Determination method for related impurities of glucosamine sulfate capsule

Abstract

The invention relates to a method for determining related impurities of glucosamine sulfate capsules, which can effectively evaluate the quality and process level of preparations and ensure the effectiveness of drugs, and adopts the technical scheme that the method comprises the following steps: S1, preparing a test solution; s2, preparing a reference substance solution; s3, chromatographic condition setting; s4, determining related impurities; according to the related impurity determination method, the minimum separation degree of the impurities is 1.74 and larger than 1.5, the separation effect is good, it is indicated that specificity is high, fructosazine, 2, 5-deoxyfructosazine, pyrazine, 5-hydroxymethylfurfural, furfuryl alcohol, 2-methylpyrazine, furfural, pyrrole-2-formaldehyde and 5-methylfurfural can be accurately and quantitatively detected, and the content of fructosazine, 2, 5-deoxyfructosazine, pyrazine, 5-hydroxymethylfurfural, furfuryl alcohol, 2-methylpyrazine, furfural, pyrrole-2-formaldehyde and 5-methylfurfural can be accurately and quantitatively detected. According to the present invention, the 5-deoxyfructosazine, the pyrazine, the 5-hydroxymethylfurfural, the furfuryl alcohol, the 2-methylpyrazine, the furfural, the pyrrole-2-formaldehyde and the 5-methylfurfural can be sensitively detected, and the large influence on the experiment result due to the laboratory change can be avoided.

Description

I. Technical Field

[0001] The present invention relates to the field of pharmaceutical analysis, and particularly to a method for determining related impurities in glucosamine sulfate capsules. II. Background Art

[0002] During the production process of capsules, impurities may be introduced due to factors such as raw material quality, synthesis process, and storage conditions. These impurities may affect the safety and effectiveness of drugs. Therefore, it is necessary to accurately determine related impurities to control the quality of drugs. For example, in the capsule preparations of some chemically synthesized drugs, there may be impurities such as unreacted raw materials, intermediates, and degradation products. By determining related impurities, problems in the production process can be detected in a timely manner to ensure the quality stability and safety of drugs.

[0003] Drug regulatory agencies in various countries have strict regulations on the limits of related impurities in capsules. For example, the Chinese Pharmacopoeia stipulates the limit requirements for related impurities in capsules of different categories of drugs. Enterprises must conduct inspections and controls in accordance with the corresponding standards to ensure that drugs meet the legal quality standards, which is also an important basis for drug marketing and supervision. Therefore, it is imperative to invent a method for determining related impurities in glucosamine sulfate capsules. III. Summary of the Invention

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

[0005] One of the technical solutions provided by the present invention is a method for determining related impurities in glucosamine sulfate capsules, including the following steps:

[0006] S1: Preparation of test solution

[0007] Take the content of glucosamine sulfate capsules, mix evenly, accurately weigh an amount equivalent to 1 g of glucosamine sulfate sodium chloride, place it in a volumetric flask, dissolve it with water and dilute to 200 ml, shake well, filter, and take the subsequent filtrate;

[0008] S2: Preparation of reference solution

[0009] Take 2-methylpyrazine reference substance, dissolve it with water and dilute to a solution containing 5 μg per 1 ml;

[0010] Blank excipient solution: Take 0.3 g of blank excipient, place it in a volumetric flask, dissolve it with water and dilute to 200 ml, shake well, filter, and take the subsequent filtrate;

[0011] S3: Chromatographic conditions

[0012] Using octadecylsilane chemically bonded silica gel as the filler (GL Inertsil ODS-3, 4.6 mm × 250 mm, 5 μm or a chromatographic column with equivalent performance), a trapping column (ES Ghost Trapping column 50 × 4.6 mm or a trapping column with equivalent performance), 0.01 mol / L potassium dihydrogen phosphate solution (adjusted to pH 2.5 with phosphoric acid) as mobile phase A, 0.01 mol / L potassium dihydrogen phosphate solution (adjusted to pH 2.5 with phosphoric acid)-acetonitrile (50:50) as mobile phase B, gradient elution was performed according to the following table; the flow rate was 1.0 ml per minute; the column temperature was 30 °C; the detection wavelength was 195 nm; the injection volume was 20 μl;

[0013]

[0014] S4: Determination of related impurities

[0015] Take the test solution and the reference solution, and inject them into the liquid chromatograph respectively, and record the chromatogram;

[0016] In the chromatogram of the test solution, after deducting the chromatographic peaks before fructosazine, if there are chromatographic peaks with the same retention time as the above-mentioned known impurity peaks, they are all calculated by the external standard method of 2-methylpyrazine with a correction factor. The contents of 2,5-deoxyfructosazine and 5-hydroxymethylfurfural impurities shall not exceed 0.20% of the labeled amount of glucosamine sulfate sodium chloride; the contents of fructosazine, pyrazine, sugar alcohol, 2-methylpyrazine, furfural, pyrrole-2-carboxaldehyde and 5-methylfurfural impurities shall not exceed 0.10% of the labeled amount of glucosamine sulfate sodium chloride; other single unknown impurities are calculated by the external standard method based on the peak area of 2-methylpyrazine, and the content shall not exceed 0.10% of the labeled amount of glucosamine sulfate sodium chloride, and the total amount of impurities shall not exceed 0.50% of the labeled amount of glucosamine sulfate sodium chloride;

[0017] Correction factor table

[0018]

[0019] Preferably, in S1, the contents of glucosamine sulfate and sodium chloride are respectively: glucosamine sulfate: 0.6 g to 0.7 g, sodium chloride: 0.3 g to 0.4 g.

[0020] Preferably, in S2, the blank excipient composition is: take 60 g of corn starch, 28.5 g of lactose, 2.5 g of talc powder, 5 g of magnesium stearate, and mix well.

[0021] Preferably, in S3, the system suitability solution is prepared by taking fructosazine, 2,5 - deoxyfructosazine, 2 - methylpyrazine, pyrazine, 5 - hydroxymethylfurfural, furfuryl alcohol, furfural, pyrrole - 2 - carbaldehyde and 5 - methylfurfural impurity reference substances, dissolving in water and diluting to make a mixed solution containing 10 μg of 2,5 - deoxyfructosazine and 5 - hydroxymethylfurfural and 5 μg of other impurities per 1 ml.

[0022] Furthermore, in S3, the system suitability requirements are as follows: in the solution chromatogram, fructosazine, 2,5 - deoxyfructosazine, pyrazine, 5 - hydroxymethylfurfural, furfuryl alcohol, 2 - methylpyrazine, furfural, pyrrole - 2 - carbaldehyde and 5 - methylfurfural elute in sequence, and the resolution between each impurity peak should be greater than 1.5.

[0023] In S4, the calculation formula for related impurities is as follows:

[0024]

[0025] Total impurity content (%) = Σ content of each known impurity (%) + Σ content of unknown impurities (%)

[0026] In the formula:

[0027] F: correction factor of each impurity; A1: peak area of known impurity in the test solution;

[0028] A2: peak area of unknown impurity in the test solution; A 对 : peak area of 2 - methylpyrazine in the reference solution;

[0029] C 对 : concentration of 2 - methylpyrazine in the reference solution (mg / ml);

[0030] W: weighed amount of the test sample, mg; W1: average filling quantity of this product, g;

[0031] V: dilution volume of the test sample, ml;

[0032] 0.314: specification of this product, calculated as glucosamine sulfate sodium chloride, g.

[0033] Application of the method for determining related impurities of the glucosamine sulfate capsules described in the present invention in the drug quality standard and quality control of glucosamine sulfate capsules.

[0034] The beneficial technical effects of the present invention:

[0035] The minimum resolution between impurities in the determination method of related impurities of the present invention is 1.74, which is greater than 1.5, indicating good separation effect and relatively high specificity. Fructosazine, 2,5 - deoxyfructosazine, pyrazine, 5 - hydroxymethylfurfural, furfuryl alcohol, 2 - methylpyrazine, furfural, pyrrole - 2 - carbaldehyde and 5 - methylfurfural can all be accurately quantitatively detected, and fructosazine, 2,5 - deoxyfructosazine, pyrazine, 5 - hydroxymethylfurfural, furfuryl alcohol, 2 - methylpyrazine, furfural, pyrrole - 2 - carbaldehyde and 5 - methylfurfural can all be sensitively detected, and the experimental results will not be greatly affected by changes in the laboratory. IV. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 It is the HPLC chromatogram of the reference substance solution of 2,5 - deoxyfructosazine of the present invention.

[0037] Figure 2 It is the HPLC chromatogram of the reference substance solution of 2 - methylpyrazine of the present invention.

[0038] Figure 3 It is the HPLC chromatogram of the reference substance solution of 5 - methylfurfural of the present invention.

[0039] Figure 4 It is the HPLC chromatogram of the reference substance solution of 5 - hydroxymethylfurfural of the present invention.

[0040] Figure 5 It is the HPLC chromatogram of the reference substance solution of pyrrole - 2 - carbaldehyde of the present invention.

[0041] Figure 6 It is the HPLC chromatogram of the reference substance solution of pyrazine of the present invention.

[0042] Figure 7 It is the HPLC chromatogram of the test solution of the present invention.

[0043] Figure 8 It is the HPLC chromatogram of the reference substance solution of fructosazine of the present invention.

[0044] Figure 9 It is the HPLC chromatogram of the reference substance solution of furfuryl alcohol of the present invention.

[0045] Figure 10 It is the HPLC chromatogram of the reference substance solution of furfural of the present invention.

[0046] Figure 11 It is the HPLC chromatogram of the blank excipient solution of the present invention.

[0047] Figure 12 It is the HPLC chromatogram of the system suitability solution of the present invention. V. SPECIFIC EMBODIMENTS

[0048] The following detailed description of the specific embodiments of the present invention is provided in conjunction with the embodiments.

[0049] Example 1

[0050] A method for the determination of related impurities in glucosamine sulfate capsules, comprising the following steps:

[0051] S1: Preparation of the test solution

[0052] Take the content of glucosamine sulfate capsules, mix evenly, accurately weigh an amount equivalent to 1 g of glucosamine sulfate sodium chloride, place it in a volumetric flask, dissolve with water and dilute to 200 ml, shake well, filter, and take the subsequent filtrate;

[0053] S2: Preparation of the reference solution

[0054] Take the reference substance of 2-methylpyrazine, accurately weigh it, dissolve with water and dilute to prepare a solution containing 5 μg per 1 ml;

[0055] Blank excipients: Take 60 g of corn starch, 28.5 g of lactose, 2.5 g of talc powder, and 5 g of magnesium stearate, and mix evenly;

[0056] Blank excipient solution: Take 0.3 g of blank excipients, place it in a volumetric flask, dissolve with water and dilute to 200 ml, shake well, filter, and take the subsequent filtrate;

[0057] S3: Chromatographic conditions

[0058] Use octadecylsilane-bonded silica gel as the filler (GL Inertsil ODS-3, 4.6 mm × 250 mm 5 μm or a chromatographic column with equivalent efficiency), a trapping column (ES Ghost Trapping column 50 × 4.6 mm or a trapping column with equivalent efficiency), use 0.01 mol / L potassium dihydrogen phosphate solution (adjust the pH value to 2.5 with phosphoric acid) as mobile phase A, 0.01 moI / L potassium dihydrogen phosphate solution (adjust the pH value to 2.5 with phosphoric acid)-acetonitrile (50:50) as mobile phase B, and perform gradient elution according to the following table; the flow rate is 1.0 ml per minute; the column temperature is 30 °C; the detection wavelength is 195 nm; the injection volume is 20 μl;

[0059]

[0060] System suitability solution: Take impurity reference substances of fructosazine, 2,5-deoxyfructosazine, 2-methylpyrazine, pyrazine, 5-hydroxymethylfurfural, furfuryl alcohol, furfural, pyrrole-2-carboxaldehyde, and 5-methylfurfural, dissolve with water and dilute to prepare a mixed solution containing 10 μg of 2,5-deoxyfructosazine and 5-hydroxymethylfurfural and 5 μg of other impurities per 1 ml;

[0061] System suitability requirements: In the solution chromatogram, fructosazine, 2,5-deoxyfructosazine, pyrazine, 5-hydroxymethylfurfural, furfuryl alcohol, 2-methylpyrazine, furfural, pyrrole-2-carboxaldehyde, and 5-methylfurfural shall elute in sequence, and the resolution between each impurity peak shall be greater than 1.5;

[0062] S4: Determination of related impurities

[0063] Take the test solution and the reference solution, and inject them into the liquid chromatograph respectively, and record the chromatogram;

[0064] In the chromatogram of the test solution, deduct the chromatographic peaks before fructosazine. If there are chromatographic peaks with the same retention time as the above-known impurity peaks, calculate them all by the external standard method of 2-methylpyrazine with a correction factor. The contents of 2,5-deoxyfructosazine and 5-hydroxymethylfurfural impurities shall not exceed 0.20% of the labeled amount of glucosamine sulfate and sodium chloride; the contents of fructosazine, pyrazine, sugar alcohol, 2-methylpyrazine, furfural, pyrrole-2-carboxaldehyde, and 5-methylfurfural impurities shall not exceed 0.10% of the labeled amount of glucosamine sulfate and sodium chloride; other individual unknown impurities shall be calculated by the external standard method based on the peak area of 2-methylpyrazine, and the content shall not exceed 0.10% of the labeled amount of glucosamine sulfate and sodium chloride, and the total impurity content shall not exceed 0.50% of the labeled amount of glucosamine sulfate and sodium chloride;

[0065] Correction factor table

[0066]

[0067] Calculation formula:

[0068]

[0069] Total impurity content (%) = Σ content of each known impurity (%) + Σ content of unknown impurities (%)

[0070] Where:

[0071] F: Correction factor of each impurity; A1: Peak area of known impurity in the test solution;

[0072] A2: Peak area of unknown impurity in the test solution; A 对 : Peak area of 2-methylpyrazine in the reference solution;

[0073] C 对 : Concentration of 2-methylpyrazine in the reference solution (mg / ml);

[0074] W: Sampling amount of the test sample, mg; W1: Average filling amount of this product, g;

[0075] V: Dilution volume of the test sample, ml;

[0076] 0.314: Specification of this product, calculated as glucosamine sulfate and sodium chloride, g.

[0077] I. Results and analysis of the determination method for related impurities in glucosamine sulfate capsules using the present invention are as follows: (1) Related impurities

[0078] Table - 1 Summary of related impurity results

[0079]

[0080]

[0081]

[0082] Based on the data and spectrum analysis obtained in the above experiments (as Figure 1 - 12 shown), the determination method for related impurities can be used for the determination of glucosamine sulfate capsules and is feasible.

[0083] II. Data collection and result recording

[0084] (1) Summary of detailed information on instruments, reagents, and test samples

[0085] Table - 2 Sample information

[0086] Name Batch Number Manufacturer Glucosamine Sulfate Capsules 240130 Henan Lingrui Pharmaceutical Co., Ltd.

[0087] Table - 3 Reference substance information

[0088] Fructosazine 140853-202302 96.9％ National Institutes for Food and Drug Control 2 - Methylpyrazine 140849-202102 99.8％ National Institutes for Food and Drug Control 2,5 - Deoxyfructosazine 140859-202101 95.1％ National Institutes for Food and Drug Control 5 - Hydroxymethylfurfural 111626-202215 99.5％ National Institutes for Food and Drug Control 5 - Hydroxymethylfurfural 111626-202316 98.4％ National Institutes for Food and Drug Control Pyrazine 13185 99.88％ Shenzhen Zhenqiang Biotechnology Co., Ltd. Furfuryl Alcohol 46842 95.83％ Shenzhen Zhenqiang Biotechnology Co., Ltd. 5 - Methylfurfural 46188 97.51％ Shenzhen Zhenqiang Biotechnology Co., Ltd. Pyrrole - 2 - Carboxaldehyde 39724 99.96％ Shenzhen Zhenqiang Biotechnology Co., Ltd. Furfural 43230 98.72％ Shenzhen Zhenqiang Biotechnology Co., Ltd.

[0089] Table - 4 Column information

[0090] GL Inertsil ODS - 3, 4.6mm×250mm, 5μm Shimadzu Related Substances ES Ghost Trapping column, 50×4.6mm ES Related Substances

[0091] Table - 5 Instrument and equipment information

[0092] Electronic Balance METTLER TOLEDO XP1203S Electronic Balance METTLER TOLEDO XA205DU Electronic Balance METTLER TOLEDO XSR205DU / A pH Meter METTLER TOLEDO FE20K High Performance Liquid Chromatograph Agilent 1260 Water Bath Thermostatic Oscillator Changzhou Yitong Analytical Instrument Manufacturing Co., Ltd. SHA - B

[0093] Table - 6 Reagent and reagent information

[0094] Name Grade Manufacturer Potassium Dihydrogen Phosphate Analytical Reagent Tianjin Kemiou Chemical Reagent Co., Ltd. Acetonitrile Chromatographic Reagent Fisher Phosphoric Acid Chromatographic Reagent Tianjin Kemiou Chemical Reagent Co., Ltd. Water / Watsons Purified Water / Self - made Borax Analytical Reagent Shanghai Macklin Biochemical Co., Ltd. 3 - Mercaptopropionic Acid Analytical Reagent Shanghai Macklin Biochemical Co., Ltd. O - Phthalaldehyde Analytical Reagent Shanghai Macklin Biochemical Co., Ltd. Anhydrous Methanol Analytical Reagent Tianjin Kemiou Chemical Reagent Co., Ltd. Sodium Hydroxide Analytical Reagent Sinopharm Chemical Reagent Co., Ltd. Sodium Acetate Analytical Reagent Tianjin Kemiou Chemical Reagent Co., Ltd. Glacial Acetic Acid Analytical Reagent Nanjing Chemical Reagent Co., Ltd. Methanol Chromatographic Reagent Tianjin Concord Technology Co., Ltd. Sulfuric Acid GR Nanjing Chemical Reagent Co., Ltd. Sodium Chloride Chromatographic Reagent Tianjin Kemiou Chemical Reagent Co., Ltd.

[0095] III. Related impurities

[0096] (1) The results of specificity are shown in the following table:

[0097] Table - 7 Specificity test results

[0098]

[0099] Conclusion: The blank solvent, sulfuric acid, and sodium chloride do not interfere with the detection of each known impurity. In the system suitability solution, the minimum resolution between each impurity is 1.74, which is greater than 1.5, indicating good separation effect and relatively high specificity.

[0100] (2) The results of the limit of quantitation are shown in Tables - 10 to - 12:

[0101] Table - 8 Results of the limit of quantitation

[0102]

[0103]

[0104] Table - 9 Results of the precision of the limit of quantitation

[0105]

[0106] Table - 10 Results of the signal - to - noise ratio (S / N) of the limit of quantitation

[0107]

[0108] Conclusion: From the above data, it can be seen that the quantitation limit of fructosazine is 2.9400 ng, the quantitation limit concentration is 0.147 μg / ml, which is equivalent to 0.0030% of the test sample concentration. The quantitation limit solution was injected continuously for 6 times, and the RSD of the peak area was 3.0%, less than 10%, and the S / N was in the range of 13.3 - 14.2; the quantitation limit of 2,5-deoxyfructosazine is 5.9400 ng, the quantitation limit concentration is 0.297 μg / ml, which is equivalent to 0.0060% of the test sample concentration. The quantitation limit solution was injected continuously for 6 times, and the RSD of the peak area was 2.7%, less than 10%, and the S / N was in the range of 13.9 - 14.5; the quantitation limit of pyrazine is 2.4200 ng, the quantitation limit concentration is 0.121 μg / ml, which is equivalent to 0.0025% of the test sample concentration. The quantitation limit solution was injected continuously for 6 times, and the RSD of the peak area was 2.2%, less than 10%, and the S / N was in the range of 12.4 - 12.7; the quantitation limit of 5-hydroxymethylfurfural is 2.0000 ng, the quantitation limit concentration is 0.100 μg / ml, which is equivalent to 0.0020% of the test sample concentration. The quantitation limit solution was injected continuously for 6 times, and the RSD of the peak area was 1.2%, less than 10%, and the S / N was in the range of 13.2 - 13.4; the quantitation limit of furfuryl alcohol is 4.7000 ng, the quantitation limit concentration is 0.235 μg / ml, which is equivalent to 0.0048% of the test sample concentration. The quantitation limit solution was injected continuously for 6 times, and the RSD of the peak area was 3.2%, less than 10%, and the S / N was in the range of 13.5 - 14.2; the quantitation limit of 2-methylpyrazine is 1.2860 ng, the quantitation limit concentration is 0.0643 μg / ml, which is equivalent to 0.0013% of the test sample concentration. The quantitation limit solution was injected continuously for 6 times, and the RSD of the peak area was 6.0%, less than 10%, and the S / N was in the range of 13.0 - 13.6; the quantitation limit of furfural is 3.7800 ng, the quantitation limit concentration is 0.189 μg / ml, which is equivalent to 0.0038% of the test sample concentration. The quantitation limit solution was injected continuously for 6 times, and the RSD of the peak area was 5.2%, less than 10%, and the S / N was in the range of 10.1 - 11.1; the quantitation limit of pyrrole-2-carboxaldehyde is 2.1400 ng, the quantitation limit concentration is 0.107 μg / ml, which is equivalent to 0.0022% of the test sample concentration. The quantitation limit solution was injected continuously for 6 times, and the RSD of the peak area was 3.1%, less than 10%, and the S / N was in the range of 12.8 - 13.3; the quantitation limit of 5-methylfurfural is 3.2200 ng, the quantitation limit concentration is 0.161 μg / ml, which is equivalent to 0.0033% of the test sample concentration. The quantitation limit solution was injected continuously for 6 times, and the RSD of the peak area was 4.4%, less than 10%, and the S / N was in the range of 12.8 - 13.6. It shows that fructosazine, 2,5-deoxyfructosazine, pyrazine, 5-hydroxymethylfurfural, furfuryl alcohol, 2-methylpyrazine, furfural, pyrrole-2-carboxaldehyde and 5-methylfurfural can all be accurately quantitated and detected.

[0109] (3) The detection limit results are shown in the following table:

[0110] Table - 11 Detection Limit Results

[0111] Name Concentration (μg / ml) Signal - to - Noise Ratio (S / N) Detection Limit (ng) Equivalent to Test Substance Concentration % Fructosazine 0.0735 3.4 1.470 0.00150 2,5 - Deoxyfructosazine 0.1480 4.0 2.960 0.0030 Pyrazine 0.0604 3.4 1.208 0.0012 5 - Hydroxymethylfurfural 0.0501 3.4 1.002 0.0010 Furfuryl Alcohol 0.1180 3.9 2.360 0.0024 2 - Methylpyrazine 0.0322 3.7 0.644 0.00066 Furfural 0.0944 3.1 1.888 0.0019 Pyrrole - 2 - Carboxaldehyde 0.0533 3.9 1.066 0.0011 5 - Methylfurfural 0.0807 3.6 1.614 0.0016

[0112] Conclusion: From the above data, it can be seen that the detection limit of fructosazine is 1.470 ng, the detection limit concentration is 0.0735 μg / ml, which is equivalent to 0.00150% of the test sample concentration, and S / N is 3.4; the detection limit of 2,5 - deoxyfructosazine is 2.960 ng, the detection limit concentration is 0.1480 μg / ml, which is equivalent to 0.0030% of the test sample concentration, and S / N is 4.0; the detection limit of pyrazine is 1.208 ng, the detection limit concentration is 0.0604 μg / ml, which is equivalent to 0.0012% of the test sample concentration, and S / N is 3.4; the detection limit of 5 - hydroxymethylfurfural is 1.002 ng, the detection limit concentration is 0.0501 μg / ml, which is equivalent to 0.0010% of the test sample concentration, and S / N is 3.4; the detection limit of furfuryl alcohol is 2.360 ng, the detection limit concentration is 0.1180 μg / ml, which is equivalent to 0.0024% of the test sample concentration, and S / N is 3.9; the detection limit of 2 - methylpyrazine is 0.644 ng, the detection limit concentration is 0.0322 μg / ml, which is equivalent to 0.0066% of the test sample concentration, and S / N is 3.7; the detection limit of furfural is 1.888 ng, the detection limit concentration is 0.0944 μg / ml, which is equivalent to 0.0019% of the test sample concentration, and S / N is 3.1; the detection limit of pyrrole - 2 - carbaldehyde is 1.066 ng, the detection limit concentration is 0.0533 μg / ml, which is equivalent to 0.0011% of the test sample concentration, and S / N is 3.9; the detection limit of 5 - methylfurfural is 1.614 ng, the detection limit concentration is 0.0807 μg / ml, which is equivalent to 0.0016% of the test sample concentration, and S / N is 3.6. It shows that fructosazine, 2,5 - deoxyfructosazine, pyrazine, 5 - hydroxymethylfurfural, furfuryl alcohol, 2 - methylpyrazine, furfural, pyrrole - 2 - carbaldehyde and 5 - methylfurfural can all be detected sensitively.

[0113] (4) Result comparison

[0114] The same batch of samples was measured by different laboratories respectively, and the results are shown in the following table:

[0115] Table - 12 Summary Table of Comparative Test Results

[0116]

[0117] Conclusion: Through the experiment, it can be known that the absolute deviations of the related substance 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.

[0118] It should be noted that the above are only the preferred embodiments of the present invention, and do not impose any formal restrictions on the present invention. Any person skilled in the relevant art can, without departing from 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 the determination of related impurities in glucosamine sulfate capsules, characterized in that, It includes the following steps: S1: Prepare the test solution Take the content of glucosamine sulfate capsules, mix evenly, accurately weigh an amount equivalent to 1 g of glucosamine sulfate sodium chloride, place it in a volumetric flask, dissolve it with water and dilute to 200 ml, shake well, filter, and take the subsequent filtrate; S2: Prepare the reference solution Take the reference substance of 2-methylpyrazine, dissolve it with water and dilute to make a solution containing 5 μg per 1 ml; Blank excipient solution: Take 0.3 g of blank excipients, place it in a volumetric flask, dissolve it with water and dilute to 200 ml, shake well, filter, and take the subsequent filtrate; S3: Chromatographic conditions Use octadecylsilane chemically bonded silica gel as the filler, the trapping column ES Ghost Trapping column 50×4.6 mm, use 0.01 mol / L potassium dihydrogen phosphate solution as mobile phase A, 0.01 moI / L potassium dihydrogen phosphate solution - acetonitrile = 50:50 as mobile phase B, and perform gradient elution according to the following table; the flow rate is 1.0 ml per minute; the column temperature is 30 °C; the detection wavelength is 195 nm; the injection volume is 20 μl; S4: Determination of related impurities Take the test solution and the reference solution, inject them into the liquid chromatograph respectively, and record the chromatogram; In the chromatogram of the test solution, deduct the chromatographic peaks before fructosazine. If there are chromatographic peaks with the same retention time as the above-known impurity peaks, calculate them all by the external standard method of 2-methylpyrazine with correction factor. The contents of 2,5-deoxyfructosazine and 5-hydroxymethylfurfural impurities shall not exceed 0.20% of the labeled amount of glucosamine sulfate sodium chloride; the contents of fructosazine, pyrazine, sugar alcohol, 2-methylpyrazine, furfural, pyrrole-2-carboxaldehyde and 5-methylfurfural impurities shall not exceed 0.10% of the labeled amount of glucosamine sulfate sodium chloride; other single unknown impurities are calculated by the external standard method with the peak area of 2-methylpyrazine, and the content shall not exceed 0.10% of the labeled amount of glucosamine sulfate sodium chloride, and the total amount of impurities shall not exceed 0.50% of the labeled amount of glucosamine sulfate sodium chloride; Correction factor table 2. The determination method for related impurities of glucosamine sulfate capsules according to claim 1, characterized in that, In S1 described above, the contents of glucosamine sulfate and sodium chloride are respectively: glucosamine sulfate: 0.6 g - 0.7 g, sodium chloride: 0.3 g - 0.4 g; in S2 described above, the composition of the blank excipients is: take 60 g of corn starch, 28.5 g of lactose, 2.5 g of talc powder, 5 g of magnesium stearate, and mix evenly.

3. The method for determining related impurities in glucosamine sulfate capsules according to claim 1, wherein, In S3 described above, the system suitability solution is to take the impurity reference substances of fructosazine, 2,5-deoxyfructosazine, 2-methylpyrazine, pyrazine, 5-hydroxymethylfurfural, furfuryl alcohol, furfural, pyrrole-2-carboxaldehyde and 5-methylfurfural, dissolve them with water and dilute to make a mixed solution containing 10 μg of 2,5-deoxyfructosazine and 5-hydroxymethylfurfural and 5 μg of other impurities per 1 ml. System suitability requirements: In the chromatogram of the solution, fructosazine, 2,5-deoxyfructosazine, pyrazine, 5-hydroxymethylfurfural, furfuryl alcohol, 2-methylpyrazine, furfural, pyrrole-2-carboxaldehyde and 5-methylfurfural elute in turn, and the resolution between each impurity peak should be greater than 1.

5.

4. The method for determining related impurities in glucosamine sulfate capsules according to claim 1, characterized in that, In S4 described above, the calculation formula for related impurities is: Total impurity content (%) = Σ content of each known impurity (%) + Σ content of unknown impurities (%) Wherein: F: correction factor of each impurity; A1: peak area of known impurity in the test solution A2: Peak area of the unknown impurity in the test solution; A 对 : Peak area of 2-methylpyrazine in the reference solution; C 对 : Concentration of 2-methylpyrazine in the reference solution (mg / ml); W: weighed amount of the test sample, mg; W1: average filling quantity of this product, g V: dilution volume of the test sample, ml 0.314: specification of this product, calculated as glucosamine sulfate and sodium chloride, g 5. Application of the method for determining related impurities of glucosamine sulfate capsules described in claim 1 in the drug quality standard and quality control of glucosamine sulfate capsules

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