Method for determining content of malic acid in auxiliary material in fudosteine oral solution
The determination of malic acid content in fodosteine oral solution by ion chromatography solves the problem of inaccuracy in existing methods, achieves highly sensitive and specific excipient analysis, and ensures drug quality and R&D efficiency.
Patent Information
- Application Number
- CN202511082836.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-08-04
- Publication Date
- 2025-10-21
AI Technical Summary
The lack of a highly sensitive, specific, and accurate method for determining the malic acid content of the excipient in fodosteine oral solution makes drug quality analysis difficult and affects drug safety and research and development efficiency.
Ion chromatography was employed using Dionex IonPac™ AS18 and Dionex IonPac™ AG18 columns, with potassium hydroxide aqueous solution as the eluent. The flow rate was 1.0 ml/min, the column temperature was 30 °C, and a conductivity detector with a current of 62 mA was used. The injection volume was 25 μl, and isocratic elution was performed. The results were then determined using conductivity detection.
This method enables accurate determination of malic acid content in fudostein oral solution, improving the sensitivity and specificity of the determination method, ensuring the accuracy and safety of drug quality control, reducing R&D workload, and shortening development time.
Smart Images
Figure CN120820652A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of analytical chemistry, and particularly relates to a method for determining the content of malic acid, an auxiliary material, in a fudosteine oral solution. Background Art
[0002] Fudosteine is an expectorant used to treat chronic respiratory diseases such as bronchial asthma, bronchiectasis, chronic bronchitis, tuberculosis, pneumoconiosis, and emphysema.
[0003] Fudosteine oral solution can reduce sputum purulent and sticky sputum, reduce coughing frequency, improve suppressed respiratory function, and alleviate local inflammation. It is primarily used clinically to treat and eliminate phlegm in chronic respiratory diseases such as bronchial asthma, chronic bronchitis, bronchiectasis, tuberculosis, pneumoconiosis, emphysema, atypical mycobacterial infection, and diffuse bronchiolitis.
[0004] The prescription of the fudosteine oral solution disclosed by the invention comprises: fudosteine, caramel, malic acid, essence, ethanol, sodium benzoate, glycerin and sorbitol.
[0005] Currently, there is no method for determining the content of malic acid, an excipient in fudosteine oral solution, recorded in pharmacopoeias, patents and literatures of various countries.
[0006] In order to effectively analyze drug quality and ensure drug safety, it is necessary to develop a method for determining the content of excipients that is highly sensitive, specific, durable, convenient, and effective. Summary of the Invention
[0007] The technical problem to be solved by the present invention is to provide a method for determining the content of malic acid, an excipient in fudosteine oral solution, to analyze the prescription of fudosteine oral solution. The method has high sensitivity, good specificity, and high accuracy, can reduce the difficulty of formulation development, reduce the workload of prescription development, shorten the research and development time, and thus improve the BE pass rate, which has practical significance.
[0008] In order to solve the above technical problems, the technical solution adopted by the present invention is:
[0009] A method for determining the content of malic acid in fudosteine oral solution using ion chromatography using a Dionex IonPac TM AS18 column; guard column Dionex IonPac TM AG18 column; potassium hydroxide aqueous solution as eluent, flow rate of 1.0 ml / min; column temperature of 30°C; conductivity detector, detection method of suppressed conductivity detection, current value of 62 mA, conductivity cell temperature of 35°C; injection volume of 25 μl; isocratic elution.
[0010] Preferably, the isocratic elution procedure is as follows:
[0011]
[0012] Preferably, the chromatographic column is an anion exchange column. In the embodiment of the present invention, an anion exchange column Dionex IonPac TM AS18 column, size 4mm×250mm, provided by Thermo Fisher Scientific; the guard column is an anion guard column, preferably Dionex IonPac TM AG18 column, size 4 mm × 50 mm, provided by Thermo Fisher Scientific.
[0013] Preferably, the flow rate is 0.5 ml / min to 1.5 ml / min, more preferably 1.0 ml / ml.
[0014] Preferably, the column temperature is 25°C to 35°C, more preferably 30°C.
[0015] Preferably, the conductivity cell temperature is 30°C to 40°C, more preferably 35°C.
[0016] A conductivity detector was used, the detection method was suppressed conductivity detection, and the current value was 62 mA.
[0017] Preferably, the injection volume is 25 μl.
[0018] Preferably, the method comprises the following steps:
[0019] (1) Test solution: Accurately measure 0.3 ml of the product into a 100 ml volumetric flask, dilute to the mark with water, and shake well;
[0020] (2) Reference solution: Take an appropriate amount of malic acid, accurately weigh it, dissolve it in water, and quantitatively dilute it to make a solution containing approximately 8 μg of malic acid per 1 ml;
[0021] (3) using ion chromatography to measure the reference solution and the test solution respectively;
[0022] The detection conditions of the ion chromatography method are as follows: chromatographic column Dionex IonPac TM AS18 column; guard column Dionex IonPac TM AG18 column; potassium hydroxide aqueous solution as eluent, flow rate of 1.0 ml / min; column temperature of 30°C; conductivity detector, detection method of suppressed conductivity detection, current value of 62 mA, conductivity cell temperature of 35°C; injection volume of 25 μl; isocratic elution.
[0023] Calculation of results: Calculated according to the external standard method.
[0024] Due to the adoption of the above technical solution, the present invention has the following advantages compared with the prior art:
[0025] 1. The determination method of the present invention comprehensively analyzes the formulation of Fudosteine oral solution and can accurately determine the content of malic acid as an excipient;
[0026] 2. The determination method of the present invention has high sensitivity, strong specificity and good accuracy, providing a simple and convenient detection method for the prescription analysis of Fudosteine oral solution. It can strictly control the quality of Fudosteine oral solution, ensure the safety and reliability of Fudosteine oral solution, and has practical significance. BRIEF DESCRIPTION OF THE DRAWINGS
[0027] Figure 1 is a chromatogram of a blank solution in the present invention;
[0028] Figure 2 is a chromatogram of the blank matrix solution in the present invention;
[0029] Figure 3 It is the chromatogram of the reference substance solution in the present invention;
[0030] Figure 4 It is the chromatogram of the test solution in the present invention; DETAILED DESCRIPTION
[0031] To make the technical solutions and beneficial effects of the present invention more clearly understood, the following is a detailed description with reference to the accompanying drawings and specific embodiments. It should be understood that these embodiments are intended only to illustrate the present invention and are not intended to limit the scope of the present invention. Any product identical or similar to the present invention that is derived by anyone under the guidance of the present invention or by combining the features of the present invention with those of other prior arts shall fall within the scope of protection of the present invention.
[0032] The experimental methods in the following examples without specific conditions are generally based on conventional experimental conditions.
[0033] The fudosteine oral solution, malic acid and equipment used in the specific embodiments of the present invention are all known products, and the fudosteine oral solution and malic acid are obtained by purchasing commercially available products.
[0034] Table 1 Equipment information
[0035] Device Name model factory Ion Chromatograph ICS2000 Thermo Fisher Scientific electronic balance 125SM-FR Prissex Weighing Equipment Co., Ltd. Water Purifier EUE-10UV Shanghai Haisi Instrument Technology Co., Ltd.
[0036] Table 2 Material information
[0037] name source batch number Potassium hydroxide eluent Thermo Fisher Scientific 074532 Blank matrix Made by the pharmaceutical preparation R&D department JT002-240530-2 Fudosteine oral solution Made by the pharmaceutical preparation R&D department JT002-240228-1
[0038] Table 3 Reference substance information
[0039] name factory batch number content Malic acid Changmao Biochemical Engineering Co., Ltd. FA20230508 99.54％
[0040] Example 1
[0041] Methodological research on the detection and analysis method of the present invention
[0042] Various tests in this embodiment all adopt the following conditions:
[0043] Column: Dionex IonPac TM AS18 column (4mm×250mm);
[0044] Guard column: Dionex IonPac TM AG18 column (4 mm × 50 mm);
[0045] Flow rate: 1.0 ml / min;
[0046] Column temperature: 30°C;
[0047] Detector: conductivity detector;
[0048] Detection method: suppressed conductivity detection;
[0049] Current value: 62mA;
[0050] Injection volume: 25 μl
[0051] The isocratic elution procedure is as follows:
[0052]
[0053] Testing steps:
[0054] Test solution: Accurately measure 0.3 ml of the product, place it in a 100 ml volumetric flask, dilute to the scale with water, and shake well.
[0055] Reference solution: Take an appropriate amount of malic acid, accurately weigh it, dissolve it in water and quantitatively dilute it to make a solution containing approximately 8 μg of malic acid per 1 ml.
[0056] 1. Specificity test
[0057] Solution preparation:
[0058] Blank solvent: ultrapure water.
[0059] Blank matrix solution: Accurately measure 0.3 ml of blank matrix, place it in a 100 ml volumetric flask, dilute to the scale with water, and shake well.
[0060] Reference solution: Take 20 mg of malic acid, accurately weigh it, place it in a 50 ml volumetric flask, dissolve it with water and dilute it to the scale, shake it well, then accurately measure 2 ml, place it in a 100 ml volumetric flask, dilute it to the scale with water, and shake it well.
[0061] Test solution: Accurately measure 0.3 ml of the product, place it in a 100 ml volumetric flask, dilute to the scale with water, and shake well.
[0062] Accurately measure 25 μl of blank solvent, blank matrix solution, malic acid reference solution and test solution and inject them into the ion chromatograph respectively, record the chromatogram, and the test results are shown in the attached Figure 1 ~Attachment Figure 4 .
[0063] From the results in the attached figure, it can be seen that the unknown peaks in the blank solution, blank matrix solution and test solution have no interference with the detection of malic acid, and the method has good specificity.
[0064] 2. Precision test
[0065] Solution preparation:
[0066] Reference solution: Take 20 mg of malic acid, accurately weigh it, place it in a 50 ml volumetric flask, dissolve it with water and dilute it to the scale, shake it well, then accurately measure 2 ml, place it in a 100 ml volumetric flask, dilute it to the scale with water, and shake it well.
[0067] Take the reference solution and inject it 5 times continuously, record the peak area and retention time, and calculate the RSD values of the peak area and retention time. The test results are shown in Table 4 below.
[0068] Table 4 Malic acid injection precision test results
[0069]
[0070] From the above results, we can see that after 5 consecutive injections of the reference solution, the RSD value of the malic acid peak area is 0.35%; the RSD value of the malic acid retention time is 0.02%, both of which are no more than 2.0%. This method has good injection precision.
[0071] 3. Linearity and range test
[0072] Solution preparation:
[0073] Linear stock solution: Take 20 mg of malic acid, accurately weigh it, place it in a 50 ml volumetric flask, dissolve it in water and dilute it to the scale, and shake well.
[0074] Take an appropriate amount of linear stock solution and prepare linear solutions of various concentrations according to the table below.
[0075] Linear concentration Volume of linear stock solution pipetted (ml) Dilute to (ml) 25％ 0.5 100 50％ 1 100 75％ 1.5 100 100％ 2 100 150％ 3 100 200％ 4 100
[0076] Accurately measure 25 μl of each linear solution, inject it into the ion chromatograph, record the chromatogram, and perform linear regression analysis on the concentration based on the peak area. The test results are shown in Table 5 below.
[0077] Table 5 Malic acid linearity test results
[0078]
[0079] From the above results, we can see that the correlation coefficient R 2 It is 0.9992 and not less than 0.998. The method has a good linear relationship.
[0080] 4. Detection limit test
[0081] Solution preparation:
[0082] Take each linear point solution prepared under 3. Linear range test and dilute it step by step. The solution with a signal-to-noise ratio of not less than 10:1 is used as the quantification limit solution, and the solution with a signal-to-noise ratio of not less than 3:1 is used as the detection limit solution.
[0083] The above diluted solutions were respectively injected into the ion chromatograph, and the chromatograms were recorded. The test results are shown in Table 6 below.
[0084] Table 6 Results of detection limits of malic acid
[0085]
[0086] From the above results, it can be seen that the ratio of the detection limit concentration of malic acid to the concentration of the reference substance is 0.05%, which is no more than 10% of the concentration of the reference substance solution; the ratio of the quantitative limit concentration of malic acid to the concentration of the reference substance solution is 0.10%, which is no more than 20% of the concentration of the reference substance. This method has high detection sensitivity.
[0087] 5. Repeatability test
[0088] Solution preparation:
[0089] Blank matrix solution: Accurately measure 0.3 ml of blank matrix, place it in a 100 ml volumetric flask, dilute to the scale with water, and shake well.
[0090] Reference solution: Take 20mg of malic acid, accurately weigh it, place it in a 50ml volumetric flask, dissolve it with water and dilute it to the mark, shake it well, then accurately measure 2ml, place it in a 100ml volumetric flask, dilute it to the mark with water, and shake it well. (Prepare 2 copies in parallel)
[0091] Test solution: Accurately measure 0.3 ml of the product into a 100 ml volumetric flask, dilute to the mark with water, and shake well. (Prepare 6 portions in parallel)
[0092] Accurately measure 25 μl of blank matrix solution, reference solution, and test solution respectively and inject them into the ion chromatograph. Record the chromatograms and calculate the measured amount of malic acid in the 6 test samples using the external standard method. The test results are shown in Table 7 below.
[0093] Table 7 Malic acid repeatability test results
[0094]
[0095] From the above results, we can see that the RSD value of the measured amount of malic acid in the 6 test solutions is 0.62%, which is no more than 2.0%, and the method has good repeatability.
[0096] 6. Accuracy test
[0097] Solution preparation:
[0098] Blank matrix solution: Accurately measure 0.3 ml of blank matrix, place it in a 100 ml volumetric flask, dilute to the scale with water, and shake well.
[0099] Malic acid reference stock solution: Take 20 mg of malic acid, accurately weigh it, place it in a 50 ml volumetric flask, dissolve it in water and dilute it to the scale, and shake well.
[0100] Reference solution: Accurately measure 2 ml of malic acid reference stock solution, place it in a 100 ml volumetric flask, dilute to the mark with water, and shake well.
[0101] Prepare two replicates.
[0102] 50% recovery solution: Accurately measure 0.3 ml of blank matrix and place it in a 100 ml volumetric flask. Accurately add 1 ml of malic acid reference stock solution, dilute to the mark with water, shake well, and prepare 3 replicates.
[0103] 100% recovery solution: Accurately measure 0.3 ml of blank matrix and place it in a 100 ml volumetric flask. Accurately add 2 ml of malic acid reference stock solution, dilute to the mark with water, shake well, and prepare 3 replicates.
[0104] 150% recovery solution: Accurately measure 0.3 ml of blank matrix and place it in a 100 ml volumetric flask. Accurately add 3 ml of malic acid reference stock solution, dilute to the mark with water, shake well, and prepare three replicates.
[0105] Accurately measure 25 μl of blank matrix solution, reference solution and recovery solution and inject them respectively. Record the chromatogram and calculate by external standard method. Compare the measured amount with the added amount and calculate the recovery. The test results are shown in Table 8 below.
[0106] Table 8 Malic acid accuracy test results
[0107]
[0108] From the above results, we can see that the average value of 9 recovery rate data at various malic acid concentrations is 100.3%, between 95.0% and 105.0%, and the RSD value of the 9 recovery rate data is 2.65%, which is no more than 5.0%. This method has good accuracy.
[0109] 7. Solution stability test
[0110] Solution preparation:
[0111] Take the reference solution (prepared in the same way as in 1. Specificity test) and place them at room temperature.
[0112] 25 μl of each of the above solutions was injected into an ion chromatograph, and the chromatogram was recorded. The test results are shown in Table 9 below.
[0113] Table 9 Reference solution storage stability test results
[0114]
[0115] From the above results, it can be seen that the ratio of the malic acid concentration of the reference solution after being placed at room temperature for 4 days to the concentration on day 0 is 99.7, which meets the requirement between 98.0% and 102.0%. Therefore, the reference solution is stable at room temperature for at least 4 days.
[0116] In summary, the present invention provides a method suitable for determining the content of malic acid, an excipient in fudosteine oral solution. The method has good specificity, and baseline separation can be achieved between malic acid and the main component; the system applicability is good; the method is extremely sensitive, and the detection limit of malic acid can reach 0.004119 μg / ml; the method has good repeatability and accuracy, and provides a convenient detection method for the prescription analysis of fudosteine oral solution, so as to prepare a product with quality and efficacy consistent with the original drug.
[0117] It should be understood that the above embodiments are exemplary and are not intended to encompass all possible implementations of the claims. Various modifications and variations may be made to the above embodiments without departing from the scope of this disclosure. Similarly, the various technical features of the above embodiments may be arbitrarily combined to form additional embodiments of the present invention that may not be explicitly described. Therefore, the above embodiments merely illustrate several implementations of the present invention and do not limit the scope of protection of the patent of this invention.
Claims
1. A method for determining the content of malic acid in a fudosteine oral solution, characterized in that: Ion chromatography was used, an anion exchange column was used as the chromatographic column, and potassium hydroxide aqueous solution was used as the eluent for isocratic elution.
2. The method for determining the content of malic acid as an auxiliary material in the fudosteine oral solution according to claim 1, characterized in that: The chromatographic column is an anion exchange column, preferably Dionex IonPac TM AS18 column, size 4mm×250mm, provided by Thermo Fisher Scientific; the guard column is an anion guard column, preferably Dionex IonPac TM AG18 column, size 4 mm × 50 mm, provided by Thermo Fisher Scientific.
3. The measuring method according to claim 1, wherein The flow rate is 1.0 ml / ml; preferably, the flow rate is 0.5 ml / min to 1.5 ml / min, and more preferably 1.0 ml / ml.
4. The measuring method according to claim 1, wherein The column temperature is 30°C; preferably, the column temperature is 25°C to 35°C, more preferably 30°C.
5. The measuring method according to claim 1, wherein The temperature of the conductivity cell is 35°C; preferably, the temperature of the conductivity cell is 30°C to 40°C, more preferably 35°C.
6. The measuring method according to claim 1, wherein A conductivity detector was used, the detection method was suppressed conductivity detection, and the current value was 62 mA.
7. The measuring method according to claim 1, wherein The injection volume was 25 μl.
8. The method for determining the content of malic acid as an auxiliary material in the fudosteine oral solution according to any one of claims 1 to 7, characterized in that: The following steps are involved: (1) Test solution: Accurately measure 0.3 ml of the product into a 100 ml volumetric flask, dilute to the mark with water, and shake well; (2) Reference solution: Take an appropriate amount of malic acid, accurately weigh it, dissolve it in water, and quantitatively dilute it to make a solution containing approximately 8 μg of malic acid per 1 ml; (3) using ion chromatography to measure the reference solution and the test solution respectively; The detection conditions of the ion chromatography method are as follows: chromatographic column Dionex IonPac TM AS18 column; guard column Dionex IonPac TM AG18 column; potassium hydroxide aqueous solution as eluent, flow rate of 1.0 ml / min; The column temperature was 30°C; a conductivity detector was used, the detection method was suppressed conductivity detection, the current value was 62 mA, the conductivity cell temperature was 35°C; the injection volume was 25 μl; and isocratic elution was performed.