Method for detecting the content of avicel in budesonide nasal spray

Abstract

The application relates to the technical field of Avicel content detection, and discloses a detection method of the Avicel content in budesonide nasal spray, which comprises the following steps: mixing diphenylamine, glacial acetic acid and concentrated hydrochloric acid, stirring uniformly to obtain a derivatization reagent, mixing the derivatization reagent and a detection aid to obtain a composite derivatization reagent; uniformly mixing the budesonide nasal spray and a solvent, heating and dissolving until completely clear, cooling to room temperature, adding deionized water to dilute to obtain a budesonide nasal spray solution; mixing the budesonide nasal spray solution and the composite derivatization reagent, carrying out a derivatization reaction, cooling to room temperature to obtain a sample solution to be detected, and using an ultraviolet-visible spectrophotometry method to detect the absorbance of the sample solution to be detected; and recording the Avicel content according to a linear equation of the absorbance and the Avicel concentration. The method has the advantages of high accuracy, high sensitivity, strong specificity, good linearity and good accuracy.

Description

Technical Field

[0001] This invention relates to the field of Avicel (microcrystalline cellulose-carboxymethyl cellulose sodium co-processed product) content detection technology, specifically to a method for detecting Avicel content in budesonide nasal spray. Background Technology

[0002] Allergic rhinitis is a non-infectious inflammatory disease of the nasal mucosa that seriously affects patients' quality of life. Budesonide is a glucocorticoid with highly effective local anti-inflammatory effects. It has anti-inflammatory and anti-allergic effects and has become a first-line drug for the treatment of moderate to severe allergic rhinitis. The acetal group in its molecular structure gives it a high affinity for glucocorticoid receptors and a first-pass metabolism effect. Therefore, when used locally, it can exert its full therapeutic effect with minimal systemic side effects, making it very suitable for local treatment via nasal administration.

[0003] Budesonide nasal spray is a convenient and non-invasive method of drug delivery that delivers the medication directly to the nasal mucosa lesions. It has a rapid onset of action and high patient compliance. Therefore, budesonide nasal spray has become one of the core drugs for the clinical treatment of allergic rhinitis and is suitable for treating seasonal and perennial allergic rhinitis as well as perennial non-allergic rhinitis.

[0004] Avicel (microcrystalline cellulose-carboxymethyl cellulose sodium co-processed compound) is a key excipient in budesonide nasal spray. Accurate determination of the Avicel content in budesonide nasal spray is crucial for guiding the formulation process. Currently, methods for determining Avicel content include titration, size exclusion chromatography, near-infrared chemometrics, and centrifugation-drying-weighing. However, titration is easily affected by interference from other components in the formulation, leading to inaccurate determination; size exclusion chromatography yields peaks that are not sharp Gaussian peaks, thus hindering accurate quantification; near-infrared chemometrics has low sensitivity and requires specialized instruments; and centrifugation-drying-weighing can only determine the content of microcrystalline cellulose and is prone to over-accuracy due to interference from other components. Neither domestic nor international pharmacopoeias record a method for detecting Avicel content in budesonide nasal spray.

[0005] Therefore, it is necessary to develop a method that can accurately detect the Avicel content in budesonide nasal spray in order to better control the quality of budesonide nasal spray products. The derivatization reagent formed by diphenylamine can be used to detect the Avicel content in budesonide nasal spray. The diphenylamine derivatization reagent can specifically bind with glucose in budesonide nasal spray to produce a color reaction and form a blue complex. It has high specificity and sensitive color reaction and can avoid interference from other excipients. However, the formed blue complex is prone to aggregation and precipitation in the colorimetric solution, which seriously affects the accuracy, precision and reliability of the analytical results and leads to detection errors. Summary of the Invention

[0006] Technical problems to be solved To address the shortcomings of existing technologies, this invention provides a method for detecting the Avicel content in budesonide nasal spray, which solves the problem that the formed blue complex easily aggregates and precipitates in the colorimetric solution, seriously affecting the accuracy, precision, and reliability of the analytical results and leading to detection errors.

[0007] Technical solution To achieve the above objectives, the present invention provides the following technical solution: A method for detecting the Avicel content in budesonide nasal spray, comprising the following steps: S1. Mix diphenylamine, glacial acetic acid, and concentrated hydrochloric acid, stir until homogeneous to obtain a derivatization reagent, and mix the derivatization reagent with a detection aid to obtain a composite derivatization reagent; S2. Mix the budesonide nasal spray and solvent evenly, heat to dissolve until completely clear, cool to room temperature, dilute with deionized water to obtain budesonide nasal spray solution; S3. Mix the budesonide nasal spray solution and the compound derivatization reagent, carry out the derivatization reaction, and cool to room temperature to obtain the test sample solution. The absorbance of the test sample solution is determined by ultraviolet-visible spectrophotometry. S4. Record the Avicel content based on the linear equation between absorbance and Avicel concentration; The detection aid is obtained by mixing modified sheet-like porous carbon and polyethylene glycol octylphenyl ether; Modified lamellar porous carbon is obtained by modifying the surface of lamellar porous carbon with aminosilane and then reacting it with sodium diphenylamine-4-sulfonate.

[0008] Furthermore, the detection aid is prepared by the following steps: A1. Add sheet-like porous carbon to ethanol and deionized water, stir until homogeneous, add aminosilane, stir until the reaction is complete, cool to room temperature, filter, wash and dry to obtain aminated sheet-like porous carbon. A2. Sodium diphenylamine-4-sulfonate was added to ethanol and stirred until completely dissolved. Aminated sheet-like porous carbon was added and stirred and mixed. The mixture was cooled to room temperature, filtered, washed, and dried to obtain modified sheet-like porous carbon. A3. Add polyethylene glycol octylphenyl ether and modified sheet-like porous carbon to deionized water, stir evenly, heat to 50-60℃, stir and mix for 20-30 minutes, filter, wash, and dry to obtain the testing additive.

[0009] Furthermore, during the A1 reaction described above, the silanol groups generated by the hydrolysis of aminosilane can chemically bond with the hydroxyl groups on the surface of the layered porous carbon, thereby grafting aminosilane onto the surface of the layered porous carbon to obtain aminated layered porous carbon.

[0010] Furthermore, during the A2 reaction described above, the amino groups in the aminated lamellar porous carbon carry positive charges, while the sulfonic acid groups in sodium diphenylamine-4-sulfonate carry negative charges. This allows the aminated lamellar porous carbon and sodium diphenylamine-4-sulfonate to be electrostatically bonded, thereby loading sodium diphenylamine-4-sulfonate onto the surface of the aminated lamellar porous carbon and obtaining modified lamellar porous carbon.

[0011] Furthermore, in the A3 reaction process described above, polyethylene glycol octylphenyl ether, as a surfactant, contains polar functional groups that can bind with the sulfonic acid groups and amino groups contained in the modified lamellar porous carbon through hydrogen bonds, thereby grafting polyethylene glycol octylphenyl ether onto the surface of the modified lamellar porous carbon to obtain a composite derivatization reagent.

[0012] Further, in step A1, the mass ratio of the sheet-like porous carbon, ethanol, deionized water and aminosilane is (2-3):(60-70):(30-40):(1-1.5).

[0013] Further, in step A2, the mass ratio of sodium diphenylamine-4-sulfonate, ethanol, and aminated sheet-like porous carbon is (1.2-1.5):(50-60):(2.5-3).

[0014] Further, in step A3, the mass ratio of polyethylene glycol octylphenyl ether, modified sheet porous carbon, and deionized water is (1.3-1.5):(2.8-3.2):(60-70).

[0015] Further, in step S1, the mass ratio of diphenylamine, glacial acetic acid and concentrated hydrochloric acid is (2-4):(100-130):(80-100).

[0016] Furthermore, in step S1, the concentrated hydrochloric acid has a mass fraction of 40-45%.

[0017] Further, in step S1, the mass ratio of the derivatization reagent to the detection aid is 1:(0.5-0.8).

[0018] Further, in step S2, the mass ratio of the budesonide nasal spray, solvent, and deionized water is (0.3-0.5):(16-20):(1.5-1.8).

[0019] Furthermore, in step S2, the solvent is selected from concentrated phosphoric acid or concentrated sulfuric acid.

[0020] Furthermore, in step S2, the temperature for heating and melting is 80-100℃.

[0021] Further, in step S3, the mass ratio of the budesonide nasal spray solution to the compound derivatizing reagent is (0.4-0.5):(14-16).

[0022] Furthermore, in step S3, the derivatization reaction temperature is 80-110℃, and the derivatization reaction time is 80-100min.

[0023] Furthermore, in step S3, the detection wavelength of the ultraviolet-visible spectrophotometry is 620-640 nm.

[0024] Further, in step S4, the linear equation between absorbance and Avicel concentration is: absorbance = 0.0675 × Avicel concentration - 0.0019.

[0025] Beneficial technical effects (1) In the technical solution of the present invention, aminosilane is grafted onto the surface of sheet-like porous carbon. The sheet-like porous carbon has a large specific surface area and porous structure, and has good adsorption performance. It can adsorb the blue complex (diphenylamine-glucose blue complex) generated in the derivatization reaction onto the surface of the sheet-like porous carbon. This pre-enrichment effect makes the product concentration in the detection area much higher than that in the bulk solution, thereby significantly enhancing the absorbance signal of the final detection and improving the detection sensitivity. Moreover, the sheet-like porous carbon, as a carrier of the blue complex, avoids the formation of the blue complex from easily agglomerating and precipitating in the colorimetric solution, which seriously affects the accuracy, precision and reliability of the analysis results and leads to detection errors. In addition, the grafting of aminosilane onto the surface of the sheet-like porous carbon imparts a large number of amino functional groups to the surface of the sheet-like porous carbon, which is beneficial for grafting sodium diphenylamine-4-sulfonate onto the sheet-like porous carbon. This allows the sheet-like porous carbon to be uniformly dispersed in the diphenylamine derivatization reagent, improving the accuracy and precision of the detection and accurately determining the content of Avicel in budesonide nasal spray.

[0026] (2) In the technical solution of the present invention, sodium diphenylamine-4-sulfonate is loaded onto the surface of aminated sheet-like porous carbon. The diphenylamine group contained in sodium diphenylamine-4-sulfonate has good compatibility with the diphenylamine derivatizing reagent, so that the sheet-like porous carbon can be uniformly dispersed in the diphenylamine derivatizing reagent. The diphenylamine can interact with sodium diphenylamine-4-sulfonate through the benzene ring, so that the diphenylamine derivatizing reagent is adsorbed on the surface of the aminated sheet-like porous carbon. During the derivatization reaction, the blue complex formed is deposited on the surface of the aminated sheet-like porous carbon, preventing the blue complex from agglomerating and forming a precipitate, which would lead to detection errors and make it impossible to accurately determine the content of Avicel in budesonide nasal spray.

[0027] (3) In the technical solution of the present invention, polyethylene glycol octylphenyl ether is grafted onto the surface of modified sheet porous carbon to form a detection aid. Polyethylene glycol octylphenyl ether is used as a surfactant. The detection aid formed on the surface of modified sheet porous carbon can be uniformly dispersed in the derivatization reagent, so that the composite derivatization reagent can fully react with the budesonide nasal spray solution to improve the detection efficiency and more accurately detect the content of Avicel in the budesonide nasal spray. In addition, polyethylene glycol octylphenyl ether can be wrapped in the hydrophobic blue complex formed during the derivatization reaction to prevent the blue complex from agglomerating with each other, improve the accuracy and precision of the detection, and accurately determine the content of Avicel in the budesonide nasal spray.

[0028] (4) In the technical solution of the present invention, the budesonide nasal spray sample is first dissolved in solvent at high temperature to a clear solution, then derivatized at high temperature with a composite derivatizing reagent, and finally determined by ultraviolet-visible spectrophotometry. This method is applicable to a variety of formulations containing Avicel. The solvent used is a concentrated acid, which catalyzes the hydrolysis of glucose in Avicel in budesonide nasal spray to form small molecule sugars or oligosaccharides, providing active sites for subsequent derivatization reactions. During the high-temperature derivatization reaction, glucose in Avicel can undergo a colorimetric reaction with diphenylamine to generate a blue complex with maximum absorption around 635 nm. By measuring the absorbance of this complex and recording the Avicel content according to the linear equation between absorbance and Avicel concentration, this method has the advantages of high accuracy, high sensitivity, strong specificity, and good linearity and accuracy. Detailed Implementation

[0029] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are described clearly and completely. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0030] The raw materials used in the embodiments of this invention are shown below, and all reagents used are analytical grade.

[0031] The diphenylamine was GC grade, batch number K2418028, and was purchased from Aladdin Biochemical Technology Co., Ltd.

[0032] The glacial acetic acid was grade AR, batch number 20250520, and was purchased from Jiangsu Qiangsheng Functional Chemical Co., Ltd.

[0033] The concentrated hydrochloric acid was grade AR, batch number 250626A1, and was purchased from Xilong Scientific.

[0034] The concentrated phosphoric acid was grade AR, batch number 20250621, and purchased from Sinopharm Group.

[0035] The water bath model is HH-8J, and the manufacturer is Jintan City Jerell Electric Appliance Co., Ltd.

[0036] The UV-Vis spectrophotometer is model HS-8, manufactured by Beijing Haotianhui Instrument Co., Ltd.

[0037] The lamellar porous carbon has a pore size of 100 nm and a particle size of 5 μm, and was purchased from Xi'an Qiyue Biotechnology Co., Ltd.

[0038] The aminosilane is KH550 (γ-aminopropyltriethoxysilane).

[0039] The polyethylene glycol octylphenyl ether, catalog number 303135-5G, was purchased from Merck Reagents.

[0040] Example 1 A method for detecting the Avicel content in budesonide nasal spray, comprising the following steps: S1. Mix diphenylamine, glacial acetic acid, and concentrated hydrochloric acid, and stir until homogeneous to obtain a derivatization reagent. Mix the derivatization reagent with the detection aid to obtain a composite derivatization reagent. The mass ratio of diphenylamine, glacial acetic acid, and concentrated hydrochloric acid is 2:100:80; the mass fraction of concentrated hydrochloric acid is 40%; and the mass ratio of the derivatization reagent to the detection aid is 1:0.5. S2. Mix budesonide nasal spray and concentrated phosphoric acid evenly, heat in a water bath until completely clear, cool to room temperature, dilute with deionized water to obtain budesonide nasal spray solution; the mass ratio of budesonide nasal spray, solvent and deionized water is 0.3:16:1.5; the heating temperature for dissolving is 80℃; S3. The budesonide nasal spray solution and the compound derivatization reagent were mixed and subjected to a derivatization reaction. After cooling to room temperature, the sample solution was obtained. The absorbance of the sample solution was determined by ultraviolet-visible spectrophotometry. The mass ratio of budesonide nasal spray solution to compound derivatization reagent was 0.4:14. The derivatization reaction temperature was 80℃ and the derivatization reaction time was 80 min. The detection wavelength of ultraviolet-visible spectrophotometry was 620 nm. S4. Record the Avicel content according to the linear equation between absorbance and Avicel concentration; the linear equation between absorbance and Avicel concentration is: absorbance = 0.0675 × Avicel concentration - 0.0019.

[0041] The testing aid is prepared by the following steps: A1. Add sheet-like porous carbon to ethanol and deionized water, stir until homogeneous, add KH550, stir and react at 70℃ for 1.5h, cool to room temperature, filter, wash 3 times with ethanol and 3 times with deionized water, and dry in an oven at 70℃ for 10min to obtain aminated sheet-like porous carbon; the mass ratio of sheet-like porous carbon, ethanol, deionized water and KH550 is 2:60:30:1. A2. Sodium diphenylamine-4-sulfonate was added to ethanol and stirred at 60°C until completely dissolved. Aminated sheet-like porous carbon was then added, and the mixture was stirred for another 30 minutes. The mixture was cooled to room temperature, filtered, washed three times with ethanol, and dried in an oven at 70°C for 10 minutes to obtain modified sheet-like porous carbon. The mass ratio of sodium diphenylamine-4-sulfonate, ethanol, and aminated sheet-like porous carbon was 1.2:50:2.5. A3. Polyethylene glycol octylphenyl ether and modified sheet-like porous carbon were added to deionized water, stirred evenly, heated to 50°C, stirred and mixed for 20 min, filtered, washed 3 times with deionized water, and dried in an oven at 70°C for 10 min to obtain the test additive; the mass ratio of polyethylene glycol octylphenyl ether, modified sheet-like porous carbon and deionized water was 1.3:2.8:60.

[0042] Example 2 A method for detecting the Avicel content in budesonide nasal spray, comprising the following steps: S1. Mix diphenylamine, glacial acetic acid, and concentrated hydrochloric acid, stir until homogeneous to obtain a derivatization reagent. Mix the derivatization reagent and the detection aid to obtain a composite derivatization reagent. The mass ratio of diphenylamine, glacial acetic acid, and concentrated hydrochloric acid is 3:120:90; the mass fraction of concentrated hydrochloric acid is 43%; and the mass ratio of the derivatization reagent to the detection aid is 1:0.7. S2. Mix budesonide nasal spray and concentrated phosphoric acid evenly, heat in a water bath until completely clear, cool to room temperature, dilute with deionized water to obtain budesonide nasal spray solution; the mass ratio of budesonide nasal spray, solvent and deionized water is 0.4:18:1.7; the heating temperature for dissolving is 90℃; S3. The budesonide nasal spray solution and the compound derivatization reagent were mixed and subjected to a derivatization reaction. After cooling to room temperature, the sample solution was obtained. The absorbance of the sample solution was determined by ultraviolet-visible spectrophotometry. The mass ratio of budesonide nasal spray solution to compound derivatization reagent was 0.45:15; the derivatization reaction temperature was 100℃ and the derivatization reaction time was 90 min; the detection wavelength of ultraviolet-visible spectrophotometry was 630 nm. S4. Record the Avicel content according to the linear equation between absorbance and Avicel concentration; the linear equation between absorbance and Avicel concentration is: absorbance = 0.0675 × Avicel concentration - 0.0019.

[0043] The testing aid is prepared by the following steps: A1. Add sheet-like porous carbon to ethanol and deionized water, stir evenly, add KH550, stir and react at 70℃ for 1.5h, cool to room temperature, filter, wash 3 times with ethanol and 3 times with deionized water, and dry in an oven at 70℃ for 10min to obtain aminated sheet-like porous carbon; the mass ratio of sheet-like porous carbon, ethanol, deionized water and KH550 is 2.5:65:35:1.3; A2. Sodium diphenylamine-4-sulfonate was added to ethanol and stirred at 60°C until completely dissolved. Aminated sheet-like porous carbon was then added, and the mixture was stirred for another 30 minutes. After cooling to room temperature, the mixture was filtered, washed three times with ethanol, and dried in an oven at 70°C for 10 minutes to obtain modified sheet-like porous carbon. The mass ratio of sodium diphenylamine-4-sulfonate, ethanol, and aminated sheet-like porous carbon was 1.3:55:2.8. A3. Polyethylene glycol octylphenyl ether and modified sheet-like porous carbon were added to deionized water, stirred evenly, heated to 55℃, stirred and mixed for 25 min, filtered, washed 3 times with deionized water, and dried in an oven at 70℃ for 10 min to obtain the testing additive; the mass ratio of polyethylene glycol octylphenyl ether, modified sheet-like porous carbon and deionized water was 1.4:3:65.

[0044] Example 3 A method for detecting the Avicel content in budesonide nasal spray, comprising the following steps: S1. Mix diphenylamine, glacial acetic acid, and concentrated hydrochloric acid, stir until homogeneous to obtain a derivatization reagent. Mix the derivatization reagent and the detection aid to obtain a composite derivatization reagent. The mass ratio of diphenylamine, glacial acetic acid, and concentrated hydrochloric acid is 4:130:100; the mass fraction of concentrated hydrochloric acid is 45%; and the mass ratio of the derivatization reagent to the detection aid is 1:0.8. S2. Mix budesonide nasal spray and concentrated phosphoric acid evenly, heat in a water bath until completely clear, cool to room temperature, dilute with deionized water to obtain budesonide nasal spray solution; the mass ratio of budesonide nasal spray, solvent and deionized water is 0.5:20:1.8; the heating temperature for dissolving is 100℃; S3. The budesonide nasal spray solution and the compound derivatization reagent were mixed and subjected to a derivatization reaction. After cooling to room temperature, the sample solution was obtained. The absorbance of the sample solution was determined by ultraviolet-visible spectrophotometry. The mass ratio of budesonide nasal spray solution to compound derivatization reagent was 0.5:16; the derivatization reaction temperature was 110℃ and the derivatization reaction time was 100 min; the detection wavelength of ultraviolet-visible spectrophotometry was 640 nm. S4. Record the Avicel content according to the linear equation between absorbance and Avicel concentration; the linear equation between absorbance and Avicel concentration is: absorbance = 0.0675 × Avicel concentration - 0.0019.

[0045] The testing aid is prepared by the following steps: A1. Add sheet-like porous carbon to ethanol and deionized water, stir evenly, add KH550, stir and react at 70℃ for 1.5h, cool to room temperature, filter, wash 3 times with ethanol and 3 times with deionized water, and dry in an oven at 70℃ for 10min to obtain aminated sheet-like porous carbon; the mass ratio of sheet-like porous carbon, ethanol, deionized water and KH550 is 3:70:40:1.5; A2. Sodium diphenylamine-4-sulfonate was added to ethanol and stirred at 60°C until completely dissolved. Aminated sheet-like porous carbon was then added, and the mixture was stirred for another 30 minutes. After cooling to room temperature, the mixture was filtered, washed three times with ethanol, and dried in an oven at 70°C for 10 minutes to obtain modified sheet-like porous carbon. The mass ratio of sodium diphenylamine-4-sulfonate, ethanol, and aminated sheet-like porous carbon was 1.5:60:3. A3. Polyethylene glycol octylphenyl ether and modified sheet-like porous carbon were added to deionized water, stirred evenly, heated to 60℃, stirred and mixed for 30 min, filtered, washed 3 times with deionized water, and dried in an oven at 70℃ for 10 min to obtain the test additive; the mass ratio of polyethylene glycol octylphenyl ether, modified sheet-like porous carbon and deionized water was 1.5:3.2:70.

[0046] Comparative Example 1 The only difference between this comparative example and Example 3 is the preparation of the detection aid, as detailed below: The testing aid is prepared by the following steps: A1. Graphene oxide was added to ethanol and deionized water, stirred until homogeneous, KH550 was added, and the mixture was stirred at 70°C for 1.5 h. After cooling to room temperature, the mixture was filtered, washed three times with ethanol and three times with deionized water, and dried in an oven at 70°C for 10 min to obtain aminated graphene oxide. The mass ratio of graphene oxide, ethanol, deionized water and KH550 was 3:70:40:1.5. A2. Sodium diphenylamine-4-sulfonate was added to ethanol and stirred at 60°C until completely dissolved. Aminated sheet-like porous carbon was added, and stirring was continued for 30 min. After cooling to room temperature, the mixture was filtered, washed three times with ethanol, and dried in an oven at 70°C for 10 min to obtain modified graphene oxide. The mass ratio of sodium diphenylamine-4-sulfonate, ethanol, and aminated graphene oxide was 1.5:60:3. A3. Polyethylene glycol octylphenyl ether and modified graphene oxide were added to deionized water, stirred evenly, heated to 60°C, stirred and mixed for 30 min, filtered, washed 3 times with deionized water, and dried in an oven at 70°C for 10 min to obtain the testing additive; the mass ratio of polyethylene glycol octylphenyl ether, modified graphene oxide and deionized water was 1.5:3.2:70.

[0047] Comparative Example 2 The only difference between this comparative example and Example 3 is the preparation of the detection aid, as detailed below: The testing aid is prepared by the following steps: A1. Sodium diphenylamine-4-sulfonate was added to ethanol and stirred at 60°C until completely dissolved. Then, sheet-like porous carbon was added, and the mixture was stirred for another 30 minutes. After cooling to room temperature, the mixture was filtered, washed three times with ethanol, and dried in an oven at 70°C for 10 minutes to obtain modified sheet-like porous carbon. The mass ratio of sodium diphenylamine-4-sulfonate, ethanol, and sheet-like porous carbon was 1.5:60:3. A2. Polyethylene glycol octylphenyl ether and modified sheet-like porous carbon were added to deionized water, stirred evenly, heated to 60℃, stirred and mixed for 30 min, filtered, washed 3 times with deionized water, and dried in an oven at 70℃ for 10 min to obtain the test additive; the mass ratio of polyethylene glycol octylphenyl ether, modified sheet-like porous carbon and deionized water was 1.5:3.2:70.

[0048] Comparative Example 3 The only difference between this comparative example and Example 3 is the preparation of the detection aid, as detailed below: The testing aid is prepared by the following steps: A1. Add sheet-like porous carbon to ethanol and deionized water, stir evenly, add KH550, stir and react at 70℃ for 1.5h, cool to room temperature, filter, wash 3 times with ethanol and 3 times with deionized water, and dry in an oven at 70℃ for 10min to obtain aminated sheet-like porous carbon; the mass ratio of sheet-like porous carbon, ethanol, deionized water and KH550 is 3:70:40:1.5; A2. Polyethylene glycol octylphenyl ether and aminated sheet-like porous carbon were added to deionized water, stirred evenly, heated to 60°C, stirred and mixed for 30 min, filtered, washed 3 times with deionized water, and dried in an oven at 70°C for 10 min to obtain the testing additive; the mass ratio of polyethylene glycol octylphenyl ether, aminated sheet-like porous carbon and deionized water was 1.5:3.2:70.

[0049] Comparative Example 4 The only difference between this comparative example and Example 3 is the preparation of the detection aid, as detailed below: The testing aid is prepared by the following steps: A1. Add sheet-like porous carbon to ethanol and deionized water, stir evenly, add KH550, stir and react at 70℃ for 1.5h, cool to room temperature, filter, wash 3 times with ethanol and 3 times with deionized water, and dry in an oven at 70℃ for 10min to obtain aminated sheet-like porous carbon; the mass ratio of sheet-like porous carbon, ethanol, deionized water and KH550 is 3:70:40:1.5; A2. Sodium diphenylamine-4-sulfonate was added to ethanol and stirred at 60°C until completely dissolved. Aminated sheet-like porous carbon was then added, and the mixture was stirred for another 30 minutes. After cooling to room temperature, the mixture was filtered, washed three times with ethanol, and dried in an oven at 70°C for 10 minutes to obtain modified sheet-like porous carbon. The mass ratio of sodium diphenylamine-4-sulfonate, ethanol, and aminated sheet-like porous carbon was 1.5:60:3. A3. Add the modified sheet-like porous carbon to deionized water, stir evenly, heat to 60℃, stir and mix for 30 min, filter, wash 3 times with deionized water, and dry in a 70℃ oven for 10 min to obtain the detection additive; the mass ratio of modified sheet-like porous carbon to deionized water is 4.7:70.

[0050] The detection method for Avicel content in budesonide nasal sprays in Examples 1-3 and Comparative Examples 1-4 is now tested. The specific test method is as follows: 1. Linear equation between absorbance and Avicel concentration: Avicel was mixed thoroughly with 45% concentrated phosphoric acid and heated at 100°C until completely clear. After cooling to room temperature, the mixture was diluted with deionized water to obtain Avicel solutions. The concentrations of the Avicel solutions were 2.104 μg / ml, 3.156 μg / ml, 5.050 μg / ml, 6.313 μg / ml, and 9.469 μg / ml, respectively. Mix 2.5g of diphenylamine and 110mL of glacial acetic acid, then add 90mL of concentrated hydrochloric acid and mix well to obtain a derivatization reagent. Mix Avicel solution and derivatization reagent to carry out derivatization reaction, then cool to room temperature to obtain the sample solution to be tested. The absorbance of the sample solution to be tested is determined by ultraviolet-visible spectrophotometry. The data and linear relationship between Avicel concentration and absorbance are as follows:

[0051]

[0052] It can be seen that the linear equation between absorbance and Avicel concentration is: absorbance = 0.0675 × Avicel concentration - 0.0019.

[0053] 2. The Avicel content in budesonide nasal spray was tested using the methods described in Examples 1-3 and Comparative Examples 1-4. The specific test methods are as follows: S1. Mix diphenylamine, glacial acetic acid, and concentrated hydrochloric acid, stir until homogeneous to obtain a derivatization reagent. Mix the derivatization reagent and the detection aid to obtain a composite derivatization reagent. The mass ratio of diphenylamine, glacial acetic acid, and concentrated hydrochloric acid is 4:130:100; the mass fraction of concentrated hydrochloric acid is 45%; and the mass ratio of the derivatization reagent to the detection aid is 1:0.8. S2. Mix budesonide nasal spray and concentrated phosphoric acid evenly, heat to dissolve until completely clear, cool to room temperature, and dilute with deionized water to obtain budesonide nasal spray solution; the mass ratio of budesonide nasal spray, solvent and deionized water is 0.5:20:1.8; the heating and dissolving temperature is 100℃; S3. The budesonide nasal spray solution and the compound derivatization reagent were mixed and subjected to a derivatization reaction. After cooling to room temperature, the sample solution was obtained. The absorbance of the sample solution was determined by ultraviolet-visible spectrophotometry. The mass ratio of budesonide nasal spray solution to compound derivatization reagent was 0.5:16; the derivatization reaction temperature was 110℃ and the derivatization reaction time was 100 min; the detection wavelength of ultraviolet-visible spectrophotometry was 640 nm. S4. Record the Avicel content according to the linear equation between absorbance and Avicel concentration; the linear equation between absorbance and Avicel concentration is: absorbance = 0.0675 × Avicel concentration - 0.0019.

[0054] The concentration of Avicel in the budesonide nasal spray solution of this invention is 5.06 μg / mL.

[0055] The Avicel content in budesonide nasal spray was determined as shown in Table 3.

[0056] As shown in Tables 1, 2, and 3, in all examples, the budesonide nasal spray samples were dissolved in solvent at high temperature to obtain a clear solution, then derivatized at high temperature using a complex derivatizing reagent, and finally determined by UV-Vis spectrophotometry. The Avicel content was recorded based on the linear equation between absorbance and Avicel concentration. This method has the advantages of high accuracy, high sensitivity, strong specificity, and good linearity and accuracy.

[0057] Comparative Example 1 showed that replacing the layered porous carbon with graphene oxide to prepare a detection aid for detecting Avicel content in budesonide nasal spray resulted in a decrease in detection performance. This demonstrates that the layered porous carbon has a large specific surface area and porous structure, exhibiting good adsorption performance. It can adsorb the blue complex (diphenylamine-glucose blue complex) generated in the derivatization reaction onto the surface of the layered porous carbon. This pre-enrichment effect makes the product concentration in the detection area much higher than that in the bulk solution, thereby significantly enhancing the absorbance signal of the final detection and improving the detection sensitivity. Furthermore, the layered porous carbon, as a carrier of the blue complex, avoids the formation of aggregates and precipitation of the blue complex in the colorimetric solution, which would seriously affect the accuracy, precision, and reliability of the analytical results and lead to detection errors.

[0058] Comparative Example 2 showed that replacing the aminated lamellar porous carbon with a detection aid prepared from lamellar porous carbon resulted in a decrease in the detection performance of Avicel in budesonide nasal spray. This demonstrates that the grafting of aminosilane onto the surface of lamellar porous carbon imparts a large number of amino functional groups, which is beneficial for grafting sodium diphenylamine-4-sulfonate onto the lamellar porous carbon. This allows the lamellar porous carbon to be uniformly dispersed in the diphenylamine derivatization reagent, improving the accuracy and precision of the detection and accurately determining the Avicel content in budesonide nasal spray.

[0059] Comparative Example 3 showed that the detection aid prepared by replacing the modified layered porous carbon with aminated layered porous carbon was used to detect the Avicel content in budesonide nasal spray. The detection performance decreased, which proved that the diphenylamine group contained in sodium diphenylamine-4-sulfonate has good compatibility with the diphenylamine derivatization reagent. This allows the layered porous carbon to be uniformly dispersed in the diphenylamine derivatization reagent. Furthermore, the diphenylamine can interact with sodium diphenylamine-4-sulfonate through the benzene ring, causing the diphenylamine derivatization reagent to be adsorbed on the surface of the aminated layered porous carbon. During the derivatization reaction, the blue complex formed is deposited on the surface of the aminated layered porous carbon, preventing the blue complex from agglomerating and forming a precipitate. This prevents the detection error and makes it impossible to accurately determine the Avicel content in budesonide nasal spray.

[0060] Comparative Example 4 replaced polyethylene glycol octylphenyl ether with a detection aid prepared from modified sheet-like porous carbon to detect the Avicel content in budesonide nasal spray. The detection performance decreased, demonstrating that polyethylene glycol octylphenyl ether, as a surfactant, can be uniformly dispersed in the derivatization reagent when grafted onto the surface of the modified sheet-like porous carbon. This allows the composite derivatization reagent to fully react with the budesonide nasal spray solution, improving detection efficiency and more accurately determining the Avicel content in the budesonide nasal spray. Furthermore, polyethylene glycol octylphenyl ether can encapsulate the hydrophobic blue complex formed during the derivatization reaction, preventing the blue complex from agglomerating and improving the accuracy and precision of the detection, thus accurately determining the Avicel content in the budesonide nasal spray.

[0061] It should be noted that, in this document, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Unless otherwise specified, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes said element.

[0062] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

[0063] Those skilled in the art should understand that the above descriptions are merely several specific embodiments of the present invention, and not all embodiments.

Claims

1. A method for detecting the Avicel content in budesonide nasal spray, characterized in that, Includes the following steps: S1. Mix diphenylamine, glacial acetic acid, and concentrated hydrochloric acid, stir until homogeneous to obtain a derivatization reagent, and mix the derivatization reagent with a detection aid to obtain a composite derivatization reagent; S2. Mix the budesonide nasal spray and solvent evenly, heat to dissolve until completely clear, cool to room temperature, dilute with deionized water to obtain budesonide nasal spray solution; S3. Mix the budesonide nasal spray solution and the compound derivatization reagent, carry out the derivatization reaction, and cool to room temperature to obtain the test sample solution. The absorbance of the test sample solution is determined by ultraviolet-visible spectrophotometry. S4. Record the Avicel content based on the linear equation between absorbance and Avicel concentration; The detection aid is obtained by mixing modified sheet-like porous carbon and polyethylene glycol octylphenyl ether; Modified sheet-like porous carbon is obtained by modifying the surface of sheet-like porous carbon with aminosilane and then reacting it with sodium diphenylamine-4-sulfonate. The detection aid is prepared by the following steps: A1. Add sheet-like porous carbon to ethanol and deionized water, stir until homogeneous, add aminosilane, stir until the reaction is complete, cool to room temperature, filter, wash and dry to obtain aminated sheet-like porous carbon; the mass ratio of sheet-like porous carbon, ethanol, deionized water and aminosilane is (2-3):(60-70):(30-40):(1-1.5); A2. Sodium diphenylamine-4-sulfonate is added to ethanol and stirred until completely dissolved. Aminated sheet-like porous carbon is then added, and the mixture is stirred and mixed. The mixture is cooled to room temperature, filtered, washed, and dried to obtain modified sheet-like porous carbon. The mass ratio of sodium diphenylamine-4-sulfonate, ethanol, and aminated sheet-like porous carbon is (1.2-1.5):(50-60):(2.5-3). A3. Polyethylene glycol octylphenyl ether and modified sheet-like porous carbon are added to deionized water, stirred evenly, heated to 50-60℃, stirred and mixed for 20-30 minutes, filtered, washed, and dried to obtain the detection additive; the mass ratio of polyethylene glycol octylphenyl ether, modified sheet-like porous carbon and deionized water is (1.3-1.5):(2.8-3.2):(60-70).

2. The method for detecting Avicel content in budesonide nasal spray according to claim 1, characterized in that, In step S1, the mass ratio of diphenylamine, glacial acetic acid, and concentrated hydrochloric acid is (2-4):(100-130):(80-100); In step S1, the concentrated hydrochloric acid has a mass fraction of 40-45%. In step S1, the mass ratio of the derivatization reagent to the detection aid is 1:(0.5-0.8).

3. The method for detecting Avicel content in budesonide nasal spray according to claim 1, characterized in that, In step S2, the mass ratio of the budesonide nasal spray, solvent, and deionized water is (0.3-0.5):(16-20):(1.5-1.8).

4. The method for detecting Avicel content in budesonide nasal spray according to claim 1, characterized in that, In step S2, the solvent is selected from concentrated phosphoric acid or concentrated sulfuric acid; In step S2, the heating and melting temperature is 80-100℃.

5. The method for detecting Avicel content in budesonide nasal spray according to claim 1, characterized in that, In step S3, the mass ratio of the budesonide nasal spray solution to the compound derivatizing reagent is (0.4-0.5):(14-16); In step S3, the derivatization reaction temperature is 80-110℃ and the derivatization reaction time is 80-100 min; In step S3, the detection wavelength of the ultraviolet-visible spectrophotometry is 620-640 nm.

6. The method for detecting Avicel content in budesonide nasal spray according to claim 1, characterized in that, In step S4, the linear equation between absorbance and Avicel concentration is: absorbance = 0.0675 × Avicel concentration - 0.0019.

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