Method for determining component content of ebastine oral solution

Abstract

The invention discloses a method for determining the component content of an ebastine oral solution. The method comprises the following steps: step 1, determining chromatographic conditions; 2, preparing a solvent; 3, preparing a reference substance solution; step 4, preparing a test solution; 5, sample introduction analysis; step 6, calculating and judging a result; in the step 1, a preparation method of the phosphate buffer solution specifically comprises the following steps: weighing 2.0 g of monopotassium phosphate, adding water for dissolving and diluting to 1000ml, and adjusting the pH value to 3.5 by using 50% phosphoric acid; according to the present invention, by using the chromatography principle, the ebastine, the methylparaben and the propylparaben in the ebastine oral solution are separated through the specific chromatographic conditions, and then the quantitative analysis is performed according to the peak areas of the reference substance solution and the test substance solution in the chromatogram and other parameters so as to simultaneously determine the contents of the three components, therefore, the analysis efficiency is improved, and the experiment time and cost are reduced.

Description

Technical Field

[0001] The invention relates to the technical field of drug detection, in particular to a method for determining the content of components of ebastine oral solution. Background Art

[0002] Ebastine is a potent, highly selective, specific H1 receptor blocker, suitable for the symptomatic treatment of allergic rhinitis and chronic idiopathic urticaria with or without allergic conjunctivitis. Oral solution has the advantages of fast absorption, rapid efficacy, easy dose division, convenient administration and wide application to various groups of people. Methylparaben sodium and propylparaben sodium are used as antibacterial agents in ebastine oral solution, which are common additives in non-solid preparations and play an important role in preventing microbial contamination of drugs and ensuring drug quality. Since antibacterial agents have certain toxicity, the scope of application and dosage need to be strictly controlled, so it is necessary to measure the content of the components of ebastine oral solution. In the prior art, potentiometric titration and high performance liquid chromatography are generally used as detection methods, but the above method cannot simultaneously measure the content of ebastine, methylparaben sodium and propylparaben sodium in ebastine oral solution, thus resulting in low detection efficiency. Summary of the invention

[0003] The object of the present invention is to provide a method for determining the content of components of ebastine oral solution to solve the problems raised in the above background technology.

[0004] To achieve the above object, the present invention provides the following technical solution: a method for determining the content of components of ebastine oral solution, comprising the following steps: step 1, determining chromatographic conditions; step 2, preparing solvents; step 3, preparing reference solution; step 4, preparing test solution; step 5, sample injection analysis; step 6, result calculation and judgment;

[0005] In the above step 1, the chromatographic conditions are determined as follows: using octadecylsilane bonded silica gel as a filler, phosphate buffer as a mobile phase A, methanol as a mobile phase B for gradient elution, setting the column temperature to 30°C, the flow rate to 1.0 ml / min, the detection wavelength to 255 nm, and the injection volume to 20 μl;

[0006] Wherein in the above step 2, mobile phase A and mobile phase B are mixed in a ratio of 35:65 to obtain a solvent;

[0007] Wherein in the above step 3, under light-proof conditions, prepare methylparaben reference substance stock solution, propylparaben reference substance stock solution and ebastine reference substance stock solution, take 1 ml of each of the three reference substance stock solutions, put them in a 20 ml volumetric flask, add solvent to dilute to the scale, and obtain a reference substance solution containing 20 μg of ebastine, 20 μg of methylparaben and 5 μg of propylparaben per 1 ml, and prepare two in parallel;

[0008] In the above step 4, under light-proof conditions, accurately measure 1 ml of the test sample, place it in a 50 ml volumetric flask, add solvent to dilute to the scale, shake well to obtain the test sample solution, and prepare two copies in parallel;

[0009] In the above step 5, the solvent obtained in step 2, the reference solution obtained in step 3, and the test solution obtained in step 4 are taken, and the chromatographic conditions determined in step 1 are used for analysis, and a chromatogram is recorded;

[0010] In the above step six, according to the peak areas of ebastine, methylparaben and propylparaben in the test and reference solutions obtained in the chromatogram, as well as the sample weight, content and dilution multiple of the test and reference solutions, the concentrations of the components in the reference solution and the peak area ratios are calculated, and then the content of ebastine in the test sample and the content of sodium methylparaben and sodium propylparaben in the test sample are calculated to determine whether the contents meet the limit requirements.

[0011] Preferably, in step 1, the preparation method of phosphate buffer is specifically as follows: weigh 2.0 g of potassium dihydrogen phosphate, dissolve it in water and dilute it to 1000 ml, and adjust the pH to 3.5 with 50% phosphoric acid.

[0012] Preferably, in step three, preparing the methylparaben reference substance stock solution is specifically as follows: weighing 10 mg of the methylparaben reference substance into a 25 ml volumetric flask, adding a solvent to dissolve and dilute to the scale, and shaking to obtain the methylparaben reference substance stock solution.

[0013] Preferably, in step three, the preparation of the propylparaben reference substance stock solution is specifically as follows: weigh 10 mg of the propylparaben reference substance into a 100 ml volumetric flask, add a solvent to dissolve and dilute to the scale, and shake well to obtain the propylparaben reference substance stock solution.

[0014] Preferably, in step 3, the preparation of the Ebastine reference substance stock solution is specifically as follows: weigh 10 mg of the Ebastine reference substance into a 25 ml volumetric flask, add 16 ml of methanol and ultrasonicate for 30 seconds to dissolve, then add mobile phase A to dilute to the scale to obtain the Ebastine reference substance stock solution.

[0015] Preferably, in step 6, the calculation formula for the concentration of each component and the peak area ratio of the reference solution is as follows:

[0016]

[0017] Among them, A 对 is the peak area of ​​ebastine, methylparaben and propylparaben in the reference solution, M 对 is the sample weight of the reference substance, in mg, V 对 is the dilution factor of the reference substance, C 对is the concentration of each component of the reference solution, the unit is mg / ml, a% is the content of each reference substance, and f is the ratio of the concentration of each component of the reference solution to the peak area.

[0018] Preferably, in step 6, the calculation formula for the content of ebastine in the test sample is as follows:

[0019]

[0020] Among them, A 样 is the peak area of ​​ebastine in the test sample, V 样 is the dilution multiple of the test sample, f is the ratio of the concentration of each component in the reference solution to the peak area, and the average value is used for calculation.

[0021] Preferably, in step 6, the calculation formula for the content of sodium methylparaben and sodium propylparaben in the test sample is as follows:

[0022] Antibacterial agent content (mg / ml) = k × A 样 ×f×V 样

[0023] Among them, k is the conversion coefficient, the conversion coefficient of sodium methylparaben is 1.144, the conversion coefficient of sodium propylparaben is 1.122, and A 样 is the peak area of ​​methylparaben and propylparaben in the test sample, V 样 is the dilution multiple of the test sample, f is the ratio of the concentration of each component in the reference solution to the peak area, and the average value is used for calculation.

[0024] Preferably, in step six, the limit requirements are: calculated by external standard method based on peak area, ebastine should be 95.0% to 105.0% of the labeled amount; calculated by external standard method by multiplying peak area by conversion factor, sodium methylparaben should be 0.96 mg / ml to 1.32 mg / ml; sodium propylparaben should be 0.24 mg / ml to 0.33 mg / ml.

[0025] Compared with the prior art, the present invention has the following beneficial effects: the present invention utilizes the principle of chromatography to separate ebastine, methylparaben and propylparaben in the ebastine oral solution under specific chromatographic conditions, and then performs quantitative analysis based on the peak areas of the reference solution and the test solution in the chromatogram and other parameters, thereby simultaneously determining the contents of the three components, thereby improving the analysis efficiency and reducing the experimental time and cost. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 is a flow chart of the method of the present invention;

[0027] Figure 2 is a solvent chromatogram of an experimental example of the present invention;

[0028] Figure 3 is a chromatogram of a reference solution of an experimental example of the present invention;

[0029] Figure 4 It is the chromatogram of the test solution of the experimental example of the present invention. DETAILED DESCRIPTION

[0030] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0031] Please refer to the attached Figure 1 -Attached Figure 4 , a technical solution provided by the present invention:

[0032] Example:

[0033] A method for determining the content of components of an ebastine oral solution comprises the following steps: step 1, determining chromatographic conditions; step 2, preparing a solvent; step 3, preparing a reference solution; step 4, preparing a test solution; step 5, sample injection and analysis; step 6, result calculation and judgment;

[0034] In the above step 1, the chromatographic conditions are determined as follows: using octadecylsilane bonded silica gel as a filler, using phosphate buffer as a mobile phase A and methanol as a mobile phase B for gradient elution, setting the column temperature to 30°C, the flow rate to 1.0 ml / min, the detection wavelength to 255 nm, and the injection volume to 20 μl; the elution gradient is shown in Table 1; wherein the preparation method of phosphate buffer is specifically as follows: weighing 2.0 g of potassium dihydrogen phosphate, dissolving it in water and diluting it to 1000 ml, and adjusting the pH to 3.5 with 50% phosphoric acid;

[0035] Wherein in the above step 2, mobile phase A and mobile phase B are mixed in a ratio of 35:65 to obtain a solvent;

[0036] Wherein, in the above step three, under light-proof conditions, prepare methylparaben reference substance stock solution, propylparaben reference substance stock solution and ebastine reference substance stock solution, specifically: weigh 10 mg of methylparaben reference substance and place it in a 25 ml volumetric flask, add a solvent to dissolve and dilute to the scale, shake well to obtain methylparaben reference substance stock solution, weigh 10 mg of propylparaben reference substance and place it in a 100 ml volumetric flask, add a solvent to dissolve and dilute to the scale, shake well to obtain propylparaben reference substance stock solution, weigh 10 mg of ebastine reference substance and place it in a 25 ml volumetric flask, add 16 ml of methanol and ultrasonically dissolve for 30 seconds, add mobile phase A and dilute to the scale to obtain ebastine reference substance stock solution, take 1 ml of each of the above three reference substance stock solutions, place them in a 20 ml volumetric flask, add a solvent to dilute to the scale, obtain reference substance solutions containing 20 μg of ebastine, 20 μg of methylparaben and 5 μg of propylparaben per 1 ml, and prepare two in parallel;

[0037] In the above step 4, under light-proof conditions, accurately measure 1 ml of the test sample, place it in a 50 ml volumetric flask, add solvent to dilute to the scale, shake well to obtain the test sample solution, and prepare two copies in parallel;

[0038] In the above step 5, the solvent obtained in step 2, the reference solution obtained in step 3, and the test solution obtained in step 4 are taken, and the chromatographic conditions determined in step 1 are used for analysis, and a chromatogram is recorded;

[0039] Wherein, in the above step six, according to the peak areas of ebastine, methylparaben and propylparaben in the test and reference solutions obtained in the chromatogram, as well as the sample weight, content and dilution multiple of the reference solution, the concentration of each component of the reference solution and the peak area ratio are calculated, and then the content of ebastine in the test sample, the content of sodium methylparaben and sodium propylparaben in the test sample are calculated to determine whether the content meets the limit requirements. The limit requirements are: calculated by external standard method based on peak area, ebastine should be 95.0% to 105.0% of the labeled amount; calculated by external standard method by multiplying peak area by conversion coefficient, sodium methylparaben should be 0.96 mg / ml to 1.32 mg / ml; sodium propylparaben should be 0.24 mg / ml to 0.33 mg / ml; wherein, the calculation formulas for the concentration of each component of the reference solution and the peak area ratio are as follows:

[0040]

[0041] Among them, A 对 is the peak area of ​​ebastine, methylparaben and propylparaben in the reference solution, M 对 is the sample weight of the reference substance, in mg, V 对 is the dilution factor of the reference substance, C 对is the concentration of each component of the reference solution, in mg / ml, a% is the content of each reference substance, and f is the ratio of the concentration of each component of the reference solution to the peak area;

[0042] The calculation formula of the content of Ebastine in the test sample is as follows:

[0043]

[0044] Among them, A 样 is the peak area of ​​ebastine in the test sample, V 样 is the dilution factor of the test sample, f is the ratio of the concentration of each component of the reference solution to the peak area, and the average value is used for calculation;

[0045] The calculation formula for the content of sodium methylparaben and sodium propylparaben in the test sample is as follows:

[0046] Antibacterial agent content (mg / ml) = k × A 样 ×f×V 样

[0047] Among them, k is the conversion coefficient, the conversion coefficient of sodium methylparaben is 1.144, the conversion coefficient of sodium propylparaben is 1.122, and A 样 is the peak area of ​​methylparaben and propylparaben in the test sample, V 样 is the dilution multiple of the test sample, f is the ratio of the concentration of each component in the reference solution to the peak area, and the average value is used for calculation.

[0048] Experimental example:

[0049] The experiment was carried out using the method provided in the embodiment. The parameters of the reference solution are shown in Table 2, and the parameters of the test solution are shown in Table 3-Table 4. It was calculated that the test sample contained 98.5% of the labeled amount of ebastine, 1.15 mg / ml of sodium methylparaben, and 0.29 mg / ml of sodium propylparaben, which met the limit requirements.

[0050] Table 1 Elution gradient

[0051]

[0052] Table 2 Reference solution parameters

[0053]

[0054]

[0055] Table 3 Parameters of sodium methylparaben and sodium propylparaben in the test solution

[0056]

[0057] Table 4 Parameters of Ebastine in the test solution

[0058]

[0059] Based on the above, the advantage of the present invention is that when the present invention is used, by adopting specific chromatographic conditions and strict solution preparation and calculation methods, the contents of ebastine, sodium methylparaben and sodium propylparaben in the ebastine oral solution can be accurately determined, providing reliable data support for drug quality control. Combined with clear limit requirements, it can ensure that the content of each component in the drug is within the specified range, ensuring the safety and effectiveness of the drug. The method has detailed operating steps and calculation formulas, which is convenient for experimenters to perform standardized operations, improve the repeatability and comparability of the experiment, and because the contents of the three components can be determined simultaneously, the analysis efficiency can be effectively improved, reducing the experimental time and cost.

[0060] It will be apparent to those skilled in the art that the invention is not limited to the details of the exemplary embodiments described above and that the invention can be implemented in other specific forms without departing from the spirit or essential features of the invention. Therefore, the embodiments should be considered exemplary and non-limiting in all respects, and the scope of the invention is defined by the appended claims rather than the foregoing description, and it is intended that all variations falling within the meaning and range of equivalent elements of the claims be included in the invention. Any reference numeral in a claim should not be considered as limiting the claim to which it relates.

Claims

1. A method for determining the content of components of ebastine oral solution, comprising the following steps: Step 1, determine the chromatographic conditions; Step 2, prepare the solvent; Step 3, prepare the reference solution; Step 4, prepare the test solution; Step 5, sample analysis; Step 6, result calculation and judgment; It is characterized by: In the above step 1, the chromatographic conditions are determined as follows: using octadecylsilane bonded silica gel as a filler, phosphate buffer as a mobile phase A, methanol as a mobile phase B for gradient elution, setting the column temperature to 30°C, the flow rate to 1.0 ml / min, the detection wavelength to 255 nm, and the injection volume to 20 μl; Wherein in the above step 2, mobile phase A and mobile phase B are mixed in a ratio of 35:65 to obtain a solvent; Wherein in the above step 3, under light-proof conditions, prepare methylparaben reference substance stock solution, propylparaben reference substance stock solution and ebastine reference substance stock solution, take 1 ml of each of the three reference substance stock solutions, put them in a 20 ml volumetric flask, add solvent to dilute to the scale, and obtain a reference substance solution containing 20 μg of ebastine, 20 μg of methylparaben and 5 μg of propylparaben per 1 ml, and prepare two in parallel; In the above step 4, under light-proof conditions, accurately measure 1 ml of the test sample, place it in a 50 ml volumetric flask, add solvent to dilute to the scale, shake well to obtain the test sample solution, and prepare two copies in parallel; In the above step 5, the solvent obtained in step 2, the reference solution obtained in step 3, and the test solution obtained in step 4 are taken, and the chromatographic conditions determined in step 1 are used for analysis, and a chromatogram is recorded; In the above step six, according to the peak areas of ebastine, methylparaben and propylparaben in the test and reference solutions obtained in the chromatogram, as well as the sample weight, content and dilution multiple of the test and reference solutions, the concentrations of the components in the reference solution and the peak area ratios are calculated, and then the content of ebastine in the test sample and the content of sodium methylparaben and sodium propylparaben in the test sample are calculated to determine whether the contents meet the limit requirements.

2. A method for determining the content of components of ebastine oral solution according to claim 1, characterized in that: In the step 1, the preparation method of the phosphate buffer is specifically as follows: weigh 2.0 g of potassium dihydrogen phosphate, dissolve it in water and dilute it to 1000 ml, and adjust the pH to 3.5 with 50% phosphoric acid.

3. A method for determining the content of components of ebastine oral solution according to claim 1, characterized in that: In the step 3, the preparation of the methylparaben reference substance stock solution is specifically as follows: 10 mg of the methylparaben reference substance is weighed and placed in a 25 ml volumetric flask, a solvent is added to dissolve and dilute to the scale, and shaken to obtain the methylparaben reference substance stock solution.

4. A method for determining the content of components of ebastine oral solution according to claim 1, characterized in that: In the step 3, the preparation of the propylparaben reference substance stock solution is specifically as follows: 10 mg of the propylparaben reference substance is weighed and placed in a 100 ml volumetric flask, a solvent is added to dissolve and dilute to the scale, and shaken to obtain the propylparaben reference substance stock solution.

5. A method for determining the content of components of ebastine oral solution according to claim 1, characterized in that: In step 3, the preparation of the Ebastine reference substance stock solution is specifically as follows: 10 mg of the Ebastine reference substance is weighed and placed in a 25 ml volumetric flask, 16 ml of methanol is added and ultrasonicated for 30 seconds to dissolve, and then mobile phase A is added to dilute to the scale to obtain the Ebastine reference substance stock solution.

6. A method for determining the content of components of ebastine oral solution according to claim 1, characterized in that: In step 6, the concentration of each component and the peak area ratio of the reference solution are calculated as follows: Among them, A 对 is the peak area of ​​ebastine, methylparaben and propylparaben in the reference solution, M 对 is the sample weight of the reference substance, in mg, V 对 is the dilution factor of the reference substance, C 对 is the concentration of each component of the reference solution, the unit is mg / ml, a% is the content of each reference substance, and f is the ratio of the concentration of each component of the reference solution to the peak area.

7. A method for determining the content of components of ebastine oral solution according to claim 1, characterized in that: In step 6, the calculation formula for the content of ebastine in the test sample is as follows: Among them, A 样 is the peak area of ​​ebastine in the test sample, V 样 is the dilution multiple of the test sample, f is the ratio of the concentration of each component in the reference solution to the peak area, and the average value is used for calculation.

8. A method for determining the content of components of ebastine oral solution according to claim 1, characterized in that: In the step 6, the calculation formula for the content of sodium methylparaben and sodium propylparaben in the test sample is as follows: Antibacterial agent content (mg / ml) = k × A 样 ×f×V 样 Among them, k is the conversion coefficient, the conversion coefficient of sodium methylparaben is 1.144, the conversion coefficient of sodium propylparaben is 1.122, and A 样 is the peak area of ​​methylparaben and propylparaben in the test sample, V 样 is the dilution multiple of the test sample, f is the ratio of the concentration of each component in the reference solution to the peak area, and the average value is used for calculation.

9. A method for determining the content of components of ebastine oral solution according to claim 1, characterized in that: In step six, the limit requirements are: calculated by external standard method based on peak area, ebastine should be 95.0% to 105.0% of the labeled amount; calculated by external standard method by multiplying peak area by conversion factor, sodium methylparaben should be 0.96 mg / ml to 1.32 mg / ml; sodium propylparaben should be 0.24 mg / ml to 0.33 mg / ml.

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