CN113984929A - Abamebactam sodium substance analysis method - Google Patents
Abamebactam sodium substance analysis method Download PDFInfo
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- CN113984929A CN113984929A CN202111240892.9A CN202111240892A CN113984929A CN 113984929 A CN113984929 A CN 113984929A CN 202111240892 A CN202111240892 A CN 202111240892A CN 113984929 A CN113984929 A CN 113984929A
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- avibactam
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- 239000000126 substance Substances 0.000 title claims abstract description 22
- 238000004458 analytical method Methods 0.000 title claims abstract description 15
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 title claims description 24
- 229910052708 sodium Inorganic materials 0.000 title claims description 24
- 239000011734 sodium Substances 0.000 title claims description 24
- 238000001514 detection method Methods 0.000 claims abstract description 38
- RTCIKUMODPANKX-JBUOLDKXSA-M avibactam sodium Chemical compound [Na+].NC(=O)[C@@H]1CC[C@H]2N(OS([O-])(=O)=O)C(=O)N1C2 RTCIKUMODPANKX-JBUOLDKXSA-M 0.000 claims abstract description 33
- 229960001496 avibactam sodium Drugs 0.000 claims abstract description 33
- 238000010828 elution Methods 0.000 claims abstract description 13
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims abstract description 9
- 235000019796 monopotassium phosphate Nutrition 0.000 claims abstract description 9
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims abstract description 9
- 230000035945 sensitivity Effects 0.000 claims abstract description 9
- QGPGUZIKJKOKRF-UHFFFAOYSA-M potassium;acetonitrile;dihydrogen phosphate Chemical compound [K+].CC#N.OP(O)([O-])=O QGPGUZIKJKOKRF-UHFFFAOYSA-M 0.000 claims abstract description 6
- 239000000243 solution Substances 0.000 claims description 51
- IBSREHMXUMOFBB-JFUDTMANSA-N 5u8924t11h Chemical compound O1[C@@H](C)[C@H](O)[C@@H](OC)C[C@@H]1O[C@@H]1[C@@H](OC)C[C@H](O[C@@H]2C(=C/C[C@@H]3C[C@@H](C[C@@]4(O3)C=C[C@H](C)[C@@H](C(C)C)O4)OC(=O)[C@@H]3C=C(C)[C@@H](O)[C@H]4OC\C([C@@]34O)=C/C=C/[C@@H]2C)/C)O[C@H]1C.C1=C[C@H](C)[C@@H]([C@@H](C)CC)O[C@]11O[C@H](C\C=C(C)\[C@@H](O[C@@H]2O[C@@H](C)[C@H](O[C@@H]3O[C@@H](C)[C@H](O)[C@@H](OC)C3)[C@@H](OC)C2)[C@@H](C)\C=C\C=C/2[C@]3([C@H](C(=O)O4)C=C(C)[C@@H](O)[C@H]3OC\2)O)C[C@H]4C1 IBSREHMXUMOFBB-JFUDTMANSA-N 0.000 claims description 21
- 239000005660 Abamectin Substances 0.000 claims description 21
- 229950008167 abamectin Drugs 0.000 claims description 21
- 238000000034 method Methods 0.000 claims description 20
- 229960002379 avibactam Drugs 0.000 claims description 13
- 238000007865 diluting Methods 0.000 claims description 13
- 239000000523 sample Substances 0.000 claims description 13
- 239000012085 test solution Substances 0.000 claims description 13
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 12
- 238000002360 preparation method Methods 0.000 claims description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 9
- NDCUAPJVLWFHHB-UHNVWZDZSA-N avibactam Chemical compound C1N2[C@H](C(N)=O)CC[C@@]1([H])N(OS(O)(=O)=O)C2=O NDCUAPJVLWFHHB-UHNVWZDZSA-N 0.000 claims description 9
- -1 [ (2S,5R) -7-oxo-2- (sulfonylcarbamoyl) -1, 6-diazabicyclo [3.2.1] oct-6-yl ] sulfuric acid Chemical compound 0.000 claims description 7
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 6
- 230000007062 hydrolysis Effects 0.000 claims description 5
- 238000006460 hydrolysis reaction Methods 0.000 claims description 5
- 239000012488 sample solution Substances 0.000 claims description 5
- 238000004587 chromatography analysis Methods 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 238000000926 separation method Methods 0.000 claims description 4
- OWIVQKMLPQEEGV-NEPJUHHUSA-N (2s,5r)-5-(phenylmethoxyamino)piperidine-2-carboxamide Chemical compound C1N[C@H](C(=O)N)CC[C@H]1NOCC1=CC=CC=C1 OWIVQKMLPQEEGV-NEPJUHHUSA-N 0.000 claims description 3
- HYTSWLKLRKLRHK-NEPJUHHUSA-N (2s,5r)-7-oxo-6-phenylmethoxy-1,6-diazabicyclo[3.2.1]octane-2-carboxamide Chemical compound C([C@]1(CC[C@H]2C(N)=O)[H])N2C(=O)N1OCC1=CC=CC=C1 HYTSWLKLRKLRHK-NEPJUHHUSA-N 0.000 claims description 3
- QXNVGIXVLWOKEQ-UHFFFAOYSA-N Disodium Chemical class [Na][Na] QXNVGIXVLWOKEQ-UHFFFAOYSA-N 0.000 claims description 3
- 239000006227 byproduct Substances 0.000 claims description 3
- 230000006324 decarbonylation Effects 0.000 claims description 3
- 238000006606 decarbonylation reaction Methods 0.000 claims description 3
- PYUXATUBICTSNB-DFQHDRSWSA-N ethyl (2s,5r)-5-(phenylmethoxyamino)piperidine-2-carboxylate;oxalic acid Chemical compound OC(=O)C(O)=O.C1N[C@H](C(=O)OCC)CC[C@H]1NOCC1=CC=CC=C1 PYUXATUBICTSNB-DFQHDRSWSA-N 0.000 claims description 3
- 238000006386 neutralization reaction Methods 0.000 claims description 3
- 239000002243 precursor Substances 0.000 claims description 3
- 230000035502 ADME Effects 0.000 claims description 2
- 210000000692 cap cell Anatomy 0.000 claims description 2
- 125000004093 cyano group Chemical group *C#N 0.000 claims description 2
- 238000002013 hydrophilic interaction chromatography Methods 0.000 claims description 2
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 2
- 159000000000 sodium salts Chemical class 0.000 claims description 2
- 239000003814 drug Substances 0.000 abstract description 9
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- 230000009286 beneficial effect Effects 0.000 description 1
- 125000003460 beta-lactamyl group Chemical group 0.000 description 1
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Images
Classifications
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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
-
- 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
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/30—Control of physical parameters of the fluid carrier of temperature
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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
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
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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
- G01N30/26—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
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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
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
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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
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
- G01N2030/324—Control of physical parameters of the fluid carrier of pressure or speed speed, flow rate
Abstract
The invention discloses an avibactam sodium analysis method, relates to the field of medicine detection, and aims to provide an avibactam sodium related substance analysis method with high sensitivity and good stability. The chromatographic conditions are as follows: the chromatographic column adopts a hydrophilic C18 column; gradient elution is carried out on a mobile phase A which is 10 mmol/L-50 mmol/L potassium dihydrogen phosphate solution and a mobile phase B which is 10 mmol/L-50 mmol/L potassium dihydrogen phosphate-acetonitrile (80: 20-30: 70); the column temperature is 25-40 ℃; the flow rate is 0.5mL/min to 1.2 mL/min; the sample volume is 20 mu L; the sample concentration is 1 mg/mL-10 mg/mL. The invention has the advantages of high detection sensitivity, good reproducibility, high stability and the like, and is suitable for the detection of related substances of the avibactam sodium.
Description
Technical Field
The invention relates to the field of medicine detection, and particularly relates to an avibactam sodium substance analysis method.
Background
Abamebactam sodium is a beta-lactamase inhibitor. To date, the most successful of the antibacterial drug potentiators have been beta-lactamase inhibitors. Beta-lactam drugs have good characteristics of high efficiency and low toxicity, and are widely used in antibacterial therapy, but with the use of the beta-lactam drugs, bacteria can resist the drugs by mechanisms such as beta-lactamase generation, cell wall or outer membrane permeability change, target protein mutation, efflux pump expression and the like, and the most important mechanism is drug hydrolysis inactivation caused by beta-lactamase generation, so that the drug sensitivity can be enhanced by selecting the combination with the beta-lactamase inhibitor.
Classical beta-lactamase inhibitors all belong to beta-lactam compounds, form covalent bond Michaelis complexes with beta-lactamase, serine nucleophilically attacks amide bonds, beta-lactam rings are opened, enzymes are inactivated through rearrangement and the like, and the self structures are also damaged. The action mechanism of the avibactam is essentially different from that of a classical beta-lactamase inhibitor, beta-lactamase serine is nucleophilic to attack an avibactam amide bond, a covalent conjugate is formed by ring opening, an enzyme-inhibitor complex is obtained, the enzyme-inhibitor complex is in an enzyme inhibition form, hydrolysis does not occur, and then a lactam ring is formed by cyclization to obtain the avibactam. The nucleophilic rate is higher than that of cyclization, so that the beta-lactamase is basically in an inhibition state, and in the process, the structure of avibactam can be restored through an inverse reaction, so that the avibactam has a long-acting enzyme inhibition effect.
The detection of related substances is an important detection item for quality control in the process of drug development, and related reports are not found for the detection method of the related substances of the avibactam sodium at present, due to the structural characteristics, the main difficulty of separation is that the retention time is too short, the retention time is within 5 minutes under the conventional chromatographic conditions, the separation of the related substances from other impurities is difficult, and when organic solvents with different proportions are used as a mobile phase, the phenomenon that a main peak is changed into a double peak occurs, so that the development of the method for detecting the related substances of the avibactam has important significance in the process of drug quality control.
The abamectin sodium as a chemical drug can generate a certain amount of impurities in the synthesis process, namely the related substances, and about more than dozens of related process impurities and degradation impurities can be consulted at present.
Disclosure of Invention
The invention aims to provide an avibactam sodium analysis method with high sensitivity and good stability.
The invention relates to an avibactam sodium substance analysis method, which is characterized by analyzing decarbonylation avibactam sodium, avibactam sodium hydrolysis sodium salt byproducts, ([ (2S,5R) -7-oxo-2- (sulfonylcarbamoyl) -1, 6-diazabicyclo [3.2.1] oct-6-yl ] sulfuric acid monoester) disodium salt, avibactam disodium salt precursor, (2S,5R) -5-benzyloxyaminopiperidine-2-carboxylic acid ethyl ester oxalate, (2S,5R) -5-benzyloxyaminopiperidine-2-formamide and (2S,5R) -6- (benzyloxy) -7-oxo-1, 6-diazabicyclo [3.2.1] octane-2-formamide by using an HPLC method, the specific operation is as follows:
step one, preparing a test solution
Placing the avibactam sodium into a measuring flask, adding the mobile phase A for dissolving, and quantitatively diluting to a scale to obtain a test solution with the concentration of 1-10 mg/mL;
step two, preparation of control solution
Placing the sample solution prepared in the step one in a measuring flask, quantitatively diluting the sample solution to a scale with the mobile phase A, and shaking up to obtain a control solution with the concentration of 0.01-0.1 mg/mL;
the chromatographic conditions are as follows: the chromatographic column adopts a hydrophilic C18 column; the mobile phase A is 10 mmol/L-50 mmol/L potassium dihydrogen phosphate solution, the mobile phase B is 10 mmol/L-50 mmol/L potassium dihydrogen phosphate-acetonitrile, wherein the volume ratio of the potassium dihydrogen phosphate solution to the acetonitrile is 80: 20-30: 70, gradient elution; the column temperature is 25-40 ℃; the flow rate is 0.5mL/min to 1.2 mL/min; the sample volume is 20 mu L; the detection wavelength is 210 nm; the sample concentration is 1 mg/mL-10 mg/mL;
the gradient elution procedure was: 0 → 10min, the dosage of the mobile phase A is 100 percent, and the dosage of the mobile phase B is 0; 45min, wherein the dosage of the mobile phase A is 20 percent, and the dosage of the mobile phase B is 80 percent; 60 → 85min, the dosage of the mobile phase A is 100%, and the dosage of the mobile phase B is 0.
Further, the hydrophilic C18 column is AQ series column, HILIC series column, CAPCELL PAK ADME series column, amino column or cyano column.
Furthermore, the model of the chromatographic column is Aglient SB-Aq C18, the specification is 250mm multiplied by 4.6mm, 5 μm, and the model is added with a ghost peak trapping column: the chromatography is carried out by a 50mm chromatograph, wherein a mobile phase A is 40mmol/L potassium dihydrogen phosphate solution, a mobile phase B is 40mmol/L potassium dihydrogen phosphate-acetonitrile, and the volume ratio of the two is 60: 40, gradient elution; the column temperature was 35 ℃; the flow rate is 1.0 mL/min; the sample volume is 20 mul; the detection wavelength is 210 nm; the sample concentration is 2 mg/mL; the gradient elution procedure was: 0 → 10min, the dosage of the mobile phase A is 100 percent, and the dosage of the mobile phase B is 0; 45min, wherein the dosage of the mobile phase A is 20 percent, and the dosage of the mobile phase B is 80 percent; 60 → 85min, the dosage of the mobile phase A is 100%, and the dosage of the mobile phase B is 0.
Further, preparation of HPLC detection system suitability solution:
putting the abamectin sodium into a measuring flask, adding 1mL of 0.01mol/L sodium hydroxide solution, standing for 3min, adding 1mL0.01mol/L hydrochloric acid solution for neutralization, and then diluting to a scale with a mobile phase A to obtain a system applicability solution with the concentration of 2 mg/mL.
Further, after the system applicability solution is injected into a liquid chromatograph, the separation degree of the avibactam peak and the front peak and the rear peak in the chromatogram is greater than 1.5.
Further, HPLC detection system sensitivity solution preparation:
taking 5mL of the control solution, placing the control solution in a 100mL measuring flask, quantitatively diluting the control solution to a scale mark by using the mobile phase A, and shaking up to obtain a sensitivity solution with the concentration of 0.0005-0.005 mg/mL.
Further, after the sensitive solution is injected into a liquid chromatograph, the ratio of the signal to the noise of the avibactam peak in the chromatogram is greater than 10.
The invention has the following beneficial effects:
the invention separates degradation impurities and part of process impurities which may be generated in the avibactam sodium, and specifically comprises the following impurities: decarbonylation of avibactam sodium, avibactam sodium hydrolysis salt byproduct, ([ (2S,5R) -7-oxo-2- (sulfonylcarbamoyl) -1, 6-diazabicyclo [3.2.1] oct-6-yl ] sulfuric acid monoester) disodium salt, avibactam disodium salt precursor, (2S,5R) -5-benzyloxyaminopiperidine-2-carboxylic acid ethyl ester oxalate, (2S,5R) -5-benzyloxyaminopiperidine-2-carboxamide, (2S,5R) -6- (benzyloxy) -7-oxo-1, 6-diazabicyclo [3.2.1] octane-2-carboxamide, the first established analysis method of avibactam sodium, the invention has high detection sensitivity, good reproducibility, High stability, and the like, and is suitable for testing related substances of the avibactam sodium.
Drawings
FIG. 1 is a system suitability map for a 0.01molL sodium hydroxide concentration;
FIG. 2 is a detection spectrum using Symmetry chromatography column;
FIG. 3 is a detection spectrum using HP-cayno chromatographic column;
FIG. 4 is a detection spectrum of a column temperature of 25 ℃;
FIG. 5 is a detection spectrum of a column temperature of 40 ℃;
fig. 6 is a detection map of mobile phase pH 4.0;
fig. 7 is a detection map of mobile phase pH 7.0;
FIG. 8 is a detection profile of a 60min elution procedure;
FIG. 9 is a detection profile of an 80min elution procedure;
FIG. 10 is a detection spectrum at a wavelength of 200 nm;
FIG. 11 is a detection spectrum at a wavelength of 254 nm;
FIG. 12 is a detection spectrum of 10mmol/L salt concentration;
FIG. 13 is a graph showing a detection pattern of a mobile phase A concentration of 30 mmol/L.
Detailed Description
It will be understood by those of ordinary skill in the art that the foregoing embodiments are specific examples for carrying out the invention, and that various changes in form and details may be made therein without departing from the spirit and scope of the invention in practice.
To make the objects, aspects and advantages of the embodiments of the present invention more apparent, the following detailed description clearly illustrates the spirit of the disclosure, and any person skilled in the art, after understanding the embodiments of the disclosure, may make changes and modifications to the technology taught by the disclosure without departing from the spirit and scope of the disclosure.
The exemplary embodiments of the present invention and the description thereof are provided to explain the present invention and not to limit the present invention.
Example 1
1) Preparation of System suitability solution
Taking 50mg of abamectin sodium, placing the abamectin sodium in a 25mL measuring flask, adding 1mL of 0.01mol/L sodium hydroxide solution, standing for 3 minutes, adding 1mL of 0.01mol/L hydrochloric acid solution for neutralization, and then diluting the abamectin sodium to the scale with the mobile phase A.
2) Preparation of test solution
Taking 50mg of abamectin sodium, placing the abamectin sodium into a 25mL measuring flask, adding the mobile phase A for dissolving, and quantitatively diluting to a scale.
3) Preparation of control solution
Taking 1mL of the test solution, placing the test solution in a 100mL measuring flask, quantitatively diluting the test solution to a scale with the mobile phase A, and shaking up.
4) Preparation of sensitive solutions
5mL of the control solution was taken, placed in a 100mL measuring flask, quantitatively diluted to the scale with mobile phase A, and shaken up.
Example 2
This example is the optimization and determination of the chromatographic conditions adopted in the analytical method of avibactam sodium
(1) Detection wavelength determination
When the detection wavelength is 200nm, the response value is higher than that of 210nm, but because the organic solvent has terminal absorption, the concentration of the sample is increased, the detection is carried out at the wavelength of 210nm, and the signal-to-noise ratio of the sensitive solution at the concentration is more than 10, so that the requirements are met.
(2) Optimum chromatographic conditions
A chromatographic column: aglient SB-Aq C18(250 mm. times.4.6 mm, 5 μm), plus ghost trap column: chromatography Mr. Heng (50 mm); the mobile phase A is 40mmol/L potassium dihydrogen phosphate solution, the mobile phase B is 40mmol/L potassium dihydrogen phosphate-acetonitrile (60: 40), and gradient elution is carried out; the column temperature was 35 ℃; the flow rate is 1.0 mL/min; the sample volume is 20 mul; the detection wavelength is 210 nm; the sample concentration was 2 mg/mL. The elution procedure was: 0 → 10min, mobile phase A to mobile phase B (100: 0); 45min, mobile phase A-mobile phase B (20: 80); 60 → 85min, mobile phase A to mobile phase B (100: 0).
The optimal detection wavelength, and thus the optimal chromatographic conditions, can be determined by means of fig. 1 to 13.
(3) Methodology validation
The test article of example 1 was verified using the above-described optimal chromatographic conditions.
1) The detection limit and the quantification limit are 50mg of abamectin sodium, the abamectin sodium is placed in a 25mL measuring flask, and the abamectin sodium is quantitatively diluted by a mobile phase until the signal-to-noise ratio of a sample solution is greater than 10 and is the quantification limit, the result is 2.112 mu g/mL, the signal-to-noise ratio is greater than 3 and is the detection limit, and the result is 0.528 mu g/mL.
2) Taking 50mg of avibactam sodium linearly, placing the avibactam sodium into a 25mL measuring flask, diluting the avibactam sodium to a scale by using a mobile phase, precisely measuring 1.4mL, 1.2mL, 1.0mL, 0.8mL, 0.6mL and 0.4mL, respectively placing the avibactam sodium into a 100mL measuring flask, diluting the avibactam sodium to the scale by using the mobile phase to serve as a linear solution, measuring by adopting the chromatographic condition, performing linear regression by taking the concentration C as an abscissa and the peak area A as an ordinate, wherein the regression equation is that A is 11085C +1.798, and the correlation coefficient R is 0.9999.
3) The precision is measured according to the detection conditions by different analysts and different equipment on different dates respectively, 6 times of measurement are carried out, and the relative standard deviation of single impurities and total impurities of the test solution is calculated for 12 times, and the result is shown in table 1, and the precision is less than 5.0%.
4) The resolution is determined according to the requirement of the system applicability test under the chromatographic condition, and the resolution of the main peak and the front and rear peaks is more than 1.5.
5) The stability of the solution at room temperature is under the chromatographic condition, the abamectin sodium test solution is prepared by using the mobile phase A, is placed at room temperature, is subjected to sample injection inspection at 0 and 2 hours, and the result is shown in table 2, and the table 2 shows that the absolute value of the change of single impurity and the absolute value of the change of total impurity of the abamectin sodium solution exceed 0.1 percent after the abamectin sodium solution is placed at room temperature for 2 hours, which indicates that the abamectin sodium solution is unstable in placement at room temperature and needs to be prepared in a new way for next use.
6) Under the chromatographic condition, the stability of the solution at 4 ℃ is characterized in that a test solution of the abamectin sodium is prepared by using the mobile phase A, the test solution is placed at 4 ℃, and the sample injection inspection is carried out at 0, 2 and 4 hours, the result is shown in table 3, and the table 3 shows that the absolute value of the change of single impurity and total impurity of the abamectin sodium solution is not more than 0.1 percent after the abamectin sodium solution is placed at 4 ℃ for 9 hours, which indicates that the abamectin sodium solution is stable when placed at 4 ℃ for 9 hours.
7) Durability test solution of the abamectin sodium is taken, the durability of the chromatographic conditions is inspected by slightly changing the column temperature, the detection wavelength, the concentration of potassium dihydrogen phosphate, the acetonitrile proportion and the flow rate in the mobile phase B and the brand of the chromatographic column under the chromatographic conditions, and the result shows that the detection of related substances of the abamectin sodium is not influenced by slight change of the chromatographic conditions.
TABLE 1 precision results
TABLE 2 Room temperature solution stability results
| Time (h) | Single impurity (%) | Total impurities (%) |
| 0 | 2.32 | 2.53 |
| 1 | 3.29 | 3.87 |
| Rate of change of impurity (%) | 0.97 | 1.34 |
TABLE 34 ℃ solution stability results
| Time (h) | Single impurity (%) | Total impurities (%) |
| 0 | 2.32 | 2.53 |
| 1 | 2.30 | 2.51 |
| 2 | 2.33 | 2.54 |
| 3 | 2.33 | 2.57 |
| 4 | 2.33 | 2.56 |
| 5 | 2.28 | 2.59 |
| 6 | 2.32 | 2.60 |
| 7 | 2.35 | 2.62 |
| 8 | 2.29 | 2.61 |
| 9 | 2.30 | 2.62 |
| Rate of change of impurity (%) | 0.03 | 0.09 |
。
Claims (7)
1. An avibactam sodium substance analysis method is characterized in that the avibactam sodium substance analysis method is to analyze decarbonylation avibactam sodium, avibactam sodium hydrolysis sodium salt byproducts, ([ (2S,5R) -7-oxo-2- (sulfonylcarbamoyl) -1, 6-diazabicyclo [3.2.1] oct-6-yl ] sulfuric acid monoester) disodium salt, avibactam disodium salt precursor, (2S,5R) -5-benzyloxyaminopiperidine-2-carboxylic acid ethyl ester oxalate, (2S,5R) -5-benzyloxyaminopiperidine-2-formamide and (2S,5R) -6- (benzyloxy) -7-oxo-1, 6-diazabicyclo [3.2.1] octane-2-formamide by using an HPLC method, the specific operation is as follows:
step one, preparing a test solution
Placing the avibactam sodium into a measuring flask, adding the mobile phase A for dissolving, and quantitatively diluting to a scale to obtain a test solution with the concentration of 1-10 mg/mL;
step two, preparation of control solution
Placing the sample solution prepared in the step one in a measuring flask, quantitatively diluting the sample solution to a scale with the mobile phase A, and shaking up to obtain a control solution with the concentration of 0.01-0.1 mg/mL;
the chromatographic conditions are as follows: the chromatographic column adopts a hydrophilic C18 column; the mobile phase A is 10 mmol/L-50 mmol/L potassium dihydrogen phosphate solution, the mobile phase B is 10 mmol/L-50 mmol/L potassium dihydrogen phosphate-acetonitrile, wherein the volume ratio of the potassium dihydrogen phosphate solution to the acetonitrile is 80: 20-30: 70, gradient elution; the column temperature is 25-40 ℃; the flow rate is 0.5mL/min to 1.2 mL/min; the sample volume is 20 mu L; the detection wavelength is 210 nm; the sample concentration is 1 mg/mL-10 mg/mL;
the gradient elution procedure was: 0 → 10min, the dosage of the mobile phase A is 100 percent, and the dosage of the mobile phase B is 0; 45min, wherein the dosage of the mobile phase A is 20 percent, and the dosage of the mobile phase B is 80 percent; 60 → 85min, the dosage of the mobile phase A is 100%, and the dosage of the mobile phase B is 0.
2. The method for analyzing avibactam sodium substances according to claim 1, wherein the hydrophilic C18 column is an AQ series chromatographic column, an HILIC series chromatographic column, a CAPCELL PAK ADME series chromatographic column, an amino column or a cyano column.
3. The avibactam sodium analysis method according to claim 1, an avibactam sodium substance analysis method, characterized in that the type of the chromatographic column is aglent SB-Aq C18, the specification is 250mm x 4.6mm, 5 μm, ghost peak trapping column: the chromatography is carried out by a 50mm chromatograph, wherein a mobile phase A is 40mmol/L potassium dihydrogen phosphate solution, a mobile phase B is 40mmol/L potassium dihydrogen phosphate-acetonitrile, and the volume ratio of the two is 60: 40, gradient elution; the column temperature was 35 ℃; the flow rate is 1.0 mL/min; the sample volume is 20 mul; the detection wavelength is 210 nm; the sample concentration is 2 mg/mL; the gradient elution procedure was: 0 → 10min, the dosage of the mobile phase A is 100 percent, and the dosage of the mobile phase B is 0; 45min, wherein the dosage of the mobile phase A is 20 percent, and the dosage of the mobile phase B is 80 percent; 60 → 85min, the dosage of the mobile phase A is 100%, and the dosage of the mobile phase B is 0.
4. The avibactam sodium substance analysis method according to claim 1, characterized in that the preparation of the HPLC detection system applicability solution:
putting the abamectin sodium into a measuring flask, adding 1mL of 0.01mol/L sodium hydroxide solution, standing for 3min, adding 1mL0.01mol/L hydrochloric acid solution for neutralization, and then diluting to a scale with a mobile phase A to obtain a system applicability solution with the concentration of 2 mg/mL.
5. The method for analyzing avibactam sodium substances according to claim 4, wherein the separation degree of the avibactam peak and the front and rear peaks in the chromatogram is greater than 1.5 after the system applicability solution is injected into the liquid chromatograph.
6. The avibactam sodium substance analysis method according to claim 1, characterized in that the preparation of the HPLC detection system sensitivity solution:
taking 5mL of the control solution, placing the control solution in a 100mL measuring flask, quantitatively diluting the control solution to a scale mark by using the mobile phase A, and shaking up to obtain a sensitivity solution with the concentration of 0.0005-0.005 mg/mL.
7. The method for analyzing avibactam sodium substances according to claim 6, wherein after the sensitive solution is injected into the liquid chromatograph, the ratio of the signal to the noise of the avibactam peak in the chromatogram is greater than 10.
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| CN107894471A (en) * | 2017-11-07 | 2018-04-10 | 中山奕安泰医药科技有限公司 | A kind of detection method of AVM hereinafter Batan sodium intermediate |
| CN108362789A (en) * | 2018-01-19 | 2018-08-03 | 珠海优润医药科技有限公司 | A kind of high-efficiency liquid chromatography method for detecting of AVM hereinafter Batan sodium optical isomer |
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| CN107894471A (en) * | 2017-11-07 | 2018-04-10 | 中山奕安泰医药科技有限公司 | A kind of detection method of AVM hereinafter Batan sodium intermediate |
| CN108362789A (en) * | 2018-01-19 | 2018-08-03 | 珠海优润医药科技有限公司 | A kind of high-efficiency liquid chromatography method for detecting of AVM hereinafter Batan sodium optical isomer |
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| CN115267025A (en) * | 2022-08-10 | 2022-11-01 | 温州大学 | Method for determining drug loading amount and drug loading condition of tetramethyl heptaphyllum elegans ghost |
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