Quality detection method of montelukast sodium chewable tablets

Abstract

The invention discloses a quality detection method of montelukast sodium chewable tablets, and belongs to the technical field of medicine detection. The quality detection comprises content detection and related substance detection, and the quality detection adopts high performance liquid chromatography; chemical substances detected in the content detection comprise montelukast sodium, and chemical substances detected in the related substance detection comprise sulfoxide, montelukast ketone and a cis-isomer. Different chromatographic detection conditions and detection parameters are set through a high performance liquid chromatograph by adopting a high performance liquid chromatography method, the content and the amount of related substances can be accurately detected respectively, the product quality can be better controlled, and the detection method is high in sensitivity and good in separation degree.

Description

Technical Field

[0001] The invention relates to a method for detecting chemical substances, in particular to a method for detecting the quality of montelukast sodium chewable tablets, and belongs to the technical field of drug detection. Background Art

[0002] Montelukast sodium chewable tablets are used for the prevention and long-term treatment of asthma in adults and children 2 years of age and older, including prevention of daytime and nighttime asthma symptoms, treatment of patients with aspirin-sensitive asthma, and prevention of exercise-induced bronchoconstriction; they are also used to relieve symptoms caused by seasonal allergic rhinitis.

[0003] The main ingredient of montelukast sodium chewable tablets is montelukast sodium, and its chemical name is: (+)-1-[[[(1R)-1-[3-[(1E)-2-(7-chloro-2-quinolyl)-vinyl]phenyl]-3-[2-(1-hydroxy-1-methylethyl)phenyl]propyl]thio]methyl]cyclopropaneacetic acid monosodium salt. Molecular formula: C 35 H 35 ClNNaO3S molecular weight: 608.18.

[0004] After the preparation of montelukast sodium chewable tablets is completed, the quality of the montelukast sodium chewable tablets needs to be tested. The quality test materials include the content of the main component and the amount of related substances. The main component is montelukast sodium, and the related substances are sulfoxides, cis-isomers, montelukast ketone, etc. The related substances are also called impurities. The content of the main component and the amount of impurities are related to the quality and effect of the product.

[0005] At present, various detection methods for the content of main components and the amount of impurities in montelukast sodium chewable tablets have not been able to accurately and sensitively detect the content of main components and the amount of impurities in montelukast sodium chewable tablets, and it is necessary to ensure the production of high-quality qualified lornoxicam injection products. Summary of the invention

[0006] The technical problem to be solved by the present invention is to provide a quality detection method for montelukast sodium chewable tablets, so as to solve the problem that the quality of montelukast sodium chewable tablets cannot be accurately detected in the prior art. The content and the amount of related substances in montelukast sodium chewable tablets can be accurately detected by using a high performance liquid chromatography (HPLC) to set different chromatographic detection conditions and detection parameters through a high performance liquid chromatograph, thereby ensuring better control of product quality.

[0007] In order to solve the above technical problems, the technical solution adopted by the present invention is:

[0008] A quality testing method for montelukast sodium chewable tablets, the quality testing comprising dissolution testing, content testing and related substance testing, the quality testing being performed by high performance liquid chromatography; the chemical substances tested in the content testing comprise: montelukast sodium, and the chemical substances tested in the related substance testing comprise: sulfoxide, cis-isomer, montelukast ketone;

[0009] The assay includes the following chromatographic conditions:

[0010] Phenyl bonded silica gel was used as filler; a mixed solution of 0.2% trifluoroacetic acid aqueous solution-0.2% trifluoroacetic acid acetonitrile solution was used as mobile phase for isocratic elution; the flow rate was 1.5 ml / min; the column temperature was 50° C.; and the detection wavelength was 358 nm.

[0011] The relevant substance detection includes the following chromatographic conditions:

[0012] Phenyl bonded silica gel was used as filler; 0.2% trifluoroacetic acid aqueous solution was used as mobile phase A; methanol-acetonitrile was used as mobile phase B; linear gradient elution was performed, column temperature was 50°C; detection wavelength was 250nm; flow rate was 1.0ml / min.

[0013] Furthermore, the volume ratio of 0.2% trifluoroacetic acid aqueous solution to 0.2% trifluoroacetic acid acetonitrile solution in the mobile phase solution is 3:1.

[0014] Furthermore, the specific operation of content detection includes the following steps:

[0015] A1. Test solution: Take 10 montelukast sodium chewable tablets, place in a 200ml brown volumetric bottle, add 40ml water, leave for 5 minutes or until the tablets are completely disintegrated, add 120ml methanol, ultrasonically treat for 20 minutes, shake, dilute to scale with methanol-water, shake well, filter, accurately measure 2ml of the filtrate, place in a 20ml brown volumetric bottle, dissolve and dilute to scale with methanol-water, shake well, and obtain;

[0016] A2. Reference substance solution: Take 17 mg of montelukast dicyclohexylamine reference substance, accurately weigh it, place it in a 50 ml brown volumetric flask, add an appropriate amount of methanol-water, ultrasonically dissolve it and dilute it to the scale, shake well; accurately measure 2 ml, place it in a 20 ml brown volumetric flask, dilute it to the scale with methanol-water, shake well, and obtain;

[0017] A3. Content determination: Take 10 μl of reference solution and test solution respectively, inject them into liquid chromatograph and record the chromatogram.

[0018] Further, in step A3, the content is calculated by the formula:

[0019]

[0020] Where:

[0021] W S The sample weight of montelukast dicyclohexylamine reference substance, mg;

[0022] AT is the peak area of ​​the main peak in the chromatogram of the test solution;

[0023] AS is the peak area of ​​the main peak in the chromatogram of the reference solution;

[0024] DT is the dilution factor of the test solution;

[0025] DS is the dilution factor of the reference solution;

[0026] n is the number of montelukast sodium chewable tablets sampled in the test solution;

[0027] L is the labeled amount;

[0028] 0.7638 is the ratio of montelukast (M: 586.18) to montelukast dicyclohexylamine (M: 767.50).

[0029] Furthermore, the content limit: containing montelukast sodium (C 35 H 35 ClNNaO3S) with montelukast (C 35 H 35 ClNNaO3S) should be 96.0% to 104.0% of the labeled amount.

[0030] Furthermore, the volume ratio of methanol:acetonitrile in mobile phase B is 85:15.

[0031] Further, the gradient elution program was: 0 min, 63% A, 37% B; 10 min, 43% A, 57% B; 15 min, 63% A, 37% B; 25 min, 63% A, 37% B; 35 min, 70% A, 30% B; 53 min, 70% A, 30% B; 55 min, 40% A, 60% B; 60 min, 40% A, 60% B.

[0032] Furthermore, the specific operation of the substance detection includes the following steps:

[0033] C1. Mixed reference solution: Take 2.5 mg of montelukast peak identification impurity reference, accurately weigh it, put it in a 10 ml brown volumetric flask, add 5 ml of methanol-water, then add 1 ml of montelukast ketone impurity reference solution, dissolve it by ultrasonic, dilute it to the mark with methanol-water, shake well, and use it as the mixed impurity reference solution; take 5 ml of the mixed impurity reference solution, put it in a 10 ml colorless volumetric flask, and place it under natural light for 1 hour to obtain;

[0034] C2. Test sample 3 solution: Take 10 montelukast sodium chewable tablets, place in a 200ml brown volumetric flask, add 40ml water, leave for 5 minutes or until the tablets are completely disintegrated, add 120ml methanol, ultrasonically treat for 20 minutes, shake, dilute to scale with methanol-water, shake well, filter, discard at least 1ml of the initial filtrate, take the subsequent filtrate, accurately measure 1ml, place in a 100ml brown volumetric flask, dilute to scale with methanol-water, shake well, and obtain;

[0035] C3. Sensitivity solution: accurately measure 5 ml of the reference substance mixed solution, place it in a 100 ml brown volumetric flask, dilute to the mark with methanol-water, shake well, and obtain;

[0036] C4. Determination of related substances: Take 20 μl each of the sensitivity solution, test sample 3 solution and reference sample mixed solution, inject them into the liquid chromatograph respectively, and record the chromatogram.

[0037] Further, in step C 5, the relevant substance calculation formula is:

[0038]

[0039] Where:

[0040] AT1 is the peak area of ​​sulfoxide in the chromatogram of test sample 3 solution;

[0041] AT2 is the peak area of ​​montelukast ketone in the chromatogram of test sample 3 solution;

[0042] AT3 is the peak area of ​​the cis isomer in the chromatogram of the solution of test sample 3;

[0043] AT4 is the peak area of ​​the other single largest impurity peak in the chromatogram of the sample 3 solution;

[0044] The total impurities of AT are the sum of the peak areas of each impurity in the chromatogram of the solution of test sample 3;

[0045] AS 1％ The peak area of ​​the main peak in the chromatogram of the reference mixed solution;

[0046] 0.6 is the correction factor for the montelukast ketone impurity.

[0047] Furthermore, the percentage content of each relevant substance is calculated according to the formula, and the content of sulfoxide, cis-isomer, and montelukast ketone shall not exceed 0.1%, 0.1%, and 0.1%, respectively.

[0048] Due to the adoption of the above technical solution, the technical progress achieved by the present invention is:

[0049] The method of the present invention can accurately detect the content and related substances, ensure product quality, and the detection method has high sensitivity and good separation. Specifically, the present invention mainly uses phenyl bonded silica gel as a filler, uses different mobile phases, different elution methods, and different detection wavelengths for different detection substances, and detects them in combination, and can accurately and sensitively detect the amount of montelukast sodium, sulfoxide, montelukast ketone, and cis-isomer in montelukast sodium chewable tablets.

[0050] First, the mobile phase material and mobile phase combination used in the present invention can better separate the montelukast sodium, sulfoxide, montelukast ketone, and cis-isomer in montelukast sodium chewable tablets, detect the amount of the substance, and thus obtain the corresponding chromatogram with high accuracy. Secondly, the gradient elution mode and elution procedure used in the present invention can better separate the montelukast sodium, sulfoxide, montelukast ketone, and cis-isomer in montelukast sodium chewable tablets, avoid interference, accurately detect the amount of the substance, and thus obtain the corresponding chromatogram, baseline separation, and good separation. Finally, the present invention accurately detects montelukast sodium, sulfoxide, montelukast ketone, and cis-isomer substances in montelukast sodium chewable tablets by setting the corresponding detection wavelength, avoids interference of substances outside the detection wavelength, and thus obtains the corresponding chromatogram, baseline separation, and good separation. BRIEF DESCRIPTION OF THE DRAWINGS

[0051] Figure 1 This is a chromatogram of a reference substance solution for content detection of montelukast sodium chewable tablets in Example 1 of the present invention;

[0052] Figure 2 This is a chromatogram of the test solution for content detection of montelukast sodium chewable tablets in Example 1 of the present invention;

[0053] Figure 3 This is a chromatogram of a mixed solution of a reference substance for detection of related substances in montelukast sodium chewable tablets according to Example 1 of the present invention;

[0054] Figure 4 This is a chromatogram of the test sample solution for the detection of related substances in the montelukast sodium chewable tablets in Example 1 of the present invention;

[0055] In the accompanying drawings, the ordinate is the peak height (mAU); the abscissa is the retention time (min). DETAILED DESCRIPTION

[0056] In order to enable those skilled in the art to better understand the solutions of the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described in combination with the embodiments of the present invention and the related drawings. 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.

[0057] Below is a combination of the embodiments and Figures 1 to 4 The present invention is described in further detail:

[0058] A quality detection method for montelukast sodium chewable tablets, the quality detection includes content detection and related substance detection, and the quality detection is detected by high performance liquid chromatography; the chemical substances detected in the content detection include: montelukast sodium, and the chemical substances detected in the related substance detection include: sulfoxide, montelukast ketone, and cis-isomer. High performance liquid chromatography requires the use of a high performance liquid chromatograph for detection operations, and different chromatographic conditions need to be set when operating the high performance liquid chromatograph to detect different substances in order to accurately detect the amount of the substance to be tested.

[0059] The present invention uses phenyl bonded silica gel as a filler, uses different mobile phases, different elution methods, and different detection wavelengths for detection of different detection substances, and can accurately and sensitively detect the amounts of montelukast, sulfoxide, montelukast ketone, and cis-isomer in montelukast sodium chewable tablets.

[0060] (1) The content test includes the following chromatographic conditions:

[0061] Phenyl bonded silica gel is used as the filler (Agilent ZORBAX SB-Phenyl 4.6mm×250mm, 3.5μm or a chromatographic column with equivalent performance); a mixed solution of 0.2% trifluoroacetic acid aqueous solution-0.2% trifluoroacetic acid acetonitrile solution is used as the mobile phase, and isocratic elution is performed; the flow rate is 1.5ml / min; the column temperature is 50℃; and the detection wavelength is 358nm.

[0062] The volume ratio of 0.2% trifluoroacetic acid aqueous solution to 0.2% trifluoroacetic acid acetonitrile solution in the mobile phase is 3:1.

[0063] Specific steps for content detection:

[0064] Preparation before operation: Take 500 ml of methanol, add 100 ml of water and mix well for later use to obtain a methanol-water solution.

[0065] A1. Test solution: Take 10 montelukast sodium chewable tablets, place them in a 200ml brown volumetric flask, add 40ml water, let stand for 5 minutes or until the tablets are completely disintegrated, add 120ml methanol, ultrasonically treat for 20 minutes, and shake, dilute to the scale with methanol-water solution, shake well, filter, accurately measure 2ml of the filtrate, place it in a 20ml brown volumetric flask, dissolve and dilute to the scale with methanol-water solution, shake well, and the test solution is obtained.

[0066] A2. Reference substance solution: accurately weigh 17 mg of montelukast dicyclohexylamine reference substance, place in a 50 ml brown volumetric flask, add an appropriate amount of methanol-water solution, sonicate to dissolve and dilute to the scale, shake well; accurately measure 2 ml, place in a 20 ml brown volumetric flask, dilute to the scale with methanol-water solution, shake well to obtain the reference substance solution.

[0067] A3. Content determination: Take 10 μl of reference solution and test solution respectively, inject them into liquid chromatograph and record the chromatogram.

[0068] The specification of the above-mentioned montelukast sodium chewable tablet is 5 mg (calculated as montelukast);

[0069] The volume ratio of methanol to water in the above methanol-water solution is 5:1;

[0070] The filter is made of Nylon, with a specification of 32mm and 0.45um.

[0071] Content calculation formula:

[0072]

[0073] Where:

[0074] W S The sample weight of montelukast dicyclohexylamine reference substance, mg;

[0075] AT is the peak area of ​​the main peak in the chromatogram of the test solution;

[0076] AS is the peak area of ​​the main peak in the chromatogram of the reference solution;

[0077] DT is the dilution factor of the test solution;

[0078] DS is the dilution factor of the reference solution;

[0079] n is the number of montelukast sodium chewable tablets sampled in the test solution;

[0080] L is the labeled amount;

[0081] 0.7638 is the ratio of montelukast (M: 586.18) to montelukast dicyclohexylamine (M: 767.50).

[0082] Contains montelukast sodium (C 35 H 35 ClNNaO3S) with montelukast (C 35 H 35 ClNNaO3S) should be 96.0% to 104.0% of the labeled amount.

[0083] (2) The relevant substance detection includes the following chromatographic conditions:

[0084] Phenyl bonded silica gel was used as the filler (Agilent ZORBAX SB-Phenyl 4.6×250mm, 3.5μm or a chromatographic column with equivalent performance); 0.2% trifluoroacetic acid aqueous solution was used as the mobile phase A; methanol-acetonitrile was used as the mobile phase B; linear gradient elution was performed, the column temperature was 50°C; the detection wavelength was 250nm; and the flow rate was 1.0ml / min.

[0085] The volume ratio of methanol to acetonitrile in mobile phase B was 85:15.

[0086] Gradient elution program: 0 min, 63% A, 37% B; 10 min, 43% A, 57% B; 15 min, 63% A, 37% B; 25 min, 63% A, 37% B; 35 min, 70% A, 30% B; 53 min, 70% A, 30% B; 55 min, 40% A, 60% B; 60 min, 40% A, 60% B.

[0087] Specific steps for substance testing:

[0088] C1. Reference substance mixed solution: Take 2.5 mg of montelukast peak identification impurity reference substance, accurately weigh it, place it in a 10 ml brown volumetric bottle, add 5 ml of methanol-water, then add 1 ml of the above montelukast ketone impurity reference substance solution, dissolve it by ultrasonic, dilute it to the scale with methanol-water, shake it well, and use it as the mixed impurity reference solution. Take 5 ml of the mixed impurity reference solution, place it in a 10 ml colorless volumetric bottle, and place it under natural light for 1 hour.

[0089] C2. Solution of test sample 3: Take 10 montelukast sodium chewable tablets, place in a 200ml brown volumetric flask, add 40ml of water, leave for 5 minutes or until the tablets are completely disintegrated, add 120ml of methanol, ultrasonically treat for 20 minutes, and shake, dilute to the scale with methanol-water, shake well, filter, discard at least 1ml of the initial filtrate and take the subsequent filtrate, accurately measure 1ml, place in a 100ml brown volumetric flask, dilute to the scale with methanol-water, shake well, and obtain.

[0090] C3. Sensitivity solution: Accurately measure 5 ml of the reference substance mixed solution, place it in a 100 ml brown volumetric flask, dilute to the scale with methanol-water, shake well, and obtain.

[0091] C4. Determination of related substances: Take 20 μl each of the sensitivity solution, test sample 3 solution and reference sample mixed solution, inject them into the liquid chromatograph respectively, and record the chromatogram.

[0092] Calculation formula for relevant substances:

[0093]

[0094] Where:

[0095] AT1 is the peak area of ​​sulfoxide in the chromatogram of test sample 3 solution;

[0096] AT2 is the peak area of ​​montelukast ketone in the chromatogram of test sample 3 solution;

[0097] AT3 is the peak area of ​​the cis isomer in the chromatogram of the solution of test sample 3;

[0098] AT4 is the peak area of ​​the other single largest impurity peak in the chromatogram of the sample 3 solution;

[0099] The total impurities of AT are the sum of the peak areas of each impurity in the chromatogram of the solution of test sample 3 (corrected peak area; chromatographic peaks consistent with the impurity peaks of montelukast methyl ketone and methylstyrene are not included in the related substances of this product; peaks smaller than the main peak area of ​​the sensitivity solution are ignored);

[0100] AS 1％ The peak area of ​​the main peak in the chromatogram of the reference mixed solution;

[0101] 0.6 is the correction factor for the montelukast ketone impurity.

[0102] Result determination: If there are impurity peaks in the chromatogram of the test sample 3 solution (except for the solvent peak and excipient peak before 0.15 times the retention time of the main peak), and the impurity peaks have the same retention time as the montelukast methyl ketone peak and methylstyrene peak, they are not included in the related substances of this product. These impurities are impurities controlled by the montelukast sodium raw material and are not potential degradation products of the tablets. The peak area of ​​the chromatographic peak consistent with the retention time of sulfoxide shall not be greater than 0.5 times (0.5%) the main peak area of ​​the reference mixed solution, the peak area of ​​the chromatographic peak consistent with the retention time of the cis isomer shall not be greater than 0.1 times (0.1%) the main peak area of ​​the reference mixed solution, the peak area of ​​the chromatographic peak consistent with the retention time of montelukast ketone shall not be greater than 0.1 times (0.1%) the main peak area of ​​the reference mixed solution after multiplying by the correction factor 0.6, the peak area of ​​other individual impurities shall not be greater than 0.1 times (0.1%) the main peak area of ​​the reference mixed solution, and the sum of the peak areas of each impurity (calculated according to the corrected peak area) shall not be greater than 0.8 times (0.8%) the main peak area of ​​the reference mixed solution. Peaks in the chromatogram of the test sample 3 solution that are smaller than the main peak area of ​​the sensitivity solution are ignored.

[0103] Example 1

[0104] 1. Preparation before the experiment

[0105] Prepare the instruments, equipment, reagents, consumables, samples and reference materials needed for the experiment, as follows:

[0106] Table 1: Instrument information

[0107] Instruments and Equipment Device Model Equipment manufacturers Chromatography University Liquid Chromatograph 1260 Series Agilent Chromatographic columns Agilent ZORBAX SB-Phenyl 4.6×250mm, 3.5μm Agilent Analytical Balance / /

[0108] Table 2: Reagent and consumables information

[0109]

[0110]

[0111] Table 3: Sample and control information

[0112] name supplier batch number Content / concentration Montelukast Sodium Chewable Tablets Hainan Jinrui Pharmaceutical Co., Ltd. 230503 N / A Montelukast TLC 1909-040A4 97.2％ Montelukast ketone TLL 3616-094A4 99.8％ Montelukast Dicyclohexylamine EP 4.0 100％

[0113] Note: N / A means “not applicable”.

[0114] 2. Experimental testing

[0115] The quality test of Montelukast Sodium Chewable Tablets is carried out twice. The first test is to test the content of the main component and the content of montelukast sodium in the Montelukast Sodium Chewable Tablets. The second test is to test the related substances and the amount of sulfoxide, montelukast ketone, cis-isomer impurities, and other single impurities and total impurities in the Montelukast Sodium Chewable Tablets, so as to achieve the purpose of monitoring and controlling the product quality of Montelukast Sodium Chewable Tablets.

[0116] 2.1 Content detection

[0117] Table 4: Instrumental testing parameter information for content testing

[0118]

[0119] Specific steps for content detection:

[0120] Preparation of methanol-water solution: Take 500 ml of methanol, add 100 ml of water and mix well for later use to obtain the methanol-water solution.

[0121] A1. Test solution: Take 10 montelukast sodium chewable tablets, place in a 200ml brown volumetric flask, add 40ml water, let stand for 5 minutes or until the tablets are completely disintegrated, add 120ml methanol, ultrasonically treat for 20 minutes, and shake, dilute to the scale with methanol-water solution, shake well, filter (filter material is Nylon, specification 32mm, 0.45um), accurately measure 2ml of the filtrate, place in a 20ml brown volumetric flask, dissolve and dilute to the scale with methanol-water solution, shake well, and obtain.

[0122] A2. Reference substance solution: accurately weigh 17 mg of montelukast dicyclohexylamine reference substance, place in a 50 ml brown volumetric flask, add an appropriate amount of methanol-water solution, sonicate to dissolve and dilute to the scale, shake well; accurately measure 2 ml, place in a 20 ml brown volumetric flask, dilute to the scale with methanol-water solution, shake well, and obtain.

[0123] A3. Content determination: Take 10 μl of reference solution and test solution respectively, inject them into liquid chromatograph and record the chromatogram.

[0124] Content calculation formula:

[0125]

[0126] Where:

[0127] W S The sample weight of montelukast dicyclohexylamine reference substance, mg;

[0128] AT is the peak area of ​​the main peak in the chromatogram of the test solution;

[0129] AS is the peak area of ​​the main peak in the chromatogram of the reference solution;

[0130] DT is the dilution factor of the test solution;

[0131] DS is the dilution factor of the reference solution;

[0132] n is the number of montelukast sodium chewable tablets sampled in the test solution;

[0133] L is the labeled amount;

[0134] 0.7638 is the ratio of montelukast (M: 586.18) to montelukast dicyclohexylamine (M: 767.50).

[0135] Contains montelukast sodium (C 35 H 35 ClNNaO3S) with montelukast (C 35 H 35 ClNNaO3S) should be 96.0% to 104.0% of the labeled amount.

[0136] Content test results: According to Figure 1 and Figure 2 The calculated content of montelukast in montelukast sodium chewable tablets is 100.7%, the content value is within the prescribed range, and the content control complies with the regulations.

[0137] 2.2 Related substance detection

[0138] Table 5: Information on instrument testing parameters for substance testing

[0139]

[0140] Specific steps for substance testing:

[0141] C1. Mixed reference solution: Take 2.5 mg of montelukast peak identification impurity reference, weigh accurately, place in a 10 ml brown volumetric flask, add 5 ml of methanol-water solution, then add 1 ml of montelukast ketone impurity reference solution, dissolve by ultrasonication, dilute to scale with methanol-water solution, shake well, and use as mixed impurity reference solution. Take 5 ml of mixed impurity reference solution, place in a 10 ml colorless volumetric flask, and place under natural light for 1 hour.

[0142] C2. Solution of test sample 3: Take 10 montelukast sodium chewable tablets, place in a 200ml brown volumetric flask, add 40ml water, leave for 5 minutes or until the tablets are completely disintegrated, add 120ml methanol, ultrasonically treat for 20 minutes, shake, dilute to the scale with methanol-water solution, shake well, filter, discard at least 1ml of the initial filtrate and take the subsequent filtrate; accurately measure 1ml of the test sample solution, place in a 100ml brown volumetric flask, dilute to the scale with methanol-water solution, shake well, and obtain.

[0143] C3. Sensitivity solution: Accurately measure 5 ml of the reference substance mixed solution, place it in a 100 ml brown volumetric flask, dilute to the scale with methanol-water solution, shake well, and obtain.

[0144] C4. Determination of related substances: Take 20 μl each of the sensitivity solution, test sample 3 solution and reference sample mixed solution, inject them into the liquid chromatograph respectively, and record the chromatogram.

[0145] Calculation formula for relevant substances:

[0146]

[0147] Where:

[0148] AT1 is the peak area of ​​sulfoxide in the chromatogram of test sample 3 solution;

[0149] AT2 is the peak area of ​​montelukast ketone in the chromatogram of test sample 3 solution;

[0150] AT3 is the peak area of ​​the cis isomer in the chromatogram of the solution of test sample 3;

[0151] AT4 is the peak area of ​​the other single largest impurity peak in the chromatogram of the sample 3 solution;

[0152] The total impurities of AT are the sum of the peak areas of each impurity in the chromatogram of the solution of test sample 3 (corrected peak area; chromatographic peaks consistent with the impurity peaks of montelukast methyl ketone and methylstyrene are not included in the related substances of this product; peaks smaller than the main peak area of ​​the sensitivity solution are ignored);

[0153] AS 1％ The peak area of ​​the main peak in the chromatogram of the reference mixed solution;

[0154] 0.6 is the correction factor for the montelukast ketone impurity.

[0155] Related substance test results: According to Figure 3 and Figure 4 The sulfoxide content of montelukast sodium chewable tablets was calculated to be 0.1%, the cis-isomer was not detected, the montelukast ketone was not detected, and the total impurities were 0.1%. The amounts of each impurity were lower than the standard value, and the control of relevant substances complied with the regulations.

[0156] 3. Method Validation

[0157] 3.1 System adaptability

[0158] Acceptance criteria: (1) The system is clean and stable, with no interference at the peak position of each component. If there is interference, the peak area of ​​the interfering peak should not be greater than the peak area of ​​the main peak in the sensitivity solution. (2) In the sensitivity solution, the signal-to-noise ratio (S / N) of the target peak should not be less than 10. (3) The RSD of the peak area of ​​5 consecutive target peaks should not be greater than 10.0%, and the RSD of the retention time should not be greater than 1.0%.

[0159] Prepare a 100% limit reference solution and inject it continuously for 5 times, use a 20% limit reference solution as the sensitivity solution, and use the solvent as the blank solution, and then test it on the instrument.

[0160] The results show that the system is clean and stable, there is no obvious interference at the peak position of each target peak, the signal-to-noise ratio of the sensitivity solution test result is not less than 10, the RSD of the peak area of ​​5 consecutive target peaks is not more than 10.0%, and the RSD of the retention time is not more than 1.0%. The system adaptability meets the requirements, and the specific results are shown in the following table:

[0161] Table 6: System suitability test results 1

[0162]

[0163] Table 7: System suitability test results 2

[0164]

[0165]

[0166] Note: The concentrations in the table (STD ug / mL) are the concentrations of the actual prepared injection solutions.

[0167] 3.2 Exclusivity

[0168] Acceptance criteria: (1) There is no interference at the target peak in the blank solution chromatogram. If there is interference, the peak area of ​​the interference peak should not be greater than the main peak area in the sensitivity solution. (2) The target peak is shown in the 100% limit reference solution. (3) The target peak is shown in the 100% limit spiked sample solution, and the resolution of all adjacent peaks greater than the sensitivity solution should be no less than 1.5 with the target peak.

[0169] Prepare 100% limit reference solution and 100% limit spiked sample solution, that is, add 100% limit reference solution to sample solution and mix well to obtain 100% limit spiked sample solution, use solvent as blank solution, and test on the machine. The results are shown in Table 8.

[0170] The results showed that: (1) There was no interference in the blank solution of montelukast sodium, sulfoxide, montelukast ketone, and cis-isomer. (2) The 100% limit reference solution showed the target peaks of montelukast sodium RT = 15.737min, sulfoxide RT = 6.899min, montelukast ketone = 12.249min, and cis-isomer = 14.422min. (3) The target peaks were shown in the spiked sample solutions of montelukast sodium, sulfoxide, and montelukast ketone at the 100% limit, and there were no adjacent peaks; the target peaks were shown in the spiked sample solutions at the cis-isomer limit, and there were adjacent peaks. The separation between the adjacent peaks and the target peak was greater than 1.5, which met the requirements. The specificity test met the requirements, and this method is specific.

[0171] Table 8: Specificity test results

[0172]

[0173]

[0174] 3.3 Detection limit

[0175] Acceptance criteria: The signal-to-noise ratio (S / N) of each injection of LOD solution is not less than 3.

[0176] Prepare 1 portion of 10% limit reference solution as LOD solution (refer to the preparation method of sensitivity solution), and test it on the machine, injecting 3 times. The results show that the signal-to-noise ratio S / N of the test results is greater than 3. The detection limit test meets the requirements, and the specific results are shown in the following table, the concentration of the test compound and the relative content in the sample:

[0177] Table 9: Concentration of the tested compounds and their relative content in the samples

[0178] name Concentration (ug / mL) Relative content in sample (ppm) Montelukast Sodium 2.5 0.3804ng Sulfoxide 15 0.0038ng Montelukast ketone 2.5 0.1509ng Cis isomer 2.5 0.3745ng

[0179] Note: The concentration is the concentration of the actual prepared injection solution.

[0180] The signal-to-noise ratio (S / N) results of the test results are shown in Table 10:

[0181] Table 10: Test results of signal-to-noise ratio (S / N) data

[0182] name Montelukast Sodium Sulfoxide Montelukast ketone Cis isomer LOD-1 3.2 4.2 4.0 5.6 LOD-2 3.5 4.7 4.6 5.2 LOD-3 3.0 4.3 4.2 5.5

[0183] 3.4 Limit of quantification

[0184] Acceptance criteria: The signal-to-noise ratio (S / N) of each injection of LOQ solution is ≥10; the RSD of the peak area of ​​the target peak in 6 consecutive injections of LOQ is not greater than 15.0%.

[0185] Prepare 1 portion of 20% limit reference solution as LOQ solution (refer to the preparation method of sensitivity solution), and test it on the machine, injecting 6 times. The results show that the signal-to-noise ratio of the test results is greater than 10; the RSD of the peak area of ​​the target peak in 6 consecutive injections of LOQ is not greater than 15.0%. The quantitative limit test meets the requirements, and the verification results are as follows in Table 11:

[0186] Table 11: Concentration of the tested compounds and their relative content in the samples

[0187]

[0188]

[0189] Note: The concentrations in Table 11 are the concentrations of the actual prepared injection solutions.

[0190] The quantitative limit results are shown in Table 12:

[0191] Table 12: Data of target peak areas

[0192]

[0193] 3.5 Linearity and range

[0194] Acceptance criteria: regression coefficient (r) ≥ 0.990.

[0195] A series of linear test solutions (20%, 50%, 100%, 150%, 200%) from LOQ to 200% limit concentration were prepared, and each solution was sampled and analyzed once. The equation and regression coefficient (r) were calculated. The results showed that the regression coefficient (r) was ≥ 0.990. The linear results met the requirements, and the verification results are shown in Table 13 below:

[0196] Table 13: Linear data information

[0197]

[0198]

[0199] Note: The concentrations in Table 13 are the concentrations of the actual prepared injection solutions.

[0200] 5.12 Conclusion

[0201] Based on the above tests, it can be seen that: the system adaptability meets the requirements, so the system used in the method of the present invention is effective and applicable; the specificity test meets the requirements, indicating that the method of the present invention can accurately and selectively analyze and detect residual substances; the detection limit test meets the requirements, indicating that the detection limit of the method of the present invention is reasonably set and the detection method has high sensitivity; the quantitative limit test meets the requirements, indicating that the method of the present invention can accurately determine the residual amount of residual substances and has sensitive quantitative detection capabilities.

[0202] All of the above verification items meet the requirements, and the method of the present invention can be used to detect the amounts of montelukast sodium, sulfoxide, montelukast ketone, and cis-isomer in montelukast sodium chewable tablets.

[0203] Comparative Example 1

[0204] Take 1 ml of each of the reference solutions of montelukast sodium, sulfoxide, montelukast ketone and cis-isomer in Example 1 and perform determination and analysis by high performance liquid chromatography. The detection conditions are as follows:

[0205] Chromatographic column: phenyl bonded silica gel;

[0206] Mobile phase A: 0.1% formic acid in water;

[0207] Mobile phase B: 0.1% formic acid in acetonitrile;

[0208] Flow rate: 1.0ml / min;

[0209] Injection volume: 10 μl;

[0210] Detection wavelength: 250nm;

[0211] Gradient elution program: 0 min, 63% A, 37% B; 10 min, 43% A, 57% B; 15 min, 63% A, 37% B; 25 min, 63% A, 37% B; 35 min, 70% A, 30% B; 53 min, 70% A, 30% B; 55 min, 40% A, 60% B; 60 min, 40% A, 60% B.

[0212] Conclusion: Comparative Example 1 is a comparative test of Example 1. Comparative Example 1 and Example 1 both use high performance liquid chromatography for detection. The difference is that the mobile phase materials and mobile phase combinations in the chromatographic detection conditions for the detection of related substances are different. Result of Comparative Example 1: The chromatogram of the substance to be tested could not be detected, and the chromatographic detection conditions were not applicable. Comparison of the mobile phase materials and mobile phase combinations used in Example 1 with the test results of Comparative Example 1 shows that: in the same detection method, different mobile phase materials and mobile phase combinations are used, and the compounds that can be detected are different, and the substance to be tested cannot be detected. In contrast, the mobile phase materials and mobile phase combinations used in Example 1 can detect the amount of montelukast sodium, sulfoxide, montelukast ketone, and cis-isomer in montelukast sodium chewable tablets, with high sensitivity, baseline separation, and good resolution.

[0213] Comparative Example 2

[0214] Take 1 mL of each of the reference solutions of montelukast sodium, sulfoxide, montelukast ketone, and cis-isomer in Example 1, and perform determination and analysis by high performance liquid chromatography. The detection conditions are:

[0215] Chromatographic column: phenyl bonded silica gel;

[0216] Mobile phase A: 0.2% trifluoroacetic acid in water;

[0217] Mobile phase B: methanol-acetonitrile (volume ratio 85:15);

[0218] Flow rate: 1.0ml / min;

[0219] Injection volume: 10 μl;

[0220] Detection wavelength: 250nm;

[0221] Gradient elution program: 0 min, 65% A, 35% B; 10 min, 75% A, 25% B; 30 min, 85% A, 15% B; 60 min, 75% A, 25% B; 70 min, 65%, 35% B.

[0222] Conclusion: Comparative Example 2 is a comparative test of Example 1. Comparative Example 2 and Example 1 both use high performance liquid chromatography for detection, the difference being that the gradient elution procedures in the chromatographic detection conditions for the detection of the relevant substances are different. Detection result of Comparative Example 2: Not detected. By comparing the detection results of Comparative Example 2 with those of Example 1, it can be seen that: in the same detection method, by changing the gradient elution procedure, the substances detected are significantly different. In comparison, the gradient elution procedure used in Example 1 can better separate montelukast sodium, sulfoxide, montelukast ketone, and cis-isomer substances in montelukast sodium chewable tablets, accurately detect the substances to be tested, obtain the corresponding chromatograms, and have baseline separation and good separation degree.

[0223] Comparative Example 3

[0224] Take 1 mL of each of the reference solutions of montelukast sodium, sulfoxide, montelukast ketone, and cis-isomer in Example 1, and perform determination and analysis by high performance liquid chromatography. The detection conditions are:

[0225] Chromatographic column: phenyl bonded silica gel;

[0226] Mobile phase A: 0.2% trifluoroacetic acid in water;

[0227] Mobile phase B: methanol-acetonitrile (volume ratio 85:15);

[0228] Flow rate: 1.0ml / min;

[0229] Injection volume: 10 μl;

[0230] Detection wavelength: 330nm;

[0231] Gradient elution program: 0 min, 63% A, 37% B; 10 min, 43% A, 57% B; 15 min, 63% A, 37% B; 25 min, 63% A, 37% B; 35 min, 70% A, 30% B; 53 min, 70% A, 30% B; 55 min, 40% A, 60% B; 60 min, 40% A, 60% B.

[0232] Conclusion: Comparative Example 3 is a comparative test of Example 1. Comparative Example 3 and Example 1 both use high performance liquid chromatography for detection, the difference being that the detection wavelengths in the chromatographic detection conditions for the detection of the relevant substances are different. Detection result of Comparative Example 3: Not detected. According to the comparison of the detection results of Comparative Example 3 with those of Example 1, it can be seen that different detection wavelengths can detect the corresponding substances within the wavelength range, while substances outside the wavelength cannot be detected. In contrast, Example 1 accurately detects montelukast sodium, sulfoxide, montelukast ketone, and cis-isomer substances in montelukast sodium chewable tablets by setting the corresponding detection wavelength, avoiding interference from substances outside the detection wavelength, thereby obtaining the corresponding chromatogram with baseline separation and good separation.

[0233] Comparative Example 4

[0234] Take 1 mL of each of the reference solutions of montelukast sodium, sulfoxide, montelukast ketone, and cis-isomer in Example 1, and perform determination and analysis by high performance liquid chromatography. The detection conditions are:

[0235] Chromatographic column: phenyl bonded silica gel;

[0236] Mobile phase A: 1.0% trifluoroacetic acid in water;

[0237] Mobile phase B: methanol-acetonitrile (volume ratio 50:50);

[0238] Flow rate: 1.0ml / min;

[0239] Injection volume: 10 μl;

[0240] Detection wavelength: 250nm;

[0241] Gradient elution program: 0 min, 63% A, 37% B; 10 min, 43% A, 57% B; 15 min, 63% A, 37% B; 25 min, 63% A, 37% B; 35 min, 70% A, 30% B; 53 min, 70% A, 30% B; 55 min, 40% A, 60% B; 60 min, 40% A, 60% B.

[0242] Conclusion: Comparative Example 4 is a comparative test of Example 1. Comparative Example 4 and Example 1 both use high performance liquid chromatography for detection, the difference being that the concentration of the mobile phase substance is different. The detection result of Comparative Example 4: interference is detected in the chromatogram. According to the comparison of the detection result of Comparative Example 4 with that of Example 1, it can be seen that different mobile phase concentrations result in different elution abilities during the elution process, and the substance to be tested cannot be completely eluted. Elution interference will also occur in the substance to be tested, resulting in interference in the final detected chromatogram, and the chromatogram of the substance to be tested cannot be accurately detected. In contrast, Example 1 sets a suitable mobile phase and substance concentration to match, and the chromatogram of the substance to be tested accurately detected does not have interference, and the obtained chromatogram has baseline separation, good separation, and high accuracy.

[0243] In summary, the difference between the same chromatographic column and the combination of mobile phase and elution mode is that the same mobile phase combined with different elution modes can detect different substances; the same elution mode combined with different mobile phases can also detect different substances. Therefore, for different detection substances, only a specific mobile phase and elution mode can be used in combination, and the appropriate detection wavelength and concentration ratio of the mobile phase substance can be adjusted to accurately detect the specified substance.

[0244] From the above comparative examples 1-4, it can be seen that:

[0245] First, in the same detection method, different compounds can be detected by changing the mobile phase material and the mobile phase combination. The mobile phase material and the mobile phase combination used in the present invention can better separate the montelukast sodium, sulfoxide, montelukast ketone, and cis-isomer in the montelukast sodium chewable tablets, detect the amount of the substance, and thus obtain the corresponding chromatogram with high accuracy.

[0246] Second, in the same detection method, by changing the gradient elution program, different compounds can be detected, and interference in the detection can be avoided by elution. The gradient elution method and elution program adopted in the present invention can better separate montelukast sodium, sulfoxide, montelukast ketone, and cis isomers in montelukast sodium chewable tablets, avoid interference, accurately detect the amount of the substance, thereby obtaining the corresponding chromatogram, baseline separation, and good separation.

[0247] Third, in the same detection method, by changing the detection wavelength, the corresponding substances can be detected within different wavelength ranges, while substances outside the wavelength cannot be detected. The present invention accurately detects montelukast sodium, sulfoxide, montelukast ketone, and cis-isomer substances in montelukast sodium chewable tablets by setting the corresponding detection wavelength, avoiding interference from substances outside the detection wavelength, thereby obtaining the corresponding chromatogram with baseline separation and good separation.

[0248] Fourth, in the same detection method, the substance concentration ratio of the mobile phase is changed. Different substance concentrations have different elution abilities during the elution process. During the detection process, the elution results of substances with different concentrations will be different, leading to interference problems. The present invention sets the corresponding mobile phase concentration ratio to completely elute the substance to be tested, accurately detect montelukast sodium, sulfoxide, montelukast ketone, and cis-isomer substances in montelukast sodium chewable tablets, avoid interference in the elution process, and thereby obtain the corresponding chromatogram with baseline separation, good separation, and high accuracy.

[0249] In summary, this example confirms that the use of high performance liquid chromatography, by combining the mobile phase and the elution mode in the limited chromatographic conditions, and adjusting the appropriate detection wavelength and the substance concentration ratio of the mobile phase, can accurately detect the content and related substances in the montelukast sodium chewable tablets, that is, accurately detect the amount of montelukast sodium, sulfoxide, montelukast ketone, and cis-isomer, and the detection method has high sensitivity and good separation.

[0250] The embodiments described above are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, a person skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. Such modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention, and all of them belong to the protection scope of the present invention.

Claims

1. A quality detection method for montelukast sodium chewable tablets, characterized in that: The quality inspection includes content inspection and related substance inspection, and the quality inspection is carried out by high performance liquid chromatography; Chemical substances tested in the content test include: montelukast sodium, and chemical substances tested in the related substance test include: sulfoxide, montelukast ketone, cis isomer; The content detection includes the following chromatographic conditions: phenyl bonded silica gel is used as filler; a mixed solution of 0.2% trifluoroacetic acid aqueous solution-0.2% trifluoroacetic acid acetonitrile solution is used as mobile phase, and isocratic elution is performed; the flow rate is 1.5 ml / min; the column temperature is 50°C; the detection wavelength is 358 nm; The detection of related substances includes the following chromatographic conditions: phenyl bonded silica gel as filler; 0.2% trifluoroacetic acid aqueous solution as mobile phase A; methanol-acetonitrile as mobile phase B; linear gradient elution, column temperature 50°C; detection wavelength 250nm; flow rate 1.0ml / min.

2. The quality detection method of montelukast sodium chewable tablets according to claim 1, characterized in that: The volume ratio of 0.2% trifluoroacetic acid aqueous solution to 0.2% trifluoroacetic acid acetonitrile solution in the mobile phase solution was 3:

1.

3. The quality detection method of montelukast sodium chewable tablets according to claim 1, characterized in that: The specific operation of content detection includes the following steps: A1. Test solution: Take 10 montelukast sodium chewable tablets, place in a 200ml brown volumetric bottle, add 40ml water, leave for 5 minutes or until the tablets are completely disintegrated, add 120ml methanol, ultrasonically treat for 20 minutes, shake, dilute to scale with methanol-water, shake well, filter, accurately measure 2ml of the filtrate, place in a 20ml brown volumetric bottle, dissolve and dilute to scale with methanol-water, shake well, and obtain; A2. Reference substance solution: Take 17 mg of montelukast dicyclohexylamine reference substance, accurately weigh it, place it in a 50 ml brown volumetric flask, add an appropriate amount of methanol-water, ultrasonically dissolve it and dilute it to the scale, shake well; accurately measure 2 ml, place it in a 20 ml brown volumetric flask, dilute it to the scale with methanol-water, shake well, and obtain; A3. Content determination: Take 10 μl of reference solution and test solution respectively, inject them into liquid chromatograph and record the chromatogram.

4. The quality detection method of montelukast sodium chewable tablets according to claim 3, characterized in that: In step A3, the content is calculated by the formula: Where: W S The sample weight of montelukast dicyclohexylamine reference substance, mg; AT is the peak area of ​​the main peak in the chromatogram of the test solution; AS is the peak area of ​​the main peak in the chromatogram of the reference solution; DT is the dilution factor of the test solution; DS is the dilution factor of the reference solution; n is the number of montelukast sodium chewable tablets sampled in the test solution; L is the labeled amount; 0.7638 is the ratio of montelukast (M: 586.18) to montelukast dicyclohexylamine (M: 767.50).

5. The quality detection method of montelukast sodium chewable tablets according to claim 3 or 4, characterized in that: Content limit: Contains montelukast sodium (C 35 H 35 ClNNaO3S) with montelukast (C 35 H 35 ClNNaO3S) should be 96.0% to 104.0% of the labeled amount.

6. The quality detection method of montelukast sodium chewable tablets according to claim 1, characterized in that: The volume ratio of methanol:acetonitrile in mobile phase B was 85:

15.

7. The quality detection method of montelukast sodium chewable tablets according to claim 1, characterized in that: Gradient elution program: 0 min, 63% A, 37% B; 10 min, 43% A, 57% B; 15min, 63%A, 37%B; 25min, 63%A, 37%B; 35min, 70%A, 30%B; 53min, 70%A, 30%B; 55min, 40%A, 60%B; 60min, 40%A, 60%B.

8. The quality inspection method for montelukast sodium chewable tablets according to claim 1, characterized in that: The specific operation of substance detection includes the following steps: C1. Mixed reference solution: Take 2.5 mg of montelukast peak identification impurity reference, accurately weigh it, put it in a 10 ml brown volumetric flask, add 5 ml of methanol-water, then add 1 ml of montelukast ketone impurity reference solution, dissolve it by ultrasonic, dilute it to the mark with methanol-water, shake well, and use it as the mixed impurity reference solution; take 5 ml of the mixed impurity reference solution, put it in a 10 ml colorless volumetric flask, and place it under natural light for 1 hour to obtain; C2. Test sample 3 solution: Take 10 montelukast sodium chewable tablets, place in a 200ml brown volumetric flask, add 40ml water, leave for 5 minutes or until the tablets are completely disintegrated, add 120ml methanol, ultrasonically treat for 20 minutes, shake, dilute to scale with methanol-water, shake well, filter, discard at least 1ml of the initial filtrate, take the subsequent filtrate, accurately measure 1ml, place in a 100ml brown volumetric flask, dilute to scale with methanol-water, shake well, and obtain; C3. Sensitivity solution: accurately measure 5 ml of the reference substance mixed solution, place it in a 100 ml brown volumetric flask, dilute to the mark with methanol-water, shake well, and obtain; C4. Determination of related substances: Take 20 μl each of the sensitivity solution, test sample 3 solution and reference sample mixed solution, inject them into the liquid chromatograph respectively, and record the chromatogram.

9. The quality detection method of montelukast sodium chewable tablets according to claim 8, characterized in that: In step C4, the relevant substances are calculated using the formula: Where: AT1 is the peak area of ​​sulfoxide in the chromatogram of test sample 3 solution; AT2 is the peak area of ​​montelukast ketone in the chromatogram of test sample 3 solution; AT3 is the peak area of ​​the cis isomer in the chromatogram of the solution of test sample 3; AT4 is the peak area of ​​the other single largest impurity peak in the chromatogram of the sample 3 solution; The total impurities of AT are the sum of the peak areas of each impurity in the chromatogram of the solution of test sample 3; AS1% is the peak area of ​​the main peak in the chromatogram of the reference mixed solution; 0.6 is the correction factor for the montelukast ketone impurity.

10. The quality detection method of montelukast sodium chewable tablets according to claim 8 or 9, characterized in that: The percentage content of each relevant substance is calculated according to the formula. The content of sulfoxide, cis-isomer and montelukast ketone shall not exceed 0.1%, 0.1% and 0.1% respectively.

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