Method for detecting content of hydrogen sulfide in acetylcysteine injection
By preparing sodium sulfide standard solution, test products and blank solutions, and using spectrophotometer to determine the absorbance of hydrogen sulfide, the problem of difficulty in detecting the content of hydrogen sulfide in acetylcysteine injection in the prior art is solved, and the sulfide content is achieved The accurate determination of hydrogen content ensures the safety of the drug solution.
Patent Information
- Application Number
- CN202411420499.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-12
- Publication Date
- 2025-01-14
AI Technical Summary
The prior art is difficult to effectively detect and control the content of hydrogen sulfide in acetylcysteine injection. Hydrogen sulfide, as a strong neurotoxin, has a huge impact on human health and a reliable detection method is needed to ensure the safety of the drug solution.
By preparing sodium sulfide standard solution, test samples and blank solutions, the absorbance of hydrogen sulfide is measured using a spectrophotometer, and combined with specific chemical reactions and dilution steps, the content of hydrogen sulfide is calculated to ensure the accuracy and reliability of the detection.
Accurate detection of the content of hydrogen sulfide in acetylcysteine injection is achieved, ensuring the safety of the drug solution, and providing a reliable method to control the concentration of hydrogen sulfide and avoid potential harm to the human body.
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Figure CN119310026A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of drug detection and relates to a method for detecting the content of hydrogen sulfide in acetylcysteine injection. Background Art
[0002] Acetylcysteine, also known as acetylcysteine, has a molecular formula of C5H9NO3S and is a white or off-white crystalline powder with a garlic-like odor. It is mainly used as a mucolytic agent and has a strong mucolytic effect. The sulfhydryl group contained in its molecules can break the disulfide bonds in the glycoprotein polypeptide chains in the sputum, thereby reducing the viscosity of the sputum and making the sputum liquid and easier to cough up. It can also break the DNA fibers in purulent sputum, so it can dissolve not only white sticky sputum, but also purulent sputum. It is mainly used clinically to treat respiratory diseases with excessive thick mucus secretions, such as acute bronchitis, chronic bronchitis and its exacerbation, emphysema, muco-obstructive disease, bronchiectasis, etc.
[0003] Injections enter the blood circulation directly, so that they can quickly enter human tissues and exert their therapeutic effects. Compared with oral medications that need to be absorbed into the blood through organs such as the stomach, injections are faster and more direct.
[0004] Hydrogen sulfide, the degradation product of acetylcysteine, is a colorless gas with the smell of rotten eggs. It is a strong neurotoxin that has a strong irritating effect on the mucous membranes and has a huge impact on the eyes, respiratory system, and brain nerves. Therefore, the content of hydrogen sulfide in the injection solution should be strictly controlled. Summary of the invention
[0005] In view of the deficiencies of the prior art, the object of the present invention is to provide a method for detecting the hydrogen sulfide content in acetylcysteine injection.
[0006] In order to achieve the above purpose, after a lot of time and experiments, the following scheme was finally obtained: a method for determining the content of hydrogen sulfide in acetylcysteine injection, the method comprising the following steps:
[0007] (1) Prepare 20 ppm hydrogen sulfide standard solution using sodium sulfide standard solution
[0008] (2) Preparation of test sample and blank solution: Accurately pipette acetylcysteine injection solution into a three-necked round-bottom flask and add water. The flask contains an air inlet tube leading to the bottom of the flask, a separatory funnel filled with hydrochloric acid, and an outlet tube connected to a volumetric flask containing 5 mol / L sodium hydroxide and water. Pass a steady stream of nitrogen through the air inlet tube and add hydrochloric acid at the same time. Allow to react for 30 minutes and disconnect the device. Add N, N-dimethyl-p-phenylenediamine dihydrochloride solution to the volumetric flask, then add 5% ferric chloride hydrochloric acid solution, keep it away from light for 20 minutes, dilute it to the scale with water, shake it well, and use it as the test sample solution. Substitute water for acetylcysteine injection and prepare a blank solution in the same way.
[0009] (3) Substituting the hydrogen sulfide standard solution obtained in step (1) for the acetylcysteine injection solution, prepare the reference solution in the same manner.
[0010] (4) Using the blank solution as the blank control, the absorbance of the test solution and the reference solution was measured by a spectrophotometer and recorded as A 供 , A 对 ; Calculate the content of hydrogen sulfide according to the following formula:
[0011]
[0012] in:
[0013] A 供 : Absorbance of the solution to be tested
[0014] A 对 : Absorbance of reference substance
[0015] C 对 : The concentration of hydrogen sulfide in the reference substance
[0016] C 供 : The concentration of acetylcysteine in the test solution
[0017] 1000: Unit conversion factor
[0018] Further optimization, as the detection method of hydrogen sulfide content in the above-mentioned acetylcysteine injection, the specific detection method of step (2) is: accurately pipette 2ml of the test solution, put it in a three-necked round-bottom flask, add 40ml of water, the flask contains an air inlet pipe leading to the bottom of the flask, a separating funnel equipped with hydrochloric acid, and an outlet pipe, the outlet pipe is connected to a 100ml measuring bottle, and the bottle contains 5mol / L sodium hydroxide 1ml and 50ml water. A stable nitrogen flow is introduced through the air inlet pipe, and 10ml of hydrochloric acid is added at the same time. Keep the nitrogen flow rate at 600ml / min, fully react for 30min, and disconnect the device. Add 10ml of N, N-dimethyl-p-phenylenediamine dihydrochloride solution to the measuring bottle, add 5ml of 5% ferric chloride hydrochloric acid solution, keep it in dark and let it stand for 20min, dilute it with water to the scale, shake it well, and use it as the test solution.
[0019] Further optimization, such as the above-mentioned method for detecting the hydrogen sulfide content in acetylcysteine injection, wherein the method for preparing the N,N-dimethyl-p-phenylenediamine dihydrochloride solution in step (2) is as follows: accurately weigh 0.1 g of N,N-dimethyl-p-phenylenediamine dihydrochloride, dissolve it in hydrochloric acid-water (45:55) and dilute it to 100 ml; the method for preparing the 5% ferric chloride hydrochloric acid solution is as follows: dilute 5 g of ferric chloride hexahydrate with 1 mol / L hydrochloric acid to 100 ml.
[0020] Further optimization, such as the above-mentioned method for detecting the hydrogen sulfide content in acetylcysteine injection, wherein the preparation method of N, N-dimethyl-p-phenylenediamine dihydrochloride solution in step (2) is as follows: accurately weigh 0.1 g of N, N-dimethyl-p-phenylenediamine dihydrochloride, dissolve it in 45 ml of hydrochloric acid, dilute it with 55 ml of water, and then add activated carbon for decolorization; the preparation method of 5% ferric chloride hydrochloric acid solution is as follows: accurately weigh 5 g of ferric chloride hexahydrate and place it in a 100 ml volumetric flask, then add 9 ml of hydrochloric acid, and dilute it to the scale with water.
[0021] Further optimization, such as the above-mentioned method for detecting the hydrogen sulfide content in acetylcysteine injection, changes the dosage of the reference substance and the test substance, and detects the solution. BRIEF DESCRIPTION OF THE DRAWINGS
[0022] Figure 1 :Diagram of hydrogen sulfide collection device
[0023] Figure 2 : Absorbance curve of hydrogen sulfide
[0024] Figure 2 The horizontal axis is the concentration of hydrogen sulfide (ug / ml), and the vertical axis is the absorbance. When hydrogen sulfide is in the range of 8ug / ml-120ug / ml, its linear equation is y=0.0075x+0.0099, and the linear correlation coefficient is R2=0.9994. Specific implementation methods
[0026] Embodiment 1: the detection of hydrogen sulfide content in acetylcysteine injection
[0027] Prepare the test sample and blank solution: accurately pipette 2 ml of the test sample solution into a three-necked round-bottom flask, add 40 ml of water, the flask contains an air inlet leading to the bottom of the flask, a separatory funnel filled with hydrochloric acid, and an outlet tube connected to a 100 ml volumetric flask containing 1 ml of 5 mol / L sodium hydroxide and 50 ml of water. Pass a steady stream of nitrogen through the air inlet tube and add 10 ml of hydrochloric acid at the same time. Keep the nitrogen flow rate at 600 ml / min, react fully for 30 minutes, and disconnect the device. Add 10 ml of N, N-dimethyl-p-phenylenediamine dihydrochloride solution (precisely weigh 0.1 g of N, N-dimethyl-p-phenylenediamine dihydrochloride, dissolve it with hydrochloric acid-water (45:55) and dilute it to 100 ml) to the measuring bottle, then add 5 ml of 5% ferric chloride hydrochloric acid solution (dilute 5 g of ferric chloride hexahydrate with 1 mol / L hydrochloric acid to 100 ml), keep it in the dark for 20 minutes, dilute it to the scale with water, shake it well, and use it as the test solution. Replace acetylcysteine injection with water and prepare a blank solution in the same way.
[0028] Reference solution: dilute 0.1 mol / L standard sodium sulfide solution 5000 times, accurately pipette 2 ml, and replace acetylcysteine injection to prepare reference solution in the same way.
[0029] Determination of hydrogen sulfide content in the test sample: Using the blank solution as the blank control, the hydrogen sulfide absorbance of the reference sample and the test sample was measured by a spectrophotometer at a wavelength of 665 nm, and the absorbance was calculated as A 对 , A 供 , calculate the content of hydrogen sulfide according to the following formula; the results are shown in Table 1
[0030]
[0031] in:
[0032] A 供 : Absorbance of the solution to be tested
[0033] A 对 : Absorbance of reference substance
[0034] C 对 : The concentration of hydrogen sulfide in the reference substance
[0035] C 供 : The concentration of acetylcysteine in the test solution
[0036] 1000: Unit conversion factor
[0037] Table 1
[0038] Solution name Absorbance blank solution 0.000 Reference solution 0.012 Test solution 0.034
[0039] Conclusion: The absorbance of the test sample should be smaller than that of the reference sample. It can be seen that the dilution factor of the reference sample is too large and the absorbance measured by this method does not meet the standard requirements.
[0040] Example 2: Detection of hydrogen sulfide content in acetylcysteine injection
[0041] Prepare the test sample and blank solution: accurately pipette 4 ml of the test sample solution into a three-necked round-bottom flask, add 40 ml of water, the flask contains an air inlet leading to the bottom of the flask, a separatory funnel filled with hydrochloric acid, and an outlet tube connected to a 100 ml volumetric flask containing 1 ml of 5 mol / L sodium hydroxide and 50 ml of water. Pass a steady stream of nitrogen through the air inlet tube and add 10 ml of hydrochloric acid at the same time. Keep the nitrogen flow rate at 600 ml / min, fully react for 30 minutes, and disconnect the device. Add 10 ml of N, N-dimethyl-p-phenylenediamine dihydrochloride solution (accurately weigh 0.1 g of N, N-dimethyl-p-phenylenediamine dihydrochloride, dissolve it in 45 ml of hydrochloric acid, dilute it with 55 ml of water, and then add activated carbon for decolorization) to the measuring bottle, and then add 5 ml of 5% ferric chloride hydrochloric acid solution (accurately weigh 5 g of ferric chloride hexahydrate and put it in a 100 ml volumetric flask, then add 9 ml of hydrochloric acid and dilute it to the scale with water), keep it in the dark for 20 minutes, dilute it to the scale with water, shake it well, and use it as the test solution. Replace acetylcysteine injection with water and prepare a blank solution in the same way.
[0042] Reference solution: dilute 1000 mg / L standard sodium sulfide solution 10 times, accurately pipette 2 ml, and replace acetylcysteine injection to prepare reference solution in the same way.
[0043] Determination of hydrogen sulfide content in the test sample: Using the blank solution as the blank control, the hydrogen sulfide absorbance of the reference sample and the test sample was measured by a spectrophotometer at a wavelength of 665 nm, and the absorbance was calculated as A 对 , A 供 , calculate the content of hydrogen sulfide according to the following formula; the results are shown in Table 2
[0044]
[0045] in:
[0046] A 供 : Absorbance of the solution to be tested
[0047] A 对 : Absorbance of reference substance
[0048] C 对 : The concentration of hydrogen sulfide in the reference substance
[0049] C 供 : The concentration of acetylcysteine in the test solution
[0050] 1000: Unit conversion factor
[0051] Table 2
[0052] Solution name Absorbance Hydrogen sulfide content blank solution 0.000 / Reference solution 0.444 / Test solution 0.019 0.0009％
[0053] Example 3: Detection of hydrogen sulfide content in acetylcysteine injection
[0054] Prepare the test sample and blank solution: accurately pipette 8 ml of the test sample solution into a three-necked round-bottom flask, add 40 ml of water, the flask contains an air inlet leading to the bottom of the flask, a separatory funnel filled with hydrochloric acid, and an outlet tube connected to a 100 ml volumetric flask containing 1 ml of 5 mol / L sodium hydroxide and 50 ml of water. Pass a steady stream of nitrogen through the air inlet tube and add 10 ml of hydrochloric acid at the same time. Keep the nitrogen flow rate at 600 ml / min, fully react for 30 minutes, and disconnect the device. Add 10 ml of N, N-dimethyl-p-phenylenediamine dihydrochloride solution (accurately weigh 0.1 g of N, N-dimethyl-p-phenylenediamine dihydrochloride, dissolve it in 45 ml of hydrochloric acid, dilute it with 55 ml of water, and then add activated carbon for decolorization) to the measuring bottle, and then add 5 ml of 5% ferric chloride hydrochloric acid solution (accurately weigh 5 g of ferric chloride hexahydrate and put it in a 100 ml volumetric flask, then add 9 ml of hydrochloric acid and dilute it to the scale with water), keep it in the dark for 20 minutes, dilute it to the scale with water, shake it well, and use it as the test solution. Replace acetylcysteine injection with water and prepare a blank solution in the same way.
[0055] Reference solution: dilute 1000 mg / L standard sodium sulfide solution 20 times, accurately pipette 4 ml, and replace acetylcysteine injection to prepare reference solution in the same way.
[0056] Determination of hydrogen sulfide content in the test sample: Using the blank solution as the blank control, the hydrogen sulfide absorbance of the reference sample and the test sample was measured by a spectrophotometer at a wavelength of 665 nm, and the absorbance was calculated as A 对 , A 供 , calculate the content of hydrogen sulfide according to the following formula; the results are shown in Table 3
[0057]
[0058] in:
[0059] A 供 : Absorbance of the solution to be tested
[0060] A 对 : Absorbance of reference substance
[0061] C 对 : The concentration of hydrogen sulfide in the reference substance
[0062] C 供 : The concentration of acetylcysteine in the test solution
[0063] 1000: Unit conversion factor
[0064] Table 3
[0065] Solution name Absorbance Hydrogen sulfide content blank solution 0.000 / Reference solution 0.404 / Test solution 0.204 0.005％
[0066] Conclusion: This method is better than Example 2.
[0067] Example 4: Linearity and range study
[0068] Preparation of hydrogen sulfide standard solution: Take 1 ml of 1000 mg / L standard sodium sulfide solution and place it in a 20 ml volumetric flask, add water to dilute to the scale, and shake well.
[0069] Accurately pipette 0.4ml, 1ml, 2ml, 4ml, and 6ml of sodium sulfide standard solution into a three-necked round-bottom flask, and add 40ml of water. The flask contains an air inlet leading to the bottom of the flask, a separatory funnel filled with hydrochloric acid, and an outlet pipe connected to a 100ml volumetric flask containing 1ml of 5mol / L sodium hydroxide and 50ml of water. A steady stream of nitrogen is introduced through the air inlet pipe, and 10ml of hydrochloric acid is added at the same time. Maintain a nitrogen flow rate of 600ml / min, fully react for 30min, and disconnect the device. Add 10 ml of N, N-dimethyl-p-phenylenediamine dihydrochloride solution (accurately weigh 0.1 g of N, N-dimethyl-p-phenylenediamine dihydrochloride, dissolve it in 45 ml of hydrochloric acid, dilute it with 55 ml of water, and then add activated carbon for decolorization) to the measuring bottle, and then add 5 ml of 5% ferric chloride hydrochloric acid solution (accurately weigh 5 g of ferric chloride hexahydrate and put it in a 100 ml volumetric flask, then add 9 ml of hydrochloric acid and dilute it to the scale with water), keep it away from light for 20 minutes, dilute it to the scale with water, shake it well, and you have it. Replace acetylcysteine injection with water and prepare a blank solution in the same way.
[0070] Accurately pipette 1ml, 2ml, 4ml and 6ml of hydrogen sulfide standard solution respectively and repeat the above operation.
[0071] Absorbance determination: The above blank solution was used as a blank control, and the absorbance of the solution was measured by a spectrophotometer at a wavelength of 665 nm; the results are shown in Table 4 and Attached Figure 2
[0072] Table 4
[0073] Amount of hydrogen sulfide (ug / ml) Absorbance 8 0.061 20 0.164 40 0.311 80 0.621 120 0.901
[0074] Results: According to Figure 2 The absorbance curve of hydrogen sulfide and Table 4 show that the linear equation of hydrogen sulfide in the range of 8ug / ml-120ug / ml is y=0.0075x+0.0099, and the linear correlation coefficient is R 2 =0.9994, the linear range meets the requirements and the linear relationship is good.
Claims
1. A method for detecting the content of hydrogen sulfide in acetylcysteine injection, comprising the following steps: (1) Prepare a 20 ppm hydrogen sulfide standard solution; (2) Preparation of test sample and blank solution: Accurately pipette acetylcysteine injection into a three-necked round-bottom flask and add water. The flask contains an air inlet leading to the bottom of the flask, a separatory funnel filled with hydrochloric acid, and an outlet tube connected to a volumetric flask containing 5 mol / L sodium hydroxide and water. Pass a steady stream of nitrogen through the air inlet tube and add hydrochloric acid at the same time. Allow to react for 30 minutes and disconnect the device. Add N, N-dimethyl-p-phenylenediamine dihydrochloride solution to the volumetric flask, then add 5% ferric chloride hydrochloric acid solution, keep it away from light for 20 minutes, dilute to the scale with water, and shake well. Use as the test sample solution. Substitute water for acetylcysteine injection and prepare a blank solution in the same manner. (3) Preparation of reference solution: Substitute the hydrogen sulfide standard solution obtained in step (1) for the acetylcysteine injection solution and prepare the reference solution in the same manner. (4) Using the blank solution as the blank control, the absorbance of the test solution and the reference solution was measured by a spectrophotometer and recorded as A 供 , A 对 ; Calculate the content of hydrogen sulfide according to the following formula in: A 供 : Absorbance of the solution to be tested A 对 : Absorbance of reference substance C 对 : The concentration of hydrogen sulfide in the reference substance C 供 : The concentration of acetylcysteine in the test solution 1000: Unit conversion factor.
2. The method for detecting the hydrogen sulfide content in the acetylcysteine injection according to claim 1, characterized in that: The nitrogen flow rate in step (2) is 1.5-2.5 L / min.
3. The method for detecting the hydrogen sulfide content in the acetylcysteine injection according to claim 1, characterized in that: Step (2) The specific steps of preparing the test sample include: accurately pipetting 2 ml of the test sample solution, placing it in a three-necked round-bottom flask, adding 40 ml of water, the flask containing an air inlet pipe leading to the bottom of the flask, a separatory funnel filled with hydrochloric acid, and an outlet pipe, the outlet pipe is connected to a 100 ml volumetric flask, and the bottle contains 1 ml of 5 mol / L sodium hydroxide and 50 ml of water. A stable nitrogen flow is introduced through the air inlet pipe, and 10 ml of hydrochloric acid is added at the same time. The nitrogen flow rate is maintained at 600 ml / min, fully reacted for 30 minutes, and the device is disconnected. 10 ml of N, N-dimethyl-p-phenylenediamine dihydrochloride solution is added to the volumetric flask, and then 5 ml of 5% ferric chloride hydrochloric acid solution is added. The solution is kept away from light and allowed to stand for 20 minutes, diluted to the scale with water, and shaken well to serve as the test sample solution.
4. The method for detecting the hydrogen sulfide content in the acetylcysteine injection according to claim 3, characterized in that: The preparation method of the N, N-dimethyl-p-phenylenediamine dihydrochloride solution is as follows: accurately weigh 0.1 g of N, N-dimethyl-p-phenylenediamine dihydrochloride, dissolve it in 45 ml of hydrochloric acid, and dilute it with 55 ml of water.
5. The method for detecting the hydrogen sulfide content in the acetylcysteine injection according to claim 3, characterized in that: The preparation method of the 5% ferric chloride hydrochloric acid solution is as follows: accurately weigh 5g of ferric chloride hexahydrate and place it in a 100ml volumetric flask, then add 9ml of hydrochloric acid, and dilute with water to the scale.
6. The method for detecting the hydrogen sulfide content in acetylcysteine injection according to claim 1, characterized in that: In step (4), the wavelength of the test sample and the reference sample detected by spectrophotometry is 665 nm.