239000012488 sample solution Substances 0.000 description 5
- AAEQXEDPVFIFDK-UHFFFAOYSA-N 3-(4-fluorobenzoyl)-2-(2-methylpropanoyl)-n,3-diphenyloxirane-2-carboxamide Chemical compound C=1C=CC=CC=1NC(=O)C1(C(=O)C(C)C)OC1(C=1C=CC=CC=1)C(=O)C1=CC=C(F)C=C1 AAEQXEDPVFIFDK-UHFFFAOYSA-N 0.000 description 4
- OUCSEDFVYPBLLF-KAYWLYCHSA-N 5-(4-fluorophenyl)-1-[2-[(2r,4r)-4-hydroxy-6-oxooxan-2-yl]ethyl]-n,4-diphenyl-2-propan-2-ylpyrrole-3-carboxamide Chemical compound C=1C=CC=CC=1C1=C(C=2C=CC(F)=CC=2)N(CC[C@H]2OC(=O)C[C@H](O)C2)C(C(C)C)=C1C(=O)NC1=CC=CC=C1 OUCSEDFVYPBLLF-KAYWLYCHSA-N 0.000 description 4
- 238000007865 diluting Methods 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- 239000012088 reference solution Substances 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 229920000715 Mucilage Polymers 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 230000006793 arrhythmia Effects 0.000 description 1
- 206010003119 arrhythmia Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 239000007857 degradation product Substances 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 229960003462 dyclonine hydrochloride Drugs 0.000 description 1
- KNZADIMHVBBPOA-UHFFFAOYSA-N dyclonine hydrochloride Chemical compound [Cl-].C1=CC(OCCCC)=CC=C1C(=O)CC[NH+]1CCCCC1 KNZADIMHVBBPOA-UHFFFAOYSA-N 0.000 description 1
- 238000002692 epidural anesthesia Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 229940004954 lidocaine hydrochloride 5 mg Drugs 0.000 description 1
- 239000003589 local anesthetic agent Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000011003 system suitability test Methods 0.000 description 1
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
- G01N30/26—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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
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)
- Spectroscopy & Molecular Physics (AREA)
- Engineering & Computer Science (AREA)
- Library & Information Science (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention discloses a method for determining related substances of lidocaine hydrochloride by high performance liquid chromatography, which relates to the field of drug detection, and has the following chromatographic conditions: the stationary phase selects octadecylsilane chemically bonded silica bonded with polar groups as a filling agent; the particle size of the chromatographic column is 5 mu m; detection wavelength: 225-235 nm; flow rate: 0.8-1.2 ml/min; column temperature: 30-40 ℃, mobile phase: the buffer solution is composed of 50-80 parts by mass of 0.035mol/L phosphate buffer solution and 20-50 parts by mass of organic solvent; the adopted detector is a high performance liquid chromatograph, the method can detect more impurities, and the detection result is accurate and reliable through methodology verification.
Description
Technical Field
The invention relates to the field of medicine detection, in particular to a method for determining related substances of lidocaine hydrochloride by high performance liquid chromatography.
Background
Lidocaine Hydrochloride (Lidocaine Hydrochloride) is an amide local anesthetic, and is mainly used for block anesthesia and epidural anesthesia, and also used for resisting arrhythmia. The substances concerned, also referred to as impurities, are mainly starting materials, intermediates, polymers, side reaction products brought in during production, degradation products during storage, etc. At present, lidocaine hydrochloride can be prepared into various preparation forms, such as injection, mucilage, gel and the like, the existing disclosed determination method for related substances cannot realize determination of the lidocaine hydrochloride related substances, and in order to strictly control the quality of the medicine, improve the safety of the medicine and reduce the harm to human bodies and the environment, the lidocaine hydrochloride related substance determination method is needed to be provided.
Disclosure of Invention
The invention aims to solve the problem that the existing lidocaine hydrochloride related substances are difficult to measure, and provides a method for measuring lidocaine hydrochloride related substances by using a high performance liquid chromatography.
In order to achieve the above purpose, the invention adopts the technical scheme that: a method for measuring related substances of lidocaine hydrochloride by high performance liquid chromatography comprises the following steps:
the stationary phase selects octadecylsilane chemically bonded silica bonded with polar groups as a filling agent;
the particle size of the chromatographic column is 5 mu m;
detection wavelength: 225-235 nm;
flow rate: 0.8-1.2 ml/min;
column temperature: 30-40 ℃,
mobile phase: the buffer solution is composed of 50-80 parts by mass of 0.035mol/L phosphate buffer solution and 20-50 parts by mass of organic solvent;
the detector used is a high performance liquid chromatograph.
In order to further optimize the present invention, the following technical solutions may be preferably selected:
preferably, the chromatographic column is one of Agilent 5 HC-C18 chromatographic column, Utimate XB-C18 chromatographic column, Waters Xterra C18 chromatographic column and German MN NUCLEOSIL 100-5C 18 chromatographic column.
Preferably, the mobile phase of the method consists of 70 parts of potassium dihydrogen phosphate solution and 30 parts of acetonitrile in parts by mass.
Preferably, the pH of the potassium dihydrogen phosphate solution is 8.0, and the pH value is adjusted by 10% sodium hydroxide solution.
Preferably, the flow rate of the process is 1.0 ml/min.
Preferably, the column temperature of the process is 35 ℃.
The invention has the beneficial effects that: the method can detect 11 impurities, the separation degree of each impurity peak, the separation degree of the impurity peak and the dyclonine hydrochloride peak is larger than 1.5, the correlation coefficient R2 of each impurity is larger than 0.9990, and the average recovery rate of each impurity is 90-108%.
Drawings
FIG. 1 is a spectrum diagram of the applicability of a lidocaine hydrochloride system according to the present invention;
FIG. 2 is a structural view of impurity A;
FIG. 3 is a structural view of impurity B;
FIG. 4 is a structural view of impurity C;
FIG. 5 is a structural view of impurity D;
FIG. 6 is a structural view of impurity E;
FIG. 7 is a structural view of impurity F;
FIG. 8 is a structural view of an impurity G;
FIG. 9 is a structural view of impurity H;
FIG. 10 is a structural view of impurity I;
FIG. 11 is a structural view of impurity J;
fig. 12 is a structural view of the impurity K.
Detailed Description
Example 1:
a method for measuring related substances of lidocaine hydrochloride by high performance liquid chromatography comprises the following steps:
the stationary phase selects octadecylsilane chemically bonded silica bonded with polar groups as a filling agent; the particle size of the chromatographic column is 5 mu m; detection wavelength: 225 nm; flow rate: 0.8 ml/min; column temperature: at a temperature of 30 c,
mobile phase: the buffer solution consists of 50ml of 0.035mol/L phosphate buffer solution and 20ml of organic solvent in parts by mass; the pH value of the potassium dihydrogen phosphate solution is adjusted to 8.0 by using 10 percent sodium hydroxide solution, and a high performance liquid chromatograph is adopted as a detector.
Example 2:
a method for measuring related substances of lidocaine hydrochloride by high performance liquid chromatography comprises the following steps:
the stationary phase selects octadecylsilane chemically bonded silica bonded with polar groups as a filling agent; the particle size of the chromatographic column is 5 mu m; detection wavelength: 230 nm; flow rate: 1.0 ml/min; column temperature: 35 ℃, mobile phase: the device consists of 70ml of 0.035mol/L potassium dihydrogen phosphate solution and 30ml of acetonitrile in parts by mass, and the adopted detector is a high performance liquid chromatograph.
Example 3:
a method for measuring related substances of lidocaine hydrochloride by high performance liquid chromatography comprises the following steps:
the stationary phase selects octadecylsilane chemically bonded silica bonded with polar groups as a filling agent; the particle size of the chromatographic column is 5 mu m; detection wavelength: 235 nm; flow rate: 1.2 ml/min; column temperature: at a temperature of 40 c,
mobile phase: the composite material consists of 80ml of 0.035mol/L phosphate buffer solution and 50ml of organic solvent in parts by mass; the detector used is a high performance liquid chromatograph.
Experiment one: system suitability test
Preparing a stock solution of each impurity solution: taking appropriate amount of each of impurities A-K, adding mobile phase to dissolve and dilute respectively to obtain solution containing about 500 μ g of each impurity in 1ml, and using the solution as stock solution of each impurity.
Preparing an impurity positioning solution: precisely measuring 1ml of the impurity solution stock solutions, respectively placing into different 100ml measuring bottles, adding mobile phase to dilute into solutions containing about 5 μ g of the impurities in each 1ml, and using the solutions as impurity positioning solutions.
Preparation of mixed control solution: precisely measuring 1ml of the impurity solution stock solutions, putting the impurity solution stock solutions into a same 100ml measuring flask, putting a proper amount of lidocaine hydrochloride into the same 100ml measuring flask, adding the mobile phase for dissolving, and diluting to obtain a solution containing about 5 mu g of impurities A-K and lidocaine hydrochloride in each 1ml, wherein the solution is used as a mixed control solution.
Preparing a test solution: precisely weighing appropriate amount of lidocaine hydrochloride, adding mobile phase for dissolving and diluting to obtain solution containing lidocaine hydrochloride 5mg per 1ml, and shaking.
Preparation of a control solution: precisely measuring 1ml of the test solution, placing into a 10ml measuring flask, adding mobile phase to dilute to scale, and shaking.
And (3) determination: octadecylsilane chemically bonded silica is used as a filler for the chromatographic column; the mobile phase is potassium dihydrogen phosphate solution (taking 4.85g of potassium dihydrogen phosphate, adding 1000ml of water for dissolution, and adjusting the pH value to 8.0 by using 10% sodium hydroxide solution): methanol: acetonitrile =60:10: 30; the column temperature was 35 ℃; the detection wavelength is 230 nm; the flow rate was 1.0 ml/min.
Precisely measuring 20 μ l of each solution, injecting into high performance liquid chromatograph, and recording chromatogram. The results are shown in Table 1, and the system suitability chromatogram is shown in FIG. 1.
TABLE 1 results of the materials methodology-specificity-mix control
And (4) conclusion: the blank diluent does not interfere the detection of related substances in the test solution, and the separation degree between each peak and the adjacent chromatographic peak in the mixed control solution meets the specification, which shows that the method for determining related substances of lidocaine hydrochloride by high performance liquid chromatography provided by the invention has good specificity.
Experiment two: linear and range tests and detection limit and quantification limit tests
Preparing a stock solution of each impurity solution: precisely weighing 10mg of each of the impurities A to K, respectively placing the impurities A to K into different measuring bottles, adding a mobile phase for dissolving and diluting to prepare a solution containing about 50 mu g/ml of the impurity A and about 500 mu g/ml of the impurity B to K per 1ml of the solution, and respectively using the solution as a stock solution of the solution containing the impurities A to K.
Preparation of a linear solution: precisely measuring 1ml of the impurity solution stock solutions respectively, placing the impurity solution stock solutions in a same 50ml measuring flask, adding a mobile phase to dilute the impurity solution stock solutions to a scale, and shaking up the impurity solution stock solutions to obtain a linear 200% solution; stepwise dilution made linear 150%, linear 100%, linear 40%, linear 20%, linear 10%, linear 4%, linear 2%, linear 1%, linear 0.5%, linear 0.125% solution.
Precisely measuring 20 μ l of each solution, injecting into high performance liquid chromatograph, and recording chromatogram. The results are shown in tables 2 and 3.
TABLE 2 materials methodology-Linear results
TABLE 3 materials methodology-detection Limit and quantitation Limit results
| Name of impurity | Detection limit (mu g/ml) | Limit of quantitation (ug/ml) |
| Impurity A | 0.00625 | 0.0125 |
| Impurity B | 0.05 | 0.1 |
| Impurity C | 0.025 | 0.05 |
| Impurity D | 0.003125 | 0.00625 |
| Impurity E | 0.025 | 0.1 |
| Impurity F | 0.05 | 0.2 |
| Impurity G | 0.025 | 0.1 |
| Impurity H | 0.025 | 0.05 |
| Impurity I | 0.05 | 0.2 |
| Impurity J | 0.05 | 0.2 |
| Impurity K | 0.025 | 0.1 |
And (4) conclusion: the concentration of the impurity A is within the range of 0.0125-1.0 mu g/ml (equivalent to 2.5% -200% of the limit concentration), the linear equation is y =85054x-1381.5, the correlation coefficient R2 is 0.9993>0.9990, and the linear relation between the peak area and the concentration is good.
The concentration of the impurity B is within the range of 0.1-10.0 mug/ml (equivalent to 2% -200% of the limit concentration), the linear equation is y =19675x-1425.2, the correlation coefficient R2 is 0.9993>0.9990, and the linear relation between the peak area and the concentration is good.
The concentration range of the impurity C is 0.05-10.0 mug/ml (equivalent to 1% -200% of the limit concentration), the linear equation is y =21838x-3521.9, the correlation coefficient R2 is 0.9989>0.990, and the linear relation between the peak area and the concentration is good.
The impurity D is in a concentration range of 0.00625-10.0 mug/ml (equivalent to 0.125% -200% of the limit concentration), the linear equation is y =14535x +216.62, the correlation coefficient R2 is 1>0.9990, and the linear relation between the peak area and the concentration is good.
The concentration range of the impurity E is 0.1-10.0 mu g/ml (equivalent to 2% -200% of the limit concentration), the linear equation is y =18466x-1756.2, the correlation coefficient R2 is 0.9997>0.9990, and the linear relation between the peak area and the concentration is good.
The concentration of the impurity F is within the range of 0.2-10.0 mug/ml (equivalent to 4% -200% of the limit concentration), the linear equation is y =27460x-3523.2, the correlation coefficient R2 is 0.9998>0.9990, and the linear relation between the peak area and the concentration is good.
The concentration of the impurity G is within the range of 0.05-10.0 mug/ml (equivalent to 1% -200% of the limit concentration), the linear equation is y =15029x-620.06, the correlation coefficient R2 is 0.9999>0.9990, and the linear relation between the peak area and the concentration is good.
The concentration of the impurity H is within the range of 0.05-10.0 mug/ml (equivalent to 1% -200% of the limit concentration), the linear equation is y =17147x-852.24, the correlation coefficient R2 is 0.9999>0.9990, and the linear relation between the peak area and the concentration is good.
The concentration range of the impurity I is 0.2-10.0 mu g/ml (equivalent to 4% -200% of the limit concentration), the linear equation is y =33885x-4640.5, the correlation coefficient R2 is 0.9997>0.9990, and the linear relation between the peak area and the concentration is good.
The concentration of the impurity J is within the range of 0.2-10.0 mug/ml (equivalent to 4% -200% of the limit concentration), the linear equation is y =26859x-3157.1, the correlation coefficient R2 is 0.9999>0.9990, and the linear relation between the peak area and the concentration is good.
The impurity K is in a concentration range of 0.1-10.0 mu g/ml (equivalent to 2% -200% of the limit concentration), the linear equation is y =16565x-185.88, the correlation coefficient R2 is 1>0.9990, and the linear relation between the peak area and the concentration is good.
Experiment three: recovery test
Sample solution stock solution: a proper amount of lidocaine hydrochloride is precisely weighed, and is dissolved and diluted by adding a mobile phase to prepare a solution containing 50mg of lidocaine hydrochloride per 1 ml.
Test solution: precisely measuring 1ml of the sample solution storage solution, placing into a 10ml measuring flask, adding mobile phase to dilute to scale, and shaking.
Stock solutions of various impurity solutions: precisely weighing 10mg of each of the impurities A to K, respectively placing the impurities A to K into different measuring bottles, adding a mobile phase for dissolving and diluting to prepare a solution containing about 50 mu g/ml of the impurity A and about 500 mu g/ml of the impurity B to K per 1ml of the solution, and respectively using the solution as a stock solution of the solution containing the impurities A to K.
Mixed impurity control solution stock solution: precisely measuring the impurity solutions respectively, storing 1ml of the impurity solutions in the same measuring flask, and shaking up to obtain the final product.
Recovery control solution: precisely measuring the mixed impurity reference solution stock solutions of 0.5ml, 1ml and 1.5ml, placing the stock solutions in different 10ml measuring bottles, adding mobile phase to dilute to scale, and shaking up to obtain reference solution with recovery rates of 50%, 100% and 150%.
Recovery of test solution: precisely measuring 1ml of the sample solution stock solution, putting the sample solution stock solution into a 10ml measuring flask, parallelly measuring 9 parts, equally dividing into 3 groups, precisely adding 0.5ml, 1ml and 1.5ml of the mixed impurity control solution stock solution respectively, adding a mobile phase to dilute to a scale, and shaking uniformly to obtain the sample solution with the recovery rates of 50%, 100% and 150%.
Precisely measuring 20 μ l of each solution, injecting into high performance liquid chromatograph, and recording chromatogram. The results are shown in Table 4.
TABLE 4 materials methodology-recovery results
| Name (R) | Corresponding to the limit of 50% | Corresponding to the limit of 100% | Equivalent to the limit of 150% | Average recovery rate% |
| Impurity A | 97.5 | 98.4 | 101.8 | 99.2 |
| Impurity B | 100.5 | 100.5 | 98.9 | 100.0 |
| Impurity C | 102.0 | 98.6 | 101.0 | 100.5 |
| Impurity D | 98.5 | 99.1 | 100.0 | 99.2 |
| Impurity E | 102.1 | 101.0 | 98.6 | 100.6 |
| Impurity F | 99.7 | 98.1 | 100.8 | 99.5 |
| Impurity G | 96.9 | 106.1 | 102.1 | 101.7 |
| Impurity H | 97.8 | 104.4 | 99.9 | 100.7 |
| Impurity I | 99.2 | 97.9 | 99.4 | 98.8 |
| Impurity J | 98.5 | 99.3 | 101.3 | 99.7 |
| Impurity K | 99.7 | 101.1 | 100.3 | 100.4 |
And (4) conclusion: under the concentration of 50%, 100% and 150%, the average recovery rate in the impurity A-K groups and the average recovery rate among the groups are between 90% and 108%, and the method is used for detecting the impurities A-K and has good accuracy.
Experiment four: durability test
Test solution: an appropriate amount of lidocaine hydrochloride is precisely weighed, and is dissolved and diluted by adding a mobile phase to prepare a solution containing about 5mg of lidocaine hydrochloride per 1 ml.
Precisely measuring 20 μ l of each solution, injecting into high performance liquid chromatograph, and recording chromatogram. The results are shown in Table 5.
TABLE 5 materials methodology-durability results
And (4) conclusion: when the chromatographic condition has small variation (the column temperature changes +/-5 ℃, the flow rate changes +/-0.05 ml/min, the wavelength changes +/-2 nm, the pH of the water phase changes +/-0.2, and the flow phase ratio changes +/-2%), the impurity separation condition is good, and the detected amount of impurities has no obvious difference, which indicates that the chromatographic condition has good durability.
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments or portions thereof without departing from the spirit and scope of the invention.
Claims (6)
1. A method for measuring lidocaine hydrochloride related substances by high performance liquid chromatography is characterized in that the chromatographic conditions are as follows: the stationary phase selects octadecylsilane chemically bonded silica bonded with polar groups as a filling agent;
the particle size of the chromatographic column is 5 mu m;
detection wavelength: 225-235 nm;
flow rate: 0.8-1.2 ml/min;
column temperature: 30-40 ℃,
mobile phase: the buffer solution is composed of 50-80 parts by mass of 0.035mol/L phosphate buffer solution and 20-50 parts by mass of organic solvent;
the adopted detector is a high performance liquid chromatograph.
2. The method for determining lidocaine hydrochloride according to claim 1, wherein: the chromatographic column is one of Agilent 5 HC-C18 chromatographic column, Utimate XB-C18 chromatographic column, Waters Xterra C18 chromatographic column, and Germany MN NUCLEOSIL 100-5C 18 chromatographic column.
3. The method for determining lidocaine hydrochloride according to claim 1, wherein: the mobile phase of the method comprises 70 parts of potassium dihydrogen phosphate solution and 30 parts of acetonitrile in parts by mass.
4. The method for determining lidocaine hydrochloride according to claim 3, wherein: the pH of the potassium dihydrogen phosphate solution was 8.0, and the pH was adjusted with 10% sodium hydroxide solution.
5. The method for determining lidocaine hydrochloride according to claim 1, wherein: the flow rate of the process was 1.0 ml/min.
6. The method for determining lidocaine hydrochloride according to claim 1, wherein: the column temperature of the process was 35 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010148004.XA CN111208236A (en) | 2020-03-05 | 2020-03-05 | Method for measuring related substances of lidocaine hydrochloride by high performance liquid chromatography |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010148004.XA CN111208236A (en) | 2020-03-05 | 2020-03-05 | Method for measuring related substances of lidocaine hydrochloride by high performance liquid chromatography |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111208236A true CN111208236A (en) | 2020-05-29 |
Family
ID=70788694
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010148004.XA Pending CN111208236A (en) | 2020-03-05 | 2020-03-05 | Method for measuring related substances of lidocaine hydrochloride by high performance liquid chromatography |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111208236A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112557569A (en) * | 2020-12-21 | 2021-03-26 | 南京海纳医药科技股份有限公司 | Method for detecting related substances in lidocaine |
| CN113219081A (en) * | 2021-03-26 | 2021-08-06 | 天圣制药集团股份有限公司 | HPLC detection method for lidocaine hydrochloride and degradation impurities in preparation of lidocaine hydrochloride |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105663035A (en) * | 2014-11-18 | 2016-06-15 | 上海朝晖药业有限公司 | Lidocaine hydrochloride injection and preparation method thereof |
| CN106248819A (en) * | 2016-07-13 | 2016-12-21 | 浙江景嘉医疗科技有限公司 | The detection method of lidocaine hydrochloride content in a kind of medical cross-linking sodium hyaluronate gel |
-
2020
- 2020-03-05 CN CN202010148004.XA patent/CN111208236A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105663035A (en) * | 2014-11-18 | 2016-06-15 | 上海朝晖药业有限公司 | Lidocaine hydrochloride injection and preparation method thereof |
| CN106248819A (en) * | 2016-07-13 | 2016-12-21 | 浙江景嘉医疗科技有限公司 | The detection method of lidocaine hydrochloride content in a kind of medical cross-linking sodium hyaluronate gel |
Non-Patent Citations (2)
| Title |
|---|
| 周雅茹 等: "利多卡因气雾剂有关物质的HPLC法测定", 《药物分析杂志》 * |
| 邓朝晖 等: "盐酸利多卡因注射液质量标准提高研究", 《药学实践杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112557569A (en) * | 2020-12-21 | 2021-03-26 | 南京海纳医药科技股份有限公司 | Method for detecting related substances in lidocaine |
| CN113219081A (en) * | 2021-03-26 | 2021-08-06 | 天圣制药集团股份有限公司 | HPLC detection method for lidocaine hydrochloride and degradation impurities in preparation of lidocaine hydrochloride |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110579542B (en) | Method for measuring ranitidine hydrochloride related substances by high performance liquid chromatography | |
| CN111208236A (en) | Method for measuring related substances of lidocaine hydrochloride by high performance liquid chromatography | |
| CN114264747A (en) | High performance liquid chromatography detection method for empagliflozin intermediate and related substances thereof | |
| CN109060973B (en) | Method for detecting ethylenediamine in lipoic acid injection | |
| CN109900830B (en) | Method for separating and determining sulfonamide impurities in celecoxib by adopting HPLC (high performance liquid chromatography) and application | |
| CN110672734A (en) | Analysis method of related substances in amiodarone hydrochloride injection | |
| CN113533578A (en) | Quality control method of related substances in bromhexine hydrochloride tablets | |
| CN113514565A (en) | Detection method of residual ethylenediamine and piperazine in homopiperazine | |
| CN114354810B (en) | Method for detecting impurity N in clindamycin phosphate and method for separating impurity | |
| CN114264765B (en) | Analytical method for determining related substances in glimepiride intermediate by utilizing HPLC | |
| CN115616133A (en) | Method for detecting cysteine in compound amino acid injection and application thereof | |
| CN111983054B (en) | Method for separating and measuring related substances of empagliflozin intermediate by using HPLC (high performance liquid chromatography) | |
| CN114166982A (en) | Method for simultaneously determining dimer, trimer and caprolactam in amino caproic acid injection | |
| CN113030328A (en) | Method for detecting genotoxic impurities in ivabradine hydrochloride | |
| CN115372489B (en) | Detection method of tizanidine hydrochloride related substances | |
| CN114235998B (en) | Method for measuring related substances in carteolol hydrochloride and eye drops thereof by ultra-performance liquid chromatography | |
| CN113406236B (en) | Method for detecting impurities in 1- (3-pyridyl) -3- (dimethylamino) -2-propylene-1-ketone | |
| CN115201342B (en) | Method for detecting diethylamine in milnacipran hydrochloride | |
| CN115436525B (en) | LLTS-M3 and detection method and application of related substances thereof | |
| CN114062567B (en) | Separation and detection method of (1R, 2S) -2- (3, 4-difluorophenyl) cyclopropylamine hydrochloride and related substances thereof | |
| CN116183759A (en) | Detection method and application of related substances of pezopanib hydrochloride intermediate PZP-M1 | |
| CN109633046B (en) | Method for detecting dimethylamine from duloxetine hydrochloride | |
| CN107843670B (en) | Method for determining related substances of tetracaine hydrochloride | |
| CN115356411A (en) | Method for detecting related substances of L-piperidinecarboxylic acid hydrochloride by high performance liquid chromatography | |
| CN114544849A (en) | Detection method of 1H-1,2, 4-triazole-1-formamidine monohydrochloride and impurities |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200529 |
|
| RJ01 | Rejection of invention patent application after publication |