Method for determining dissolution rate and content of glucosamine sulfate capsule

Abstract

The invention relates to a method for determining the dissolution rate and content of glucosamine sulfate capsules by a derivatization method, which can effectively evaluate the quality and process level of preparations and ensure the effectiveness of drugs, and adopts the technical scheme that the dissolution rate determination method comprises the following steps: S1, preparing a dissolution solution; s2, preparing a test solution; s3, preparing a reference substance solution; s4, preparing a derivatization reagent; s5, chromatographic condition determination; s6, determining the dissolution rate; the content determination method comprises the following steps: S1, preparing a test solution; s2, preparing a reference substance solution; s3, preparing a derivatization reagent; s4, chromatographic condition setting; s5, content determination; according to the method, the dissolution rate and content determination repeatability is good, absolute deviations of dissolution rate determination results in different laboratories all meet requirements, and experimental results cannot be greatly influenced due to changes of the laboratories.

Description

1. Technical Field

[0001] The present invention relates to the field of pharmaceutical analysis, and particularly to a method for determining the dissolution and content of glucosamine sulfate capsules by derivatization method. 2. Background Art

[0002] For the same drug capsule preparations produced by different manufacturers, or products of the same manufacturer in different batches, even if the content is the same, their dissolution behaviors may be different. The dissolution determination can effectively distinguish these differences, and then reflect the bioavailability of the product in vivo, ensuring the clinical efficacy of the drug. For example, for some capsule preparations of poorly soluble drugs, if the dissolution is poor, it may lead to incomplete absorption of the drug in vivo and affect the therapeutic effect.

[0003] In the production process of capsules, content determination is a key link in quality control, which can be used to monitor the stability and consistency of the production process. At the same time, in the research on the storage and shelf life of drugs, by regularly measuring the content, the stability of the drug under different conditions can be understood, providing a basis for determining the shelf life and storage conditions of the drug. Therefore, it is imperative to invent a method for determining the dissolution and content of glucosamine sulfate capsules. 3. Summary of the Invention

[0004] In view of the above situation, the purpose of the present invention is to provide a method for determining the dissolution and content of glucosamine sulfate capsules, which can effectively evaluate the quality of the preparation and the level of the process, and ensure the effectiveness of the drug.

[0005] One of the technical solutions provided by the present invention is a method for determining the dissolution of glucosamine sulfate capsules, which includes the following steps:

[0006] S1: Prepare the dissolution solution

[0007] Take glucosamine sulfate capsules, put them into the sedimentation basket, use water as the dissolution medium, take samples, and obtain the dissolution solution;

[0008] S2: Prepare the test solution

[0009] Take the dissolution solution in step S1, filter it, and take the subsequent filtrate;

[0010] S3: Prepare the reference solution

[0011] Take glucosamine hydrochloride reference substance, dissolve it in water and dilute it to make a solution containing 0.26 mg per 1 ml;

[0012] S4: Prepare the derivatization reagent

[0013] Take 50 mg of o-phthalaldehyde, dissolve it in 1.25 ml of anhydrous methanol, add 50 μl of 3-mercaptopropionic acid and 11.2 ml of 0.2 mol / L borate buffer solution (take 7.63 g of borax, add about 80 ml of water, heat it slowly to dissolve, let it cool to slightly warm at room temperature, quickly adjust the pH value to 9.5 with 10% sodium hydroxide solution, dilute it to 100 ml with water, store it at room temperature, if crystals precipitate, heat it to dissolve or take the supernatant), mix well, and let the solution stand in the dark for 30 minutes before use (add 10 μl of 3-mercaptopropionic acid every two days to maintain a certain concentration, store it at room temperature in the dark, and use it within two weeks);

[0014] S5: Chromatographic conditions

[0015] Use octadecylsilane-bonded silica gel as the filler (Welch XB-C18, 4.6 mm × 150 mm, 5 μm or a chromatographic column with equivalent efficiency); use acetate buffer solution (take 6.8 g of sodium acetate, dissolve it in 700 ml of water, adjust the pH value to 5.9 with 50% acetic acid solution, add water to 1000 ml) - methanol (75:25) as the mobile phase; the detection wavelength is 344 nm; the flow rate is 1.5 ml per minute; the column temperature is 30 °C;

[0016] S6: Dissolution determination

[0017] Precisely measure 4 μl each of the derivatization solvent and the test solution, 16 μl of 0.2 mol / L borate buffer solution, mix well, derivatize for 1.5 minutes, inject it into the liquid chromatograph, record the chromatogram, and take the reference solution and determine it in the same way.

[0018] Preferably, in S1, the amount of water used is 900 ml, the rotation speed is 50 revolutions per minute, and the sample is taken after 45 minutes.

[0019] Preferably, in S5, the system suitability requirements are as follows: in the chromatogram of the reference solution, the number of theoretical plates calculated based on the main peak of glucosamine hydrochloride (α-glucosamine peak) should be not less than 2000, and the tailing factor should be 0.8 - 1.5.

[0020] Furthermore, in S6, the dissolution calculation formula is as follows:

[0021]

[0022] In the formula: A 供 is the peak area of α-glucosamine in the test solution;

[0023] A 对 is the peak area of α-glucosamine in the reference solution;

[0024] m is the weighed amount of the reference substance, mg;

[0025] V is the dilution factor of the test sample;

[0026] V 对 is the dilution factor of the reference substance;

[0027] P is the content of the reference substance;

[0028] M1 is the molecular weight of glucosamine sulfate, 456.42;

[0029] M2 is twice the molecular weight of glucosamine hydrochloride, 431.26;

[0030] 0.25: the specification of this product, calculated as glucosamine sulfate, g.

[0031] The second technical solution provided by the present invention is a method for determining the content of glucosamine sulfate capsules, comprising the following steps:

[0032] S1: Prepare the test sample solution

[0033] Take the content of glucosamine sulfate capsules, mix well, accurately weigh an amount equivalent to 250 mg of glucosamine sulfate, place it in a volumetric flask, add water, shake vigorously for more than 30 minutes to dissolve, dilute to 100 ml with water, shake well, accurately measure 5 ml of the solution, place it in a volumetric flask, dilute to 50 ml with water, filter, and take the subsequent filtrate;

[0034] S2: Prepare the reference substance solution

[0035] Take the reference substance of glucosamine hydrochloride, accurately weigh it, dissolve and dilute it with water to make a solution containing 0.25 mg per 1 ml;

[0036] S3: Prepare the derivatization reagent

[0037] Take 50 mg of o-phthalaldehyde, dissolve it with 1.25 ml of anhydrous methanol, add 50 μl of 3-mercaptopropionic acid and 11.2 ml of 0.2 mol / L borate buffer solution (take 7.63 g of borax, add about 80 ml of water, slowly heat to dissolve, cool to slightly warm at room temperature, quickly adjust the pH value to 9.5 with 10% sodium hydroxide solution, dilute to 100 ml with water, store at room temperature, if crystals precipitate, heat to dissolve or take the supernatant), mix well, and let the solution stand in the dark for 30 minutes before use (add 10 μl of 3-mercaptopropionic acid every two days to maintain a certain concentration, store at room temperature in the dark, and use within no more than two weeks);

[0038] S4: Chromatographic conditions

[0039] Use octadecylsilane chemically bonded silica gel as the filler (Welch XB-C18, 4.6 mm × 150 mm, 5 μm or a chromatographic column with equivalent efficiency); use acetate buffer solution (take 6.8 g of sodium acetate, add 700 ml of water to dissolve it, adjust the pH value to 5.9 with 50% acetic acid solution, and add water to 1000 ml) - methanol (75:25) as the mobile phase; the detection wavelength is 344 nm; the flow rate is 1.5 ml per minute; the column temperature is 30 °C;

[0040] S5: Assay

[0041] Precisely measure 4 μl each of the derivatization solvent and the test solution, 16 μl of 0.2 mol / L borate buffer solution, mix well, derivatize for 1.5 minutes, inject into the liquid chromatograph, record the chromatogram, and take the reference solution and determine in the same way.

[0042] Preferably, in S4, the system suitability requirements are as follows: in the chromatogram of the reference solution, the number of theoretical plates calculated based on the main peak of glucosamine hydrochloride (α-glucosamine peak) should be not less than 2000, and the tailing factor should be 0.8 - 1.5.

[0043] Preferably, in S5, the limit is: calculate the content of (C6H 14 NO5)2SO4 by the external standard method based on the peak area of α-glucosamine (the molecular weight of glucosamine hydrochloride doubled is 431.26, and the molecular weight of glucosamine sulfate is 456.22).

[0044] Furthermore, in S5, the content calculation formula is as follows:

[0045]

[0046] In the formula: A 供 is the peak area of α-glucosamine in the test solution;

[0047] A 对 is the peak area of α-glucosamine in the reference solution;

[0048] m is the weighed amount of the reference substance, mg;

[0049] W is the weighed amount of the test sample, mg;

[0050] W1: The average filling amount of this product, g;

[0051] V is the dilution factor of the test sample;

[0052] V 对 is the dilution factor of the reference substance;

[0053] P is the content of the reference substance;

[0054] M1 is the molecular weight of glucosamine sulfate, 456.42;

[0055] M2 is twice the molecular weight of glucosamine hydrochloride, 431.26;

[0056] 0.25: The specification of this product, calculated as glucosamine sulfate, g.

[0057] The application of the method for determining the dissolution and content of the glucosamine sulfate capsules described in the present invention in the drug quality standard and quality control of glucosamine sulfate capsules.

[0058] The beneficial technical effects of the present invention:

[0059] 1) In the dissolution determination of the present invention, the blank solvent and the blank excipient solution have no interference with the detection of the main component, and the theoretical plate numbers are all greater than 2000, indicating high specificity. The RSD of the dissolution of the main component in 6 test solution samples is 0.6%, less than 2.0%, indicating good repeatability. The absolute deviations of the dissolution determination results in different laboratories all meet the requirements, and the experimental results will not be greatly affected by the change of laboratories.

[0060] 2) In the content determination of the present invention, the blank solvent, the blank excipient solution, and the mixed impurity solution have no interference with the detection of the main component, and the theoretical plate numbers are all greater than 2000, indicating high specificity. The RSD of the content of the main component in 6 test solution samples is 0.7%, less than 2.0%, indicating good repeatability. The absolute deviations of the content determination results in different laboratories all meet the requirements, and the experimental results will not be greatly affected by the change of laboratories. IV. Description of the Drawings

[0061] Figure 1 It is the HPLC chromatogram of the repeatability of the content determination of the present invention - D-glucosamine hydrochloride reference substance 1.

[0062] Figure 2 It is the HPLC chromatogram of the repeatability of the content determination of the present invention - D-glucosamine hydrochloride reference substance 2.

[0063] Figure 3 It is the HPLC chromatogram of the dissolution determination of the present invention - D-glucosamine hydrochloride reference substance.

[0064] Figure 4 It is the HPLC chromatogram of the dissolution determination of the present invention - test sample.

[0065] Figure 5 It is the HPLC chromatogram of the dissolution specificity of the present invention - D-glucosamine hydrochloride reference substance.

[0066] Figure 6 It is the HPLC chromatogram of the dissolution specificity of the present invention - test sample.

[0067] Figure 7 This is the HPLC chromatogram of the dissolution specificity - blank excipients of the present invention. V. Specific Embodiments

[0068] The following further elaborates on the specific embodiments of the present invention in conjunction with examples.

[0069] Example 1

[0070] A method for determining the dissolution of glucosamine sulfate capsules, comprising the following steps:

[0071] S1: Prepare the dissolution solution

[0072] Take glucosamine sulfate capsules, place them in a sedimentation basket, use 900 ml of water as the dissolution medium, rotate at 50 revolutions per minute, operate according to law, and take a sample after 45 minutes to obtain the dissolution solution;

[0073] S2: Prepare the test solution

[0074] Take the dissolution solution from step S1, filter it, and take the subsequent filtrate;

[0075] S3: Prepare the reference solution

[0076] Take glucosamine hydrochloride reference substance, dissolve it in water and dilute to make a solution containing 0.26 mg per 1 ml;

[0077] S4: Prepare the derivatization reagent

[0078] Take 50 mg of o-phthalaldehyde, dissolve it in 1.25 ml of anhydrous methanol, add 50 μl of 3-mercaptopropionic acid and 11.2 ml of 0.2 mol / L borate buffer solution (take 7.63 g of borax, add about 80 ml of water, slowly heat to dissolve, let it cool to slightly warm at room temperature, quickly adjust the pH value to 9.5 with 10% sodium hydroxide solution, dilute to 100 ml with water, store at room temperature, if crystals precipitate, heat to dissolve or take the supernatant), mix well, and let this solution stand in the dark for 30 minutes before use (add 10 μl of 3-mercaptopropionic acid every two days to maintain a certain concentration, store in the dark at room temperature, and use within no more than two weeks);

[0079] S5: Chromatographic conditions

[0080] Use octadecylsilyl silica gel as the filler (Welch XB-C18, 4.6 mm × 150 mm, 5 μm or a chromatographic column with equivalent efficiency); use acetate buffer solution (take 6.8 g of sodium acetate, add 700 ml of water to dissolve it, adjust the pH value to 5.9 with 50% acetic acid solution, and add water to 1000 ml) - methanol (75:25) as the mobile phase; the detection wavelength is 344 nm; the flow rate is 1.5 ml per minute; the column temperature is 30 °C.

[0081] System suitability requirements: In the chromatogram of the reference solution, the number of theoretical plates calculated based on the main peak of glucosamine hydrochloride (α-glucosamine peak) should not be less than 2000, and the tailing factor should be 0.8 - 1.5;

[0082] S6: Dissolution determination

[0083] Precisely measure 4 μl each of the derivatization solvent and the test solution, 16 μl of 0.2 mol / L borate buffer solution, mix well, derivatize for 1.5 minutes, inject into the liquid chromatograph, record the chromatogram, and take another reference solution and determine in the same way;

[0084] Agilent derivatization procedure

[0085] A. Vial number and name

[0086] Vial P1-A1: Water;

[0087] Vial P1-A2: 0.2 mol / L borate buffer solution;

[0088] Vial P1-A3: OPA (derivatization reagent);

[0089] sample: Test solution.

[0090] B. Autosampler program

[0091]

[0092]

[0093] The dissolution calculation formula is as follows:

[0094]

[0095] In the formula: A 供 is the peak area of α-glucosamine in the test solution;

[0096] A 对 is the peak area of α-glucosamine in the reference solution;

[0097] m is the weighed amount of the reference substance, mg;

[0098] V is the dilution factor of the test sample;

[0099] V 对 is the dilution factor of the reference substance;

[0100] P is the content of the reference substance;

[0101] M1 is the molecular weight of glucosamine sulfate, 456.42;

[0102] M2 is twice the molecular weight of glucosamine hydrochloride, 431.26;

[0103] 0.25: the specification of this product, calculated as glucosamine sulfate, g.

[0104] Example 2

[0105] A method for the determination of the content of glucosamine sulfate capsules, comprising the following steps:

[0106] S1: Prepare the test sample solution

[0107] Take the content of glucosamine sulfate capsules, mix well, accurately weigh an amount equivalent to 250 mg of glucosamine sulfate, place it in a volumetric flask, add water, shake vigorously for more than 30 minutes to dissolve, dilute to 100 ml with water, shake well, accurately measure 5 ml of the solution, place it in a volumetric flask, dilute to 50 ml with water, filter, and take the subsequent filtrate;

[0108] S2: Prepare the reference substance solution

[0109] Take the reference substance of glucosamine hydrochloride, accurately weigh it, dissolve and dilute it with water to make a solution containing 0.25 mg per 1 ml;

[0110] S3: Prepare the derivatization reagent

[0111] Take 50 mg of o-phthalaldehyde, dissolve it in 1.25 ml of anhydrous methanol, add 50 μl of 3-mercaptopropionic acid and 11.2 ml of 0.2 mol / L borate buffer solution (take 7.63 g of borax, add about 80 ml of water, slowly heat to dissolve, let it cool to slightly warm at room temperature, quickly adjust the pH value to 9.5 with 10% sodium hydroxide solution, dilute to 100 ml with water, store at room temperature, if crystals precipitate, heat to dissolve or take the supernatant), mix well, and let this solution stand in the dark for 30 minutes before use (add 10 μl of 3-mercaptopropionic acid every two days to maintain a certain concentration, store at room temperature in the dark, and use within no more than two weeks);

[0112] S4: Chromatographic conditions

[0113] Use octadecylsilane chemically bonded silica gel as the filler (Welch XB-C18, 4.6 mm × 150 mm, 5 μm or a chromatographic column of equivalent efficiency); use acetate buffer solution (weigh 6.8 g of sodium acetate, add 700 ml of water to dissolve it, adjust the pH value to 5.9 with 50% acetic acid solution, and add water to 1000 ml) - methanol (75:25) as the mobile phase; the detection wavelength is 344 nm; the flow rate is 1.5 ml per minute; the column temperature is 30 °C;

[0114] System suitability requirements: In the chromatogram of the reference solution, the number of theoretical plates calculated based on the main peak of glucosamine hydrochloride (α-glucosamine peak) should be not less than 2000, and the tailing factor should be 0.8 - 1.5;

[0115] S5: Content determination

[0116] Precisely measure 4 μl each of the derivatization solvent and the test solution, 16 μl of 0.2 mol / L borate buffer solution, mix well, derivatize for 1.5 minutes, inject into the liquid chromatograph, record the chromatogram, and take another reference solution and determine in the same way;

[0117] Limit: Calculate the content of (C6H 14 NO5)2SO4 by the external standard method based on the peak area of the α-glucosamine peak (the molecular weight of glucosamine hydrochloride doubled is 431.26, and the molecular weight of glucosamine sulfate is 456.22);

[0118] Agilent derivatization procedure

[0119] A. Vial number and name

[0120] Vial P1-A1: Water;

[0121] Vial P1-A2: 0.2 mol / L borate buffer solution;

[0122] Vial P1-A3: OPA (derivatization reagent);

[0123] sample: Test solution.

[0124] B. Autosampler program

[0125]

[0126] The content calculation formula is as follows:

[0127]

[0128] In the formula: A 供 is the peak area of α-glucosamine in the test solution;

[0129] A 对is the peak area of α-glucosamine in the reference solution;

[0130] m is the weighed amount of the reference substance, mg;

[0131] W is the weighed amount of the test sample, mg;

[0132] W1: the average filling quantity of this product, g;

[0133] V is the dilution factor of the test sample;

[0134] V 对 is the dilution factor of the reference substance;

[0135] P is the content of the reference substance;

[0136] M1 is the molecular weight of glucosamine sulfate 456.42;

[0137] M2 is twice the molecular weight of glucosamine hydrochloride 431.26;

[0138] 0.25: the specification of this product, calculated as glucosamine sulfate, g.

[0139] The results and analysis obtained by using the method for determining the dissolution and content of glucosamine sulfate capsules of the present invention are as follows:

[0140] I. Dissolution

[0141] (1) The results of specificity are shown in the following table:

[0142] The HPLC chromatogram of D-glucosamine hydrochloride reference substance is as Figure 5 shown, the HPLC chromatogram of the test sample is as Figure 6 shown, and the HPLC chromatogram of the blank excipient is as Figure 7 shown. The results of the specificity test are shown in the following table:

[0143] Table - 1 Results of specificity test

[0144]

[0145] Conclusion: The blank solvent and the blank excipient solution do not interfere with the detection of the main component, and the theoretical plate numbers are all greater than 2000, indicating high specificity.

[0146] (2) The results of repeatability are shown in the following table:

[0147] The HPLC chromatogram of D-glucosamine hydrochloride reference substance is as Figure 3 shown, the HPLC chromatogram of the test sample is as Figure 4 shown. The results of the repeatability test of the test sample are shown in the following table:

[0148] Table - 2 Results of repeatability test

[0149]

[0150] Conclusion: The RSD of the dissolution of the main component in 6 test solution samples was 0.6%, less than 2.0%, indicating good repeatability of this method.

[0151] (3) Result comparison

[0152] The same batch of samples was determined by different laboratories, and the results are shown in the following table:

[0153] Table - 3 Summary table of comparative test results

[0154]

[0155] Conclusion: It can be seen from the experiment that the absolute deviations of the dissolution determination results of the two laboratories all meet the requirements, and the experimental results will not be greatly affected by the change of the laboratory.

[0156] II. Content

[0157] (1) The specificity results are shown in the following table:

[0158] Table - 4 Specificity test results

[0159]

[0160] Conclusion: The blank solvent, blank excipient solution, and mixed impurity solution all have no interference on the detection of the main component, and the theoretical plate numbers are all greater than 2000, indicating high specificity.

[0161] (2) The repeatability results are shown in the following table:

[0162] The HPLC chromatograms of D - glucosamine hydrochloride reference substances 1 and 2 are as Figure 1 、 2 shown, and the repeatability test results of the test samples are shown in the following table:

[0163] Table - 5 Repeatability test results

[0164]

[0165] Conclusion: The RSD of the main component content in 6 test solution samples was 0.7%, less than 2.0%, indicating good repeatability.

[0166] (3) Result comparison

[0167] The same batch of samples was determined by different laboratories, and the results are shown in the following table:

[0168] Table - 6 Summary table of comparative test results

[0169]

[0170] Conclusion: It can be seen from the experiments that the absolute deviations of the assay results in the two laboratories all meet the requirements, and the experimental results will not be greatly affected by the change of the laboratory.

[0171] It should be noted that the above are only the preferred embodiments of the present invention, and do not impose any form of limitation on the present invention. Any person skilled in the relevant art can, without departing from the scope of the technical solution of the present invention, make changes or modifications to equivalent embodiments with equivalent changes by using the technical content disclosed above, and all fall within the protection scope of the present invention.

Claims

1. A method for determining the dissolution of glucosamine sulfate capsules, characterized in that: The following steps are involved: S1: Preparation of dissolution solution Take glucosamine sulfate capsules, put them into a sinker, use water as the dissolution medium, take samples, and obtain the dissolution solution; S2: Preparation of test solution Take the dissolution solution of step S1, filter it, and take the filtrate; S3: Preparation of reference solution Take the glucosamine hydrochloride reference substance, dissolve it in water and dilute it to make a solution containing 0.26 mg per 1 ml; S4: Preparation of derivatization reagents Take 50 mg of o-phthalaldehyde, add 1.25 ml of anhydrous methanol to dissolve it, add 50 μl of 3-mercaptopropionic acid and 11.2 ml of 0.2 mol / L borate buffer, mix well, and let the solution stand in the dark for 30 minutes before use; S5: Chromatographic conditions Octadecylsilane bonded silica gel was used as filler; acetate buffer solution-methanol = 75:25 was used as mobile phase; the detection wavelength was 344nm; the flow rate was 1.5ml per minute; the column temperature was 30℃; S6: Dissolution Assay Accurately measure 4 μl of derivatization solvent and test solution, 16 μl of 0.2 mol / L borate buffer, mix well, derivatize for 1.5 minutes, inject into liquid chromatograph, record the chromatogram, and take another reference solution and determine in the same way.

2. The method for determining the dissolution rate of glucosamine sulfate capsules according to claim 1, characterized in that: In the S1, the amount of water used is 900 ml, the rotation speed is 50 revolutions per minute, and sampling is performed after 45 minutes.

3. The method for determining the dissolution rate of glucosamine sulfate capsules according to claim 1, characterized in that: In the above-mentioned S5, the system suitability requirement is: in the chromatogram of the reference solution, the number of theoretical plates calculated based on the main peak of glucosamine hydrochloride (α-glucosamine peak) should be no less than 2000, and the tailing factor should be 0.8 to 1.

5.

4. The method for determining the dissolution rate of glucosamine sulfate capsules according to claim 1, characterized in that: In the S6, the dissolution calculation formula is as follows: Where: A 供 is the peak area of ​​α-glucosamine in the test solution; A 对 is the peak area of ​​α-glucosamine in the reference solution; m is the weight of the reference substance, mg; V is the dilution factor of the test product; V 对 is the dilution multiple of the reference substance; P is the content of the reference substance; M1 is the molecular weight of glucosamine sulfate, 456.42; M2 is twice the molecular weight of glucosamine hydrochloride, 431.26; 0.25: Specification of this product, calculated as glucosamine sulfate, g.

5. A method for determining the content of glucosamine sulfate capsules, characterized in that: The following steps are involved: S1: Preparation of test solution Take the contents of glucosamine sulfate capsules, mix well, accurately weigh the equivalent of 250 mg of glucosamine sulfate, put it in a volumetric bottle, add water, shake vigorously for more than 30 minutes to dissolve, dilute to 100 ml with water, shake well, accurately measure 5 ml of the solution, put it in a volumetric bottle, dilute to 50 ml with water, filter, and take the filtrate; S2: Preparation of reference solution Take the glucosamine hydrochloride reference substance, weigh it accurately, dissolve it in water and dilute it to make a solution containing 0.25 mg per 1 ml; S3: Preparation of derivatization reagents Take 50 mg of o-phthalaldehyde, add 1.25 ml of anhydrous methanol to dissolve it, add 50 μl of 3-mercaptopropionic acid and 11.2 ml of 0.2 mol / L borate buffer, mix well, and let the solution stand in the dark for 30 minutes before use; S4: Chromatographic conditions Octadecylsilane bonded silica gel was used as filler; acetate buffer solution-methanol = 75:25 was used as mobile phase; the detection wavelength was 344nm; the flow rate was 1.5ml per minute; the column temperature was 30℃; S5: Assay Accurately measure 4 μl of derivatization solvent and test solution, 16 μl of 0.2 mol / L borate buffer, mix well, derivatize for 1.5 minutes, inject into liquid chromatograph, record the chromatogram, and take another reference solution and determine in the same way.

6. The method for determining the content of glucosamine sulfate capsules according to claim 5, characterized in that: In the above-mentioned S4, the system suitability requirement is: in the chromatogram of the reference solution, the theoretical plate number calculated based on the main peak of glucosamine hydrochloride (α-glucosamine peak) should be no less than 2000, and the tailing factor should be 0.8 to 1.

5.

7. The method for determining the content of glucosamine sulfate capsules according to claim 5, characterized in that: In the above S5, the limit is: calculated by external standard method using the α-glucosamine peak area (C6H 14 NO5)2SO4 content.

8. The method for determining the content of glucosamine sulfate capsules according to claim 5, characterized in that: In the S5, the content calculation formula is as follows: Where: A 供 is the peak area of ​​α-glucosamine in the test solution; A 对 is the peak area of ​​α-glucosamine in the reference solution; m is the weight of the reference substance, mg; W is the sample weight of the test product, mg; W1: average packing volume of the product, g; V is the dilution factor of the test product; V 对 is the dilution multiple of the reference substance; P is the content of the reference substance; M1 is the molecular weight of glucosamine sulfate, 456.42; M2 is twice the molecular weight of glucosamine hydrochloride, 431.26; 0.25: Specification of this product, calculated as glucosamine sulfate, g.

9. Application of the method for determining the dissolution rate of glucosamine sulfate capsules according to claims 1 to 4 in the drug quality standard and quality control of glucosamine sulfate capsules.

10. Application of the method for determining the content of glucosamine sulfate capsules according to claims 5-7 in the drug quality standard and quality control of glucosamine sulfate capsules.

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