CN117269349A - Method for measuring content of norepinephrine sulfonate and sodium metabisulfite in norepinephrine bitartrate injection - Google Patents
Method for measuring content of norepinephrine sulfonate and sodium metabisulfite in norepinephrine bitartrate injection Download PDFInfo
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- CN117269349A CN117269349A CN202311173571.0A CN202311173571A CN117269349A CN 117269349 A CN117269349 A CN 117269349A CN 202311173571 A CN202311173571 A CN 202311173571A CN 117269349 A CN117269349 A CN 117269349A
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- HRZFUMHJMZEROT-UHFFFAOYSA-L sodium disulfite Chemical compound [Na+].[Na+].[O-]S(=O)S([O-])(=O)=O HRZFUMHJMZEROT-UHFFFAOYSA-L 0.000 title claims abstract description 56
- 229940001584 sodium metabisulfite Drugs 0.000 title claims abstract description 56
- 235000010262 sodium metabisulphite Nutrition 0.000 title claims abstract description 56
- BDHFUVZGWQCTTF-UHFFFAOYSA-M sulfonate Chemical compound [O-]S(=O)=O BDHFUVZGWQCTTF-UHFFFAOYSA-M 0.000 title claims abstract description 53
- 238000002347 injection Methods 0.000 title claims abstract description 47
- 239000007924 injection Substances 0.000 title claims abstract description 47
- SFLSHLFXELFNJZ-QMMMGPOBSA-N (-)-norepinephrine Chemical compound NC[C@H](O)C1=CC=C(O)C(O)=C1 SFLSHLFXELFNJZ-QMMMGPOBSA-N 0.000 title claims abstract description 43
- 229960002748 norepinephrine Drugs 0.000 title claims abstract description 43
- SFLSHLFXELFNJZ-UHFFFAOYSA-N norepinephrine Natural products NCC(O)C1=CC=C(O)C(O)=C1 SFLSHLFXELFNJZ-UHFFFAOYSA-N 0.000 title claims abstract description 43
- LNBCGLZYLJMGKP-LUDZCAPTSA-N 4-[(1r)-2-amino-1-hydroxyethyl]benzene-1,2-diol;(2r,3r)-2,3-dihydroxybutanedioic acid;hydrate Chemical compound O.OC(=O)[C@H](O)[C@@H](O)C(O)=O.NC[C@H](O)C1=CC=C(O)C(O)=C1 LNBCGLZYLJMGKP-LUDZCAPTSA-N 0.000 title claims abstract description 36
- 229960001695 norepinephrine bitartrate Drugs 0.000 title claims abstract description 36
- 238000000034 method Methods 0.000 title claims abstract description 25
- 239000000243 solution Substances 0.000 claims abstract description 54
- 238000001514 detection method Methods 0.000 claims abstract description 25
- 239000013558 reference substance Substances 0.000 claims abstract description 15
- 239000012071 phase Substances 0.000 claims abstract description 12
- 238000012360 testing method Methods 0.000 claims abstract description 11
- 239000000126 substance Substances 0.000 claims abstract description 8
- 239000007788 liquid Substances 0.000 claims abstract description 7
- 239000007791 liquid phase Substances 0.000 claims abstract description 7
- FWFGVMYFCODZRD-UHFFFAOYSA-N oxidanium;hydrogen sulfate Chemical compound O.OS(O)(=O)=O FWFGVMYFCODZRD-UHFFFAOYSA-N 0.000 claims abstract description 7
- CHRJZRDFSQHIFI-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;styrene Chemical class C=CC1=CC=CC=C1.C=CC1=CC=CC=C1C=C CHRJZRDFSQHIFI-UHFFFAOYSA-N 0.000 claims abstract description 4
- 239000000945 filler Substances 0.000 claims abstract description 4
- 238000010829 isocratic elution Methods 0.000 claims abstract description 4
- 239000000523 sample Substances 0.000 claims description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- 239000012488 sample solution Substances 0.000 claims description 6
- 238000002360 preparation method Methods 0.000 claims description 3
- 229940079593 drug Drugs 0.000 abstract description 4
- 239000003814 drug Substances 0.000 abstract description 4
- 238000004458 analytical method Methods 0.000 abstract description 3
- 230000002349 favourable effect Effects 0.000 abstract description 2
- 229940090044 injection Drugs 0.000 description 31
- 230000006378 damage Effects 0.000 description 13
- 239000011259 mixed solution Substances 0.000 description 7
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 230000001066 destructive effect Effects 0.000 description 6
- 239000012535 impurity Substances 0.000 description 6
- 239000012085 test solution Substances 0.000 description 6
- 238000000926 separation method Methods 0.000 description 5
- 208000001953 Hypotension Diseases 0.000 description 4
- 230000036543 hypotension Effects 0.000 description 4
- 238000004811 liquid chromatography Methods 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 230000014759 maintenance of location Effects 0.000 description 3
- 230000001590 oxidative effect Effects 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 239000011550 stock solution Substances 0.000 description 3
- 239000002253 acid Substances 0.000 description 2
- 239000002585 base Substances 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 238000005286 illumination Methods 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- 230000004807 localization Effects 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000006386 neutralization reaction Methods 0.000 description 2
- 239000003381 stabilizer Substances 0.000 description 2
- 230000001502 supplementing effect Effects 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- FEWJPZIEWOKRBE-UHFFFAOYSA-M 3-carboxy-2,3-dihydroxypropanoate Chemical compound OC(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-M 0.000 description 1
- 208000010496 Heart Arrest Diseases 0.000 description 1
- 206010021137 Hypovolaemia Diseases 0.000 description 1
- 206010000891 acute myocardial infarction Diseases 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 230000002490 cerebral effect Effects 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 239000007857 degradation product Substances 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- OKKJLVBELUTLKV-UHFFFAOYSA-N methanol Substances OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 1
- 229940030825 norepinephrine injection Drugs 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000010412 perfusion Effects 0.000 description 1
- 208000028591 pheochromocytoma Diseases 0.000 description 1
- 230000008832 photodamage Effects 0.000 description 1
- 239000012286 potassium permanganate Substances 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 239000012088 reference solution Substances 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- REFMEZARFCPESH-UHFFFAOYSA-M sodium;heptane-1-sulfonate Chemical compound [Na+].CCCCCCCS([O-])(=O)=O REFMEZARFCPESH-UHFFFAOYSA-M 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 150000003871 sulfonates Chemical class 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid 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
- G01N2030/042—Standards
- G01N2030/047—Standards external
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- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
The invention discloses a method for measuring the content of norepinephrine sulfonate and sodium metabisulfite in norepinephrine bitartrate injection, and belongs to the technical field of medical analysis. The method comprises the following steps: 1) Preparing a reference substance solution and a test substance solution; 2) Setting high performance liquid phase detection conditions; a chromatographic column with sulfonated styrene divinylbenzene as a filler and sulfuric acid water as a mobile phase are adopted, and isocratic elution is carried out by using the mobile phase; 3) Respectively sucking the reference substance solution and the test substance solution, injecting into a liquid chromatograph, recording a chromatogram, and calculating the contents of norepinephrine sulfonate and sodium metabisulfite in the sample to be detected. The invention can rapidly, effectively, accurately and reliably separate and detect the norepinephrine sulfonate and the sodium metabisulfite in the norepinephrine bitartrate injection, is favorable for improving the product quality of the norepinephrine bitartrate injection and the medication safety of patients.
Description
Technical Field
The invention belongs to the technical field of medical analysis, and particularly relates to a method for measuring the content of norepinephrine sulfonate and sodium metabisulfite in norepinephrine bitartrate injection.
Background
Norepinephrine bitartrate injection for treating hypotension caused by acute myocardial infarction, extracorporeal circulation, etc.; for shock, hypotension or hypotension after pheochromocytoma excision caused by hypovolemia, the product is used as an auxiliary treatment for supplementing blood volume in emergency, so that the blood pressure rises, and cerebral and coronary perfusion is temporarily maintained until the treatment for supplementing blood volume takes place; can also be used for maintaining hypotension during intraspinal block and after resuscitation due to cardiac arrest.
The method under the relevant matter item of the norepinephrine bitartrate injection in the 2020 edition of Chinese pharmacopoeia adopts 0.14 percent sodium heptanesulfonate solution-methanol (80:20) to carry out isocratic elution, the retention time of the norepinephrine peak is about 10 minutes, the relative retention time of the sodium metabisulfite peak and the norepinephrine sulfonate peak is about 0.23 and 0.26 respectively, the separation degree of the sodium metabisulfite peak and the norepinephrine sulfonate peak is lower, auxiliary material peak interference exists, and the detection accuracy is lower; sodium metabisulfite is an antioxidant in bitartrate norepinephrine injection, and the existing sodium metabisulfite content detection method is a titration method or an ultraviolet spectrophotometry detection method, but the specificity is poor. Therefore, an improvement on the detection method of the norepinephrine sulfonate and the sodium metabisulfite in the norepinephrine bitartrate injection is needed, a rapid and effective detection method is established, the product quality of the norepinephrine bitartrate injection is improved, and the medication safety of patients is improved.
Disclosure of Invention
Aiming at the problems in the prior art, the technical problem to be solved by the invention is to provide a method for measuring the content of norepinephrine sulfonate and sodium metabisulfite in the norepinephrine bitartrate injection, which can quickly and effectively separate and detect the norepinephrine sulfonate and sodium metabisulfite in the norepinephrine bitartrate injection, improve the product quality of the norepinephrine bitartrate injection and improve the medication safety of patients.
In order to solve the problems, the technical scheme adopted by the invention is as follows:
a method for determining the content of norepinephrine sulfonate and sodium metabisulfite in norepinephrine bitartrate injection comprises the following specific steps:
1) Preparing a reference substance solution and a test substance solution;
2) Setting high performance liquid phase detection conditions; a chromatographic column with sulfonated styrene divinylbenzene as a filler and sulfuric acid water as a mobile phase are adopted, and isocratic elution is carried out by using the mobile phase;
3) Respectively sucking the reference substance solution and the test substance solution, injecting into a liquid chromatograph, recording a chromatogram, and calculating the contents of norepinephrine sulfonate and sodium metabisulfite in the sample to be detected.
Further, in the step 1), the sample solution to be tested is an injection sample, and the sample is directly injected; the preparation method of the reference substance solution comprises the following steps: the reference substances of norepinephrine sulfonate and sodium metabisulfite are respectively weighed and added with water to be dissolved into a reference substance solution containing 0.2mg of norepinephrine sulfonate and sodium metabisulfite per 1 mL.
Further, in the step 2), the column was Phenomenex Rezex ROA-Organic Acid H+8% and had a length of 300mm and an inner diameter of 7.8mm.
Further, in the step 2), the column temperature of the chromatographic column is 58-62 ℃.
Further, the column temperature of the chromatographic column is 60 ℃.
Further, in the step 2), the w/w of sulfuric acid water was 0.05%.
Further, in the step 2), the flow rate of the mobile phase is 0.55-0.65 mL/min.
Further, the mobile phase flow rate was 0.6mL/min.
Further, in step 2), the sample injection amounts of the sample solution and the control solution are 20. Mu.L.
Further, in the step 3), the detection wavelength of the liquid chromatograph was 276nm.
The beneficial effects are that: compared with the prior art, the invention has the advantages that:
(1) Compared with the method for measuring related substances under the norepinephrine bitartrate injection item in the Chinese pharmacopoeia 2020 edition, the method adopts a high-efficiency liquid phase method to measure the content of the norepinephrine sulfonate impurity and the content of the stabilizer sodium metabisulfite in the norepinephrine bitartrate injection, and the norepinephrine sulfonate and the sodium metabisulfite can be well separated, and other impurities, auxiliary material peaks and solvent peaks do not interfere with detection; .
(2) The auxiliary materials and degradation products generated by the test sample under various damage conditions do not interfere with the measurement of sodium metabisulfite and the sulfonate, the detection specificity of related substances is good, and meanwhile, the separation and detection results of impurities and stabilizing agents are not influenced when chromatographic condition parameters are slightly changed, so that the durability is good.
(3) The invention can rapidly, effectively, accurately and reliably separate and detect the norepinephrine sulfonate and the sodium metabisulfite in the norepinephrine bitartrate injection, is favorable for improving the product quality of the norepinephrine bitartrate injection and the medication safety of patients.
Drawings
FIG. 1 is a test chromatogram under chromatographic conditions of the present invention;
FIG. 2 is a chromatogram of the Chinese pharmacopoeia analysis method;
FIG. 3 is a graph of linear and range of sodium metabisulfite tested under the liquid chromatography conditions of the present invention;
FIG. 4 is a graph of the linearity and range of norepinephrine sulfonate tested under the liquid chromatography conditions of the present invention.
Detailed Description
The invention is further described below in connection with specific embodiments.
The conditions for high performance liquid phase detection used in the following examples were as follows:
chromatographic column: phenomenex Rezex ROA-Organic Acid H+ (8%) 7.8X100 mm
Flow rate: 0.6mL/min;
column temperature: 60 ℃;
mobile phase: 0.05% (w/w) sulfuric acid water;
detection wavelength: 276nm.
Example 1
The method for measuring the content of norepinephrine sulfonate and sodium metabisulfite in the norepinephrine bitartrate injection by adopting a high performance liquid phase method comprises the following steps of:
(1) Preparing a control solution and a test solution: the sample solution is an injection sample, and is directly injected; the norepinephrine sulfonate and sodium metabisulfite reference substance are weighed and added with water to be dissolved, and the concentration of the reference substance solution is 0.2mg/mL.
(2) Setting high performance liquid phase detection conditions: the chromatographic column is Phenomenex Rezex ROA-Organic Acid H+ (8%) with the length of 7.8X300 mm, sulfonated styrene divinylbenzene is used as a filler, the mobile phase is 0.05% (w/w) sulfuric Acid water, gradient elution is carried out at the flow rate of 0.6mL/min, and the column temperature is 60 ℃; the detection wavelength is 276nm;
(3) 20 mu L of the sample solution and the reference solution are respectively sucked precisely, injected into a liquid chromatograph, and the chromatogram is recorded, and the result is shown in figure 1.
FIG. 1 shows a test chromatogram under the chromatographic condition of the invention, and the main component can be well separated between the norepinephrine sulfonate and sodium metabisulfite under the chromatographic condition at the detection wavelength of 276nm, and the solvent peak and the blank auxiliary material peak do not interfere with the measurement, thereby conforming to the standard.
Example 2
The detection method of example 1 was subjected to method verification, and verification was performed from several aspects such as system applicability, destructive test, quantitative limit, detection limit, linear relationship, filter membrane adsorption, precision, accuracy, solution stability, and the like, respectively.
1. System suitability determination
The norepinephrine bitartrate is taken and dissolved and diluted by water to prepare a positioning solution containing about 0.1mg of norepinephrine bitartrate per 1 mL.
Dissolving sulfonate in water, and diluting to obtain a positioning solution containing about 0.2mg of norepinephrine bitartrate per 1 mL.
Sodium metabisulfite is taken, and water is added for dissolution and dilution to prepare a positioning solution containing about 0.2mg of norepinephrine bitartrate per 1 mL.
The method comprises the steps of taking norepinephrine bitartrate, sulfonate and sodium metabisulfite, and diluting with water to prepare a mixed solution containing 1mg of norepinephrine bitartrate, 0.2mg of norepinephrine sulfonate and 0.2mg of sodium metabisulfite in each 1mL, wherein the mixed solution is used as a system applicability solution.
The results are shown in Table 1 below.
TABLE 1 impurity localization and separation degree
| Name of the name | Blank space | Sodium metabisulfite | Norepinephrine sulfonate |
| Single solution retention time (minutes) | —— | 14.5 | 11.9 |
| System applicability solution (minutes) | —— | 14.6 | 11.9 |
| Degree of separation | —— | —— | 4.1 |
Table 1 shows the impurity localization and separation degree, and it is clear from the table that the sodium metabisulfite and the norepinephrine sulfonate can be effectively separated without interfering with detection.
2. Destructive test
Taking norepinephrine bitartrate injection for preparing destructive test solution:
unbroken solution: the injection was taken as a non-destructive solution (2 mg/mL).
Acid breaking solution: 10mL of the injection was taken, 1mL of a 0.1mol/L HCl solution was added thereto for 1 hour, and then 1mL of a 0.1mol/L NaOH solution was added thereto for neutralization to obtain an acid-disrupted solution.
Base destruction solution: 10mL of the injection was taken, 1mL of a 0.1mol/L NaOH solution was added thereto for 1 hour, and then 1mL of a 0.1mol/L HCl solution was added thereto for neutralization to obtain a base disruption solution.
Oxidative destruction solution: : 20mL of the injection was taken, and 0.5mL of a 0.01mol/L KMnO4 solution was added as an oxidative destruction solution.
High temperature destruction solution: taking 10mL of injection, breaking for 3 hours at 90 ℃, and cooling to room temperature to obtain a high-temperature breaking solution.
Light damage solution: 10mL of the injection is taken and is subjected to destruction under 254nm and 365nm ultraviolet light for 3 hours to be used as a preparation illumination destruction solution.
The results are shown in Table 2 below.
TABLE 2 destructive inspection results
| Impurity(s) | Before destruction | Destruction by illumination | High temperature damage | Oxidative destruction of | Acid damage | Alkali destruction |
| Sodium metabisulfite | 100.00％ | 71.96％ | 75.30％ | 90.52％ | 94.53％ | 93.89％ |
| Sulfonate compounds | 100.00％ | 97.41％ | 187.61％ | 112.61％ | 99.61％ | 100.37％ |
Table 2 shows the results of the destructive inspection, and it is clear from the results that the high temperature and oxidation conditions greatly affect the sulfonate, and the sulfonate significantly increases.
3. Quantitative limit and detection limit experiments
The sulfonate and sodium metabisulfite were diluted with water to prepare a mixed solution containing 0.2mg of norepinephrine sulfonate and 0.2mg of sodium metabisulfite per 1mL as a stock solution.
The quantitative limit (S/N is more than or equal to 10) and the detection limit (S/N is more than or equal to 3) of the sample are measured by a progressive dilution method.
The results are shown in Table 3 below.
TABLE 3 detection limits and quantification limits
Table 3 shows the results of the detection limit and the quantitative limit, and it is clear from the tables that sodium metabisulfite and the sulfonate at low concentrations can be efficiently detected under the chromatographic conditions.
4. Precision of sample injection
The sulfonate and sodium metabisulfite are diluted by water to prepare a mixed solution containing 0.2mg of norepinephrine sulfonate and 0.2mg of sodium metabisulfite in each 1mL, and the mixed solution is used as a sample injection precision solution.
Under the above chromatographic conditions, 20. Mu.L of the mixture was injected into a liquid chromatograph, and the sample was continuously introduced 6 times, and the peak area was recorded.
The results are shown in Table 4 below.
TABLE 4 sample injection precision results
Table 4 shows the results of the sample injection precision, and the sample injection precision of the chromatographic conditions is good.
5. Solution stability
The sulfonate and sodium metabisulfite are diluted with water to prepare a mixed solution containing 0.2mg of norepinephrine sulfonate and 0.2mg of sodium metabisulfite in each 1mL, and the mixed solution is used as a reference substance solution.
The injection is taken and directly injected to be used as a test solution (2 mg/mL).
Under the chromatographic conditions, sampling is carried out at 0h, 12h, 24h, 36h and 48h respectively, and the daily stability is examined.
The results are shown in tables 5 and 6 below.
TABLE 5 stability results of control solutions
TABLE 6 test solution stability results
Table 5 shows the stability results of the control solutions, and Table 6 shows the stability results of the test solutions, and it can be seen from tables 5 and 6 that the test solutions and the control solutions have good stability within 48 hours
6. Linearity and range
Sodium metabisulfite and sulfonate are taken, water is added for dissolution and dilution, and a stock solution which contains 0.4mg of sodium metabisulfite and 0.4mg of sulfonate in each 1mL is prepared as a linear stock solution. The above solutions were diluted with solvents in the following Table 7 to prepare linear solutions of respective concentrations.
TABLE 7 Linear solution formulation Table
Under the above chromatographic conditions, linear solutions were prepared at 1%,10%,50%,100%,150%,200% limits, and the results are shown in fig. 3 and 4.
FIG. 3 is a graph showing the linearity and range of sodium metabisulfite tested under the liquid chromatography condition of the present invention, and the graph shows that the linear result of sodium metabisulfite is good under the present chromatography condition.
FIG. 4 is a graph showing the linearity and range of norepinephrine sulfonate tested under the liquid chromatography conditions of the present invention, showing that sodium metabisulfite has good linearity.
Claims (10)
1. The method for measuring the content of norepinephrine sulfonate and sodium metabisulfite in norepinephrine bitartrate injection is characterized by comprising the following specific steps of:
1) Preparing a reference substance solution and a test substance solution;
2) Setting high performance liquid phase detection conditions; a chromatographic column with sulfonated styrene divinylbenzene as a filler and sulfuric acid water as a mobile phase are adopted, and isocratic elution is carried out by using the mobile phase;
3) Respectively sucking the reference substance solution and the test substance solution, injecting into a liquid chromatograph, recording a chromatogram, and calculating the contents of norepinephrine sulfonate and sodium metabisulfite in the sample to be detected.
2. The method for determining the content of norepinephrine sulfonate and sodium metabisulfite in norepinephrine bitartrate injection of claim 1, wherein in step 1), the sample solution is an injection sample, and the sample is directly injected; the preparation method of the reference substance solution comprises the following steps: the reference substances of norepinephrine sulfonate and sodium metabisulfite are respectively weighed and added with water to be dissolved into a reference substance solution containing 0.2mg of norepinephrine sulfonate and sodium metabisulfite per 1 mL.
3. The method for determining the content of norepinephrine sulfonate and sodium metabisulfite in norepinephrine bitartrate injection of claim 1, wherein in step 2), the chromatographic column is Phenomenex Rezex ROA-Organic Acid h+8%, with a length of 300mm and an inner diameter of 7.8mm.
4. The method for determining the content of norepinephrine sulfonate and sodium metabisulfite in norepinephrine bitartrate injection of claim 1, wherein in step 2), the column temperature of the chromatographic column is 58-62 ℃.
5. The method for determining the content of norepinephrine sulfonate and sodium metabisulfite in norepinephrine bitartrate injection of claim 4, wherein the column temperature is 60 ℃.
6. The method for determining the content of norepinephrine sulfonate and sodium metabisulfite in norepinephrine bitartrate injection of claim 1, wherein in step 2), the w/w of sulfuric acid water is 0.05%.
7. The method for determining the content of norepinephrine sulfonate and sodium metabisulfite in norepinephrine bitartrate injection of claim 1, wherein in step 2), the mobile phase flow rate is 0.55-0.65 mL/min.
8. The method for determining the content of norepinephrine sulfonate and sodium metabisulfite in norepinephrine bitartrate injection of claim 7, wherein the mobile phase flow rate is 0.6mL/min.
9. The method for determining the content of norepinephrine sulfonate and sodium metabisulfite in norepinephrine bitartrate injection of claim 1, wherein in step 2), the sample injection amounts of the sample solution and the control solution are 20 μl.
10. The method for determining the content of norepinephrine sulfonate and sodium metabisulfite in norepinephrine bitartrate injection of claim 1, wherein in step 3), the detection wavelength of the liquid chromatograph is 276nm.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311173571.0A CN117269349A (en) | 2023-09-12 | 2023-09-12 | Method for measuring content of norepinephrine sulfonate and sodium metabisulfite in norepinephrine bitartrate injection |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311173571.0A CN117269349A (en) | 2023-09-12 | 2023-09-12 | Method for measuring content of norepinephrine sulfonate and sodium metabisulfite in norepinephrine bitartrate injection |
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