Method for detecting edetate disodium in levosalbutamol hydrochloride aerosol inhalation solution

Abstract

The invention belongs to the technical field of drug detection, and discloses a method for detecting edetate disodium in a levosalbutamol hydrochloride aerosol inhalation solution, which comprises the following steps: detecting a test solution of a to-be-detected sample by high performance liquid chromatography, with a mobile phase being a mixed solution of a borax solution and acetonitrile, the mass ratio of sodium tetraborate to water to acetonitrile in the mobile phase is (6-9): 100: (100-120). The content of edetate disodium in each batch of samples is detected, so that the safety and effectiveness of the levosalbutamol hydrochloride aerosol inhalation solution are guaranteed.

Description

Technical Field

[0001] The invention relates to the technical field of drug detection, and specifically to a method for detecting edetate disodium in a levosalbutamol hydrochloride nebulized inhalation solution, so that the method for detecting edetate disodium is sensitive, accurate and rapid, and the accurate dosage of edetate disodium is ensured, thereby ensuring the safety and effectiveness of the levosalbutamol hydrochloride nebulized inhalation solution. Background Art

[0002] The statements herein merely provide background information related to the present invention and do not necessarily constitute prior art.

[0003] Asthma is the most common respiratory disease in the world with a high prevalence. Clinical studies have shown that compared with its racemic form, salbutamol hydrochloride has the advantages of high efficacy, mild side effects and good tolerance. Dexabutamol can cause adverse reactions such as headache, dizziness, palpitations, and finger tremors.

[0004] But the storage stability of levosalbuterol hydrochloride atomized inhalation solution is poor. In order to improve its storage stability, its preparation method can be to use nitrogen to displace oxygen from the solution in the preparation process until the dissolved oxygen is less than 1ppm, and the product headspace after the filling needs nitrogen filling protection, and then packaged in an oxygen-impermeable aluminum-plastic film to ensure that the residual oxygen content in the container is below 2%. But during production, there is a certain difficulty in nitrogen filling by a liquid preparation tank, nitrogen filling of the headspace of a low-density polyethylene ampoule, and nitrogen filling protection in an aluminum-plastic bag, and it is easy to cause damage and leakage during transportation.

[0005] In addition, disodium edetate (disodium ethylenediaminetetraacetic acid) as an antioxidant can improve the stability of salbutamol nebulized inhalation solution. However, the side effects of disodium edetate include nausea, vomiting, and joint pain. Excessive use of disodium edetate will form a water-soluble chelate with calcium in the blood and bones and be excreted from the body, which can easily cause hypocalcemia or bone calcium loss. Muscle tremors and convulsions may occur due to the rapid decrease in blood calcium. Therefore, if the content of disodium edetate is too high, it is easy to increase the medication risk of salbutamol hydrochloride nebulized inhalation solution; and if the content of disodium edetate is too low, the salbutamol hydrochloride nebulized inhalation solution cannot be guaranteed to be stable within the validity period. At present, there is no existing technology for detecting the content of disodium edetate in salbutamol nebulized inhalation solution to control its quality. Summary of the invention

[0006] In view of the shortcomings of the prior art, the purpose of the present invention is to provide a method for detecting disodium edetate in a nebulized inhalation solution of salbutamol hydrochloride. The content of disodium edetate in each batch of samples is detected, thereby ensuring the safety and effectiveness of the nebulized inhalation solution of salbutamol hydrochloride.

[0007] In order to achieve the above object, the present invention is implemented through the following technical solutions:

[0008] In a first aspect, the present invention provides a method for detecting disodium edetate in a nebulized inhalation solution of salbutamol hydrochloride, comprising the following steps: using high performance liquid chromatography to detect a test solution of a sample to be tested, adding copper ions to the test solution, and using a mobile phase that is a mixture of a borax solution and acetonitrile, wherein the mass ratio of sodium tetraborate, water, and acetonitrile in the mobile phase is 6-9:1000:100-120.

[0009] In some embodiments, the mass ratio of sodium tetraborate, water and acetonitrile in the mobile phase is 7-9:1000:110-115.

[0010] In some embodiments, the mobile phase is prepared by mixing 0.02 mol / L borax solution and acetonitrile in a volume ratio of 9:1.

[0011] In some embodiments, the salbutamol hydrochloride atomized inhalation solution is diluted 10-15 times in the test sample solution, and copper sulfate with a mass fraction of 0.001%-0.003% is added. The ultraviolet absorption of disodium edetate is very weak and cannot be tested. After complexing with copper, ethylenediamine copper sodium acetate (EDTA-Cu) is generated, which has strong absorption in the wavelength range of 200-300nm, and can effectively detect disodium edetate.

[0012] Preferably, the preparation method of the test solution of the sample to be tested is: measure a set volume of salbutamol hydrochloride nebulized inhalation solution, then add copper sulfate solution thereto in proportion, mix well and let stand for 10-30 minutes, and obtain the test solution after constant volume.

[0013] The purpose of standing still after mixing is to ensure that disodium edetate and copper ions are fully complexed.

[0014] In some embodiments, the HPLC column is an octadecylsilane bonded silica gel column.

[0015] In some embodiments, the wavelength of HPLC detection is 280-290 nm; the flow rate is 1.0-2.0 ml / min; the column temperature is 25-35° C.; and the injection volume is 20 μl.

[0016] In some embodiments, the blank solution, the reference solution and the test solution are accurately aspirated respectively, and the chromatogram is recorded; the content of disodium edetate in the levosalbutamol hydrochloride nebulized inhalation solution is calculated by the peak area according to the external standard method:

[0017] Edetate disodium content = rU÷rS×mS×P×VS;

[0018] Where:

[0019] rU: peak area of ​​disodium edetate in sample solution;

[0020] rS: peak area of ​​edetate disodium in reference solution;

[0021] mS: the amount of disodium edetate in the reference solution, mg;

[0022] VS: volume of reference solution, ml;

[0023] V: dilution multiple of the test solution;

[0024] P: Content of edetate disodium reference substance (calculated as anhydrous substance).

[0025] Preferably, the blank solution is a copper sulfate solution with a mass fraction of 0.001%-0.003%.

[0026] The beneficial effects achieved by one or more embodiments of the present invention are as follows:

[0027] Without borax solution, the ultraviolet absorption of disodium edetate is very weak, with no absorption or peak at 228nm, and cannot be tested. After complexing with copper, it generates ethylenediamine copper acetate sodium (EDTA-Cu), which has strong absorption in the wavelength range of 200-300nm, and can effectively detect disodium edetate.

[0028] The addition of borax can effectively improve the accuracy and sensitivity of detection.

[0029] Acetonitrile does not absorb ultraviolet light at 200nm to 400nm. In the present invention, a wavelength of 228nm is selected for detection, which has no solvent effect and can make the analysis sensitivity as high as 10 -9 class. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] The accompanying drawings in the specification, which constitute a part of the present invention, are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

[0031] Figure 1 is a chromatographic separation diagram of the blank solution in Example 1 of the present invention;

[0032] Figure 2 is a chromatographic separation diagram of the reference substance solution in Example 1 of the present invention;

[0033] Figure 3 is a chromatographic separation diagram of the test solution in Example 1 of the present invention;

[0034] Figure 4 is the chromatogram in Example 3 of the present invention;

[0035] Figure 5 It is the chromatogram of Comparative Example 1 of the present invention. DETAILED DESCRIPTION

[0036] It should be noted that the following detailed descriptions are illustrative and intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used in the present invention have the same meanings as those commonly understood by those skilled in the art to which the present invention belongs.

[0037] The present invention will be further described below in conjunction with the embodiments.

[0038] The batch number of the following test sample solution is: 230305, manufacturer: Shandong Dayin Jinkong Children's Pharmaceutical Co., Ltd.

[0039] Example 1

[0040] Test for the content of disodium edetate in salbutamol hydrochloride nebulized inhalation solution:

[0041] Blank solvent (diluent): water.

[0042] Blank solution: Take 5 ml of 0.01% copper sulfate solution, place it in a 25 ml volumetric flask, dilute to the scale with blank solvent, mix well, and use it as the blank solution.

[0043] Test solution: Accurately measure 2 ml of the product, place it in a 25 ml volumetric flask, add 5 ml of 0.01% copper sulfate solution, let it stand for 20 minutes, dilute to the scale with blank solvent, mix well, and use this as the test solution.

[0044] Reference substance solution: take about 10 mg of edetate disodium reference substance, accurately weigh it, place it in a 100 ml volumetric flask, add blank solvent to dissolve and dilute to the mark, mix well, and obtain the reference substance stock solution (100 μg / ml); then accurately measure 2 ml of the reference substance stock solution and place it in a 25 ml volumetric flask, add 5 ml of 0.01% copper sulfate solution, let it stand for 20 minutes, dilute to the mark with blank solvent, mix well, and obtain the reference substance solution. Prepare 2 portions in parallel.

[0045] System suitability solution: Same as reference solution. The RSD of the main peak area of ​​the system suitability solution for 5 consecutive injections is not greater than 2.0%. Check that the recovery rate of the reference solution is between 98.0% and 102.0%, and the theoretical plate number is not less than 1500 calculated based on disodium edetate.

[0046] Chromatographic conditions: a chromatographic column (4.6×250 mm, 5 μm) filled with octadecylsilane bonded silica gel, 0.02 mol / L borax solution (7.64 g of sodium tetraborate decahydrate, dissolved in 1000 ml of water)-acetonitrile (90:10) as the mobile phase, detection wavelength of 280 nm, flow rate of 2.0 ml / min, column temperature of 30°C, injection volume of 20 μl.

[0047] Accurately pipette 20 μl of blank solution, system suitability solution, reference solution, and test solution filtrate respectively, and record the chromatogram. Figure 1-Figure 3 Calculated by peak area using external standard method.

[0048] Edetate disodium content = rU÷rS×mS×P×VS;

[0049] Where:

[0050] rU: peak area of ​​disodium edetate in sample solution;

[0051] rS: peak area of ​​edetate disodium in reference solution;

[0052] mS: the amount of disodium edetate in the reference solution, mg;

[0053] VS: volume of reference solution, ml;

[0054] V: dilution multiple of the test solution;

[0055] P: content of edetate disodium reference substance (calculated as anhydrous substance);

[0056] According to the external standard method, the peak area was calculated based on the anhydrous form of disodium edetate, which should be 0.08-0.12 mg / ml. The test results are shown in Table 1.

[0057] Table 1 Test results

[0058]

[0059]

[0060] Example 2 Specificity Test

[0061] Blank solvent (diluent): water.

[0062] Blank solution: Take 5 ml of 0.01% copper sulfate solution, place it in a 25 ml volumetric flask, dilute to the scale with blank solvent, mix well, and use it as the blank solution.

[0063] Test solution: Accurately measure 2 ml of the product, place it in a 25 ml volumetric flask, add 5 ml of 0.01% copper sulfate solution, let it stand for 20 minutes, dilute to the scale with blank solvent, mix well, and use this as the test solution.

[0064] Reference substance solution: take about 10 mg of edetate disodium reference substance, accurately weigh it, place it in a 100 ml volumetric flask, add blank solvent to dissolve and dilute to the scale, mix well, and obtain the reference substance stock solution (100 μg / ml); then accurately measure 2 ml of the reference substance stock solution and place it in a 25 ml volumetric flask, add 5 ml of 0.01% copper sulfate solution, let it stand for 20 minutes, dilute to the scale with blank solvent, mix well, and obtain the reference substance solution.

[0065] Accurately pipette 20 μl of the blank solution, reference solution, and test solution filtrate respectively, and record the chromatogram.

[0066] Test results: Blank solvent and blank excipients do not interfere with the determination of edetate disodium content. The retention time of the main component chromatographic peak of the reference solution is consistent with that of the main component chromatographic peak of the test solution, and the chromatographic peak shape is good.

[0067] Acceptable criteria: No interfering peaks should be detected at the retention time of edetate disodium peak in blank solvent and blank excipient solutions.

[0068] Example 3 Investigation of the effect of copper sulfate - After adding copper sulfate, let it stand for 20 minutes and then make up to volume with water.

[0069] Table 2 Detection methods

[0070]

[0071]

[0072] Depend on Figure 4 It can be seen that when the detection method in Table 2 is used for detection, there is no interference from the blank solvent and other components in the test sample, the specificity is good, the peak shape of the EDTA main peak is good, and the sensitivity meets the requirements.

[0073] Comparative Example 1 Investigation of the Effect of Copper Sulfate - No Standing After Adding Copper Sulfate

[0074] Table 3 Detection methods

[0075]

[0076] The difference between Comparative Example 1 and Example 3 is that the step of standing is omitted. The test results are as follows: Figure 5 As shown by Figure 5 It can be seen that under the detection method in Table 3, the EDTA reference substance has a peak at about 7.2 minutes, and the chromatogram of the test solution has a bulge at about 6.2 minutes, which affects the integration of EDTA and cannot be accurately quantified.

[0077] Comparative Example 2

[0078] The difference from Example 1 is that the borax solution is omitted in the mobile phase, and only acetonitrile is used to detect the test sample. The rest is the same as Example 1.

[0079] When only acetonitrile is used in the mobile phase, the pH value of the eluent of the chromatographic column changes, elution cannot be performed, there is no absorption peak, and detection cannot be performed.

[0080] Comparative Example 3

[0081] The difference from Example 1 is that 0.01% copper sulfate is not added to the test solution, and the test solution is tested. Others are the same as in Example 1.

[0082] When copper sulfate is not added to the test solution, the UV absorption of disodium edetate is very weak, with no absorption peak, and cannot be tested.

[0083] In summary, the method for detecting the content of disodium edetate in a nebulized inhalation solution of salbutamol hydrochloride proposed by the present invention innovatively screens out the optimal chromatographic conditions for high performance liquid chromatography detection, and the detection is sensitive, accurate and rapid, thereby ensuring the effectiveness of disodium edetate as a stabilizer in the drug production process and the safety of the product levosalbutamol hydrochloride nebulized inhalation solution.

[0084] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for detecting edetate disodium in a levosalbutamol hydrochloride aerosol inhalation solution, characterized in that: The method comprises the following steps: using high performance liquid chromatography to detect a test sample solution of a sample to be tested, wherein the mobile phase is a mixed solution of a borax solution and acetonitrile, and the mass ratio of sodium tetraborate, water and acetonitrile in the mobile phase is 6-9:1000:100-120.

2. The method for detecting edetate disodium in the salbutamol hydrochloride atomized inhalation solution according to claim 1, characterized in that: The mass ratio of sodium tetraborate, water and acetonitrile in the mobile phase is 7-9:1000:110-115.

3. The method for detecting edetate disodium in the salbutamol hydrochloride atomized inhalation solution according to claim 2, characterized in that: The mobile phase is prepared by mixing 0.02 mol / L borax solution and acetonitrile in a volume ratio of 9:

1.

4. The method for detecting edetate disodium in the salbutamol hydrochloride atomized inhalation solution according to claim 1, characterized in that: In the test solution of the sample to be tested, the salbutamol hydrochloride nebulized inhalation solution is diluted 10-15 times, and copper sulfate with a mass fraction of 0.001%-0.003% is added.

5. The method for detecting edetate disodium in the salbutamol hydrochloride atomized inhalation solution according to claim 4, characterized in that: The preparation method of the test solution of the sample to be tested is as follows: measuring a set volume of salbutamol hydrochloride nebulized inhalation solution, then adding copper sulfate solution thereto in proportion, mixing well and letting it stand for 10-30 minutes, and then fixing the volume to obtain the test solution.

6. The method for detecting edetate disodium in the levosalbuterol hydrochloride atomized inhalation solution according to claim 1, characterized in that: The chromatographic column for high performance liquid chromatography is an octadecylsilane bonded silica gel filler chromatographic column.

7. The method for detecting edetate disodium in the levosalbuterol hydrochloride atomized inhalation solution according to claim 6, characterized in that: The wavelength of HPLC detection is 280-290nm.

8. The method for detecting edetate disodium in the levosalbuterol hydrochloride atomized inhalation solution according to claim 7, characterized in that: The flow rate is 1.0-2.0 ml / min; the column temperature is 25-35°C; the injection volume is 20 μl.

9. The method for detecting edetate disodium in the levosalbuterol hydrochloride atomized inhalation solution according to claim 1, characterized in that: Accurately aspirate the blank solution, reference solution and test solution respectively, and record the chromatogram; calculate the content of disodium edetate in the levosalbuterol hydrochloride nebulized inhalation solution by the peak area according to the external standard method: Edetate disodium content = rU÷rS×mS×P×VS; Where: rU: peak area of ​​disodium edetate in sample solution; rS: peak area of ​​edetate disodium in reference solution; mS: the amount of disodium edetate in the reference solution, mg; VS: volume of reference solution, ml; V: dilution factor of the test solution; P: Content of edetate disodium reference substance, calculated on anhydrous basis.

10. The method for detecting edetate disodium in the levosalbuterol hydrochloride atomized inhalation solution according to claim 9, characterized in that: The blank solution is a copper sulfate solution with a mass fraction of 0.001%-0.003%.

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