HPLC (High Performance Liquid Chromatography) detection method for sucralose, sorbitol and cane sugar in ambroxol hydrochloride oral solution

Abstract

The invention discloses an HPLC (High Performance Liquid Chromatography) detection method for sucralose, sorbitol and cane sugar in an ambroxol hydrochloride oral solution. The HPLC detection method comprises the following steps: a, preparing a reference substance solution; b, preparing a test solution; c, respectively sucking the reference solution and the test solution and injecting into an HPLC-ELSD (High Performance Liquid Chromatography-Evaporative Light Scattering Detector); chromatographic conditions are as follows: a chromatographic column: a C18 column is connected in series with an amino column; and the mobile phase is 65% acetonitrile aqueous solution. The method is rapid, efficient and high in sensitivity, sucralose, sorbitol and cane sugar in the ambroxol hydrochloride oral solution can be accurately quantified at the same time, a simple and accurate method is provided for detection of the ambroxol hydrochloride oral solution, the quality of the ambroxol hydrochloride oral solution is guaranteed, a scientific basis is provided for clinical application of the ambroxol hydrochloride oral solution, and the method is worthy of popularization and application. Good application prospects are realized.

Description

Technical Field

[0001] The invention relates to the field of drug detection and analysis, and in particular to an HPLC detection method for sucralose, sorbitol and sucrose in an ambroxol hydrochloride oral solution. Background Art

[0002] Ambroxol hydrochloride oral solution is a drug used to treat respiratory diseases, and its main ingredient is ambroxol hydrochloride. Ambroxol hydrochloride itself has a bitter taste and is irritating to the throat, which can cause patients to feel uncomfortable when taking it, and even induce vomiting. Therefore, it is necessary to add natural sweeteners (sorbitol, sucrose, etc.) or artificial sweeteners (sucralose, saccharin sodium, acesulfame potassium, aspartame, etc.) to effectively mask the bitterness of the drug and increase the aroma of the drug, thereby improving patient compliance.

[0003] There are large differences in the types and dosages of sweeteners in ambroxol hydrochloride oral solutions produced by different manufacturers. Excessive intake of sweeteners may have adverse effects on gastrointestinal function. The types and dosages of sweeteners can also affect the chemical stability of the drug. Studies have shown that sweeteners can affect the stability of ambroxol hydrochloride. According to national drug standards and quality evaluation requirements, ambroxol hydrochloride oral solutions need to strictly control and test excipients such as sweeteners to ensure that they meet statutory standards.

[0004] The sweeteners sucralose, sorbitol, and sucrose in ambroxol hydrochloride oral solution have large differences in polarity, so different chromatographic columns are required when quantifying their contents by high performance liquid chromatography. The current relevant standards generally use a C18 column to analyze sucralose, and an amino column to analyze sorbitol and sucrose. This analysis method results in low efficiency of analysis and detection, and it is impossible to simultaneously detect sucralose, sorbitol, and sucrose in large quantities of ambroxol hydrochloride oral solution, which in turn affects the effectiveness of quality control of ambroxol hydrochloride oral solution and has a negative impact on public health, the economy, and society. Therefore, improving the detection efficiency of sucralose, sorbitol, and sucrose in ambroxol hydrochloride oral solution is an important issue that needs to be urgently addressed in the current medical field. Summary of the invention

[0005] To solve the above problems, the present invention provides a HPLC detection method for sucralose, sorbitol and sucrose in ambroxol hydrochloride oral solution, which comprises the following steps:

[0006] a. Preparation of reference solution: respectively take sucralose standard and sorbitol standard, add water to dissolve to prepare sucralose standard stock solution and sorbitol standard stock solution; take sucrose standard, add water and acetonitrile to dissolve to prepare sucrose standard stock solution; take sucralose standard stock solution, sorbitol standard stock solution and sucrose standard stock solution, mix them, add water to dilute to prepare reference solution of series concentration;

[0007] b. Preparation of test solution: Take ambroxol hydrochloride oral solution, add water and mix;

[0008] c. Separately aspirate the reference solution and the test solution and inject them into the HPLC-ELSD instrument; the chromatographic conditions are as follows: chromatographic column: C18 column connected with amino column; mobile phase: 65% acetonitrile aqueous solution.

[0009] Furthermore, the mass volume ratio of sucrose, water and acetonitrile in step a is 0.1-1 g: 5-10 mL: 0-1 mL.

[0010] Furthermore, in the reference solution of step a, the concentration of sucralose is 0.13-0.54 mg / mL, the concentration of sorbitol is 0.15-0.60 mg / mL, and the concentration of sucrose is 0.13-0.53 mg / mL.

[0011] Furthermore, the ambroxol hydrochloride oral solution in step b is diluted with water to a concentration of sucralose of 0.13-0.54 mg / mL, a concentration of sorbitol of 0.15-0.60 mg / mL, and a concentration of sucrose of 0.13-0.53 mg / mL.

[0012] Furthermore, in step c, the mobile phase flows into the amino column and flows out of the C18 column.

[0013] Furthermore, the C18 column is a Waters Atlantis T3 column, 4.6×250 mm, 5 μm, and the amino column is a Waters Carbohydrate Analysis column, 3.9×300 mm, 10 μm.

[0014] Furthermore, in the chromatographic conditions of step c, the flow rate is 0.5-2 mL / min; the column temperature is 25-35° C., and the injection volume is 5-20 μL.

[0015] Furthermore, the chromatographic conditions include flow rate: 1 mL / min; column temperature: 30° C., and injection volume: 10 μL.

[0016] Furthermore, the evaporative light scattering detector parameters in the chromatographic conditions of step c are: air flow rate: 2.5 L / min, drift tube temperature: 55° C., detector gain: 1.

[0017] Furthermore, the contents of sucralose, sorbitol and sucrose in the ambroxol hydrochloride oral solution are calculated by peak area using an external standard method.

[0018] The invention discloses an HPLC detection method for sucralose, sorbitol and sucrose in an ambroxol hydrochloride oral solution. The method connects specific chromatographic columns in series in a specific manner and then, in cooperation with a specific mobile phase, enables the sucralose, sorbitol and sucrose in the ambroxol hydrochloride oral solution to be effectively separated simultaneously in a chromatogram. Methodological verification shows that the method is rapid, efficient and highly sensitive, and can accurately quantify sucralose, sorbitol and sucrose in the ambroxol hydrochloride oral solution simultaneously, thus providing a simple and accurate method for detecting the ambroxol hydrochloride oral solution, ensuring the quality of the ambroxol hydrochloride oral solution, and providing a scientific basis for its clinical application, and having a good application prospect.

[0019] Obviously, according to the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modification, replacement or change may be made.

[0020] The above contents of the present invention are further described in detail below through specific implementation methods in the form of embodiments. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 Chromatogram of blank solution plus mixed reference;

[0022] Figure 2 Chromatographic Condition Screening - Examining the Use of C 18 column;

[0023] Figure 3 Screening of chromatographic conditions - investigating the use of amino columns alone①;

[0024] Figure 4 Screening of chromatographic conditions - investigating the use of amino columns alone②;

[0025] Figure 5 Screening of chromatographic conditions - investigating the use of amino columns alone③;

[0026] Figure 6 Screening of chromatographic conditions - investigating the combined chromatographic columns①;

[0027] Figure 7 Screening of chromatographic conditions - examining the combined chromatographic column②;

[0028] Figure 8 Screening of chromatographic conditions - investigating the combined chromatographic column③;

[0029] Fig. 9 Screening of chromatographic conditions - investigating the combined chromatographic columns④;

[0030] Fig.10 Screening of chromatographic conditions—investigation of combined chromatographic columns⑤. DETAILED DESCRIPTION

[0031] The raw materials, reagents and equipment in the specific embodiments of the present invention are all known products and can be purchased commercially.

[0032] Example 1. HPLC Detection of Sucralose, Sorbitol and Sucrose in Ambroxol Hydrochloride Oral Solution

[0033] (1) Preparation of test solution

[0034] Accurately measure the ambroxol hydrochloride oral solution and dilute it with water to a concentration of 0.13-0.54 mg / mL of sucralose, 0.15-0.60 mg / mL of sorbitol, and 0.13-0.53 mg / mL of sucrose;

[0035] (2) Preparation of reference solution

[0036] Weigh the sucralose standard and dissolve it in water to prepare a sucralose standard stock solution with a concentration of 10 mg / mL;

[0037] Weigh the sorbitol standard and dissolve it in water to prepare a sorbitol standard stock solution with a concentration of 40 mg / mL;

[0038] Weigh the sucrose standard, dissolve it in water, add 0.5 mL of acetonitrile, and then dilute to volume with water to make a sucrose standard stock solution with a concentration of 20 mg / mL;

[0039] The sucralose standard stock solution, sorbitol standard stock solution and sucrose standard stock solution were measured and mixed, and diluted with water to form mixed reference solution of series concentration; the concentrations of sucralose in the mixed reference solution of series concentration were 0.13 mg / mL, 0.18 mg / mL, 0.27 mg / mL, 0.36 mg / mL, 0.45 mg / mL and 0.54 mg / mL, the concentrations of sorbitol were 0.15 mg / mL, 0.20 mg / mL, 0.30 mg / mL, 0.40 mg / mL, 0.50 mg / mL and 0.60 mg / mL, and the concentrations of sucrose were 0.13 mg / mL, 0.18 mg / mL, 0.26 mg / mL, 0.35 mg / mL, 0.44 mg / mL and 0.53 mg / mL.

[0040] (3) Pipette 10 μL of the reference solution and the test solution into the HPLC-ELSD instrument; the chromatographic conditions are as follows:

[0041] Chromatographic column: Waters Carbohydrate Analysis column (3.9×300 mm, 10 μm) in series with Waters Atlantis T3 (4.6×250 mm, 5 μm column); mobile phase: 65% acetonitrile aqueous solution, flowing in from the Waters Carbohydrate Analysis column and flowing out from the Waters Atlantis T3 (4.6×250 mm, 5 μm column); flow rate: 1 mL / min; column temperature: 30°C;

[0042] Evaporative light scattering detector parameters: air flow rate: 2.5 L / min, drift tube temperature: 55 °C, detector gain: 1.

[0043] (4) The contents of sucralose, sorbitol and sucrose in ambroxol hydrochloride oral solution were calculated by peak area using the external standard method.

[0044] The beneficial effects of the present invention are described below through test examples.

[0045] Experimental Example 1 Study on the simultaneous detection of sucralose, sorbitol and sucrose in ambroxol hydrochloride oral solution by high performance liquid chromatography evaporative light scattering detection

[0046] 1. Screening of chromatographic conditions

[0047] 1.1 Selection of chromatographic column and elution system

[0048] 1.1.1 Single-use chromatographic columns

[0049] The single use of C 18 Column: Agilent Zorbax SB-C 18 (4.6×150mm, 3.5μm), Waters Atlantis T3 column (4.6×250mm, 5μm), the results showed that under different elution intensities of the mobile phase acetonitrile-water system (11% acetonitrile, 15% acetonitrile, 85% acetonitrile), the two components of sucrose and sorbitol were basically not retained on the chromatographic column, and could not be separated and effectively detected ( Figure 2 ).

[0050] The amino columns used alone were investigated: Hypersil GoldAmino (4.6×150mm, 3μm) and WatersCarbohydrateAnalysis column (3.9×300mm, 10μm). Under the conditions of acetonitrile-water system (75% acetonitrile) as the mobile phase, sucralose was not retained on the chromatographic column and did not produce any peaks, and could not be detected ( Figures 3-4 ); Under the conditions of the mobile phase acetonitrile-water system (15% acetonitrile), sucralose peaks at about 3 minutes on the chromatogram, but sucralose and sucrose cannot be separated and completely overlap ( Figure 5).

[0051] 1.1.2 Combined chromatographic columns

[0052] The C18 column or amino column alone cannot achieve simultaneous detection and effective separation of sucralose, sorbitol, and sucrose. If further attempts to replace other types of chromatographic columns still fail to effectively separate sucralose, sorbitol, and sucrose in the liquid chromatogram, consider using a column series method to achieve simultaneous detection of sucralose, sorbitol, and sucrose in one analytical test.

[0053] Based on the analysis of the various chromatographic behavior parameters of sucralose, sorbitol, and sucrose in the chromatogram when using a single chromatographic column, a variety of chromatographic column combinations were investigated. Among them, the Agilent Zorbax SB-C18 (4.6×150mm, 3.5μm) column was combined with the Waters Carbohydrate Analysis column (3.9×300mm), and the mobile phase ratio (75% acetonitrile, 70% acetonitrile, 60% acetonitrile, 50% acetonitrile) was adjusted, but it was impossible to achieve effective separation of sucralose, sorbitol, and sucrose, and the interference of the solvent peak could not be avoided. However, when the mobile phase ratio was 65% acetonitrile, sucralose, sorbitol, and sucrose could be effectively separated, and sucralose was still interfered by the solvent peak but to a lesser extent. When the combined C18 column was replaced with a Waters Atlantis T3 column (4.6×250mm, 5μm), sucralose was well separated from the solvent peak without interference, and sucralose, sorbitol, and sucrose were well separated. That is, only when the columns used are Waters Atlantis T3 column (4.6×250mm, 5μm) and Waters Carbohydrate Analysis column (3.9×300mm) can the solvent peak, sucralose, sorbitol, and sucrose be separated well ( Figures 6 to 10 ).

[0054] When the WatersAtlantis T3 column and the Waters CarbohydrateAnalysis column are connected in series, when the WatersAtlantis T3 column is in front, the retention time of the ambroxol hydrochloride peak becomes longer, and the analysis time of sucralose, sorbitol, and sucrose in the ambroxol hydrochloride oral solution becomes longer. Therefore, in order to shorten the injection and analysis time, the WatersCarbohydrateAnalysis column (3.9×300mm) is placed in front and the WatersAtlantis T3 column is placed in the back when the chromatographic columns are connected in series.

[0055] 1.2 Screening of other chromatographic conditions

[0056] By optimizing the parameters of the evaporative light scattering detector, including air flow rate (compared 2L / min, 2.5L / min and 2.8L / min), drift tube temperature (compared 55℃, 83℃, 95℃), and detector gain (compared 1 and 4), the air flow rate was 2.5L / min, the drift tube temperature was 55℃, and the gain was 1.

[0057] The column temperature was set at 30 °C, the flow rate was 1 mL / min, and the injection volume was 10 μL.

[0058] According to the screening results, the final chromatographic conditions are:

[0059] Chromatographic column: Waters Carbohydrate Analysis column (3.9×300 mm) connected in series with Waters Atlantis T3 (4.6×250 mm, 5 μm column); mobile phase: 65% acetonitrile aqueous solution, flowing into the Waters Carbohydrate Analysis column and flowing out of the Waters Atlantis T3 (4.6×250 mm, 5 μm column); flow rate: 1 mL / min; column temperature: 30°C;

[0060] Evaporative light scattering detector parameters: air flow rate: 2.5 L / min, drift tube temperature: 55 °C, detector gain: 1.

[0061] 2. Preparation of reference solution

[0062] Prepare the sucralose reference substance stock solution by referring to the preparation method of sucralose standard stock solution in "Determination of sucralose (sucralose) in food - GB5009.298-2023", that is, weigh 0.1g (accurate to 0.0001g) of sucralose standard, dissolve it in water and transfer it to a 10mL volumetric flask, dilute to the scale, and mix well.

[0063] Refer to the preparation method of sorbitol standard stock solution in "Determination of xylitol, sorbitol, maltitol and erythritol in food - GB5009.279-2016" to prepare sorbitol reference stock solution, that is, weigh 400 mg (accurate to 0.1 mg) of sorbitol standard and add water to make up to 10 mL.

[0064] Prepare the sucrose reference stock solution by referring to the preparation method of sucrose standard stock solution in "Determination of fructose, glucose, sucrose, maltose and lactose in food--GB5009.8-2023", that is, weigh 0.2 g of sucrose standard, dissolve it in water and transfer it to a 10 mL volumetric flask, add 0.5 mL of acetonitrile, and dilute to the scale with water.

[0065] 3. Preparation of test solution

[0066] According to the sample prescription, dilute with water to prepare the test solution so that the concentration of the measured component is within the linear range.

[0067] 4. Specificity test

[0068] According to the prescription of ambroxol hydrochloride oral solution, water was used to dissolve glycerin, flavors, inorganic salts and other water-soluble excipients, 50% acetonitrile was used to dissolve other excipients and main drug components, and blank solution was prepared according to the preparation method of the test solution. Each component in the prescription of ambroxol hydrochloride oral solution had no interference with the determination of sucralose, sorbitol and sucrose. The chromatogram of blank solution plus mixed reference substance is shown in Figure 1 .

[0069] 5. Linear relationship investigation

[0070] Accurately measure appropriate amounts of sucralose reference stock solution, sorbitol reference stock solution, and sucrose reference stock solution, and use water as the solvent to prepare a linear mixed reference solution. The concentrations of sucralose are 0.13 mg / mL, 0.18 mg / mL, 0.27 mg / mL, 0.36 mg / mL, 0.45 mg / mL, and 0.54 mg / mL; the concentrations of sorbitol are 0.15 mg / mL, 0.20 mg / mL, 0.30 mg / mL, 0.40 mg / mL, 0.50 mg / mL, and 0.60 mg / mL; the concentrations of sucrose are 0.13 mg / mL, 0.18 mg / mL, 0.26 mg / mL, 0.35 mg / mL, 0.44 mg / mL, and 0.53 mg / mL. Use the common logarithm value Y of the peak area as the ordinate and the common logarithm value X of the injection volume (μg) as the abscissa to draw a standard curve. The results showed that the three sugars had good linear relationships within their respective linear ranges. The experimental results are shown in Table 1.

[0071] Table 1 Linear regression equation, range and correlation coefficient of sucralose, sorbitol and sucrose

[0072]

[0073] 6. Limit of detection and limit of quantification

[0074] Take an appropriate amount of linear mixed reference solution, use water as solvent, and prepare solutions of different concentrations. The detection limit is calculated based on 3 times the signal-to-noise ratio, and the quantification limit is calculated based on 10 times the signal-to-noise ratio. Results The detection limit of sucralose is 0.45μg, and the quantification limit is 1.34μg; the detection limit of sorbitol is 0.50μg, and the quantification limit is 1.50μg; the detection limit of sucrose is 0.44μg, and the quantification limit is 1.31μg.

[0075] 7. Accuracy test

[0076] Accurately measure the appropriate amount of sucralose, sorbitol, and sucrose reference stock solutions, use the blank solution under the specific test item as the solvent, prepare solutions at low, medium, and high concentration levels, and prepare 3 copies for each concentration level. According to the standard curve method, the average recovery rate of sucralose is 97.9%, RSD is 1.4%; the average recovery rate of sorbitol is 100.5%, RSD is 1.3%; the average recovery rate of sucrose is 100.3%, RSD% is 1.4%. The results are shown in Table 2.

[0077] Table 2 Accuracy test

[0078]

[0079] 8. Daytime precision test

[0080] Accurately measure the appropriate amount of sucralose, sorbitol, and sucrose reference stock solutions, and use the blank solution under the specific test item as the solvent to prepare sample solutions with a sucralose concentration of 0.27 mg / mL, a sorbitol concentration of 0.30 mg / mL, and a sucrose concentration of 0.26 mg / mL. The same analyst determined the results on different days (day 1, day 4, and day 6) using the standard curve method. The RSDs of the three determinations were 1.6% for sucralose, 1.9% for sorbitol, and 1.9% for sucrose.

[0081] 9. Stability test

[0082] The test solution was prepared according to the sample solution under the daily precision item, placed at room temperature for 0h, 3h, 17h, and 24h, and determined according to the standard curve method. The RSDs of the four determination results were 0.7% for sucralose, 1.3% for sorbitol, and 2.2% for sucrose. The results showed that the test solution was basically stable within 24h.

[0083] 10. Sample determination

[0084] Multiple batches of samples from different companies were tested, and the results are shown in Table 3. The results show that there is no significant difference between batches from the same company.

[0085] Table 3 Sample determination

[0086]

[0087] Note: “-” means not detected.

[0088] 11. Conclusion

[0089] The present invention compares amino columns and C18 columns of different brands. It is found that the amino column or C18 column alone cannot achieve the purpose of simultaneously and effectively separating sucralose, sorbitol and sucrose. The Waters Carbohydrate Analysis column has advantages in separating the three sugars, but due to the chromatographic column, there is a phenomenon that the column loss components peak out to form a solvent peak during the sample injection analysis, which interferes with the determination of sucralose. When the Agilent Zorbax SB-C18 chromatographic column is used in combination, the problem of solvent peak interference cannot be avoided, but when the Waters Atlantis T3 column is used in combination, the unexpected solvent peak is well separated from the sucralose peak. The order of using the Waters Carbohydrate Analysis column and the Waters Atlantis T3 column in combination has little effect on the peaks of sucralose, sorbitol and sucrose, but has a greater effect on components that are easily retained on the C18 column, such as ambroxol hydrochloride. When the C18 column is in front, the retention time of the ambroxol hydrochloride peak becomes longer. In the inspection of sucralose, sorbitol and sucrose in ambroxol hydrochloride oral solution, the injection analysis time will be prolonged. In order to shorten the analysis time, the Waters Carbohydrate Analysis column is in front and the Waters Atlantis T3 column is in the back when the columns are connected.

[0090] The three different air flow rates of the evaporative light scattering detector were compared. When the air flow rate was 2L / min, the baseline noise was slightly higher. When the air flow rate was 2.5L / min and 2.8L / min, there was no significant difference in the analysis of the mixed reference solution. In order to make the gas flow rate stable, a low flow rate was selected as much as possible, so the air flow rate of 2.5L / min was finally selected.

[0091] Comparison of three different drift tube temperatures of the evaporative light scattering detector showed no significant difference in the separation of the three component peaks of the mixed reference solution. However, when the drift tube temperature was 55°C, the peak areas of the three components increased. Therefore, the drift tube temperature of 55°C was finally selected.

[0092] Ambroxol hydrochloride oral solution from three companies was selected, and the established method can effectively detect sucralose, sorbitol, and sucrose in ambroxol hydrochloride solution. The sample test results showed that the batches from the same company were consistent, and the detected species were consistent with the instructions and labels. Compared with the prescription, the detection results were consistent.

[0093] In summary, the present invention connects specific chromatographic columns in series in a specific manner, and then with the cooperation of a specific mobile phase, the sucralose, sorbitol and sucrose in the ambroxol hydrochloride oral solution can be effectively separated in the chromatogram at the same time. Methodological verification shows that the method of the present invention is fast, efficient and highly sensitive, and can accurately quantify the sucralose, sorbitol and sucrose in the ambroxol hydrochloride oral solution at the same time, providing a simple and accurate method for the detection of the ambroxol hydrochloride oral solution, and ensuring the quality of the ambroxol hydrochloride oral solution.

Claims

1. An HPLC detection method for sucralose, sorbitol and sucrose in an ambroxol hydrochloride oral solution, characterized in that: The steps include: a. Preparation of reference solution: respectively take sucralose standard and sorbitol standard, add water to dissolve to prepare sucralose standard stock solution and sorbitol standard stock solution; take sucrose standard, add water and acetonitrile to dissolve to prepare sucrose standard stock solution; take sucralose standard stock solution, sorbitol standard stock solution and sucrose standard stock solution, mix them, add water to dilute to prepare reference solution of series concentration; b. Preparation of test solution: Take ambroxol hydrochloride oral solution, add water and mix; c. Pipette the reference solution and the test solution into the HPLC-ELSD instrument respectively; the chromatographic conditions are as follows: chromatographic column: C 18 The columns were connected in series with amino columns; the mobile phase was 65% acetonitrile aqueous solution.

2. The HPLC detection method according to claim 1, wherein: The mass volume ratio of sucrose, water and acetonitrile in step a is 0.1-1 g: 5-10 mL: 0-1 mL.

3. The HPLC detection method according to claim 1, wherein: In the reference solution of step a, the concentration of sucralose is 0.13-0.54 mg / mL, the concentration of sorbitol is 0.15-0.60 mg / mL, and the concentration of sucrose is 0.13-0.53 mg / mL.

4. The HPLC detection method according to claim 1, wherein: In step b, the ambroxol hydrochloride oral solution is diluted with water to a concentration of sucralose of 0.13-0.54 mg / mL, a concentration of sorbitol of 0.15-0.60 mg / mL, and a concentration of sucrose of 0.13-0.53 mg / mL.

5. The HPLC detection method according to claim 1, wherein: The mobile phase in step c flows into the amino column, C 18 column outflow.

6. The HPLC detection method according to claim 1, characterized in that: The C 18 The column was a Waters Atlantis T3 column, 4.6×250 mm, 5 μm, and the amino column was a Waters Carbohydrate Analysis column, 3.9×300 mm, 10 μm.

7. The HPLC detection method according to claim 1, characterized in that: The chromatographic conditions in step c include flow rate of 0.5 to 2 mL / min, column temperature of 25 to 35° C., and injection volume of 5 to 20 μL.

8. The HPLC detection method according to claim 7, characterized in that: The chromatographic conditions include flow rate: 1 mL / min; column temperature: 30° C., and injection volume: 10 μL.

9. The HPLC detection method according to claim 1, wherein: The evaporative light scattering detector parameters in the chromatographic conditions of step c are: air flow rate: 2.5 L / min, drift tube temperature: 55° C., detector gain:

1.

10. The HPLC detection method according to any one of claims 1 to 9, characterized in that: The contents of sucralose, sorbitol and sucrose in the ambroxol hydrochloride oral solution are calculated by peak area using an external standard method.

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