CN111398441A - Method for detecting diastereoisomers in arformoterol tartrate solution for inhalation - Google Patents
Method for detecting diastereoisomers in arformoterol tartrate solution for inhalation Download PDFInfo
- Publication number
- CN111398441A CN111398441A CN202010156018.6A CN202010156018A CN111398441A CN 111398441 A CN111398441 A CN 111398441A CN 202010156018 A CN202010156018 A CN 202010156018A CN 111398441 A CN111398441 A CN 111398441A
- Authority
- CN
- China
- Prior art keywords
- solution
- arformoterol
- detecting
- diastereoisomers
- diastereoisomer
- 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
- FCSXYHUNDAXDRH-OKMNHOJOSA-N (2r,3r)-2,3-dihydroxybutanedioic acid;n-[2-hydroxy-5-[(1r)-1-hydroxy-2-[[(2r)-1-(4-methoxyphenyl)propan-2-yl]amino]ethyl]phenyl]formamide Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O.C1=CC(OC)=CC=C1C[C@@H](C)NC[C@H](O)C1=CC=C(O)C(NC=O)=C1 FCSXYHUNDAXDRH-OKMNHOJOSA-N 0.000 title claims abstract description 30
- 229960000612 arformoterol tartrate Drugs 0.000 title claims abstract description 30
- 238000000034 method Methods 0.000 title claims abstract description 29
- BPZSYCZIITTYBL-YJYMSZOUSA-N R-Formoterol Chemical compound C1=CC(OC)=CC=C1C[C@@H](C)NC[C@H](O)C1=CC=C(O)C(NC=O)=C1 BPZSYCZIITTYBL-YJYMSZOUSA-N 0.000 claims abstract description 54
- 229960001692 arformoterol Drugs 0.000 claims abstract description 52
- 239000000243 solution Substances 0.000 claims abstract description 39
- 238000001514 detection method Methods 0.000 claims abstract description 26
- 239000011550 stock solution Substances 0.000 claims abstract description 17
- 239000012085 test solution Substances 0.000 claims abstract description 16
- 239000012490 blank solution Substances 0.000 claims abstract description 15
- 239000012088 reference solution Substances 0.000 claims abstract description 11
- 238000004458 analytical method Methods 0.000 claims abstract description 9
- 238000010812 external standard method Methods 0.000 claims abstract description 7
- 239000007788 liquid Substances 0.000 claims abstract description 6
- 238000012417 linear regression Methods 0.000 claims abstract description 5
- 238000005070 sampling Methods 0.000 claims abstract description 4
- 239000012488 sample solution Substances 0.000 claims abstract description 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 30
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- 238000010828 elution Methods 0.000 claims description 7
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 6
- 239000000126 substance Substances 0.000 claims description 6
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 5
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 239000012528 membrane Substances 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical compound [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 2
- 229910000404 tripotassium phosphate Inorganic materials 0.000 claims description 2
- 238000004587 chromatography analysis Methods 0.000 claims 1
- 239000000523 sample Substances 0.000 abstract description 8
- 230000035945 sensitivity Effects 0.000 abstract description 6
- 239000007791 liquid phase Substances 0.000 abstract description 3
- 238000011112 process operation Methods 0.000 abstract description 3
- 238000005220 pharmaceutical analysis Methods 0.000 abstract description 2
- 238000011084 recovery Methods 0.000 description 12
- 238000012360 testing method Methods 0.000 description 11
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 239000012535 impurity Substances 0.000 description 9
- 239000012071 phase Substances 0.000 description 9
- 238000004811 liquid chromatography Methods 0.000 description 6
- 239000003814 drug Substances 0.000 description 5
- 229940079593 drug Drugs 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 239000013558 reference substance Substances 0.000 description 4
- 238000007865 diluting Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 230000014759 maintenance of location Effects 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 239000008186 active pharmaceutical agent Substances 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000000945 filler Substances 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000004445 quantitative analysis Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000013112 stability test Methods 0.000 description 2
- HBAQYPYDRFILMT-UHFFFAOYSA-N 8-[3-(1-cyclopropylpyrazol-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-yl]-3-methyl-3,8-diazabicyclo[3.2.1]octan-2-one Chemical class C1(CC1)N1N=CC(=C1)C1=NNC2=C1N=C(N=C2)N1C2C(N(CC1CC2)C)=O HBAQYPYDRFILMT-UHFFFAOYSA-N 0.000 description 1
- 241001164604 Aphera Species 0.000 description 1
- 208000009079 Bronchial Spasm Diseases 0.000 description 1
- 208000014181 Bronchial disease Diseases 0.000 description 1
- 206010006482 Bronchospasm Diseases 0.000 description 1
- 208000006545 Chronic Obstructive Pulmonary Disease Diseases 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 description 1
- 239000000048 adrenergic agonist Substances 0.000 description 1
- 102000014974 beta2-adrenergic receptor activity proteins Human genes 0.000 description 1
- 108040006828 beta2-adrenergic receptor activity proteins Proteins 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 229920001577 copolymer Polymers 0.000 description 1
- BPZSYCZIITTYBL-UHFFFAOYSA-N formoterol Chemical compound C1=CC(OC)=CC=C1CC(C)NCC(O)C1=CC=C(O)C(NC=O)=C1 BPZSYCZIITTYBL-UHFFFAOYSA-N 0.000 description 1
- 229960002848 formoterol Drugs 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 231100000279 safety data Toxicity 0.000 description 1
- 238000011896 sensitive detection Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- -1 small molecule compounds Chemical class 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000012453 solvate Substances 0.000 description 1
- 239000011877 solvent mixture Substances 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 235000002906 tartaric acid Nutrition 0.000 description 1
- 239000011975 tartaric acid Substances 0.000 description 1
- 238000009210 therapy by ultrasound 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N2030/042—Standards
- G01N2030/047—Standards external
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/065—Preparation using different phases to separate parts of sample
Abstract
The invention relates to the technical field of pharmaceutical analysis, in particular to a method for detecting diastereoisomers in an arformoterol tartrate solution for inhalation, which comprises the following steps: preparing a blank solution, a test solution and a reference stock solution; respectively sampling the sample solution and the reference solution with a certain concentration gradient, detecting by using a high performance liquid chromatograph, and recording a chromatogram; performing linear regression analysis on each mass concentration and chromatogram peak area of the reference solution to obtain a regression equation and a correlation coefficient, and preparing a standard curve; and calculating the content of the arformoterol diastereoisomer according to an external standard method by using the peak area of the arformoterol diastereoisomer in the chromatogram of the test solution. The liquid phase detection method provided by the invention has the advantages of simple sample treatment process operation, accurate detection result, high sensitivity and good stability.
Description
Technical Field
The invention belongs to the technical field of pharmaceutical analysis, relates to a detection method of arformoterol diastereoisomer, and particularly relates to a determination method of arformoterol diastereoisomer in arformoterol tartrate solution for inhalation.
Background
Arformoterol tartrate is a tartaric acid solvate of arformoterol, the chemical name of arformoterol is N- [ 2-hydroxy-5- [ (1R) -1-hydroxy-2- [ [ (1R) -2- (4-methoxyphenyl) -1-methylethyl ] amino ] ethyl ] phenyl ] formamide, (2R,3R) -2, 3-dihydroxy, the chemical structure is shown as formula I, and the arformoterol tartrate is a selective long-acting β 2-adrenoceptor agonist and is used for treating bronchospasm of COPD patients.
To ensure medication safety, each impurity in the Active Pharmaceutical Ingredient (API) must be evaluated for safety, i.e., establish safety-ensuring impurity limits. According to the requirement of international harmonization (ICH) of the technical standard of registration of human drugs, if the amount of a single impurity in the raw material drug exceeds 0.05%, the report is required; the amount of individual impurities, for example, exceeding 0.1%, needs to be confirmed; if the single impurity content exceeds 0.15%, safety data support is required.
Arformoterol ((R, R) configuration) has two chiral centers, i.e. four configurations: (R, R), (S, S), (R, S) and (S, R). Enantiomers are in the (S, S) configuration and diastereomers are in the (R, S) and (S, R) configurations. There are many reported detection methods for the arformoterol enantiomer, but the diastereomer ((R, S) and (S, R) configuration) has no proprietary, highly sensitive detection method. Therefore, the development of a method for detecting the arformoterol diastereoisomer is essential.
The present invention provides arformoterol diastereomers of formula II.
In the research on arformoterol analysis method, arformoterol and its enantiomer are found to have more detection method reports, but arformoterol diastereomer (shown in formula II) impurities in the raw material medicaments are only reported, and the detection method of arformoterol diastereomer in part of reports has less description, and meanwhile, the problems of poor specificity, low sensitivity and the like of the detection method exist. Therefore, the development of a method for detecting the arformoterol diastereoisomer is essential.
Disclosure of Invention
The technical problem to be solved by the invention is to overcome the defects of the prior art and provide a liquid chromatography method which has the characteristics of simple operation, good precision, high recovery rate, low detection limit and the like and can be used for detecting the arformoterol diastereomer impurity in the arformoterol tartrate solution for inhalation.
In order to solve the technical problems, the invention provides a method for detecting arformoterol diastereomer by liquid chromatography, which uses high performance liquid chromatography for detection and adopts an external standard method quantitative method for analysis, and specifically comprises the following steps:
preparing a blank solution, a test solution and a reference stock solution;
respectively sampling the sample solution and the reference solution with a certain concentration gradient, detecting by using a high performance liquid chromatograph, and recording a chromatogram;
performing linear regression analysis on each mass concentration and chromatogram peak area of the reference solution to obtain a regression equation and a correlation coefficient, and preparing a standard curve; and calculating the content of the arformoterol diastereoisomer according to an external standard method by using the peak area of the arformoterol diastereoisomer in the chromatogram of the test solution.
Furthermore, gradient elution is adopted during the measurement by a high performance liquid chromatograph.
Further, the mobile phase A is acetonitrile with the volume fraction of 15 percent and K with the volume fraction of 85 percent3PO4·3H2O, and the mobile phase B is an acetonitrile solution with the volume fraction of 85 percent.
Further, gradient elution was performed as follows:
furthermore, in the chromatographic condition, the flow rate is 0.2m L/min-1.0 m L/min, the detection wavelength is 210nm-280nm, and the column temperature is 25-40 ℃.
Further, the regression equation is that y is 175.44x-2.1976, and the correlation coefficient R is 0.9963.
Further, the blank solution is water, and the concentration of the test solution is 0.2mg/m L.
Further, the preparation method of the mobile phase A comprises the steps of taking acetonitrile 50m L-150 m L and 5.3 g/L K3PO4·3H2O (850m L-950 m L), mixing uniformly, adjusting pH value to 8.0-12.5 with phosphoric acid, and filtering with 0.45 μm water system filter membrane.
Further, the concentration of the control stock solution is 0.4mg/m L.
Advantageous effects
According to the technical scheme, the arformoterol diastereoisomer is subjected to silica gel and polystyrene composite filler chromatographic column, and the chromatographic column has the advantages of the arformoterol diastereoisomer and the arformoterol. The chromatographic column using the vinyl alcohol copolymer as the matrix can keep the surface wet even if the carbon carrying amount is high. The porous structure has a large enough average pore size to produce ideal separation effects on small molecule compounds, polypeptides and small molecule proteins. The most significant feature is the elution order of the matrix alkylated fillers, whose retention capacity is directly proportional to the carbon chain length. The content of the arformoterol diastereoisomer is controlled by using the analysis method so as to meet the quality control requirement of the arformoterol tartrate solution for inhalation.
The method for detecting the arformoterol diastereoisomer by using the liquid chromatography is an arformoterol diastereoisomer detection method newly established in the field, and can effectively detect the arformoterol diastereoisomer content in an arformoterol tartrate solution for inhalation.
The liquid phase detection method provided by the invention has the advantages of simple sample treatment process operation, accurate detection result, high sensitivity and good stability, and can detect 50ppm of arformoterol diastereoisomer contained in the arformoterol tartrate solution for inhalation. According to the guidance principle of ICH for the limit of the arformoterol diastereoisomer, the method provided by the invention completely achieves the detection index, so that the method can be used for detecting the arformoterol diastereoisomer.
Drawings
Figure 1 is an arformoterol diastereomer standard curve;
FIG. 2 is a chromatogram of an empty solution in an example specificity experiment;
FIG. 3 is a chromatogram of a control solution from a specificity experiment of an example;
FIG. 4 is a chromatogram of a sample from a specific experiment of an example.
Detailed Description
The following detailed description of embodiments of the invention refers to the accompanying drawings.
The technical problem to be solved by the invention is to overcome the defects of the prior art and provide a liquid chromatography method for measuring the content of the arformoterol diastereoisomer, which has the characteristics of simple operation, good precision, high recovery rate, low detection limit and the like, and can be used for controlling the quality of arformoterol diastereoisomer impurities in an arformoterol tartrate solution for inhalation. In order to solve the technical problems, the invention provides a method for detecting arformoterol diastereoisomer by liquid chromatography, which uses high performance liquid chromatography for detection and adopts an external standard method quantitative method for analysis, and specifically comprises the following steps:
step 1) preparing a blank solution: water was taken as the blank solution.
And step 2) preparing a reference substance stock solution, namely taking 10mg of a standard substance into a 25m L volumetric flask, dissolving the standard substance by using methanol (ultrasonic if necessary), fixing the volume to a scale, and shaking up to obtain the arformoterol diastereoisomer stock solution (0.4mg/m L).
And step 3) preparing a reference substance solution, namely taking the reference substance stock solution in a volumetric flask with the volume of 1m L-500 m L, dissolving the reference substance stock solution in a blank solution, fixing the volume to a scale, and shaking up to obtain the arformoterol diastereoisomer solution (0.2 mu g/m L).
And step 4) preparing a test solution, namely taking 2mg of arformoterol tartrate, precisely weighing, placing the arformoterol tartrate in a 10m L volumetric flask, dissolving the arformoterol tartrate in a 0.2 mu g/m L reference solution, fixing the volume, and shaking up to obtain the test solution.
And 5) taking a blank solution, a test solution and a reference solution 20 mu L, and recording chromatograms.
Chromatographic conditions are as follows: the chromatographic column is as follows: ApHera C-18Polymer Column; flow ofPhase A taking acetonitrile 50m L-150 m L and 5.3 g/L (K)3PO4·3H2O)850m L-950 m L, the mixture is mixed evenly, the pH value is adjusted to 8.0-12.5 by phosphoric acid, 0.45 mu m water system filter membrane is used for filtration, the flow rate of the mobile phase B is acetonitrile, the flow rate is 0.2m L/min-1.0 m L/min, the detection wavelength is 210nm-280nm, the column temperature is 25 ℃ to 40 ℃, and the mobile phase uses the elution gradient described in the following table:
| time (min) | Mobile phase A (%) | Mobile phase B (%) | Remarks for note |
| 0-15 | 100 | 0 | Linearity |
| 5-35 | 100→71 | 0→29 | Initial conditions |
| 35-55.1 | 71→100 | 29→0 | Return to initial elution |
Step 6) carrying out linear regression analysis on each concentration data and chromatogram peak area of the reference solution with a certain concentration gradient to obtain a regression equation and a correlation coefficient, and preparing a standard curve; and calculating the content of the arformoterol diastereoisomer according to an external standard method by utilizing the peak area of the arformoterol diastereoisomer in the chromatogram of the test solution.
Preferably, in the step 2), the concentrations of the control solutions are 0.05 μ g/m L, 0.10 μ g/m L, 0.15 μ g/m L, 0.20 μ g/m L, 0.25 μ g/m L and 0.30 μ g/m L, respectively.
Preferably, in the step 4), the concentration of the test solution is 0.2 μ g/m L.
Preferably, in said step 5), the mobile phase A of the chromatographic conditions is a mixture containing 15% acetonitrile and 85% K by volume fraction3PO4·3H2O solution, and the mobile phase B is acetonitrile solution containing 85% of volume fraction.
Specifically, in the chromatographic conditions, the flow rate is 1.0m L/min, the column temperature is 40 ℃, and the sample injection amount is 20 mu L.
Specifically, in the step 4), y is 175.44x-2.1976, and the correlation coefficient R is 0.99.
The method for detecting the arformoterol diastereoisomer by using the liquid chromatography is a method for detecting the arformoterol diastereoisomer newly established in the field, is simple in sample processing process operation, accurate in detection result, high in sensitivity and good in stability, and can be used for effectively detecting the arformoterol diastereoisomer content in an arformoterol tartrate solution for inhalation. According to the liquid phase detection method provided by the invention, the method provided by the invention completely reaches the detection index according to the guidance principle of ICH on impurity limit, so that the method can be used for detecting arformoterol diastereomer.
3. Experimental part
3.1 solution preparation
3.1.1 blank solution preparation: water was taken as the blank solution.
3.1.2 reference stock solution, taking 10mg standard substance to a 25m L volumetric flask, dissolving with methanol (ultrasonic if necessary), fixing the volume to the scale, shaking up to obtain arformoterol diastereoisomer stock solution (0.4mg/m L).
3.1.3 control solution A control stock solution 1m L-500 m L volumetric flask was taken, dissolved in a blank solution, and the volume was fixed to the scale and shaken up to be used as a formoterol diastereoisomer solution (0.2. mu.g/m L).
3.1.4 preparation of test solution, 2mg of arformoterol tartrate is precisely weighed and placed in a 10m L volumetric flask, dissolved by 0.2 mu g/m L reference solution and subjected to constant volume and shaking uniformly to be used as the test solution.
3.2 methodological investigation
3.2.1 specificity test
Sampling blank solution, reference solution, and test solution, and collecting chromatogram, respectively, and the results are shown in FIGS. 1-3. Test results show that the blank solution has no interference, other peaks in the sample have no interference on the arformoterol diastereoisomer chromatographic peak, and the method has good specificity.
3.2.2 precision test
The 0.2 μ g/m L control solution was injected into the chromatograph for 6 times with a volume of 20 μ L under HPLC detection conditions, the retention time and peak area were recorded, and the results were evaluated as shown in Table 1.
TABLE 1 results of systematic precision test
The result shows that the retention time RSD% is 0.8%, the peak area RSD% is 1.3%, and the method shows that the system precision is good and completely meets the requirement of drug analysis and test.
3.2.3 Linear and Range test
A proper amount of arformoterol diastereoisomer control solution is precisely transferred and diluted by blank solution to be 0.05 mu g/m L, 0.10 mu g/m L, 0.15 mu g/m L, 0.20 mu g/m L, 0.25 mu g/m L and 0.30 mu g/m L, the solutions are respectively injected into a high performance liquid chromatograph, the injection volume is 20 mu L, linear regression is carried out according to the mass concentration and the peak area, the regression equation is 175.44x-2.1976, the correlation coefficient R is 0.99, and the results are shown in Table 2.
TABLE 2 results of the arformoterol diastereomer linearity and range tests
The results show that the linearity of the arformoterol tartrate diastereomer is good in the range of 0.05 μ g/m L to 0.30 μ g/m L, and the correlation coefficient R is 0.99.
3.2.4 detection Limit and quantitation Limit tests
Adjusting the sensitivity of the instrument, taking a proper amount of arformoterol diastereoisomer stock solution, gradually diluting and injecting samples, injecting the sample with the volume of 20 mu L to ensure that the height of a main peak is 2.3 times of the baseline noise, and recording a chromatogram map to obtain the arformoterol diastereoisomer with the minimum detection limit of 0.025 mu g/m L (12.5%).
Adjusting the sensitivity of the instrument, taking a proper amount of arformoterol diastereoisomer stock solution, gradually diluting and injecting samples, injecting the sample with the volume of 20 mu L to ensure that the height of a main peak is 10.3 times of the baseline noise, and recording a chromatogram map to obtain the arformoterol diastereoisomer with the minimum detection limit of 0.05 mu g/m L (25%).
3.2.5 recovery test
And (3) precisely weighing 10mg of arformoterol tartrate in a recovery rate stock solution 1, placing the arformoterol tartrate in a 10m L volumetric flask, adding a solvent mixture, dissolving the arformoterol tartrate by ultrasonic treatment, and shaking the arformoterol tartrate to a constant volume.
And 2, precisely weighing 1mg of arformoterol diastereoisomer, putting the arformoterol diastereoisomer in a 25m L volumetric flask, adding methanol to dissolve to a constant volume, shaking up, putting a 1m L solution in a 20m L volumetric flask, adding water to dissolve to a constant volume, and shaking up.
And (3) precisely measuring recovery rate stock solutions 1 and 2 in different proportions respectively, placing the solutions in a 10m L volumetric flask, and diluting the solutions respectively into solutions with 75%, 100% and 150% recovery rates in parallel.
The recovery ratio stock solution 2 and the recovery ratio solution were taken and injected into a high performance liquid chromatograph, the injection volume was 20 μ L, the concentration of each peak in the recovery ratio solution was calculated according to an external standard method, and the test results are shown in table 3.
TABLE 3 results of the spiked recovery test
3.2.6 solution stability test
The yield solution (100%) was taken out and left at room temperature for 36 hours, and the stability of the solution was examined by injecting the solution into a chromatograph under HPLC conditions at 0, 3, 8, and 36 hours, respectively, at an injection volume of 10-25. mu. L, and recording the chromatogram, the results are shown in Table 4.
TABLE 4 recovery rate solution stability test
| Time (hours) | 0 | 3 | 8 | 36 | Mean value of | RSD％ |
| Peak area of spiked solution | 33.536 | 32.375 | 34.088 | 32.482 | 33.120 | 2.5 |
The result shows that the recovery rate solution is placed for 36 hours at room temperature, and the stability of the solution is good.
In summary, the above embodiments are merely preferred embodiments of the present invention, and are not intended to limit the scope of the present invention, and any modifications, equivalents, improvements, etc. made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (9)
1. A method for detecting diastereoisomers in an arformoterol tartrate solution for inhalation, comprising the steps of:
preparing a blank solution, a test solution and a reference stock solution;
respectively sampling the sample solution and the reference solution with a certain concentration gradient, detecting by using a high performance liquid chromatograph, and recording a chromatogram;
performing linear regression analysis on each mass concentration and chromatogram peak area of the reference solution to obtain a regression equation and a correlation coefficient, and preparing a standard curve; and calculating the content of the arformoterol diastereoisomer according to an external standard method by using the peak area of the arformoterol diastereoisomer in the chromatogram of the test solution.
2. The method for detecting diastereoisomers of arformoterol tartrate for inhalation according to claim 1, wherein the gradient elution is performed by HPLC.
3. Method for detecting diastereoisomers of arformoterol tartrate for inhalation according to claim 2, wherein the mobile phase A comprises acetonitrile 15% and K85% in volume fraction3PO4·3H2O, and the mobile phase B is an acetonitrile solution with the volume fraction of 85 percent.
4. Method for the detection of diastereoisomers in arformoterol tartrate for inhalation according to claim 3, wherein the gradient elution is carried out as follows:
5. the method for detecting diastereoisomers of arformoterol tartrate for inhalation according to claim 1, wherein the flow rate is 0.2m L/min to 1.0m L/min, the detection wavelength is 210nm to 280nm, and the column temperature is 25 ℃ to 40 ℃ under the chromatography conditions.
6. The method for detecting diastereoisomers of arformoterol tartrate for inhalation according to claim 1, wherein the regression equation is 175.44x-2.1976 and the correlation coefficient R is 0.9963.
7. The method for detecting diastereoisomers of arformoterol tartrate for inhalation according to claim 1, wherein the blank solution is water, and the test solution has a concentration of 0.2 μ g/m L.
8. The method for detecting diastereoisomers of arformoterol tartrate for inhalation according to claim 3, wherein the mobile phase A is prepared by taking acetonitrile 50m L-150 m L and 5.3 g/L of K3PO4·3 H2O850 m L-950 m L, mixing evenly, adjusting the pH value to 8.0-12.5 by phosphoric acid, and filtering by a 0.45 mu m water system filter membrane.
9. The method for detecting diastereoisomers of arformoterol tartrate for inhalation according to claim 1, wherein the stock solution of the control substance has a concentration of 0.4mg/m L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010156018.6A CN111398441A (en) | 2020-03-09 | 2020-03-09 | Method for detecting diastereoisomers in arformoterol tartrate solution for inhalation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010156018.6A CN111398441A (en) | 2020-03-09 | 2020-03-09 | Method for detecting diastereoisomers in arformoterol tartrate solution for inhalation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111398441A true CN111398441A (en) | 2020-07-10 |
Family
ID=71432284
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010156018.6A Pending CN111398441A (en) | 2020-03-09 | 2020-03-09 | Method for detecting diastereoisomers in arformoterol tartrate solution for inhalation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111398441A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114924020A (en) * | 2022-05-30 | 2022-08-19 | 上海奥科达生物医药科技有限公司 | Quality control method for diastereoisomer in arformoterol tartrate inhalation solution |
| CN115015459A (en) * | 2022-05-30 | 2022-09-06 | 上海奥科达生物医药科技有限公司 | Method for detecting diastereoisomers in formoterol fumarate inhalation solution |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110376313A (en) * | 2019-08-20 | 2019-10-25 | 广州健康元呼吸药物工程技术有限公司 | A method of impurity in detection formoterol fumarate or its related preparations |
-
2020
- 2020-03-09 CN CN202010156018.6A patent/CN111398441A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110376313A (en) * | 2019-08-20 | 2019-10-25 | 广州健康元呼吸药物工程技术有限公司 | A method of impurity in detection formoterol fumarate or its related preparations |
Non-Patent Citations (4)
| Title |
|---|
| SAMUEL AKAPO ET AL.: "Chiral HPLC analysis of formoterol stereoisomers and thermodynamic study of their interconversion in aqueous pharmaceutical formulations", 《JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS》 * |
| 国家药典委员会: "《中华人民共和国药典 2015年版二部》", 30 June 2015, 中国医药科技出版社 * |
| 昭和电工科学仪器（上海）有限公司: "根据药典标准分析富马酸福莫特罗", 《HTTPS://IBOOK.ANTPEDIA.COM/X/586818.HTML》 * |
| 欧洲药品质量管理局: "《EUROPEAN PHARMACOPOEIA 9.0》", 31 January 2017 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114924020A (en) * | 2022-05-30 | 2022-08-19 | 上海奥科达生物医药科技有限公司 | Quality control method for diastereoisomer in arformoterol tartrate inhalation solution |
| CN115015459A (en) * | 2022-05-30 | 2022-09-06 | 上海奥科达生物医药科技有限公司 | Method for detecting diastereoisomers in formoterol fumarate inhalation solution |
| CN116165319A (en) * | 2022-05-30 | 2023-05-26 | 上海奥科达医药科技股份有限公司 | Method for detecting diastereoisomers in formoterol fumarate inhalation solution |
| CN116298046A (en) * | 2022-05-30 | 2023-06-23 | 上海奥科达医药科技股份有限公司 | Quality control method for diastereoisomers in arformoterol tartrate inhalation solution |
| CN116298046B (en) * | 2022-05-30 | 2023-11-17 | 上海奥科达医药科技股份有限公司 | Quality control method for diastereoisomers in arformoterol tartrate inhalation solution |
| CN116165319B (en) * | 2022-05-30 | 2023-11-17 | 上海奥科达医药科技股份有限公司 | Method for detecting diastereoisomers in formoterol fumarate inhalation solution |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111398441A (en) | Method for detecting diastereoisomers in arformoterol tartrate solution for inhalation | |
| CN104749269A (en) | Method for determining enantiomer impurity in alogliptin crude drug and preparation by virtue of HPLC | |
| CN115060832B (en) | Method for detecting contents of oxcarbazepine and preservatives in oxcarbazepine oral suspension | |
| CN113866337B (en) | Mass analysis method for separating and measuring oseltamivir phosphate isomer | |
| Peinado et al. | Rapid method for micro-analysis of endogenous amino acid neurotransmitters in brain perfusates in the rat by isocratic HPLC-EC | |
| CN113702514A (en) | Method for determining atorvastatin calcium related impurity I | |
| CN111220730A (en) | Analysis method of related substances in irbesartan and hydrochlorothiazide compound preparation | |
| CN114839287B (en) | Method for detecting sodium myristate in miboplatin | |
| CN113588837B (en) | Detection method of moxifloxacin hydrochloride related substance | |
| CN114689737B (en) | Analysis method of S-o-chlorophenylglycine methyl tartrate related substances | |
| CN109030668B (en) | High performance liquid phase analysis method of gadoxetic acid disodium intermediate | |
| CN114200050B (en) | HPLC detection method for content of related substances in p-bromoanisole | |
| CN115236255B (en) | Method for detecting related substances of loxoprofen sodium | |
| CN114062567B (en) | Separation and detection method of (1R, 2S) -2- (3, 4-difluorophenyl) cyclopropylamine hydrochloride and related substances thereof | |
| CN116930368B (en) | Detection method of settop alcohol isomer | |
| CN113419006B (en) | Liquid phase analysis method of aprepitant optical isomer | |
| CN115184527B (en) | Detection method of heliotropin impurity | |
| CN113960201B (en) | Method for detecting 4-dimethylaminopyridine in entecavir | |
| CN112305100B (en) | Method for detecting content of genotoxic impurity benzyl bromide in medicine | |
| CN115128184B (en) | Method for determining thiourea content in pramipexole dihydrochloride raw material by using HPLC external standard method | |
| CN106918658B (en) | axitinib raw materials and analysis method of related substances in preparation thereof | |
| CN115453033A (en) | Method for detecting content of vitamin D in vitamin D drops | |
| CN110658263A (en) | Ramelteon enantiomer detection method and quality control standard thereof | |
| CN106841405B (en) | Method for detecting related substances of ephedrine hydrochloride tablets | |
| CN116699038A (en) | Detection and analysis method for related substances in lidocaine gel plaster |
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: 20200710 |
|
| RJ01 | Rejection of invention patent application after publication |