Method for detecting enantiomer in esomeprazole sodium medicine
阅读说明：本技术 一种艾司奥美拉唑钠药物中对映异构体的检测方法 (Method for detecting enantiomer in esomeprazole sodium medicine ) 是由 佘鹏鹏 徐浩宇 蔡伟 徐春来 周培培 蔡丽娟 刘政伟 董达文 李浩冬 吴青青 于 2019-10-15 设计创作，主要内容包括：本发明公开了一种艾司奥美拉唑钠药物中对映异构体的检测方法,所述检测方法包括：采用反相高效液相色谱法,以表面有纤维素-三(3,5-二甲基苯基氨基甲酸酯)共价键合硅胶为色谱柱填料,采用紫外检测器；流动相为碳酸氢铵水溶液、水溶性非质子有机溶剂以及低级烷醇所形成的混合体系；以所述流动相进行洗脱,检测艾司奥美拉唑钠药物中的对映异构体含量。相比于现有技术,本发明的检测方法具有更高的专属性,有效地避免了稳定性放样过程中主要降解杂质(加速和长期稳定性过程中产生)及可能存在的工艺杂质和中间体对于对映异构体检测的干扰；更能适用于注射用艾司奥美拉唑钠,具有更优越的精密度、准确度和耐用性。(The invention discloses a detection method of enantiomer in esomeprazole sodium medicine, which comprises the following steps: adopting reversed-phase high performance liquid chromatography, using cellulose-tri (3, 5-dimethyl phenyl carbamate) covalent bonding silica gel on the surface as chromatographic column packing, and adopting an ultraviolet detector; the mobile phase is a mixed system formed by ammonium bicarbonate aqueous solution, water-soluble aprotic organic solvent and lower alkanol; eluting with the mobile phase, and detecting the content of enantiomer in the esomeprazole sodium medicament. Compared with the prior art, the detection method has higher specificity, and effectively avoids the main degradation impurities (generated in the acceleration and long-term stability processes) in the stability lofting process and the possible interference of process impurities and intermediates on the enantiomer detection; the injection is more suitable for esomeprazole sodium for injection, and has more excellent precision, accuracy and durability.)
1. A method for detecting enantiomers in esomeprazole sodium medicine comprises the following steps:
adopting reversed-phase high performance liquid chromatography, using cellulose-tri (3, 5-dimethyl phenyl carbamate) covalent bonding silica gel on the surface as chromatographic column packing, and adopting an ultraviolet detector; the mobile phase is a mixed system formed by ammonium bicarbonate aqueous solution, water-soluble aprotic organic solvent and lower alkanol; eluting with the mobile phase, and detecting the content of enantiomer in the esomeprazole sodium medicament.
2. The detection method according to claim 1, wherein the esomeprazole sodium drug is an esomeprazole sodium raw drug or an esomeprazole sodium injection, and preferably, the esomeprazole sodium injection is a lyophilized powder injection, i.e. esomeprazole sodium for injection.
3. The detection method according to claim 1, wherein the water-soluble aprotic organic solvent in the mobile phase is one selected from acetonitrile, tetrahydrofuran, acetone and dioxane, preferably acetonitrile.
4. The detection method according to claim 1, wherein the lower alkanol in the mobile phase is one selected from methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, and t-butanol; preferably, it is methanol.
5. The detection method according to claim 1, wherein the volume ratio of the ammonium bicarbonate aqueous solution to the water-soluble aprotic organic solvent to the lower alkanol in the mobile phase is (48-52): (29-31): (19-21), preferably 50:30: 20.
6. The assay of claim 1 wherein the mobile phase comprises an aqueous ammonium bicarbonate solution at a concentration of 10mM to 50mM, preferably 20 mM.
7. The detection method according to claim 6, wherein the mobile phase is a mixed system of 20mM ammonium bicarbonate aqueous solution, acetonitrile and methanol, wherein the volume ratio of 20mM ammonium bicarbonate aqueous solution to acetonitrile to methanol is (48-52): 29-31): 19-21, preferably 50:30: 20.
8. The detection method of any one of claims 1 to 7, the chromatographic conditions of the reverse phase high performance liquid chromatography further comprising:
the flow rate is 0.7-0.9 ml/min, preferably 0.8 ml/min;
the column temperature is 23-27 ℃, and 25 ℃ is preferred;
the detection wavelength is 298 nm-306 nm, preferably 302 nm;
the amount of the sample is 10 to 50. mu.l, preferably 20. mu.l.
9. The detection method of any one of claims 1 to 7, further comprising:
preparation of R-omeprazole solution: precisely weighing about 12mg of R-omeprazole reference substance, placing the R-omeprazole reference substance into a 20mL measuring flask, dissolving the R-omeprazole reference substance by using a diluent, diluting the R-omeprazole reference substance to a scale, and shaking up; precisely measuring 1mL of the solution, placing the solution in a 100mL measuring flask, dissolving the solution by using a diluent, diluting the solution to a scale, and shaking up; here, the diluent is a mixture of 1mM aqueous sodium hydroxide solution and acetonitrile, preferably, the volume ratio of 1mM aqueous sodium hydroxide solution to acetonitrile is 90: 10; or
Preparing a test solution: taking about 15mg of esomeprazole sodium medicine, placing the medicine in a 50mL measuring flask, dissolving the medicine by using a proper amount of diluent, diluting the medicine to a scale, and shaking up; here, the diluent is a mixture of 1mM aqueous sodium hydroxide solution and acetonitrile, preferably, the volume ratio of 1mM aqueous sodium hydroxide solution to acetonitrile is 90: 10.
10. The detection method of any one of claims 1 to 7, further comprising:
and (3) injecting the solutions into a chromatograph, recording a chromatogram, and calculating the content of the enantiomer according to the peak area by an external standard method according to the peak area of the R-omeprazole chromatographic peak.
Technical Field
The invention relates to the technical field of drug analysis, and particularly relates to a method for detecting enantiomers in an esomeprazole sodium drug.
Background
Esomeprazole sodium is a chiral Proton Pump Inhibitor (PPis), is mainly used for treating gastroesophageal reflux disease (GORD) clinically, and is also combined with antibiotics to treat duodenal ulcer caused by helicobacter pylori. It was developed by the pharmaceutical company of Aslicon, and approved by FDA 2 months in 2001 for marketing in the United states, Europe, etc.; is the S-isomer of omeprazole, which can be concentrated and converted to the active form in the highly acidic environment of parietal cells, is more potent than omeprazole and is more effective in controlling gastric acid levels, alleviating pain symptoms and promoting healing. Omeprazole is mainly metabolized by CYP2C19, individual difference is large, esomeprazole sodium is metabolized by CYP3A4 and CYP2C19, and acid inhibition effect is strong. Trace amount of R-configuration impurity may be introduced in the synthesis process to produce R-omeprazole, so that enantiomer exists in the injection to influence the product quality, and the enantiomer in the finished product needs to be controlled.
The chemical name of esomeprazole sodium is: sodium 5-methoxy-2- ((S) - ((4-methoxy-3, 5-dimethyl-2-pyridyl) methyl) sulfinyl-1H-benzimidazole having the structural formula shown below.
The Chinese pharmacopoeia 2015 edition discloses a method for measuring enantiomers in esomeprazole sodium raw material medicine, and an AGP chiral column (alpha 1-acid glycoprotein bonded silica gel column) is combined with a high performance liquid chromatography system for separation and detection.
Other patents adopt normal-phase high performance liquid chromatography (patent CN106093233A), the concentration of a used sample is large, the normal-phase method has certain limitations (the prescription contains strong-polarity auxiliary material sodium hydroxide, the retention time of a chromatographic peak is continuously fluctuated, the reproducibility of the method is poor), and the column effect of the normal-phase chromatographic column is rapidly reduced in the detection process of esomeprazole sodium for injection. Therefore, a reliable detection method for the esomeprazole sodium enantiomer for injection is newly established, and the method has important significance for quality control.
Disclosure of Invention
The following is a summary of the subject matter described in detail herein. This summary is not intended to limit the scope of the claims.
The inventor of the application develops an analysis and detection method for enantiomers in esomeprazole sodium drug, the method is good in specificity, possible process impurities, intermediates and degradation impurities can be effectively separated from main components and enantiomers, the method is high in sensitivity, good in repeatability, high in accuracy and good in durability, and the method can be used for detecting the content of the enantiomers in the esomeprazole sodium drug substance or a preparation thereof and effectively controlling the quality of products.
The invention aims to provide a method for detecting enantiomers in esomeprazole sodium medicine.
The invention provides a method for detecting enantiomers in esomeprazole sodium medicine, which comprises the following steps:
adopting reversed-phase high performance liquid chromatography, using cellulose-tri (3, 5-dimethyl phenyl carbamate) covalent bonding silica gel on the surface as chromatographic column packing, and adopting an ultraviolet detector; the mobile phase is a mixed system formed by ammonium bicarbonate aqueous solution, water-soluble aprotic organic solvent and lower alkanol; eluting with the mobile phase, and detecting the content of enantiomer in the esomeprazole sodium medicament.
In an embodiment of the present application, the esomeprazole sodium drug is an esomeprazole sodium raw drug or an esomeprazole sodium injection, and preferably, the esomeprazole sodium injection is a lyophilized powder injection, i.e., esomeprazole sodium for injection.
In an embodiment of the present application, the water-soluble aprotic organic solvent in the mobile phase is selected from one of acetonitrile, tetrahydrofuran, acetone and dioxane, preferably acetonitrile.
In an embodiment of the present application, the lower alkanol in the mobile phase is selected from one of methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol and tert-butanol; preferably, it is methanol.
In an embodiment of the application, the concentration of the aqueous ammonium bicarbonate solution in the mobile phase is between 10mM and 50mM, preferably 20 mM.
In the embodiment of the application, the volume ratio of the ammonium bicarbonate aqueous solution to the water-soluble aprotic organic solvent to the lower alkanol in the mobile phase is (48-52) to (29-31) to (19-21), and preferably is 50:30: 20.
In a preferred embodiment of the application, the mobile phase is a mixed system formed by 20mM ammonium bicarbonate aqueous solution, acetonitrile and methanol, wherein the volume ratio of the 20mM ammonium bicarbonate aqueous solution to the acetonitrile to the methanol is (48-52): 29-31): 19-21, preferably 50:30: 20.
In embodiments of the present application, the chromatographic conditions of the reverse phase high performance liquid chromatography further comprise:
the flow rate is 0.7-0.9 ml/min, preferably 0.8 ml/min;
the column temperature is 23-27 ℃, and the preferable condition is 25 ℃;
the detection wavelength is 298 nm-306 nm, preferably 302 nm;
the amount of the sample is 10 to 50. mu.l, preferably 20. mu.l.
In some examples of the present application, the chromatographic conditions of the reverse phase high performance liquid chromatography further comprise: the chromatographic column is Phenomenex chirall MZ (2) -RH, specification 4.6X 250mm, 5 μm.
In an embodiment of the present application, the detection method further comprises:
preparation of R-omeprazole solution: precisely weighing about 12mg of R-omeprazole reference substance, placing the R-omeprazole reference substance into a 20mL measuring flask, dissolving the R-omeprazole reference substance by using a diluent, diluting the R-omeprazole reference substance to a scale, and shaking up; precisely measuring 1mL of the solution, placing the solution in a 100mL measuring flask, dissolving the solution by using a diluent, diluting the solution to a scale, and shaking up; here, the diluent is a mixture of 1mM aqueous sodium hydroxide solution and acetonitrile, preferably, the volume ratio of 1mM aqueous sodium hydroxide solution to acetonitrile is 90: 10.
In an embodiment of the present application, the detection method further comprises:
preparing a test solution: taking about 15mg of esomeprazole sodium medicine, placing the medicine in a 50mL measuring flask, dissolving the medicine by using a proper amount of diluent, diluting the medicine to a scale, and shaking up; here, the diluent is a mixture of 1mM aqueous sodium hydroxide solution and acetonitrile, preferably, the volume ratio of 1mM aqueous sodium hydroxide solution to acetonitrile is 90: 10.
In an embodiment of the present application, the detection method further comprises:
injecting the solutions into chromatograph, recording chromatogram, calculating enantiomer content according to peak area by external standard method based on peak area of R-omeprazole chromatogram
In the embodiment of the invention, the method for detecting the esomeprazole sodium enantiomer is provided, wherein an external standard method is adopted for accurate quantification according to peak area when the content of the enantiomer is calculated.
In an embodiment of the present application, the detection method further comprises:
preparing a sensitivity solution: taking 2.5mL to 100mL of the R-omeprazole solution in a volumetric flask, adding a diluent for dilution, fixing the volume to a scale, and shaking up; here, the diluent is a mixture of 1mM aqueous sodium hydroxide solution and acetonitrile, preferably, the volume ratio of 1mM aqueous sodium hydroxide solution to acetonitrile is 90: 10; or
Preparing a resolution solution: weighing about 15mg of esomeprazole magnesium reference substance, precisely weighing, placing into a 50mL volumetric flask, adding 5mL of R-omeprazole solution, diluting and dissolving with a diluent, fixing the volume, and shaking up; here, the diluent is a mixture of 1mM aqueous sodium hydroxide solution and acetonitrile, preferably, the volume ratio of 1mM aqueous sodium hydroxide solution to acetonitrile is 90: 10; or
Preparation of control solution 1: precisely weighing about 12mg of R-omeprazole reference substance, precisely weighing, placing in a 20mL measuring flask, dissolving and diluting to scale with diluent, and shaking up; precisely measuring 1mL of the solution, placing the solution in a 100mL measuring flask, dissolving the solution by using a diluent, diluting the solution to a scale, and shaking up; precisely measuring 5mL of the solution, placing the solution in a 50mL measuring flask, dissolving the solution by using a diluent, diluting the solution to a scale, and shaking up; here, the diluent is a mixture of 1mM aqueous sodium hydroxide solution and acetonitrile, preferably, the volume ratio of 1mM aqueous sodium hydroxide solution to acetonitrile is 90: 10; or
Preparation of control solution 2: precisely weighing about 12mg of R-omeprazole reference substance, precisely weighing, placing in a 20mL measuring flask, dissolving and diluting to scale with diluent, and shaking up; precisely measuring 1mL of the solution, placing the solution in a 100mL measuring flask, dissolving the solution by using a diluent, diluting the solution to a scale, and shaking up; precisely measuring 5mL of the solution, placing the solution in a 50mL measuring flask, dissolving the solution by using a diluent, diluting the solution to a scale, and shaking up; here, the diluent is a mixture of 1mM aqueous sodium hydroxide solution and acetonitrile, preferably, the volume ratio of 1mM aqueous sodium hydroxide solution to acetonitrile is 90: 10.
In embodiments of the present application, the detection method, wherein the impurities that may interfere with the detection of enantiomers include, but are not limited to, the following known impurities, the chemical structures of which are as follows:
in the present application, the chemical structure of the esomeprazole sodium enantiomer is:
i.e. enantiomer (R-ompprazole) or R-Omeprazole.
The methodological verification of the method is carried out by a system applicability test, an impurity positioning test, a quantitative limit detection limit test, a linear test and an accuracy test, and the detection method of the enantiomer in the esomeprazole sodium medicament provided by the invention has the following advantages:
(1) the method has good system applicability.
(2) Good separation of enantiomers from the main peak and each impurity peak can be achieved.
(3) The method has high sensitivity, and the concentrations of the enantiomers which can be effectively detected are all lower than the report limit.
(4) The method has high accuracy.
(5) Enantiomers exhibit a good linear relationship over a range of concentrations.
Additional features and advantages of the invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. The objectives and other advantages of the invention will be realized and attained by the structure particularly pointed out in the written description and claims hereof as well as the appended drawings.
Drawings
The accompanying drawings are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the example serve to explain the principles of the invention and not to limit the invention.
Fig. 1 shows an enantiomer detection chromatogram map of interference of degradation impurity H with an enantiomer when applied to esomeprazole sodium for injection according to the determination method of enantiomer in esomeprazole sodium crude drug of chinese pharmacopoeia 2015 edition;
fig. 2 is an impurity separation chromatogram (esomeprazole mixed solution with other impurities) of example 2 of the present application;
fig. 3 is an impurity separation chromatogram (resolution of esomeprazole from enantiomer) of example 6 of the present application;
fig. 4 is an enantiomer detection chromatogram of a stable sample of esomeprazole sodium for injection of example 6 of the present application (no enantiomer was detected).
Detailed Description
Hereinafter, embodiments of the present invention will be described in detail in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be noted that the embodiments and features of the embodiments in the present application may be arbitrarily combined with each other without conflict.
Example 1
Endomprazole sodium enantiomer system suitability test
Instrument (Agilent HPLC 1260) and chromatographic conditions:
a chromatographic column: philomenchiral ME (2) -RH 4.6X 250mm 5 μm
Mobile phase: 20mM NH4HCO3Mixing the solution-acetonitrile-methanol (volume ratio is 50:30: 20) uniformly;
diluent agent: 1mM NaOH-acetonitrile (90: 10 by volume), in the following examples, the same diluent as in example 1;
flow rate: 0.8mL/min, column temperature: 25 ℃, detection wavelength: 302nm, sample size: 20 mu l of the mixture;
operating time: and (4) 45 min.
Injecting the following solutions into a chromatograph respectively, recording a chromatogram, and calculating the content of the enantiomer according to the peak area by an external standard method according to the peak area of the R-omeprazole chromatographic peak.
The experimental steps are as follows:
solution preparation (use of brown volumetric flask and brown sampling flask in dark):
r-omeprazole solution: precisely weighing about 12mg of R-omeprazole reference substance, precisely weighing, placing in a 20mL measuring flask, dissolving and diluting to scale with diluent, and shaking up; precisely measuring 1mL of the solution, placing the solution in a 100mL measuring flask, dissolving the solution with a diluent, diluting the solution to a scale, and shaking up.
Sensitivity solution: and (3) taking the R-omeprazole solution from 2.5mL to 100mL in a volumetric flask, adding a diluent for dilution, fixing the volume to a scale, and shaking up.
Resolution solution: weighing about 15mg of esomeprazole magnesium reference substance, precisely weighing, placing in a 50mL volumetric flask, adding 5mL of R-omeprazole solution, diluting and dissolving with a diluent, fixing the volume, and shaking up.
Control solution: precisely weighing about 12mg of R-omeprazole reference substance, precisely weighing, placing in a 20mL measuring flask, dissolving and diluting to scale with diluent, and shaking up; precisely measuring 1mL of the solution, placing the solution in a 100mL measuring flask, dissolving the solution by using a diluent, diluting the solution to a scale, and shaking up; precisely measure 5mL of the solution, place the solution in a 50mL measuring flask, dissolve the solution with a diluent, dilute the solution to the mark, and shake the solution.
The retention time and peak area results for esomeprazole isomer (R-omeprazole) in system applicability are given in the following table:
the blank solution has no interference on the detection of R-omeprazole (enantiomer); the signal-to-noise ratio (S/N) of R-omeprazole in the sensitivity solution is 10.43, and the separation degree of a main peak and the R-omeprazole in the separation degree solution is 6.47; 6 RSD of the peak area of R-omeprazole in the control solution 1 is 2.5 percent, and RSD of retention time is 0.2 percent; the RSD of 6 against control solutions 1 and 2 against the f-value of R-omeprazole in control solution 2 was 3.3%, and the system applicability was good.
Example 2 known impurity localization test
The chromatographic conditions were the same as in example 1
The experimental steps are as follows:
blank auxiliary material stock solution: dissolving and diluting edetate disodium 225.72mg and sodium hydroxide about 54.47mg to 20mL in a measuring flask with a diluent to a constant volume, and shaking up; precisely transferring into a measuring flask of 1mL to 100mL, diluting with the diluent to a constant volume, and shaking up. Blank adjuvant solution: precisely transferring 5mL of blank auxiliary material storage solution into a 50mL measuring flask, diluting with diluent to constant volume, and shaking up.
R-omeprazole solution: accurately weighing 12.14mg of R-omeprazole reference substance, placing the reference substance in a 20mL measuring flask, dissolving the reference substance by using a diluent, diluting the reference substance to a scale, and shaking up; precisely measuring 1mL of the solution, placing the solution in a 100mL measuring flask, dissolving the solution with a diluent, diluting the solution to a scale, and shaking up.
Resolution solution: weighing 15.03mg of esomeprazole magnesium reference substance, placing the reference substance in a 50mL volumetric flask, adding 5mL of R-omeprazole solution, diluting and dissolving the reference substance by using a diluent, fixing the volume and shaking up.
Each impurity localization solution: accurately weighing the reference substances of the impurities A-H to be about 10mg respectively, placing the reference substances into different 25mL measuring bottles, dissolving and diluting the reference substances to scales by using 50% ethanol solution, and shaking up; precisely transferring into a measuring flask of 5mL to 50mL, diluting to scale with the diluent, and shaking up.
Esomeprazole sodium localization solution: weighing 15.22mg of esomeprazole magnesium reference substance, placing the reference substance into a 50mL measuring flask, dissolving and diluting the reference substance to the scale with a diluent, and shaking up.
Test solution: taking 15.66mg of esomeprazole sodium for injection, placing the esomeprazole sodium for injection into a 50mL measuring flask, dissolving the esomeprazole sodium with a proper amount of diluent, diluting the esomeprazole sodium to a scale, and shaking up.
Known impurity separation degree solution: taking 30.31mg of esomeprazole sodium for injection, adding a proper amount of diluent to dissolve, placing in a 100mL volumetric flask, transferring 1.5mL of impurity A-H positioning solution and 5mL of R-omeprazole solution to the volumetric flask respectively, adding the diluent to a constant volume to scale, and shaking up.
Remarking: after entering an HPLC system, both esomeprazole sodium and esomeprazole magnesium show chromatographic peaks, and the esomeprazole magnesium standard substance is selected to prepare the esomeprazole sodium positioning solution because the standard substance of the esomeprazole sodium is not available.
And (3) respectively injecting the positioning solution, the resolution solution and the known impurity resolution solution of each impurity into a liquid chromatograph, and recording a chromatogram (the separation condition is shown in figure 2). The retention times of the various impurities are shown in the following table:
name (R)
Retention time (min)
Impurity F
4.780
Impurity A
5.602
Impurity G
6.229
Impurity E
10.906
Impurity D
12.225
Esomeprazole
15.692
R-omeprazole
22.254
Impurity B
23.849
Impurity H
25.812, 26.620 (double peak)
Impurity C
34.206
The results show that: the peak time of other known impurities in the solution with the known impurity separation degree is known to not interfere the detection of the enantiomer, the separation degree of the main peak and the enantiomer in the solution with the separation degree is 6.47, and the baseline separation is achieved.
Note: the separation degree of the impurity B and the R-omeprazole is 1.5, which meets the regulation. The impurity B is a process impurity which is not detected in a plurality of batches of raw material medicines, so that the detection of the enantiomer in the esomeprazole sodium for injection cannot be interfered.
Example 3 detection Limit and quantitation Limit tests
The chromatographic conditions were the same as in example 1
The experimental steps are as follows:
r-omeprazole solution: accurately weighing 12.62mg of R-omeprazole reference substance, placing the reference substance in a 20mL measuring flask, dissolving the reference substance by using a diluent, diluting the reference substance to a scale, and shaking up; precisely measuring 1mL of the solution, placing the solution in a 100mL measuring flask, dissolving the solution with a diluent, diluting the solution to a scale, and shaking up.
Esomeprazole stock solution: weighing 14.99mg of esomeprazole reference substance, placing the reference substance into a 50mL measuring flask, dissolving and diluting the reference substance to a scale by using a diluent, and shaking up; precisely measure 1mL of the solution, place it in a 50mL measuring flask, dissolve it with diluent and dilute it to the mark, shake it up.
LOQ solution: precisely measuring 5mL esomeprazole stock solution and 5mL R-omeprazole solution respectively, placing the stock solution and the 5mL R-omeprazole solution into a 200mL measuring flask, dissolving and diluting the stock solution and the 5mL R-omeprazole solution to a scale by using a diluent, and shaking up.
LOD solution: precisely measure 3mL of LOQ solution, place in a 10mL measuring flask, dissolve with diluent and dilute to the mark, shake well.
Respectively sampling the solutions, and quantitatively limiting the signal-to-noise ratio (S/N) of the R-omeprazole in the solution to be about 10; the signal-to-noise ratio (S/N) of the test line solution was about 3.
Data results are as follows:
the results show that: the quantitative limit concentration of the R-omeprazole is less than 0.1 percent (report limit) of the concentration of a test sample, and the content of the enantiomer can be accurately controlled.
Example 4 Linear test
The chromatographic conditions were the same as in example 1
The experimental steps are as follows:
r-omeprazole solution: accurately weighing 12.62mg of R-omeprazole reference substance, placing the reference substance in a 20mL measuring flask, dissolving the reference substance by using a diluent, diluting the reference substance to a scale, and shaking up; precisely measuring 1mL of the solution, placing the solution in a 100mL measuring flask, dissolving the solution with a diluent, diluting the solution to a scale, and shaking up.
The R-omeprazole linear solution was prepared as follows:
and (3) respectively sampling the solutions, recording a chromatogram, and taking the concentration as an abscissa and the peak area as an ordinate to make a linear recovery equation of the R-omeprazole.
The linearity results are as follows:
the results show that: r-omeprazole exhibits good linearity over a concentration range of 0.14671 μ g/mL (LOQ) to 1.17366 μ g/mL (200% limit level).
Example 5 accuracy test
The chromatographic conditions were the same as in example 1
The experimental steps are as follows:
r-omeprazole solution: accurately weighing 12.62mg of R-omeprazole reference substance, placing the reference substance in a 20mL measuring flask, dissolving the reference substance by using a diluent, diluting the reference substance to a scale, and shaking up; precisely measuring 1mL of the solution, placing the solution in a 100mL measuring flask, dissolving the solution with a diluent, diluting the solution to a scale, and shaking up.
Test solution: taking 15.42mg of esomeprazole sodium for injection, placing the esomeprazole sodium for injection into a 50mL measuring flask, dissolving the esomeprazole sodium with a proper amount of diluent, diluting the esomeprazole sodium to a scale, and shaking up.
Accuracy solution (0.05%): weighing about 30mg of esomeprazole sodium for injection, precisely weighing, placing in a 100mL volumetric flask, adding 2.5mL of R-omeprazole solution, diluting with a diluent, dissolving and fixing the volume. 3 parts are prepared in parallel.
Accuracy solution (0.2%) -1: weighing about 15mg of esomeprazole sodium for injection, precisely weighing, placing in a 50mL volumetric flask, adding 5mL of R-omeprazole solution, diluting with a diluent, dissolving and fixing the volume. 3 parts are prepared in parallel.
Accuracy solution (0.3%) -1: weighing about 15mg of esomeprazole sodium for injection, precisely weighing, placing in a 50mL volumetric flask, adding 7.5mL of R-omeprazole solution, diluting with a diluent, dissolving and fixing the volume. 3 parts are prepared in parallel.
And (3) respectively injecting samples and recording chromatograms, and calculating the recovery rate, wherein the results are shown in the following table:
the results show that: the recovery rate of R-omeprazole is between 95 and 120 percent within the concentration level range of the quantitative limit to 0.3 percent (the limit level of 150 percent), and the accuracy of the method is good.
Example 6 stability sample testing
According to the requirements of the general rules of the four parts of the Chinese pharmacopoeia 2015 edition, the preparation samples are subjected to stability sample retention and enantiomer detection.
The chromatographic conditions were the same as in example 1
The experimental steps are as follows:
r-omeprazole solution: precisely weighing 11.89mg of R-omeprazole reference substance, placing the reference substance in a 20mL measuring flask, dissolving the reference substance by using a diluent, diluting the reference substance to a scale, and shaking up; precisely measuring 1mL of the solution, placing in a 100mL measuring flask, dissolving with diluent, diluting to scale, and shaking. 2 parts are prepared in parallel.
Resolution solution: weighing 14.85mg of esomeprazole magnesium reference substance, placing in a 50mL volumetric flask, adding 5mL of R-omeprazole solution, diluting and dissolving with a diluent, fixing the volume, and shaking up.
Formulation sample solution (25 ℃. + -. 2 ℃ RH 60%. + -. 10%, 24 months): taking 15.78mg of esomeprazole sodium for injection, placing the esomeprazole sodium for injection into a 50mL measuring flask, dissolving the esomeprazole sodium with a proper amount of diluent, diluting the esomeprazole sodium to a scale, and shaking up.
And respectively injecting samples and recording chromatograms (the resolution solution is shown in figure 3, and the preparation sample solution is shown in figure 4), wherein enantiomers are not detected in the preparation sample solution.
Although the embodiments disclosed in the present application are described above, the descriptions are only for the convenience of understanding the present application, and are not intended to limit the present application. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the disclosure as defined by the appended claims.