Method for measuring contents of auxiliary materials sorbitol and glycerol in fudosteine oral solution

Abstract

The invention relates to a method for determining the content of sorbitol and glycerol as auxiliary materials in a fudosteine oral solution, which has the advantages of high sensitivity, strong specificity, good accuracy and simple and rapid operation, can reduce the difficulty of preparation development, reduce the workload of prescription development, shorten the research and development time and further improve the BE passing rate, and has practical significance.

Description

Technical Field

[0001] This invention belongs to the field of analytical chemistry technology, and specifically relates to a method for determining the content of excipients sorbitol and glycerol in fodosteine ​​oral solution. Background Technology

[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 reduces the purulent and adhesive nature of sputum, decreases the frequency of coughing, improves suppressed respiratory function, and alleviates local inflammation. Clinically, it is mainly used as an expectorant for chronic respiratory diseases such as bronchial asthma, chronic bronchitis, bronchiectasis, pulmonary tuberculosis, pneumoconiosis, emphysema, atypical mycobacterial infection, and diffuse bronchiolitis.

[0004] The formula of the fudostein oral solution disclosed in this invention includes: fudostein, caramel, malic acid, flavoring, ethanol, sodium benzoate, glycerin and sorbitol.

[0005] Currently, there are no records in pharmacopoeias, patents, or literature from various countries regarding the detection methods for the excipients sorbitol and glycerin in fudostein oral solution.

[0006] In order to effectively analyze drug quality and ensure medication safety, it is necessary to develop a method for determining excipient content that is highly sensitive, specific, robust, 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 excipients sorbitol and glycerol in fudostein oral solution, and to analyze the formulation of fudostein oral solution. This method has high sensitivity, good specificity and high accuracy, which can reduce the difficulty of formulation development, reduce the workload of formulation development, shorten the research and development time, and thus improve the bioequivalence (BE) success rate, which has practical significance.

[0008] To solve the above technical problems, the technical solution adopted by the present invention is as follows:

[0009] A method for determining the content of excipients sorbitol and glycerol in fodosteine ​​oral solution, using gas chromatography with a DB-624 column; initial column temperature of 160℃, maintained for 2 min, then increased to 240℃ at a rate of 5℃ / min, maintained for 15 min; injection port temperature of 280℃; FID detector temperature of 250℃; injection volume of 1 μl.

[0010] Preferably, the chromatographic column is a capillary column. In this embodiment of the invention, a medium polarity chromatographic column DB-624 is used; the column has dimensions of 30m × 0.32mm and 1.8μm, and is provided by Agilent Technologies.

[0011] Preferably, the carrier gas used is nitrogen.

[0012] Preferably, the detector used is a flame ionization detector (FID).

[0013] Preferably, the initial column temperature is 130℃~180℃, and more preferably 160℃.

[0014] Preferably, the injection port temperature is 240℃~300℃; the detector temperature is 230℃~300℃; more preferably, the injection port temperature is 280℃ and the detector temperature is 250℃.

[0015] Preferably, the injection volume is 1 μl; the split mode is 5:1 to 50:1, preferably 20:1.

[0016] Preferably, the carrier gas flow rate is 1.0 ml / min to 5.0 ml / min, and more preferably 3.5 ml / min.

[0017] Preferably, the injection method is direct injection.

[0018] Preferably, the method includes the following steps:

[0019] (1) Test solution: Accurately measure 10 ml of this product and place it in a 20 ml volumetric flask. Dilute to the mark with pyridine and shake well. Accurately measure 1 ml of this product and place it in a 10 ml volumetric flask. Dilute to the mark with acetic anhydride and shake well. Heat in a 75°C water bath for 30 minutes.

[0020] (2) Reference solution: Weigh 2000 mg of sorbitol and 100 mg of glycerol accurately, place them in a 20 ml volumetric flask, add 2 ml of water to dissolve them, dilute with pyridine to the mark, shake well, accurately measure 1 ml, place it in a 10 ml volumetric flask, dilute with acetic anhydride to the mark, shake well, and incubate in a 75°C water bath for 30 minutes.

[0021] (3) The reference solution and the test solution were determined by gas chromatography, respectively;

[0022] The gas chromatography detection conditions are as follows: DB-624 column; initial column temperature 160℃, maintained for 2 min, then increased to 240℃ at a rate of 5℃ / min, maintained for 15 min; injection port temperature 280℃; FID detector temperature 250℃; injection volume 1 μl; carrier gas flow rate 3.5 ml / ml; split ratio 20:1.

[0023] Calculation results: Calculated using the external standard method.

[0024] Due to the adoption of the above technical solutions, the present invention has the following advantages compared with the prior art:

[0025] 1. The assay method of this invention provides a comprehensive analysis of the formulation of fodosteine ​​oral solution and can effectively separate the excipients sorbitol and glycerol;

[0026] 2. The assay method of this invention has high sensitivity, strong specificity, and good accuracy, providing a simple and convenient detection method for the prescription analysis of fudostein oral solution. It can strictly control the quality of fudostein oral solution, ensuring its safety and reliability, and has practical significance. Attached Figure Description

[0027] Figure 1 This is the chromatogram of the blank solution in this invention;

[0028] Figure 2 This is the chromatogram of the blank matrix solution in this invention;

[0029] Figure 3 The chromatograms are of the positioning solutions of each excipient in this invention;

[0030] Figure 4 This is the chromatogram of the reference solution in this invention;

[0031] Figure 5 This is the chromatogram of the test solution in this invention; Detailed Implementation

[0032] To make the technical solution and beneficial effects of the present invention more apparent and understandable, a detailed description is provided below in conjunction with the accompanying drawings and specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Any product identical or similar to the present invention, derived by any person under the guidance of the present invention or by combining the features of the present invention with other prior art, falls within the protection scope of the present invention.

[0033] Experimental methods not specified in the following examples are generally performed under standard experimental conditions.

[0034] The fudosteine ​​oral solution, sorbitol, glycerin, and equipment used in the specific embodiments of the present invention are all known products, and the fudosteine ​​oral solution, sorbitol, and glycerin are obtained by purchasing commercially available products.

[0035] Table 1 Equipment Information

[0036] Equipment Name model factory Gas chromatograph TRACE1600 Thermo Fisher Scientific Electronic balance 125SM-FR Prisys Weighing Equipment Co., Ltd. water bath W-201B Jiangsu Jinyi Instrument Technology Co., Ltd. Ultrasonic instrument KM-1030B Guangzhou Kemeng Cleaning Technology Co., Ltd. Pure water machine EUE-10UV Shanghai Haisi Instrument Technology Co., Ltd.

[0037] Table 2 Material Information

[0038] name source batch number Acetic anhydride Sinopharm Chemical Reagent Co., Ltd. 20231116 Pyridine Sinopharm Chemical Reagent Co., Ltd. 20231008 Blank substrate Pharmaceutical formulation R&D department self-made JT002-240530-4

[0039] Table 3. Reference Standard Information

[0040] name batch number factory content Sorbitol F303C231201 Hubei Gedian Renfu Pharmaceutical Excipients Co., Ltd. 100.8％ glycerin GY20230701 Shandong Ruisheng Pharmaceutical Excipients Co., Ltd. 99.6％

[0041] Example 1

[0042] Methodological study of the detection and analysis method of the present invention

[0043] The following conditions were used for all experiments in this embodiment:

[0044] Column: DB-624 (30m × 0.32mm, 1.8μm);

[0045] Column temperature program: Initial temperature 160℃, hold for 2 min, then increase to 240℃ at a rate of 5℃ / min and hold for 15 min.

[0046] Inlet temperature: 280℃

[0047] FID detector temperature: 250℃

[0048] Nitrogen flow rate: 3.5 ml / min

[0049] Flow split ratio: 20:1

[0050] Injection volume: 1 μl

[0051] Testing steps:

[0052] Blank solution: Accurately measure 0.5 ml of pyridine and 0.5 ml of water, place them in a 10 ml volumetric flask, dilute to the mark with acetic anhydride, shake well, and incubate in a 75°C water bath for 30 minutes.

[0053] Test solution: Accurately measure 10 ml of this product and place it in a 20 ml volumetric flask. Dilute to the mark with pyridine and shake well. Accurately measure 1 ml of this product and place it in a 10 ml volumetric flask. Dilute to the mark with acetic anhydride and shake well. Incubate in a 75°C water bath for 30 minutes.

[0054] Reference solution: Weigh 2000 mg of sorbitol and 100 mg of glycerol accurately, place them in a 20 ml volumetric flask, add 2 ml of water to dissolve them, dilute to the mark with pyridine, shake well, accurately measure 1 ml, place it in a 10 ml volumetric flask, dilute to the mark with acetic anhydride, shake well, and incubate in a 75°C water bath for 30 minutes.

[0055] 1. Specificity test

[0056] Solution preparation:

[0057] Blank solution: Accurately measure 0.5 ml of pyridine and 0.5 ml of water, place them in a 10 ml volumetric flask, dilute to the mark with acetic anhydride, shake well, and incubate in a 75°C water bath for 30 minutes.

[0058] Blank matrix stock solution: Accurately measure 10 ml of blank matrix, place it in a 20 ml volumetric flask, dilute to the mark with pyridine, and shake well.

[0059] Blank matrix solution: Accurately measure 1 ml of blank matrix stock solution, place it in a 10 ml volumetric flask, dilute to the mark with acetic anhydride, shake well, and incubate in a 75°C water bath for 30 minutes.

[0060] Sorbitol positioning solution: Weigh 2000 mg of sorbitol accurately, place it in a 20 ml volumetric flask, add 2 ml of water to dissolve it, dilute to the mark with pyridine, shake well, accurately measure 1 ml, place it in a 10 ml volumetric flask, dilute to the mark with acetic anhydride, shake well, and incubate in a 75°C water bath for 30 minutes.

[0061] Glycerin positioning solution: Weigh 100 mg of glycerin accurately, place it in a 20 ml volumetric flask, add 2 ml of water to dissolve it, dilute to the mark with pyridine, shake well, accurately measure 1 ml, place it in a 10 ml volumetric flask, dilute to the mark with acetic anhydride, shake well, and incubate in a 75°C water bath for 30 minutes.

[0062] Reference solution: Weigh 2000 mg of sorbitol and 100 mg of glycerol accurately, place them in a 20 ml volumetric flask, add 2 ml of water to dissolve them, dilute to the mark with pyridine, shake well, accurately measure 1 ml, place it in a 10 ml volumetric flask, dilute to the mark with acetic anhydride, shake well, and incubate in a 75°C water bath for 30 minutes.

[0063] Test solution: Accurately measure 10 ml of this product and place it in a 20 ml volumetric flask. Dilute to the mark with pyridine and shake well. Accurately measure 1 ml of this product and place it in a 10 ml volumetric flask. Dilute to the mark with acetic anhydride and shake well. Incubate in a 75°C water bath for 30 minutes.

[0064] Inject 1 μl each of the blank solution, blank matrix solution, reference solution, and test solution into the gas chromatograph and record the chromatograms. The detection results are attached. Figure 1 ~Attached Figure 5 And Table 4 below.

[0065] Table 4 Results of specificity test

[0066]

[0067] The results above show that the unknown peaks in the blank solution, blank matrix solution, and test solution do not interfere with the detection of sorbitol and glycerol; the resolution between the sorbitol and glycerol peaks in the reference solution is 74.12, which meets the requirements, indicating that the method has good specificity.

[0068] 2. Precision test

[0069] Solution preparation:

[0070] Reference solution: Weigh 2000 mg of sorbitol and 100 mg of glycerol accurately, place them in a 20 ml volumetric flask, add 2 ml of water to dissolve them, dilute to the mark with pyridine, shake well, accurately measure 1 ml, place it in a 10 ml volumetric flask, dilute to the mark with acetic anhydride, shake well, and incubate in a 75°C water bath for 30 minutes.

[0071] Five consecutive injections of the reference solution were performed, and the peak area and retention time were recorded. The RSD values ​​of the peak area and retention time were calculated. The detection results are shown in Tables 5 and 6 below.

[0072] Table 5 Results of Sorbitol Injection Precision Test

[0073]

[0074] Table 6 Results of Glycerol Injection Precision Test

[0075]

[0076] The results above show that after five consecutive injections of the reference solution, the RSD values ​​of the peak areas of sorbitol and glycerol were 0.33% and 0.53%, respectively, both not exceeding 5.0%; the RSD values ​​of the retention times of sorbitol and glycerol were 0.04% and 0.03%, respectively, both not exceeding 2.0%. This method has good injection precision.

[0077] 3. Linearity and Range Tests

[0078] Solution preparation:

[0079] Linear stock solution: Weigh 2000 mg of sorbitol and 100 mg of glycerol accurately, place them in a 20 ml volumetric flask, add 2 ml of water to dissolve, dilute to the mark with pyridine, and shake well.

[0080] Take an appropriate amount of linear stock solution and prepare linear solutions of various concentrations according to the table below.

[0081] linear concentration Transfer the linear stock solution volume (ml). Adjust the volume to (ml) 25％ 0.25 10 50％ 0.5 10 75％ 0.75 10 100％ 1 10 150％ 1.5 10 200％ 2 10

[0082] Accurately measure 1 μl of each linear solution, inject it into the gas chromatograph, record the chromatogram, and perform linear regression analysis on the concentration using peak area. The detection results are shown in Tables 7 and 8 below.

[0083] Table 7 Results of the linearity test for sorbitol

[0084]

[0085]

[0086] Table 8 Results of the linearity test for glycerol

[0087]

[0088] The results above show that sorbitol has a correlation coefficient R within the concentration range of 2.524 mg / ml to 20.19 mg / ml. 2 The correlation coefficient R is 0.9999 and not less than 0.998; for glycerol in the concentration range of 0.1355 mg / ml to 1.084 mg / ml, the correlation coefficient R is... 2 The value is 0.9998, which is not less than 0.998, indicating that the method has a good linear relationship.

[0089] 4. Limit of detection test

[0090] Solution preparation:

[0091] Take the linear point solutions prepared under item 3. Linearity test and dilute them stepwise. Use a signal-to-noise ratio of not less than 10:1 as the limit of quantitation solution and not less than 3:1 as the limit of detection solution.

[0092] Each of the above diluted solutions was injected into a gas chromatograph, and the chromatograms were recorded. The detection results are shown in Tables 9 and 10 below.

[0093] Table 9 Results of Sorbitol Detection Limits

[0094]

[0095] Table 10 Results of the detection limit for glycerol

[0096]

[0097] The results above show that the ratios of the detection limit concentrations of sorbitol and glycerol to the concentrations of the reference standard are 0.02% and 0.15%, respectively, both not exceeding 10% of the reference standard concentration; the ratios of the quantitation limit concentrations of sorbitol and glycerol to the concentrations of the reference standard are 0.05% and 0.30%, respectively, both not exceeding 20% ​​of the reference standard concentration. This method has high detection sensitivity.

[0098] 5. Repeatability test

[0099] Solution preparation:

[0100] Reference solution: Accurately weigh 2000 mg of sorbitol and 100 mg of glycerol, place them in a 20 ml volumetric flask, add 2 ml of water to dissolve, dilute to the mark with pyridine, and shake well. Accurately measure 1 ml of the solution and place it in a 10 ml volumetric flask, dilute to the mark with acetic anhydride, shake well, and incubate in a 75°C water bath for 30 minutes. (Prepare two parallel solutions)

[0101] Test solution: Accurately measure 10 ml of this product into a 20 ml volumetric flask, dilute to the mark with pyridine, and shake well. Accurately measure 1 ml into a 10 ml volumetric flask, dilute to the mark with acetic anhydride, and shake well. Incubate in a 75°C water bath for 30 minutes. (Prepare 6 parallel solutions)

[0102] Inject 1 μl of each of the above solutions into the gas chromatograph, record the chromatograms, and calculate the amounts of sorbitol and glycerol in the six test solutions using the external standard method. The results are shown in Tables 11 and 12 below.

[0103] Table 11 Results of Sorbitol Repeatability Tests

[0104]

[0105] Table 12 Results of Glycerol Repeatability Tests

[0106]

[0107] The results above show that the RSD values ​​of sorbitol and glycerol in the six test solutions were 2.14% and 4.63%, respectively, both of which were no greater than 5.0%, indicating that the method has good repeatability.

[0108] 6. Accuracy Test

[0109] Solution preparation:

[0110] Reference stock solution: Accurately weigh 2000 mg of sorbitol and 100 mg of glycerol, place them in a 20 ml volumetric flask, add 2 ml of water to dissolve, dilute to the mark with pyridine, and shake well.

[0111] Reference solution: Accurately measure 1 ml of the reference stock solution, place it in a 10 ml volumetric flask, dilute to the mark with acetic anhydride, shake well, and incubate in a 75°C water bath for 30 minutes.

[0112] Prepare two portions in parallel.

[0113] Blank matrix stock solution: Accurately measure 10 ml of blank matrix, place it in a 20 ml volumetric flask, dilute to the mark with pyridine, and shake well.

[0114] Blank matrix solution: Accurately measure 1 ml of blank matrix stock solution, place it in a 10 ml volumetric flask, dilute to the mark with acetic anhydride, shake well, and incubate in a 75°C water bath for 30 minutes.

[0115] 50% recovery solution: Accurately measure 1 ml of blank matrix stock solution and place it in a 10 ml volumetric flask. Accurately add 0.5 ml of reference stock solution, dilute to the mark with acetic anhydride, shake well, and incubate in a 75°C water bath for 30 minutes. Prepare 3 parallel solutions.

[0116] 100% recovery solution: Accurately measure 1 ml of blank matrix stock solution and place it in a 10 ml volumetric flask. Accurately add 1 ml of reference stock solution, dilute to the mark with acetic anhydride, shake well, and incubate in a 75°C water bath for 30 minutes. Prepare 3 parallel solutions.

[0117] 150% recovery solution: Accurately measure 1 ml of blank matrix stock solution and place it in a 10 ml volumetric flask. Accurately add 1.5 ml of reference stock solution, dilute to the mark with acetic anhydride, shake well, and incubate in a 75°C water bath for 30 minutes. Prepare 3 parallel aliquots.

[0118] Precisely measure 1 μl of each of the above solutions and inject them into the solution. Record the chromatograms and calculate the recovery rate by comparing the measured amount with the added amount using the external standard method. The detection results are shown in Tables 13 and 14 below.

[0119] Table 13 Results of Sorbitol Accuracy Test

[0120]

[0121] Table 14 Results of Glycerin Accuracy Test

[0122]

[0123] The results show that the average recovery rate of sorbitol at various concentrations was 100.4%, ranging from 95.0% to 105.0%, with an RSD of 2.22%, not exceeding 5.0%. Similarly, the average recovery rate of glycerol at various concentrations was 100.4%, ranging from 95.0% to 105.0%, with an RSD of 2.28%, not exceeding 5.0%. The method demonstrates good accuracy.

[0124] 7. Solution stability test

[0125] Solution preparation:

[0126] Take the reference solution (prepared as described in section 1. Specificity test) and place it at room temperature.

[0127] Inject 1 μl of the above reference solution into the gas chromatograph, record the chromatogram, and the detection results are shown in Table 15 below.

[0128] Table 15 Results of the stability test of the reference solution.

[0129] Placement time 4 days at room temperature Sorbitol reference solution content 100.8％ Glycerin reference solution content 100.5％

[0130] The results above show that the contents of sorbitol and glycerol reference solutions were 100.8% and 100.5% respectively after being placed at room temperature for 4 days, both within the range of 95.0% to 105.0%. The reference solutions were stable after being placed at room temperature for at least 4 days.

[0131] In summary, this invention provides a method for determining the content of sorbitol and glycerol excipients in fodosteine ​​oral solution. This method exhibits good specificity, achieving baseline separation between the sorbitol and glycerol peaks; it also demonstrates good system suitability; the method possesses extremely high sensitivity, with detection limits for sorbitol and glycerol reaching as low as 0.8130 μg / ml; and the method exhibits good repeatability and accuracy. It provides a convenient detection method for the formulation analysis of fodosteine ​​oral solution, enabling the preparation of products with quality and efficacy consistent with the original drug.

[0132] It should be understood that the above embodiments are exemplary and are not intended to encompass all possible implementations included in the claims. Various modifications and changes can be made to the above embodiments without departing from the scope of this disclosure. Similarly, the various technical features of the above embodiments can be arbitrarily combined to form other embodiments of the present invention that may not be explicitly described. Therefore, the above embodiments only illustrate several implementations of the present invention and do not limit the scope of protection of this patent.

Claims

1. A method for determining the content of excipients sorbitol and glycerol in fodosteine ​​oral solution, characterized in that, Gas chromatography was used with a DB-624 column; the initial column temperature was 160℃, maintained for 2 min, and then increased to 240℃ at a rate of 5℃ / min, maintained for 15 min; the injection port temperature was 280℃; the FID detector temperature was 250℃; and the injection volume was 1 μl.

2. The method for determining the content of excipients sorbitol and glycerol in the fodosteine ​​oral solution according to claim 1, characterized in that, The chromatographic column is a capillary column, preferably a medium polarity chromatographic column DB-624; Specifications: 30m × 0.32mm, film thickness: 1.8μm, supplied by Agilent Technologies.

3. The determination method according to claim 1, characterized in that, The carrier gas used is nitrogen; preferably, the carrier gas flow rate is 1.0 ml / min to 5 ml / min, and more preferably 3.5 ml / min.

4. The determination method according to claim 1, characterized in that, The detector used was a hydrogen flame ionization detector.

5. The determination method according to claim 1, characterized in that, The injection volume is 1 μl, and the split mode is 5:1 to 50:1, preferably 20:

1.

6. The determination method according to claim 1, characterized in that, The initial column temperature is 130℃~180℃, and more preferably 160℃.

7. The determination method according to claim 1, characterized in that, The injection port temperature is 240℃~300℃; the detector temperature is 230℃~300℃; more preferably, the injection port temperature is 280℃ and the detector temperature is 250℃.

8. The method for determining the content of excipients sorbitol and glycerol in fodosteine ​​oral solution according to any one of claims 1 to 7, characterized in that, Includes the following steps: (1) Test solution: Accurately measure 10 ml of this product and place it in a 20 ml volumetric flask. Dilute to the mark with pyridine and shake well. Accurately measure 1 ml of this product and place it in a 10 ml volumetric flask. Dilute to the mark with acetic anhydride and shake well. Heat in a 75°C water bath for 30 minutes. (2) Reference solution: Weigh 2000 mg of sorbitol and 100 mg of glycerol accurately, place them in a 20 ml volumetric flask, add 2 ml of water to dissolve them, dilute with pyridine to the mark, shake well, accurately measure 1 ml, place it in a 10 ml volumetric flask, dilute with acetic anhydride to the mark, shake well, and incubate in a 75°C water bath for 30 minutes. (3) The reference solution and the test solution were determined by gas chromatography, respectively; The gas chromatography detection conditions are as follows: DB-624 column; initial column temperature 160℃, maintained for 2 min, then increased to 240℃ at a rate of 5℃ / min, maintained for 15 min; injection port temperature 280℃. The FID detector temperature was 250℃; the injection volume was 1 μl; the carrier gas flow rate was 3.5 ml / ml; and the split ratio was 20:1.

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