Method for detecting sodium metabisulfite content in dopamine hydrochloride injection by HPLC

Abstract

The application discloses a method for detecting the content of sodium metabisulfite in dopamine hydrochloride injection by HPLC, provides a high performance liquid chromatography mobile phase, the high performance liquid chromatography mobile phase is composed of water phase, acetonitrile and methanol with a volume ratio of 78-82:9-11:9-11; wherein, the pH value of the water phase is 6.0+ / -0.2; a high performance liquid chromatography column with cyano silane bonded silica gel as a filler is provided; a test sample solution and different concentrations of control sample solutions are provided; the flow rate is set to 0.4-0.6 mL / min, and the detection wavelength is set to 208-212 nm; the control sample solutions with different concentrations are injected, the peak time of sodium metabisulfite is determined, and a linear regression equation is made according to the peak area of the control sample solutions with different concentrations; the test sample solution is injected, the peak area is recorded, and the concentration of sodium metabisulfite in the test sample solution is calculated by using the linear regression equation and the peak area. The method has the advantages of small sample amount, high sensitivity, and after the sample is prepared, the quantitative work of sodium metabisulfite in dopamine hydrochloride injection can be completed within 9 minutes.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of chemical detection, and particularly relates to a method for detecting the content of sodium metabisulfite in dopamine hydrochloride injection by HPLC and a mobile phase thereof. BACKGROUND

[0002] Sodium metabisulfite, English name Sodium Metabisulfite, alias heavy sulfur oxygen, also known as sodium metabisulfite, sodium metabisulfite, aqueous solution is acidic, dissolves in water to generate stable sodium sulfite, is easy to decompose when wet, and is easy to slowly oxidize and release SO2 when exposed to air.

[0003] Sodium metabisulfite is used as an antioxidant in pharmaceutical preparations, with a concentration of 0.01% to 1%. Sodium metabisulfite is used as an antioxidant for acidic liquid, and sodium sulfite works better in alkaline liquid. Sodium metabisulfite also has certain antibacterial effect, and works best in acidic environment, and can also be used as a preservative in oral preparations.

[0004] Sodium metabisulfite is widely used as an antioxidant in pharmaceutical preparations and is also widely used in food processing. Like other sulfites, sodium metabisulfite can cause some serious or life-threatening adverse reactions, such as bronchospasm and allergy. After oral absorption, it has a stimulating effect on the stomach. Sodium metabisulfite can cause damage to various systems, organs and tissues of the human body to a certain extent, and its residual product SO2 can generate corrosive sulfite, sulfuric acid and sulfate on wet mucosa, aggravate the stimulating effect, damage the respiratory system, and further cause various types of respiratory inflammation.

[0005] The 2020 edition of Chinese Pharmacopoeia adopts ultraviolet method to determine the content of sodium metabisulfite, but the specificity and accuracy of the method for detecting the content of sodium metabisulfite in dopamine hydrochloride injection are relatively low. Therefore, it is necessary to develop a method for determining the content of sodium metabisulfite in dopamine hydrochloride injection to ensure the quality of the product. SUMMARY

[0006] In view of the problem that there is no method for determining the content of sodium metabisulfite in dopamine hydrochloride injection in the prior art, the present application provides a method for detecting the content of sodium metabisulfite in dopamine hydrochloride injection by HPLC and a mobile phase thereof.

[0007] The technical scheme provided by the present application is as follows:

[0008] A high performance liquid chromatography mobile phase for separating sodium metabisulfite in dopamine hydrochloride injection, the high performance liquid chromatography mobile phase is composed of water phase, acetonitrile and methanol with a volume ratio of 78-82:9-11:9-11; wherein the pH value of the water phase is 6.0±0.2.

[0009] As the preferred solution of the above technical scheme, the water phase adds one or more of potassium dihydrogen phosphate, sodium dihydrogen phosphate, ammonium dihydrogen phosphate and sodium perchlorate as a buffer salt, and ammonia is used to adjust the pH value.

[0010] As the preferred solution of the above technical scheme, the concentration of the buffer salt in the water phase is 0.005-0.2 mol / L, and contains 0.015-0.100 mol / L of tetrabutylammonium hydrogen sulfate.

[0011] As the preferred solution of the above technical scheme, the high-performance liquid chromatography mobile phase is composed of water phase, acetonitrile and methanol in a volume ratio of 78:11:11, 80:10:10 or 82:9:9.

[0012] A method for detecting the content of sodium metabisulfite in dopamine hydrochloride injection by high-performance liquid chromatography, comprising the following steps:

[0013] The above high-performance liquid chromatography mobile phase is provided;

[0014] A high-performance liquid chromatography column with cyanosilane-bonded silica gel as the filler is provided;

[0015] A test sample solution and different concentrations of control sample solutions are provided;

[0016] The flow rate is set to 0.4-0.6 mL / min, and the detection wavelength is set to 208-212 nm;

[0017] The control sample solutions of different concentrations are injected, the peak time of sodium metabisulfite is determined, and a linear regression equation is made according to the peak areas of the control sample solutions of different concentrations;

[0018] The test sample solution is injected, the peak area is recorded, and the concentration of sodium metabisulfite in the test sample solution is calculated using the linear regression equation and the peak area.

[0019] As the preferred solution of the above technical scheme, the specifications of the high-performance liquid chromatography column are 4.6 mm x 250 mm, and the particle size of the filler is 5 μm.

[0020] As the preferred solution of the above technical scheme, the column temperature is set to 25-35℃ before injection, the injection volume is set to 10 μL, and the sample tray temperature is controlled at 8℃.

[0021] As the preferred solution of the above technical scheme, the method for providing the test sample solution is to dilute the L-ascorbic acid aqueous solution with a concentration of 0.2 mg / mL by 50 times, i.e. to obtain the test sample solution.

[0022] Specifically, the peak time of sodium metabisulfite is 7.0-7.3 min from the start of injection.

[0023] Specifically, the linear regression equation is Y=100.9735X-0.4297, X is the concentration of sodium pyrosulfite, and Y is the peak area.

[0024] Compared with the prior art, the present application has the beneficial effects that:

[0025] (1) The present application provides a method for detecting the content of sodium pyrosulfite in dopamine hydrochloride injection and a mobile phase thereof, which solves the problem that the prior art does not have a specific detection method for sodium pyrosulfite in dopamine hydrochloride injection.

[0026] (2) The tetrabutylammonium hydrogen sulfate added in the mobile phase of the present application belongs to a tetraalkyl quaternary ammonium salt. + (CH2CH2CH2CH3)4 structure, which can effectively produce strong electrostatic interaction with Si-O-, prevent the contact between amino and silicon hydroxyl, and improve the peak shape. BRIEF DESCRIPTION OF DRAWINGS

[0027] Figure 1 is a chromatogram of a control diluted with freshly boiled cooling water;

[0028] Figure 2 is a chromatogram of a test solution diluted with freshly boiled cooling water;

[0029] Figure 3 is a chromatogram of a control solution diluted with 0.01% EDTA-2Na aqueous solution;

[0030] Figure 4 is a chromatogram of a test solution diluted with 0.01% EDTA-2Na aqueous solution;

[0031] Figure 5 is a chromatogram of a control solution diluted with 0.1 mg / mL L-ascorbic acid aqueous solution;

[0032] Figure 6 is a chromatogram of a test solution diluted with 0.1 mg / mL L-ascorbic acid aqueous solution;

[0033] Figure 7 is a chromatogram of a control solution diluted with 0.2 mg / mL L-ascorbic acid aqueous solution;

[0034] Figure 8 is a chromatogram of a test solution diluted with 0.2 mg / mL L-ascorbic acid aqueous solution;

[0035] Figure 9Chromatogram of the control solution diluted with 0.4 mg / mL L-ascorbic acid aqueous solution;

[0036] Figure 10 Chromatogram of the test solution diluted with 0.4 mg / mL L-ascorbic acid aqueous solution;

[0037] Figure 11 Chromatogram of the control solution with the mobile phase of water phase-acetonitrile (v / v=80:20);

[0038] Figure 12 Chromatogram of the control solution with the mobile phase of water phase-acetonitrile (v / v=85:15);

[0039] Figure 13 Chromatogram of the control solution with the mobile phase of water phase-acetonitrile-methanol (v / v / v=80:15:5);

[0040] Figure 14 Chromatogram of the control solution with the mobile phase of water phase-acetonitrile-methanol (v / v / v=80:10:10). DETAILED DESCRIPTION

[0041] In order to make the purpose, technical scheme and advantages of the present application more clear, the present application is further described in detail below in combination with the drawings and examples. It should be understood that the specific examples described herein are only used to explain the present application and do not limit the present application.

[0042] Pharmacopoeia method: an appropriate amount of the product (equivalent to 0.15 g of sodium pyrosulfite) is precisely weighed, placed in an iodine volumetric flask, 50 mL of iodine titration solution (0.05 mol / L) is precisely added, tightly sealed, shaken to dissolve, then 1 mL of hydrochloric acid is added, titrated with sodium thiosulfate titration solution (0.1 mol / L) until the blue color disappears, and 2 mL of starch indicator is added at the near end point. The result of the titration is corrected by a blank test. Each 1 mL of iodine titration solution (0.05 mol / L) is equivalent to 4.752 mg of Na2S2O2. The sample amount required by this method is large, and the sensitivity is low.

[0043] The present application provides a method for detecting the content of sodium pyrosulfite in dopamine hydrochloride injection by using HPLC as the separation and detection means, and the specific content is as follows:

[0044] The chromatographic column (WelchXtimate CN, 4.6 mm x 250 mm, 5 μm) used in the following examples is filled with cyano-silane bonded silica gel, and other chromatographic columns with equivalent performance can also be used.

[0045] Example 1

[0046] 1. Chromatographic conditions:

[0047] Column: cyanosilane bonded silica gel as the filler (Welch Xtimate CN, 4.6 mm x 250 mm, 5 μm);

[0048] Mobile phase: 0.005 mol / L potassium dihydrogen phosphate solution (containing 0.015 mol / L tetrabutylammonium hydrogen sulfate, and adjusting pH value to 6.0 with ammonia water)-acetonitrile-methanol (v / v / v = 80:10:10);

[0049] Instrument setting before sample injection: flow rate 0.5 mL / min; column temperature 30°C; detection wavelength 210 nm; sample injection volume 10 μL; sample tray temperature control 8°C.

[0050] 2. Selection of diluent solution:

[0051] Reference solution: accurately weigh a certain amount of sodium pyrosulfite reference substance, dissolve with the diluent, and quantitatively dilute with the diluent to prepare a solution containing about 0.2 mg of sodium pyrosulfite per 1 mL, which is freshly prepared before use.

[0052] Test solution: take a certain amount of dopamine hydrochloride injection, quantitatively dilute with the diluent to prepare a solution containing about 0.2 mg of sodium pyrosulfite per 1 mL, which is freshly prepared before use.

[0053] The present application attempts to use new boiling cooling water and 0.01% EDTA-2Na aqueous solution as the diluent of the reference solution and the test solution, and the obtained chromatogram is as shown in Figures 1-4 , which is not desirable according to the peak shape.

[0054] The present application further selects L-ascorbic acid aqueous solution with concentrations of 0.1 mg / mL, 0.2 mg / mL and 0.4 mg / mL as the diluent of the reference solution and the test solution, and the obtained chromatogram is as shown in Figures 5-10 , according to the solution stability of the reference solution and the test solution, the result is that when the concentration of L-ascorbic acid is 0.2 mg / mL, the solution is most stable. Table 1 shows the content change of the sample at different time points when using L-ascorbic acid aqueous solution with different concentrations as the diluent.

[0055] Table 1 Stability of L-ascorbic acid solution with different concentrations

[0056]

[0057]

[0058] Finally, an aqueous solution containing about 0.2 mg of L-ascorbic acid per 1 mL is selected as the diluent.

[0059] Example 2

[0060] 1. Chromatographic conditions:

[0061] Chromatographic column: packed with cyano-silane bonded silica gel (Welch Xtimate CN, 4.6 mm x 250 mm, 5 μm);

[0062] Instrument settings before injection: flow rate 0.5 mL / min; column temperature 30 °C; detection wavelength 210 nm; injection volume 10 μL; sample tray temperature control 8 °C.

[0063] 2. Preparation of solution:

[0064] Diluent: take L-ascorbic acid, dissolve in water, and dilute to about 0.2 mg of L-ascorbic acid per 1 mL, filter, and degas.

[0065] Reference solution: accurately weigh a certain amount of sodium pyrosulfite reference substance, dissolve in diluent, and quantitatively dilute with diluent to prepare a solution containing about 0.2 mg of sodium pyrosulfite per 1 mL, freshly prepared.

[0066] Test solution: take a certain amount of dopamine hydrochloride injection, quantitatively dilute with diluent to prepare a solution containing about 0.2 mg of sodium pyrosulfite per 1 mL, freshly prepared.

[0067] 3. Selection of mobile phase ratio:

[0068] This experiment investigated four mobile phase systems: water phase-acetonitrile (v / v = 80:20), water phase-acetonitrile (v / v = 85:15), water phase-acetonitrile-methanol (v / v / v = 80:15:5), and water phase-acetonitrile-methanol (v / v / v = 80:10:10). The results showed that under the water phase-acetonitrile-methanol (v / v / v = 80:10:10) mobile phase system, the separation degree of L-ascorbic acid peak and sodium pyrosulfite peak met the requirements, and the peak shape was better. (Note: the water phase is 0.005 mol / L potassium dihydrogen phosphate aqueous solution (containing 0.015 mol / L tetrabutylammonium hydroxylamine), and the pH value is adjusted to 6.0 with ammonia water). Figures 11-14 Reference chromatograms of different mobile phase systems.

[0069] Example 3

[0070] 1. Chromatographic conditions

[0071] Column: Silica gel bonded with cyano-silane as packing material (Welch Xtimate CN, 4.6 mm x 250 mm, 5 μm); mobile phase: 0.005 mol / L potassium dihydrogen phosphate solution (containing 0.015 mol / L tetrabutylammonium hydrogen sulfate, and the pH value is adjusted to 6.0 with ammonia water)-acetonitrile-methanol (v / v / v = 80:10:10); flow rate: 0.5 mL / min; column temperature: 30 °C; detection wavelength: 210 nm; injection volume: 10 μL; sample tray temperature: 8 °C.

[0072] 2. Preparation of solution:

[0073] Diluent: an appropriate amount of L-ascorbic acid was dissolved in water, and diluted to about 0.2 mg of L-ascorbic acid per 1 mL, filtered, and degassed to obtain.

[0074] Control solution: an appropriate amount of sodium pyrosulfite control was dissolved in diluent, and quantitatively diluted with diluent to prepare sodium pyrosulfite solutions of different concentrations, which were prepared freshly before use.

[0075] Test solution: an appropriate amount of dopamine hydrochloride injection was quantitatively diluted with diluent to prepare a solution containing about 0.2 mg of sodium pyrosulfite per 1 mL, which was prepared freshly before use.

[0076] 3. Linear test

[0077] The peak time was determined by injecting different concentrations of control solution, and the peak area of different concentrations of control solution was recorded, and the results are shown in Table 2:

[0078] Table 2 Linear test results

[0079]

[0080] Conclusion: Sodium pyrosulfite showed a good linear relationship with the peak area in the concentration range of 0.0487 mg / mL to 0.2923 mg / mL, the linear regression equation was Y = 100.9735X - 0.4297, the correlation coefficient was 1.000, and the absolute ratio of the intercept to the 100% response value was 2.2%. Among them, X is the concentration of sodium pyrosulfite in the control solution, and Y is the peak area.

[0081] 4. Test solution detection

[0082] The test solution was precisely measured, injected, and the peak area was recorded. The concentration of sodium pyrosulfite in the test solution was calculated by peak area according to the external standard method.

[0083] The foregoing is considered as illustrative only of the principles of the application. Numerous modifications and changes will readily occur to those skilled in the art, and it is intended to embrace all such modifications and changes that fall within the scope of the application. Accordingly, the application is not to be restricted in scope to the specific embodiments disclosed herein but is to be accorded the full scope that the principles and novel features request appropriately granted.

Claims

1. A high performance liquid chromatography mobile phase for separating sodium metabisulfite in a dopamine hydrochloride injection, characterized by: The high performance liquid chromatography mobile phase is composed of water phase, acetonitrile and methanol with a volume ratio of 78-82:9-11:9-11; wherein the pH value of the water phase is 6.0±0.

2.

2. The high performance liquid chromatography mobile phase for separating sodium pyrosulfite in dopamine hydrochloride injection according to claim 1, characterized by: The water phase adds one or more of potassium dihydrogen phosphate, sodium dihydrogen phosphate, ammonium dihydrogen phosphate and sodium perchlorate as buffer salt, and adjusts the pH value by using ammonia water.

3. The high performance liquid chromatography mobile phase for separating sodium pyrosulfite in dopamine hydrochloride injection according to claim 2, characterized by: The concentration of the buffer salt in the water phase is 0.005-0.2 mol / L, and contains 0.015-0.100 mol / L of tetrabutylammonium hydrogen sulfate.

4. The high performance liquid chromatography mobile phase for separating sodium pyrosulfite in dopamine hydrochloride injection according to claim 1, characterized by: The high performance liquid chromatography mobile phase is composed of water phase, acetonitrile and methanol with a volume ratio of 78:11:11, 80:10:10 or 82:9:

9.

5. A method for detecting the content of sodium metabisulfite in dopamine hydrochloride injection by high performance liquid chromatography, characterized in that, The method comprises the following steps: The high performance liquid chromatography mobile phase is provided according to any one of claims 1-4; The high performance liquid chromatography column is provided with cyano silane bonded silica gel as the filler; The test sample solution and different concentrations of control sample solutions are provided; The flow rate is set to 0.4-0.6 mL / min, and the detection wavelength is set to 208-212 nm; The control sample solutions with different concentrations are injected, the peak time of sodium pyrosulfite is determined, and a linear regression equation is made according to the peak area of the control sample solutions with different concentrations; The test sample solution is injected, the peak area is recorded, and the concentration of sodium pyrosulfite in the test sample solution is calculated by using the linear regression equation and the peak area.

6. The method for detecting the content of sodium pyrosulfite in dopamine hydrochloride injection by high performance liquid chromatography according to claim 5, characterized in that: The specification of the high performance liquid chromatography column is 4.6 mm×250 mm, and the particle size of the filler is 5 μm.

7. The method for detecting the content of sodium pyrosulfite in dopamine hydrochloride injection by high performance liquid chromatography according to claim 5, characterized in that: The column temperature is set to 25-35 ℃, the injection volume is set to 10 μL, and the sample tray temperature is controlled to 8 ℃.

8. The method for detecting the content of sodium pyrosulfite in dopamine hydrochloride injection by high performance liquid chromatography according to claim 5, characterized in that: The method for providing the test sample solution is as follows: taking dopamine hydrochloride injection, diluting 50 times with L-ascorbic acid aqueous solution with a concentration of 0.2 mg / mL to obtain the test sample solution.

9. The method for detecting the content of sodium pyrosulfite in dopamine hydrochloride injection by high performance liquid chromatography according to claim 5, characterized in that: The peak time of sodium pyrosulfite is 7.0-7.3 min from the injection.

10. The method for detecting the content of sodium pyrosulfite in dopamine hydrochloride injection by high performance liquid chromatography according to claim 5, characterized in that: The linear regression equation is Y=100.9735X-0.4297, X is the concentration of sodium pyrosulfite, and Y is the peak area.

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