CN113740476A - Method for detecting content of impurity L-2-aminobutanamide hydrochloride in brivaracetam drug - Google Patents
Method for detecting content of impurity L-2-aminobutanamide hydrochloride in brivaracetam drug Download PDFInfo
- Publication number
- CN113740476A CN113740476A CN202010477290.4A CN202010477290A CN113740476A CN 113740476 A CN113740476 A CN 113740476A CN 202010477290 A CN202010477290 A CN 202010477290A CN 113740476 A CN113740476 A CN 113740476A
- Authority
- CN
- China
- Prior art keywords
- brivaracetam
- solution
- aminobutanamide
- phase
- aminobutanamide hydrochloride
- 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
- HDBMIDJFXOYCGK-DFWYDOINSA-N (2s)-2-aminobutanamide;hydrochloride Chemical compound Cl.CC[C@H](N)C(N)=O HDBMIDJFXOYCGK-DFWYDOINSA-N 0.000 title claims abstract description 78
- 229960002161 brivaracetam Drugs 0.000 title claims abstract description 57
- MSYKRHVOOPPJKU-BDAKNGLRSA-N brivaracetam Chemical compound CCC[C@H]1CN([C@@H](CC)C(N)=O)C(=O)C1 MSYKRHVOOPPJKU-BDAKNGLRSA-N 0.000 title claims abstract description 57
- 239000012535 impurity Substances 0.000 title claims abstract description 32
- 238000000034 method Methods 0.000 title claims abstract description 19
- 239000003814 drug Substances 0.000 title abstract description 26
- 229940079593 drug Drugs 0.000 title abstract description 23
- 239000000243 solution Substances 0.000 claims abstract description 46
- 238000001514 detection method Methods 0.000 claims abstract description 35
- 239000013558 reference substance Substances 0.000 claims abstract description 25
- 239000012085 test solution Substances 0.000 claims abstract description 22
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical group CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 18
- 239000012088 reference solution Substances 0.000 claims abstract description 14
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 11
- 239000000945 filler Substances 0.000 claims abstract description 7
- HRQDCDQDOPSGBR-UHFFFAOYSA-M sodium;octane-1-sulfonate Chemical compound [Na+].CCCCCCCCS([O-])(=O)=O HRQDCDQDOPSGBR-UHFFFAOYSA-M 0.000 claims abstract description 7
- 239000007788 liquid Substances 0.000 claims abstract description 6
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims abstract description 6
- 239000000377 silicon dioxide Substances 0.000 claims abstract description 5
- 239000002131 composite material Substances 0.000 claims abstract description 3
- 238000002360 preparation method Methods 0.000 claims description 15
- 238000005303 weighing Methods 0.000 claims description 15
- HNNJFUDLLWOVKZ-UHFFFAOYSA-N 2-aminobutanamide Chemical compound CCC(N)C(N)=O HNNJFUDLLWOVKZ-UHFFFAOYSA-N 0.000 claims description 13
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 13
- 239000011550 stock solution Substances 0.000 claims description 12
- 238000004364 calculation method Methods 0.000 claims description 4
- 238000010812 external standard method Methods 0.000 claims description 4
- 238000007865 diluting Methods 0.000 claims description 3
- 150000001875 compounds Chemical class 0.000 claims description 2
- 238000002347 injection Methods 0.000 claims description 2
- 239000007924 injection Substances 0.000 claims description 2
- 239000000523 sample Substances 0.000 description 29
- 238000012360 testing method Methods 0.000 description 10
- 238000000926 separation method Methods 0.000 description 8
- 238000002474 experimental method Methods 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 239000012488 sample solution Substances 0.000 description 5
- 206010010904 Convulsion Diseases 0.000 description 4
- 230000006378 damage Effects 0.000 description 4
- 238000012417 linear regression Methods 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- XFXIJSRNFKHZFW-UHFFFAOYSA-N [Na].CCCCCCCC Chemical compound [Na].CCCCCCCC XFXIJSRNFKHZFW-UHFFFAOYSA-N 0.000 description 3
- BDHFUVZGWQCTTF-UHFFFAOYSA-M sulfonate Chemical compound [O-]S(=O)=O BDHFUVZGWQCTTF-UHFFFAOYSA-M 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 229960003965 antiepileptics Drugs 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000002964 excitative effect Effects 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 239000010413 mother solution Substances 0.000 description 2
- 239000002858 neurotransmitter agent Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 239000012086 standard solution Substances 0.000 description 2
- 206010067484 Adverse reaction Diseases 0.000 description 1
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 1
- 108010005730 R-SNARE Proteins Proteins 0.000 description 1
- 102000002215 Synaptobrevin Human genes 0.000 description 1
- 238000009098 adjuvant therapy Methods 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 239000001961 anticonvulsive agent Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 206010015037 epilepsy Diseases 0.000 description 1
- 230000001037 epileptic effect Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 229960004002 levetiracetam Drugs 0.000 description 1
- HPHUVLMMVZITSG-ZCFIWIBFSA-N levetiracetam Chemical compound CC[C@H](C(N)=O)N1CCCC1=O HPHUVLMMVZITSG-ZCFIWIBFSA-N 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000003957 neurotransmitter release Effects 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 230000003518 presynaptic effect Effects 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 238000009097 single-agent therapy Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 238000012795 verification 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
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
-
- 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
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
- G01N2030/8872—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample impurities
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)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to a method for detecting the content of impurity L-2-aminobutanamide hydrochloride in a brivaracetam drug. The method for detecting the impurity L-2-aminobutanamide hydrochloride in the brivaracetam specifically comprises the following steps of: (1) preparing a test solution; (2) preparing a reference substance solution; (3) respectively sucking the test solution and the reference solution, injecting into a high performance liquid chromatograph, wherein the chromatographic conditions are as follows: a chromatographic column: octadecylsilane chemically bonded silica is used as a filling agent; mobile phase: the composite material consists of a phase A and a phase B, wherein the phase A is a sodium octane sulfonate solution, and the phase B is acetonitrile. The detection method can accurately detect the content of the L-2-aminobutanamide hydrochloride in the brivaracetam drug, effectively separate the chromatographic peak of the brivaracetam and the impurity L-2-aminobutanamide hydrochloride, improve the quality of the brivaracetam drug and have wide application prospect.
Description
Technical Field
The invention relates to a detection method of a brivaracetam impurity, in particular to a detection method of the content of an impurity L-2-aminobutanamide hydrochloride in a brivaracetam drug, and belongs to the technical field of medicines.
Background
The European Medicine Administration (EMA) and the U.S. Food and Drug Administration (FDA) have respective trade names of Buvalracetam (BRV), a third-generation antiepileptic drug developed by CB of Belgiam, Inc., of 2016 (1/14/1/18/2/2016), and Brivaracetam
Can be used for treating partial seizure type epilepsies of 16 years old and above with or without secondary systemic seizure as adjuvant therapy. In 2017 the FDA has approved a new drug supplement application for bwaitam that would be a monotherapy for local (focal) seizures in epileptic patients over the age of 16 years. The bravaracetam has high affinity and can selectively bind synaptobrevin 2A (SV2A), and SV2A is located in a presynaptic membrane and is involved in regulating the release of neurotransmitters and vesicle circulation so as to maintain the normal function of synaptoblemmas. AED binds to SV2A to reduce excitatory neurotransmitter release and achieve seizure control by regulating the balance of excitatory and inhibitory neurotransmitters in the brain. The affinity of the brivaracetam is 15-30 times that of the levetiracetam, so that the dosage of the brivaracetam is reduced by about 10 times. The structural formula of the brivaracetam is as follows:
the L-2-aminobutanamide hydrochloride is a key intermediate for synthesizing the bulk drug of the busulfacetam, and is used as a reaction raw material in the synthesis process of the busulfacetam, if the reaction is incomplete, a small amount of the L-2-aminobutanamide hydrochloride can exist in the bulk drug of the busulfacetam as an impurity. Researches show that the impurities can cause adverse reactions after clinical patients take the brivaracetam preparation, and influence the treatment effect of the brivaracetam preparation. Therefore, it is very important to strictly control the content of L-2-aminobutanamide hydrochloride in the brivaracetam drug. The structural formula of the L-2-aminobutanamide hydrochloride is as follows:
at present, no report on how to control the content of L-2-aminobutanamide hydrochloride in the brivaracetam medicine is available.
Disclosure of Invention
In order to effectively control the quality of the brivaracetam medicament and reduce the content of the impurity L-2-aminobutanamide hydrochloride in the brivaracetam medicament, the invention provides a method for detecting the content of the impurity L-2-aminobutanamide hydrochloride in brivaracetam.
The invention provides a method for detecting impurity L-2-aminobutanamide hydrochloride in brivaracetam, which adopts high performance liquid chromatography for detection and comprises the following specific steps:
(1) preparation of a test solution: weighing the to-be-detected brivaracetam, and dissolving the brivaracetam in a mobile phase to obtain the brivaracetam;
(2) preparation of control solutions: weighing L-2-aminobutanamide hydrochloride, and dissolving with mobile phase to obtain the compound;
(3) respectively sucking the test solution and the reference solution, injecting into a high performance liquid chromatograph, wherein the chromatographic conditions are as follows:
a chromatographic column: octadecylsilane chemically bonded silica is used as a filling agent;
mobile phase: the composite material consists of a phase A and a phase B, wherein the phase A is a sodium octane sulfonate solution, and the phase B is acetonitrile.
Further, in the step (3), the volume ratio of the phase A to the phase B is (70-90): (10-30);
and/or the concentration of the sodium octane sulfonate solution is 0.005-0.02 mol/L, and the pH value is 2.0-4.0.
Further, in the step (3), the volume ratio of the phase A to the phase B is 80: 20;
and/or the concentration of the sodium octane sulfonate solution is 0.02mol/L, and the pH value is 3.0.
Further, in the step (3), the octadecylsilane chemically bonded silica is Inertsil ODS-3.
Further, the detection wavelength of the step (3) is 205-220 nm, the flow rate is 0.8-1.2 ml/min, and the column temperature is 20-40 ℃;
preferably, the detection wavelength of the step (3) is 210nm, the flow rate is 1.0ml/min, and the column temperature is 30 ℃.
Further, the injection volume of the step (3) is 10-30 μ l, and preferably 20 μ l.
Further, in the step (1), the concentration of the to-be-detected brivaracetam in the test solution is 0.5-2.0 mg/ml, and preferably 1.0 mg/ml.
Further, in the step (2), the concentration of the L-2-aminobutanamide hydrochloride in the control solution is 0.083 μ g/ml or more, preferably 0.2 to 2.0 μ g/ml, and more preferably 1.0 μ g/ml.
Further, in the step (2), the preparation method of the reference solution comprises: weighing L-2-aminobutanamide hydrochloride, adding a mobile phase for dissolving to obtain a reference substance stock solution, and adding the mobile phase for diluting to obtain the L-2-aminobutanamide hydrochloride; in the reference stock solution, the concentration of the L-2-aminobutanamide hydrochloride is 50-200 mug/ml, preferably 100 mug/ml.
Further, the method for calculating the content of the impurity L-2-aminobutanamide hydrochloride in the to-be-detected brivaracetam comprises the following steps: calculating the content of L-2-aminobutanamide hydrochloride in the brivaracetam to be detected by using an external standard method according to the obtained high performance liquid chromatography spectrogram; the calculation formula is as follows:
the content of L-2-aminobutanamide hydrochloride was (Au/As) x (Cs/Cu) x (M)1/M2)×100％，
Au: the peak area of L-2-aminobutanamide in the spectrogram obtained from the test solution,
as: the peak area of L-2-aminobutanamide in the spectrogram obtained from the reference solution,
cs: the concentration of L-2-aminobutanamide hydrochloride in the control solution,
cu: the concentration of the brivaracetam to be tested in the test solution,
M1: l-2-aminobutyrylThe molecular weight of the amine is such that,
M2: molecular weight of L-2-aminobutanamide hydrochloride.
Inertsil ODS-3 is an octadecylsilane bonded silica gel.
The detection method can accurately detect the content of the impurity L-2-aminobutanamide hydrochloride in the brivaracetam drug, and effectively separate the chromatographic peak of the brivaracetam in the brivaracetam drug from the chromatographic peak of the impurity L-2-aminobutanamide hydrochloride, thereby effectively controlling the content of the impurity L-2-aminobutanamide hydrochloride in the brivaracetam drug, improving the purity of the active ingredient brivaracetam in the brivaracetam drug and improving the quality of the brivaracetam drug.
The invention has stable and reliable measuring result and strong specificity. The result of methodology verification shows that the detection method meets the requirements of 'Chinese pharmacopoeia' 2015 edition on analysis methodology. The detection method has good specificity, precision, accuracy and sensitivity, and has wide application prospect in improving the quality of the brivaracetam drug and the pharmaceutical preparation.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Drawings
FIG. 1 is an HPLC chromatogram of a solution suitable for use in the system of example 1.
FIG. 2 is an HPLC chromatogram of the control solution of example 1.
FIG. 3 is an HPLC chromatogram of the test solution of example 1.
Detailed Description
The raw materials and equipment used in the invention are known products and are obtained by purchasing commercial products.
(1) Principal material
Sample of brivaracetam: the achievement is provided by the nation pharmaceutical industry limited company with the batch numbers of 200301 and 200302.
Brivaracetam control: the achievement is provided by Bangbang pharmaceutical industry Co., Ltd, and the batch number is 200301 WS.
L-2-aminobutanamide hydrochloride control: aladdin, batch No. C1307130.
(2) Main instrument
High Performance Liquid Chromatography (HPLC) instrument: shimadzu LC-20 AT.
A chromatographic column: inertsil ODS-3 was used as a filler, with a size of 4.6X 250mm, 5 μm.
Example 1 method for detecting the content of impurity L-2-aminobutanamide hydrochloride in a sample of Buvaracetam
(1) Preparation of a test solution: precisely weighing 25.30mg of a brivaracetam sample (batch number 200301), placing the brivaracetam sample in a 25ml volumetric flask, adding a mobile phase for dissolving, and fixing the volume to prepare a brivaracetam sample solution with the concentration of 1.012 mg/ml.
(2) Preparation of control solutions: accurately weighing 10.40mg of an L-2-aminobutanamide hydrochloride reference substance, placing the L-2-aminobutanamide hydrochloride reference substance in a 100ml volumetric flask, adding a mobile phase for dissolving, and preparing into an L-2-aminobutanamide hydrochloride reference substance stock solution with the concentration of 104 mu g/ml; then 1ml of the L-2-aminobutanamide hydrochloride control stock solution was measured precisely and placed in a 100ml volumetric flask and diluted with a mobile phase to prepare a control solution having a concentration of 1.04. mu.g/ml.
(3) Preparation of system suitability solution: precisely weighing 10.05mg of a Buvalsartan reference substance, placing the Buvalsartan reference substance in a 10ml volumetric flask, precisely adding 0.1ml of an L-2-aminobutanamide hydrochloride reference substance stock solution, dissolving by using a mobile phase, fixing the volume, and preparing a mixed solution of the Buvalsartan reference substance with the concentration of 1.005mg/ml and the L-2-aminobutanamide hydrochloride reference substance with the concentration of 1.04 mu g/ml as a system applicability solution.
(4) And respectively sucking 20 mul of the test solution, the reference solution and the system applicability solution, injecting into a high performance liquid chromatograph, and recording the chromatogram.
The chromatographic conditions were as follows:
a chromatographic column: inertsil ODS-3 was used as a filler.
Mobile phase: a is 0.01mol/L octane sodium sulfonate solution (pH3.0), B is acetonitrile; a: b volume ratio is 80: 20.
detection wavelength: 210 nm.
Flow rate: 1.0 ml/min.
Column temperature: at 30 ℃.
(5) And (3) calculating: respectively determining the peak areas of L-2-aminobutanamide in a reference solution and a test solution in a chromatogram, and calculating the content of impurity L-2-aminobutanamide hydrochloride in the test product, namely the brivaracetam according to an external standard method, wherein the calculation formula is as follows:
content of L-2-aminobutanamide hydrochloride [% ] [ (Au/As) × (Cs/Cu) × (M) ]1/M2)×100％
Au: the peak area of L-2-aminobutanamide in the test solution was 7680
As: the peak area of L-2-aminobutanamide in the control solution was 17082
Cs: the concentration of L-2-aminobutanamide hydrochloride in the control solution was 1.04. mu.g/ml
Cu: the concentration of the sample solution, i.e., the concentration of the busulfacetam is 1.012mg/ml
M1: the molecular weight of the L-2-aminobutanamide is 102.1
M2: the molecular weight of L-2-aminobutanamide hydrochloride is 138.6
And (3) calculating the result: the content of impurity L-2-aminobutanamide hydrochloride in the sample of Buvalracetam (batch No. 200301) was 0.03%.
The HPLC separation of the solution for suitability for use in the system of this example is shown in Table 1 below.
Table 1 HPLC separation data for system suitability solutions
The separation degree is 33.24 and is far more than 2.0, and the method meets the analysis requirements specified by Chinese pharmacopoeia.
Example 2 method for detecting content of impurity L-2-aminobutanamide hydrochloride in Buvalracetam sample
(1) Preparation of a test solution: precisely weighing 25.40mg of a Buvalsartan sample (batch number 200302), placing the Buvalsartan sample in a 25ml volumetric flask, adding a mobile phase for dissolution, and fixing the volume to prepare a Buvalsartan sample solution with the concentration of 1.016 mg/ml.
(2) Preparation of control solutions: accurately weighing 10.10mg of an L-2-aminobutanamide hydrochloride reference substance, placing the L-2-aminobutanamide hydrochloride reference substance in a 100ml volumetric flask, adding a mobile phase for dissolving, and preparing an L-2-aminobutanamide hydrochloride reference substance stock solution with the concentration of 101 mu g/ml; then 1ml of the L-2-aminobutanamide hydrochloride reference stock solution was measured precisely and placed in a 100ml volumetric flask and diluted with a mobile phase to prepare a reference solution having a concentration of 1.01. mu.g/ml.
(3) Preparation of system suitability solution: precisely weighing 10.23mg of a Buvalsartan reference substance, placing the Buvalsartan reference substance in a 10ml volumetric flask, precisely adding 0.1ml of an L-2-aminobutanamide hydrochloride reference substance stock solution, dissolving with a mobile phase, fixing the volume, and preparing a mixed solution of the Buvalsartan reference substance with the concentration of 1mg/ml and the L-2-aminobutanamide hydrochloride reference substance with the concentration of 1.01 mu g/ml as a system applicability solution.
(4) And respectively sucking 20 mul of the test solution, the reference solution and the system applicability solution, injecting into a high performance liquid chromatograph, and recording the chromatogram.
The chromatographic conditions were as follows:
a chromatographic column: inertsil ODS-3 is used as a filling agent;
mobile phase: a is 0.02mol/L octane sodium sulfonate solution (pH3.5), B is acetonitrile; a: b volume ratio is 80: 20.
detection wavelength: 210 nm.
Flow rate: 1.2 ml/min.
Column temperature: 35 ℃ is carried out.
(5) And (3) calculating: respectively determining the peak areas of L-2-aminobutanamide in a reference solution and a test solution in a chromatogram, and calculating the content of impurity L-2-aminobutanamide hydrochloride in the test product, namely the brivaracetam according to an external standard method, wherein the calculation formula is as follows:
content of L-2-aminobutanamide hydrochloride [% ] [ (Au/As) × (Cs/Cu) × (M) ]1/M2)×100％
Au: the peak area of L-2-aminobutanamide in the test solution was 0
As: the peak area of L-2-aminobutanamide in the control solution was 17522
Cs: concentration of L-2-aminobutanamide hydrochloride in control solution, 1.01. mu.g/ml
Cu: the concentration of the Buvalacetam in the test solution is 1.016mg/ml
M1: molecular weight of L-2-aminobutanamide 102.1
M2: molecular weight of L-2-aminobutanamide hydrochloride 138.6
And (3) calculating the result: the impurity, L-2-aminobutanamide hydrochloride, was not detected in the sample of Buvalracetam (batch No. 200302).
The HPLC separation of the solution for suitability for use in the system of this example is shown in Table 2 below.
Table 2 HPLC separation data for system adapted solutions
The separation degree is 28.46 and is far more than 2.0, and the method meets the analysis requirements specified by Chinese pharmacopoeia.
The advantageous effects of the present invention are further illustrated by way of experimental examples below.
The following experiment of the invention is supported by the science and technology bureau of Yibin City of 2019, namely introducing a high-level talent project in Yibin City (project number: 2019YG 03).
Experimental example 1 evaluation of System applicability
1. Solution preparation
(1) L-2-aminobutanamide hydrochloride stock solution: precisely weighing a 10.40 mg-100 ml volumetric flask of the L-2-aminobutanamide hydrochloride reference substance, adding a mobile phase for dissolving and diluting to a scale;
(2) the system applicability solution is as follows: accurately weighing 10.64mg of a Buvalsartan reference substance into a 10ml measuring flask, accurately adding 0.1ml of L-2-aminobutanamide hydrochloride stock solution, and adding a mobile phase to dilute to a scale to obtain the product.
2. And (3) detection:
mu.l of the system suitability solution was taken, injected into a high performance liquid chromatograph under the chromatography conditions of example 1, and 6 needles were injected to examine the retention time and the Relative Standard Deviation (RSD) of the peak area, and the results are shown in Table 3.
The chromatographic conditions were as follows:
a chromatographic column: inertsil ODS-3 was used as a filler.
Mobile phase: a is 0.01mol/L octane sodium sulfonate solution (pH3.0), B is acetonitrile; a: b volume ratio is 80: 20.
detection wavelength: 210 nm.
Flow rate: 1.0 ml/min.
Column temperature: at 30 ℃.
TABLE 3 summary of System suitability solution test results
As can be seen, the HPLC separation degree of the system applicability solution is 32.86-33.89, which is much larger than 2.0, and meets the analysis requirements specified in Chinese pharmacopoeia.
Experimental example 2 Special Property experiment
(1) The experimental method comprises the following steps:
the method comprises the steps of precisely weighing 10.12mg,10.68mg and 10.22mg of a bravaracetam sample (batch number 200301), placing the bravaracetam sample into three 10ml measuring bottles respectively, adding 1mol/L hydrochloric acid solution, 1mol/L sodium hydroxide solution and 1ml of 3% hydrogen peroxide solution respectively to obtain an acid destruction sample, an alkali destruction sample and an oxidation sample in sequence, placing the samples at room temperature for 2 hours, and neutralizing the acid destruction sample and the alkali destruction sample respectively. And (4) respectively adding mobile phases to dilute the neutralized acid-damaged sample, the neutralized alkali-damaged sample and the oxidized sample to a scale, and shaking up. According to the chromatographic conditions of the experimental example 1, samples are respectively injected, and chromatograms are recorded.
(2) The experimental results are as follows:
the separation degree between the main peak and each adjacent peak of each spectrogram is more than 1.5 percent; the main peak purity threshold is smaller than the purity similarity, and meets the requirement.
Experimental example 3 testing of detection limits and quantitation limits
(1) The experimental method comprises the following steps:
the L-2-aminobutanamide hydrochloride reference substance is precisely weighed, a mobile phase is added to dilute the reference substance into a mother solution of 10.4 mu g/ml, the mother solution is further diluted into reference substance solutions with different gradient concentrations step by step, and the quantitative limit and the detection limit are determined according to the chromatographic conditions of the experimental example 1. The detection signal-to-noise ratio S/N is more than or equal to 10 and is determined as the quantitative limit, and the S/N is about 3 and is determined as the detection limit. The test results are shown in Table 4.
(2) The experimental results are as follows:
TABLE 4 quantification and detection limits
The limit of quantitation of L-2-aminobutanamide hydrochloride is 0.208. mu.g/ml, corresponding to 0.02% (< 0.05%) of the sample concentration, the limit of detection is 0.083. mu.g/ml, corresponding to 0.008% of the sample concentration.
Experimental example 4 Linear and Range analysis
(1) The experimental method comprises the following steps:
according to the chromatographic conditions of experimental example 1, 6 points were taken in the range from the quantitative limit concentration to the index concentration of not less than 150% for the test. The linear relationship was plotted as a function of measured response signal (peak area) versus analyte concentration (i.e., L-2-aminobutanamide hydrochloride control), and linear regression was performed using the least squares method, with a linear regression coefficient R2To confirm a good linear relationship, the linear regression coefficient R is required2Should not be less than 0.990. The results are shown in Table 5.
(2) The experimental results are as follows:
TABLE 5 Linear and Range measurements
It can be seen that the linear range of the L-2-aminobutanamide hydrochloride reference substance detected by the chromatographic conditions of the invention is 0.208-2.080 mug/ml, the linear equation is y ═ 16984x +406, and the linear regression coefficient R is20.9989, meets the requirement of good linear relation (R)2＞0.990)。
EXAMPLE 5 precision measurement
(1) The experimental method comprises the following steps:
the L-2-aminobutanamide hydrochloride reference (batch No. C1307130) is precisely weighed, a mobile phase is added to prepare a reference solution of 1.0 mu g/ml, a sample to be tested (namely a Buvalacetam sample (batch No. 200301)) is weighed, a mobile phase is added to prepare a sample solution of 1.0mg/ml, 6 parts are prepared in parallel, samples are sequentially injected according to the chromatographic conditions of the experimental example 1, and the repeatability test result is shown in Table 6.
(2) The experimental results are as follows:
TABLE 6 repeatability
It can be seen that the RSD of the peak area of 6 test sample solutions is 7.44%, which meets the requirement (RSD < 15%). The detection method of the invention has high precision.
Experimental example 6 accuracy test
(1) The experimental method comprises the following steps:
adding three impurity (namely L-2-aminobutanamide hydrochloride) solutions with different concentrations (the concentrations of the impurity solutions are respectively 80%, 100% and 120% of the concentration of a standard solution) into a test sample (namely a Buvalsartan sample (batch number 200301)) solution to obtain a standard sample, and then determining the content of impurities in the standard sample according to the chromatographic conditions of experimental example 1; and calculating the recovery rate. Recovery is the measured content of impurities in the spiked sample/theoretical content of impurities in the spiked sample, expressed as a percentage%. The results are shown in Table 7.
Standard solution: 1.040 ug/ml L-2-aminobutanamide hydrochloride.
(2) The experimental results are as follows:
TABLE 7 accuracy
It can be seen that the recovery rate of L-2-aminobutanamide hydrochloride is between 87.67% and 98.82%, which meets the requirement (recovery rate: 80.0% to 120.0%). The detection method of the invention has good accuracy.
The research results of experimental examples 1-6 show that the detection method has good specificity, precision, accuracy and sensitivity.
In conclusion, the detection method can accurately detect the content of the impurity L-2-aminobutanamide hydrochloride in the brivaracetam drug, and effectively separate the chromatographic peak of the brivaracetam in the brivaracetam drug from the chromatographic peak of the impurity L-2-aminobutanamide hydrochloride, so that the content of the impurity L-2-aminobutanamide hydrochloride in the brivaracetam drug can be effectively controlled, the purity of the active ingredient brivaracetam in the brivaracetam drug can be improved, the quality of the brivaracetam drug can be improved, and the application prospect is wide.
Claims (10)
1. A method for detecting L-2-aminobutanamide hydrochloride as an impurity in brivaracetam is characterized by comprising the following steps: the detection is carried out by adopting a high performance liquid chromatography, and the method comprises the following specific steps:
(1) preparation of a test solution: weighing the to-be-detected brivaracetam, and dissolving the brivaracetam in a mobile phase to obtain the brivaracetam;
(2) preparation of control solutions: weighing L-2-aminobutanamide hydrochloride, and dissolving with mobile phase to obtain the compound;
(3) respectively sucking the test solution and the reference solution, injecting into a high performance liquid chromatograph, wherein the chromatographic conditions are as follows:
a chromatographic column: octadecylsilane chemically bonded silica is used as a filling agent;
mobile phase: the composite material consists of a phase A and a phase B, wherein the phase A is a sodium octane sulfonate solution, and the phase B is acetonitrile.
2. The detection method according to claim 1, characterized in that: in the step (3), the volume ratio of the phase A to the phase B is (70-90): (10-30);
and/or the concentration of the sodium octane sulfonate solution is 0.005-0.02 mol/L, and the pH value is 2.0-4.0.
3. The detection method according to claim 2, characterized in that: in the step (3), the volume ratio of the phase A to the phase B is 80: 20;
and/or the concentration of the sodium octane sulfonate solution is 0.02mol/L, and the pH value is 3.0.
4. The detection method according to any one of claims 1 to 3, characterized in that: in the step (3), the octadecylsilane chemically bonded silica is Inertsil ODS-3.
5. The detection method according to claim 4, characterized in that: the detection wavelength of the step (3) is 205-220 nm, the flow rate is 0.8-1.2 ml/min, and the column temperature is 20-40 ℃;
preferably, the detection wavelength of the step (3) is 210nm, the flow rate is 1.0ml/min, and the column temperature is 30 ℃.
6. The detection method according to claim 4, characterized in that: the sample injection volume of the step (3) is 10-30 mu l, and preferably 20 mu l.
7. The detection method according to any one of claims 1 to 6, characterized in that: in the step (1), the concentration of the to-be-detected brivaracetam in the test solution is 0.5-2.0 mg/ml, and preferably 1.0 mg/ml.
8. The detection method according to any one of claims 1 to 6, characterized in that: in the step (2), the concentration of the L-2-aminobutanamide hydrochloride in the reference solution is more than 0.083 mu g/ml, preferably 0.2 to 2.0 mu g/ml, and more preferably 1.0 mu g/ml.
9. The detection method according to claim 8, characterized in that: in the step (2), the preparation method of the reference solution comprises the following steps: weighing L-2-aminobutanamide hydrochloride, adding a mobile phase for dissolving to obtain a reference substance stock solution, and adding the mobile phase for diluting to obtain the L-2-aminobutanamide hydrochloride; in the reference stock solution, the concentration of the L-2-aminobutanamide hydrochloride is 50-200 mug/ml, preferably 100 mug/ml.
10. The detection method according to any one of claims 1 to 9, characterized in that: the method for calculating the content of the impurity L-2-aminobutanamide hydrochloride in the to-be-detected brivaracetam comprises the following steps: calculating the content of L-2-aminobutanamide hydrochloride in the brivaracetam to be detected by using an external standard method according to the obtained high performance liquid chromatography spectrogram; the calculation formula is as follows:
the content of L-2-aminobutanamide hydrochloride was (Au/As) x (Cs/Cu) x (M)1/M2)×100％，
Au: the peak area of L-2-aminobutanamide in the spectrogram obtained from the test solution,
as: the peak area of L-2-aminobutanamide in the spectrogram obtained from the reference solution,
cs: the concentration of L-2-aminobutanamide hydrochloride in the control solution,
cu: the concentration of the brivaracetam to be tested in the test solution,
M1: the molecular weight of L-2-aminobutanamide,
M2: molecular weight of L-2-aminobutanamide hydrochloride.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010477290.4A CN113740476A (en) | 2020-05-29 | 2020-05-29 | Method for detecting content of impurity L-2-aminobutanamide hydrochloride in brivaracetam drug |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010477290.4A CN113740476A (en) | 2020-05-29 | 2020-05-29 | Method for detecting content of impurity L-2-aminobutanamide hydrochloride in brivaracetam drug |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113740476A true CN113740476A (en) | 2021-12-03 |
Family
ID=78724863
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010477290.4A Pending CN113740476A (en) | 2020-05-29 | 2020-05-29 | Method for detecting content of impurity L-2-aminobutanamide hydrochloride in brivaracetam drug |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113740476A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114213451A (en) * | 2021-12-31 | 2022-03-22 | 南京望知星医药科技有限公司 | Synthesis method of impurities of brivaracetam |
Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080206883A1 (en) * | 2007-02-26 | 2008-08-28 | Cody Laboratories, Inc. | Hplc method for separation and detection of hydromorphone and related opioid pharmacophores |
| CN104251889A (en) * | 2013-06-26 | 2014-12-31 | 深圳海王药业有限公司 | Method for determining content of three components comprising phenylephrine hydrochloride, chlorphenamine maleate and ibuprofen in compound cold treatment tablet |
| RU2650968C1 (en) * | 2017-06-29 | 2018-04-18 | Федеральное государственное бюджетное научное учреждение "Научный центр неврологии" (ФГБНУ НЦН) | Method of quantification of amantadine in blood plasma |
| CN108548873A (en) * | 2018-03-20 | 2018-09-18 | 丽珠集团新北江制药股份有限公司 | A kind of detection method of Bu Waxitan intermediates isomers |
| CN109655557A (en) * | 2019-01-08 | 2019-04-19 | 丽珠集团新北江制药股份有限公司 | A kind of detection method of Bu Waxitan and its impurity |
| CN109781902A (en) * | 2019-03-11 | 2019-05-21 | 山东新华制药股份有限公司 | The HPLC quantitative detecting method of glycyl amide hydrochloride in a kind of Oxiracetam |
| CN109932442A (en) * | 2019-03-04 | 2019-06-25 | 成都美域高制药有限公司 | A kind of detection method of Bu Waxitan isomers |
| CN110824093A (en) * | 2018-08-13 | 2020-02-21 | 北京海晶生物医药科技有限公司 | Method for detecting brivaracetam and related substances thereof |
| CN111103372A (en) * | 2019-12-31 | 2020-05-05 | 成都惟邦药业有限公司 | Method for detecting related substances of procaterol hydrochloride intermediate |
| CN111122736A (en) * | 2019-12-30 | 2020-05-08 | 北京鑫开元医药科技有限公司海南分公司 | Method for detecting enantiomer in intermediate of brivaracetam |
| CN111579706A (en) * | 2020-07-03 | 2020-08-25 | 宜宾市南溪区红光制药有限公司 | Detection method of impurities in hydrolysis of brivaracetam |
-
2020
- 2020-05-29 CN CN202010477290.4A patent/CN113740476A/en active Pending
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080206883A1 (en) * | 2007-02-26 | 2008-08-28 | Cody Laboratories, Inc. | Hplc method for separation and detection of hydromorphone and related opioid pharmacophores |
| CN104251889A (en) * | 2013-06-26 | 2014-12-31 | 深圳海王药业有限公司 | Method for determining content of three components comprising phenylephrine hydrochloride, chlorphenamine maleate and ibuprofen in compound cold treatment tablet |
| RU2650968C1 (en) * | 2017-06-29 | 2018-04-18 | Федеральное государственное бюджетное научное учреждение "Научный центр неврологии" (ФГБНУ НЦН) | Method of quantification of amantadine in blood plasma |
| CN108548873A (en) * | 2018-03-20 | 2018-09-18 | 丽珠集团新北江制药股份有限公司 | A kind of detection method of Bu Waxitan intermediates isomers |
| CN110824093A (en) * | 2018-08-13 | 2020-02-21 | 北京海晶生物医药科技有限公司 | Method for detecting brivaracetam and related substances thereof |
| CN109655557A (en) * | 2019-01-08 | 2019-04-19 | 丽珠集团新北江制药股份有限公司 | A kind of detection method of Bu Waxitan and its impurity |
| CN109932442A (en) * | 2019-03-04 | 2019-06-25 | 成都美域高制药有限公司 | A kind of detection method of Bu Waxitan isomers |
| CN109781902A (en) * | 2019-03-11 | 2019-05-21 | 山东新华制药股份有限公司 | The HPLC quantitative detecting method of glycyl amide hydrochloride in a kind of Oxiracetam |
| CN111122736A (en) * | 2019-12-30 | 2020-05-08 | 北京鑫开元医药科技有限公司海南分公司 | Method for detecting enantiomer in intermediate of brivaracetam |
| CN111103372A (en) * | 2019-12-31 | 2020-05-05 | 成都惟邦药业有限公司 | Method for detecting related substances of procaterol hydrochloride intermediate |
| CN111579706A (en) * | 2020-07-03 | 2020-08-25 | 宜宾市南溪区红光制药有限公司 | Detection method of impurities in hydrolysis of brivaracetam |
Non-Patent Citations (2)
| Title |
|---|
| 石璐燕;于燕燕;吴范宏;: "HPLC法测定左乙拉西坦含量及其有关物质", 化学试剂, vol. 35, no. 04, 15 April 2013 (2013-04-15), pages 337 - 340 * |
| 黄春青;左红雨;曹桂红;苌玲;: "HPLC法测定盐酸纳洛酮注射液有关物质的含量", 沈阳药科大学学报, vol. 33, no. 10, 20 October 2016 (2016-10-20), pages 778 - 782 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114213451A (en) * | 2021-12-31 | 2022-03-22 | 南京望知星医药科技有限公司 | Synthesis method of impurities of brivaracetam |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103592379B (en) | Analytic method of omeprazole related substance | |
| CN112782327B (en) | Method for separating and determining carbocisteine and impurities thereof by liquid chromatography | |
| CN107543872B (en) | Method for separating and determining edoxaban tosylate hydrate and isomer impurities thereof by chiral high performance liquid chromatography | |
| CN110455944A (en) | Method that is a kind of while detecting apo- Changchun amino acid and Changchun amino acid in vinpocetine | |
| CN102375033B (en) | High performance liquid chromatographic analysis method of bendamustine hydrochloride and its related substances | |
| CN106338564B (en) | A method of for detecting enantiomter in vildagliptin intermediate | |
| CN113009003A (en) | Method for detecting related substances in itopride hydrochloride preparation | |
| CN113740476A (en) | Method for detecting content of impurity L-2-aminobutanamide hydrochloride in brivaracetam drug | |
| CN107870209A (en) | The method for determining impurity content in Li Gelieting bulk drugs | |
| CN111024831A (en) | Method for separating moxifloxacin hydrochloride and impurities thereof by high performance liquid chromatography | |
| CN110514759B (en) | Method for detecting azide in candesartan cilexetil | |
| CN113702514A (en) | Method for determining atorvastatin calcium related impurity I | |
| CN106525994A (en) | Method for determination of related substances of paracetamol and tramadol hydrochloride capsules | |
| CN105675754B (en) | The method of high effective liquid chromatography for measuring Li Gelieting enantiomter contents | |
| CN104965031B (en) | Content measuring method for compound ketoprofen and omeprazole sustained-release capsules | |
| CN109781894B (en) | Method for detecting lifuster R isomer | |
| CN110501436B (en) | Detection method of related substances in tinidazole pharmaceutical composition | |
| CN107976489B (en) | Method for determining residual pyridine in pregabalin | |
| CN110133168A (en) | HPLC method measures method of the Cabazitaxel in relation to substance | |
| CN102109501B (en) | Method for detecting related substances in quinapril hydrochloride and hydrochlorothiazide composition | |
| CN116203172A (en) | Method for detecting 2-chloro-4-bromobenzoyl chloride in dapagliflozin SM-1 | |
| CN113237976B (en) | Method for detecting impurities in diclofenac sodium | |
| CN107870210A (en) | The method for determining Li Gelieting contents in Li Gelieting bulk drugs | |
| CN113092656B (en) | Method for detecting related substances in vecuronium bromide medicine for injection | |
| CN115166073B (en) | Method for detecting isomer of I Sha Zuomi or salt thereof |
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 |