Method for detecting related substances in potassium sucrose octasulfate

Abstract

The invention relates to the technical field of drug analysis and detection, and particularly discloses a method for detecting related substances in sucrose octasulfate potassium. According to the method, a Hypersil Gold Amine (250mm * 4.6 mm, 5 [mu] m) chromatographic column is adopted, a mixed solution of an ammonium sulfate buffer solution with the pH value of 2.8-3.2 and acetonitrile is used as a mobile phase, and effective separation of impurities such as sucrose potassium heptasulfate and sucrose potassium octasulfate is realized in an isocratic elution mode through high performance liquid chromatography. The method provided by the invention is relatively good in sensitivity, accuracy and reproducibility, and provides reliable guarantee for improving and better controlling the quality of the sucrose potassium octasulfate, so that the quality and the safety of a pharmaceutical preparation containing the sucrose potassium octasulfate are favorably improved; the progress and development of the whole pharmaceutical industry in the aspects of medicine research and development, production and quality control can be powerfully promoted, and the practical value is high.

Description

Technical Field

[0001] The invention relates to the technical field of drug analysis and detection, and in particular to a method for detecting related substances in potassium sucrose octasulfate. Background Art

[0002] In the modern pharmaceutical industry, pharmaceutical excipients play a vital role in the performance, quality and stability of pharmaceutical preparations. As an emerging pharmaceutical excipient, potassium sucrose octasulfate has shown broad application prospects in the field of pharmaceutical preparations due to its unique chemical structure and physicochemical properties. It can improve the solubility and stability of drugs, regulate the release rate of drugs, and in some cases also has certain biologically active auxiliary effects, such as enhancing the affinity of drugs with the cornea and promoting drug absorption in ocular drug preparations, and helping to reduce inflammatory reactions and promote joint tissue repair in drug preparations for the treatment of joint diseases. With the increasing application of potassium sucrose octasulfate in the field of pharmaceutical excipients, the requirements for its quality control are becoming increasingly stringent. Among them, the detection of related substances is a key link to ensure its quality and safety.

[0003] The eight free hydroxyl groups in sucrose react with sulfur trioxide, and depending on the degree of reaction, sucrose heptasulfate (impurity A) may be produced. If its impurity content is higher than the safety level (0.1%), it will affect the quality of the product and the safety of medication. Therefore, it is necessary to control the content of related substances in potassium sucrose octasulfate. However, the pharmacopoeias of various countries currently do not include the detection method of related substances in potassium sucrose octasulfate. Therefore, it is urgent to develop a more accurate, sensitive, efficient and selective detection method for related substances in potassium sucrose octasulfate to ensure the quality and safety of potassium sucrose octasulfate drugs and promote its further development and application in the field of medicine. Summary of the invention

[0004] In view of the above technical problems, the present invention provides a method for detecting related substances in potassium sucrose octasulfate.

[0005] In order to solve the above technical problems, the technical solution provided by the present invention is:

[0006] A method for detecting related substances in potassium sucrose octasulfate, using high performance liquid chromatography for detection, the chromatographic conditions are:

[0007] Column: Hypersil Gold Amino, 250 mm × 4.6 mm, 5 μm;

[0008] Detector: differential refractometer;

[0009] Mobile phase: a mixed solution of ammonium sulfate buffer with a concentration of 0.55 mol / L to 0.65 mol / L and acetonitrile, wherein the pH of the ammonium sulfate buffer is 2.8 to 3.2, and the volume ratio of the ammonium sulfate buffer to acetonitrile is (78 to 82):(22 to 18);

[0010] The elution mode was isocratic.

[0011] Compared with the prior art, the detection method of related substances in potassium sucrose octasulfate provided by the present invention adopts a Hypersil Gold Amino (250mm×4.6mm, 5μm) chromatographic column, and a mixed solution of ammonium sulfate buffer and acetonitrile with a pH of 2.8 to 3.2 as the mobile phase, and isocratic elution is performed by high performance liquid chromatography to achieve effective separation of impurities such as potassium sucrose octasulfate and potassium sucrose octasulfate, and accurately detect the situation of related substances in potassium sucrose octasulfate. And the method provided by the present invention is studied and verified by methodological studies such as specificity and sensitivity, and it is found that the method of the present invention has good sensitivity, accuracy, and reproducibility, and can achieve accurate quantitative detection of related substances in potassium sucrose octasulfate with a simple and fast method, which solves the problem that the prior art cannot accurately and quantitatively detect related substances in potassium sucrose octasulfate, and provides reliable guarantee for improving and better controlling the quality of potassium sucrose octasulfate, thereby being conducive to improving the quality and safety of pharmaceutical preparations containing potassium sucrose octasulfate, and will also effectively promote the progress and development of the entire pharmaceutical industry in drug research and development, production and quality control, and has high practical value.

[0012] It should be noted that the impurity mainly detected in the present invention is potassium sucrose heptasulfate, the structural formula of which is shown below, hereinafter referred to as impurity A.

[0013]

[0014] Preferably, the volume ratio of ammonium sulfate buffer to acetonitrile in the mobile phase is 80:20.

[0015] The inventors found during the experiment that the peaks of potassium sucrose octasulfate and the adjacent impurity A potassium sucrose heptasulfate could not be effectively separated, and the main component and impurity A had serious tailing. Through creative thinking, the inventors used a mixed solution of ammonium sulfate buffer and acetonitrile at a specific ratio of pH 2.8 to 3.2 as the mobile phase, selected Hypersil Gold Amino (250mm×4.6mm, 5μm) as the detection chromatographic column, and achieved effective separation of impurity A and substances such as potassium sucrose octasulfate through isocratic elution, which is conducive to the accurate qualitative and quantitative detection of impurity A in potassium sucrose octasulfate products, and facilitated more accurate monitoring of the quality of salt potassium sucrose octasulfate products.

[0016] Preferably, the concentration of the ammonium sulfate buffer is 0.6 mol / L and the pH is 3.0.

[0017] More preferably, phosphoric acid is used to adjust the pH of the ammonium sulfate solution to 2.8-3.2.

[0018] The preferred pH value of the ammonium sulfate buffer can reduce spectral band tailing and improve peak shape, thereby facilitating improved separation between components and making the detection results more accurate and precise.

[0019] Furthermore, the preparation method of the ammonium sulfate buffer comprises the following steps: taking ammonium sulfate and dissolving it in water, and adjusting the pH to 2.8-3.2 with phosphoric acid to obtain an ammonium sulfate buffer with a concentration of 0.5 mol / L-0.7 mol / L.

[0020] Preferably, the column temperature is 35°C to 45°C.

[0021] More preferably, the column temperature is 40°C.

[0022] Preferably, the flow rate is 0.8 mL / min to 1.2 mL / min.

[0023] More preferably, the flow rate is 1.0 mL / min.

[0024] Preferably, the injection volume is 50 μL.

[0025] The preferred detection conditions can achieve a higher separation degree between potassium sucrose octasulfate and impurity A to ensure effective detection of the impurity, thereby achieving the purpose of effectively and accurately controlling the content of impurity A in potassium sucrose octasulfate.

[0026] Furthermore, the preparation of the test solution specifically includes the following steps: taking a sucrose potassium octasulfate sample, adding a mobile phase to dissolve and dilute it to obtain a test solution with a concentration of 50 mg / mL.

[0027] Furthermore, the preparation of the control solution specifically includes the following steps: taking the test solution, diluting it with the mobile phase, and obtaining a control solution with a concentration of 0.05 mg / mL.

[0028] The detection method provided by the present invention can realize the effective separation of potassium sucrose octasulfate and impurity A, accurately qualitatively and quantitatively detect related substances in potassium sucrose octasulfate, which is beneficial to effectively control the quality of potassium sucrose octasulfate, and the operation is simple and fast with high sensitivity, and provides a simple and reliable method for quality control of potassium sucrose octasulfate products. At the same time, it is also beneficial to improve the quality and safety of pharmaceutical preparations containing potassium sucrose octasulfate, and has high practical value. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1is the chromatogram of the blank solution under the system suitability item in Example 2;

[0030] Figure 2 is the chromatogram of the control solution under the system suitability item in Example 2;

[0031] Figure 3 is the chromatogram of the test solution under the system suitability item in Example 2;

[0032] Figure 4 is a chromatogram of the system suitability solution under the system suitability item in Example 2;

[0033] Figure 5 The chromatograms of the impurity A location solution and the test solution in Comparative Example 1 are shown;

[0034] Figure 6 It is the chromatogram of the test solution in Comparative Example 2. DETAILED DESCRIPTION

[0035] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0036] Example 1

[0037] Materials and Methods

[0038] Instruments: High performance liquid chromatograph, differential refractometer, volumetric flask, electronic balance.

[0039] Reagents: acetonitrile, ammonium sulfate, phosphoric acid, purified water.

[0040] Blank solvent: mobile phase.

[0041] 1.1 Solution preparation

[0042] Preparation of ammonium sulfate buffer: Take about 79.28 g of ammonium sulfate, add 1000 mL of purified water to dissolve, adjust the pH to 3.0 with phosphoric acid, shake well, filter through a 0.45 μm filter membrane, and degas by ultrasonication to obtain ammonium sulfate buffer.

[0043] Mobile phase: pH 3.0 ammonium sulfate buffer and acetonitrile were mixed evenly in a volume ratio of 80:20, filtered through a 0.45 μm filter membrane, and ultrasonically degassed to obtain the mobile phase.

[0044] Preparation of test solution: Take an appropriate amount of potassium sucrose octasulfate sample, accurately weigh it, add mobile phase to dissolve and dilute it to make a solution containing approximately 50 mg of potassium sucrose octasulfate per 1 mL to obtain the test solution.

[0045] Control solution: Accurately measure 1 mL of the test solution, place it in a 100 mL volumetric flask, dilute it to the scale with the mobile phase, and shake well; accurately measure 1 mL, place it in a 10 mL volumetric flask, dilute it to the scale with the mobile phase, and shake well to obtain the control solution.

[0046] 1.2 High performance liquid chromatography conditions:

[0047] Column: Hypersil Gold Amino, 250 mm × 4.6 mm, 5 μm;

[0048] Detector: differential refractometer;

[0049] Mobile phase: a mixed solution of 0.6 mol / L ammonium sulfate buffer (pH 3.0) and acetonitrile, wherein the volume ratio of the ammonium sulfate buffer to acetonitrile is 80:20;

[0050] Flow rate: 1.0 mL / min;

[0051] Column temperature: 40°C;

[0052] Detector temperature: 40°C;

[0053] Injection volume: 50 μL;

[0054] Isocratic elution;

[0055] Running time: 30min.

[0056] Determination method: Accurately measure the test solution and the reference solution and inject them into the high performance liquid chromatograph respectively, and record the chromatogram. The content of impurity A is calculated by the self-reference method with the correction factor added.

[0057] Example 2

[0058] Methodological validation:

[0059] 2.1 System Applicability

[0060] Sucrose potassium heptasulfate reference stock solution: Take about 40 mg of sucrose potassium heptasulfate reference, accurately weigh, place in a 20 mL volumetric flask, dissolve and dilute to the scale with mobile phase, shake well, and obtain a 2 mg / mL sucrose potassium heptasulfate reference stock solution.

[0061] Sucrose potassium octasulfate reference stock solution: Take about 40 mg of sucrose potassium octasulfate reference, accurately weigh, place in a 20 mL volumetric flask, dissolve and dilute to the scale with mobile phase, shake well, and obtain a 2 mg / mL sucrose potassium octasulfate reference stock solution.

[0062] System suitability solution: Accurately pipette 1 mL each of sucrose potassium heptasulfate reference stock solution and sucrose potassium octasulfate reference stock solution, mix well, and obtain the system suitability solution.

[0063] Take blank solution, system suitability solution, test solution and control solution for testing respectively. The separation degree between the main component and adjacent impurities in the system suitability solution is required to be no less than 1.5. In the 5-point reference solution spectrum, the RSD of the main component peak area shall not exceed 5.0%, and the RSD of the retention time shall not exceed 1.0%.

[0064] Take each solution and inject it into the sample according to the above HPLC conditions to record the chromatogram. The chromatogram of the blank solution is as follows: Figure 1 The chromatogram of the control solution is shown in Figure 2 As shown in Table 1, the chromatogram of the test solution is as follows Figure 3 The chromatogram of the system suitability solution is shown in Figure 4 And as shown in Table 2.

[0065] Table 1 Results of 5-point control solution

[0066]

[0067]

[0068] Table 2 Results of system suitability solutions

[0069]

[0070] The test results show that in the system suitability solution, the separation between the main component and the adjacent impurities is 6.68, which is greater than 1.5. The RSD of the main component peak area in the 5-point reference solution is 2.8%, which is less than 5.0%; the RSD of the retention time is 0.03%, which is less than 1.0%.

[0071] 2.2 Exclusivity

[0072] 2.2.1 Preparation of impurity localization solution

[0073] Blank solution / diluent: mobile phase.

[0074] Sucrose potassium heptasulfate stock solution: Take 40 mg of sucrose potassium heptasulfate reference substance, accurately weigh it, place it in a 20 mL volumetric flask, dilute it to the scale with the mobile phase, shake well, and obtain a 2 mg / L sucrose potassium heptasulfate stock solution.

[0075] Sucrose potassium heptasulfate fixing solution: Accurately measure 2.5 mL of sucrose potassium heptasulfate stock solution into a 100 mL volumetric flask and dilute to the mark with the mobile phase to obtain the sucrose potassium heptasulfate fixing solution (50 μg / mL).

[0076] Potassium ion positioning solution: Take about 250 mg of potassium sulfate, weigh it accurately, put it in a 10 mL volumetric flask, dilute it to the scale with the mobile phase, shake well, and use it as the potassium ion positioning solution.

[0077] 2.2.2 Forced degradation test

[0078] The forced degradation destruction solution was prepared according to the conditions shown in Table 3.

[0079] Table 3 Preparation of forced degradation destruction solution

[0080]

[0081]

[0082] After the instrument is stable, take blank solution, system suitability solution, sucrose heptasulfate potassium positioning solution, potassium ion positioning solution, undestroyed and destroyed solutions for sampling and testing, and record the color break diagram. It is required that the blank solution does not interfere with the determination, the separation degree between the peaks of sucrose heptasulfate potassium and sucrose octasulfate potassium in the system suitability solution should be greater than 1.5, the separation degree between the main peak and the adjacent impurity peak or the peak-to-valley ratio in the destroyed solution shall not be less than 1.5, and the material conservation is between 90.0% and 110.0%. The test results are shown in Tables 4 to 6.

[0083] Table 4 Specificity test results of impurity localization solution

[0084]

[0085] Note: N / A means not applicable.

[0086] Table 5 Destruction test results

[0087]

[0088] Note: “ND” stands for not detected and “N / A” stands for not applicable.

[0089] Table 6 Destruction test results

[0090]

[0091]

[0092] Note: * indicates peak-to-valley ratio.

[0093] The results show that the blank solution does not interfere with the determination; in the system suitability solution, the separation between the potassium sucrose heptasulfate peak and the potassium sucrose octasulfate peak is 6.68, which is greater than 1.5. In the chromatograms of undestroyed, alkali-destroyed, oxidatively-destroyed and light-destroyed solutions, the separation between the main peak and the adjacent impurities is greater than 1.5, which meets the requirements. In the chromatograms of acid-destroyed and high-temperature-destroyed solutions, the peak-to-valley ratio needs to be calculated because the peak width of the main component is too large. In the chromatogram of the high-temperature-destroyed solution, the peak-to-valley ratio is 4.08, which meets the requirements; in the acid-destroyed chromatogram, the main component and the adjacent peaks have reached baseline separation, there is no peak-to-valley value, and the software cannot calculate the peak-to-valley ratio. This product is unstable under acid-destroyed and high-temperature conditions, and can be degraded into sucrose hexasulfate and its isomers, sucrose heptasulfate and its isomers, and the impurity detection amount increases from 0.04% to 1.76% and 0.43%, respectively. It is relatively stable under other destruction conditions. The material conservation is 98.3% to 104.2%, between 90.0% and 110.0%. This shows that the method has good specificity.

[0094] 2.3 Linear range

[0095] Blank solution / diluent: mobile phase.

[0096] Sucrose potassium heptasulfate stock solution: Take about 40 mg of sucrose potassium heptasulfate reference substance, accurately weigh it, put it in a 20 mL volumetric flask, dilute it to the scale with mobile phase, shake well, and obtain a 2 mg / L sucrose potassium heptasulfate stock solution.

[0097] Sucrose potassium heptasulfate linear solution: Accurately measure 12.5 ml of sucrose potassium heptasulfate stock solution, place it in a 50 mL volumetric flask, dilute to the scale with the mobile phase, shake well, and use this as the sucrose potassium heptasulfate linear solution (0.5 mg / mL).

[0098] Sucrose potassium octasulfate stock solution: Take about 40 mg of sucrose potassium octasulfate reference substance, accurately weigh it, put it in a 20 mL volumetric flask, add the mobile phase to dissolve and dilute to the scale, shake well, and obtain a 2 mg / mL sucrose potassium octasulfate stock solution.

[0099] Sucrose octasulfate linear potassium solution: Accurately measure 12.5 mL of sucrose octasulfate potassium stock solution, place it in a 50 mL volumetric flask, dilute to the scale with the mobile phase, shake well, and use this as the sucrose octasulfate potassium linear solution (0.5 mg / mL).

[0100] Prepare the linear solution according to Table 7.

[0101] Table 7 Linear solution preparation table

[0102]

[0103] The blank solution and the linear solution prepared above were sampled and tested respectively. The peak area was used as the ordinate and the concentration was used as the abscissa to perform linear regression and calculate the linear equation and correlation coefficient. The test results are shown in Table 8.

[0104] Table 8 Sucrose potassium heptasulfate linear results

[0105]

[0106]

[0107] Table 8 Sucrose octasulfate potassium linear results

[0108]

[0109] Conclusion: Potassium sucrose heptasulfate is linear in the range of 15.26-254.25 μg / ml (30%-500%), the linear equation is y=0.0008x+0.0036, the correlation coefficient r is 0.9994, which is greater than 0.990, and the ratio of the y-axis intercept to the peak area at the limit 100% concentration is 7.8%, which is less than 25.0%.

[0110] Sucrose potassium octasulfate is linear in the range of 14.75-245.84 μg / ml (30%-500%), the linear equation is y=0.0022x+0.0014, the correlation coefficient r is 0.9995, which is greater than 0.990, and the ratio of the y-axis intercept to the peak area at the limit 100% concentration is 1.1%, which is less than 25.0%, indicating that the method of the present invention has a good linear relationship.

[0111] 2.4 Limit of quantitation

[0112] Blank solution / diluent: mobile phase.

[0113] Quantitation limit solution: Take the 100% limit concentration solution under the linear range and dilute it step by step with the mobile phase until the signal-to-noise ratio is not less than 10, which is used as the quantitation limit solution.

[0114] Take the quantitative limit solution and inject 6 injections continuously. The signal-to-noise ratio of each impurity should not be less than 10; the RSD of the peak area should not exceed 15.0%; and the RSD of the retention time should not exceed 1.0%. The results are shown in Table 9.

[0115] Table 9 Limit of quantitation results (n=6)

[0116]

[0117] Note: “N / A” means not applicable.

[0118] Conclusion: The limits of quantification of potassium sucrose heptasulfate and potassium sucrose octasulfate are 15.0 μg / mL (0.03%) and 15.0 μg / mL (0.03%), respectively. The minimum signal-to-noise ratio is 10.1, and the signal-to-noise ratio of each chromatographic peak is greater than 10.

[0119] After 6 consecutive injections, the peak area RSDs of potassium sucrose heptasulfate and potassium sucrose octasulfate were 8.1% and 9.7%, respectively, both less than 15.0%, and the retention time RSDs were 0.1% and 0.1%, respectively, both less than 1.0%.

[0120] 2.5 Detection limit

[0121] Blank solution / diluent: mobile phase.

[0122] Detection limit solution: Take the solution under the quantitative limit and dilute it step by step with the mobile phase until the signal-to-noise ratio is not less than 3, which is used as the detection limit solution. The detection limit solution is injected three times continuously, and the signal-to-noise ratio of each chromatographic peak should be not less than 3. The test results are shown in Table 10.

[0123] Table 10 Detection limit results (n=3)

[0124]

[0125] Conclusion: The detection limits of potassium sucrose heptasulfate and potassium sucrose octasulfate are 5.0 μg / mL (0.01%) and 5.0 μg / mL (0.01%), respectively. The minimum signal-to-noise ratio is 3.6, and the signal-to-noise ratio of each chromatographic peak is greater than 3.

[0126] 2.6 Accuracy

[0127] Blank solution / diluent: mobile phase.

[0128] Sucrose potassium heptasulfate stock solution: under the same linear range.

[0129] Sucrose potassium heptasulfate linear solution: Accurately measure 12.5 mL of sucrose potassium heptasulfate stock solution, place it in a 50 mL volumetric flask, dilute to the scale with the mobile phase, shake well, and use this as the sucrose potassium heptasulfate linear solution (0.5 mg / mL).

[0130] Test solution: Take about 500 mg of the product, accurately weigh it, put it in a 10 mL volumetric flask, add mobile phase to dissolve and dilute to the scale, shake well, and prepare 2 portions (50 mg / mL) in parallel.

[0131] Control solution: Accurately measure 1 mL of the test solution, place it in a 50 mL volumetric flask, and dilute it to the mark with the mobile phase; accurately measure 1 mL of the test solution, place it in a 20 mL volumetric flask, dilute it to the mark with the diluent, and shake well (0.05 mg / mL).

[0132] 50% horizontal accuracy solution: Take about 500 mg of this product, weigh accurately, put it in a 10mL volumetric flask, add 0.5mL of sucrose potassium heptasulfate linear solution, and then dilute to the scale with mobile phase, shake well, as the 50% horizontal accuracy solution, prepare 3 copies in parallel.

[0133] 100% horizontal accuracy solution: Take about 500 mg of this product, weigh accurately, put it in a 10mL volumetric flask, add 1mL of sucrose potassium heptasulfate linear solution, and then dilute to the scale with diluent, shake well, as the 100% horizontal accuracy solution, prepare 3 copies in parallel.

[0134] 150% horizontal accuracy solution: Take about 500 mg of this product, weigh accurately, put it in a 10mL volumetric flask, add 1.5mL of sucrose potassium heptasulfate linear solution, and then dilute to the scale with diluent, shake well, as the 150% horizontal accuracy solution, prepare 3 copies in parallel.

[0135] Blank solution, control solution, test solution and accuracy solution of each level were tested and calculated by self-control method. The recovery rates at 50%, 100% and 150% concentration levels should be between 80.0% and 120.0%, and the RSD of the recovery rate should not exceed 10.0%. The results are shown in Table 11.

[0136] Table 11 Accuracy results (n=9)

[0137]

[0138]

[0139] Note: “ND” stands for not detected.

[0140] Conclusion: The average recovery rate of potassium sucrose heptasulfate calculated by the self-control method with correction factor added was 107.2% at a concentration of 50% to 150%, within the range of 80.0% to 120.0%; the RSD of the 9 recoveries was 3.9%, less than 10.0%, which met the requirements. This shows that the method has good accuracy.

[0141] 2.6 Durability

[0142] Blank solution / diluent: mobile phase.

[0143] Durability solution: 100% horizontal accuracy solution and control solution under the same accuracy item.

[0144] Durability test:

[0145] Whether the measurement results change when there are slight changes in chromatographic conditions such as flow rate ±0.2mL / min, column temperature ±5℃, mobile phase ratio change ±2%, mobile phase pH ±0.2, replacement of chromatographic column, etc.

[0146] The blank solution and the durability solution were tested and the chromatograms were recorded. The results are shown in Table 12.

[0147] Table 12 Durability test results

[0148]

[0149]

[0150] Note: “N / A” means not applicable.

[0151] Conclusion: When the chromatographic conditions such as flow rate, column temperature, mobile phase ratio and mobile phase pH change slightly, the minimum separation between the main component and its adjacent impurities in the durable solution is 6.42, which is greater than 1.5 and meets the requirements; the ratio of the detection amount of potassium sucrose heptasulfate to the standard conditions ranges from 84.2% to 95.0%, and the ratio of the detection amount of unknown impurities to the standard conditions ranges from 95.2% to 111.9%, both between 80.0% and 120.0%, indicating that the method has good durability.

[0152] The optimal method was used to test the test solution and forced destruction samples. The results are shown in Table 13.

[0153] Table 13

[0154] name Impurity A(%) Maximum single impurity (%) Total impurities (%) Test solution ND 0.04％ 0.04％ Acid destruction of samples 0.099 0.903％ 1.76％ Alkali damaged samples ND 0.057％ 0.11％ Oxidative damage to samples ND 0.04％ 0.04％ High temperature damage sample 0.162 0.296％ 2.88％

[0155] Note: ND means not detected.

[0156] Comparative Example 1

[0157] This comparative example provides a method for detecting related substances in potassium sucrose octasulfate, which is different from that in Example 1 only in that the chromatographic column is NH2LC Column, specification is 4.6mm×250mm, 5μm, and other detection conditions are exactly the same.

[0158] Take the sucrose potassium heptasulfate positioning solution and the test solution in Example 2 for testing, and record the chromatogram. Figure 5 shown.

[0159] As can be seen from the figure, the peaks of potassium sucrose heptasulfate and potassium sucrose octasulfate have serious tailing and do not meet the requirements for accurate quantitative detection.

[0160] Comparative Example 2

[0161] This comparative example provides a method for detecting related substances in potassium sucrose octasulfate, which is different from that in Example 1 only in that the mobile phase is replaced with a 0.1 mol / L calcium chloride solution, and the other detection conditions are exactly the same.

[0162] Take the test solution in Example 2 for testing, and the results are as follows Figure 6 As shown in the figure, it can be seen that there is no peak of sucrose octasulfate potassium in the test solution.

[0163] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modification, equivalent substitution or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for detecting related substances in potassium sucrose octasulfate, characterized in that: High performance liquid chromatography was used for detection, and the chromatographic conditions were: Column: Hypersil Gold Amino, 250 mm × 4.6 mm, 5 μm; Detector: differential refractive index detector; Mobile phase: a mixed solution of ammonium sulfate buffer with a concentration of 0.5 mol / L to 0.7 mol / L and acetonitrile, wherein the pH of the ammonium sulfate buffer is 2.8 to 3.2, and the volume ratio of the ammonium sulfate buffer to acetonitrile is (78 to 82):(22 to 18); The elution mode was isocratic.

2. The method for detecting related substances in potassium sucrose octasulfate as claimed in claim 1, characterized in that: The volume ratio of ammonium sulfate buffer to acetonitrile in the mobile phase is 80:

20.

3. The method for detecting related substances in potassium sucrose octasulfate as claimed in claim 2, characterized in that: The concentration of the ammonium sulfate buffer is 0.6 mol / L, and the pH is 3.

0.

4. The method for detecting related substances in potassium sucrose octasulfate according to claim 1 or 2, characterized in that: Phosphoric acid is used to adjust the pH of the ammonium sulfate solution to 2.8-3.

2.

5. The method for detecting related substances in potassium sucrose octasulfate as claimed in claim 1, characterized in that: The column temperature is 35℃~45℃.

6. The method for detecting related substances in potassium sucrose octasulfate as claimed in claim 5, characterized in that: The column temperature was 40°C.

7. The method for detecting related substances in potassium sucrose octasulfate as claimed in claim 1, characterized in that: The flow rate is 0.8mL / min~1.2mL / min.

8. The method for detecting related substances in potassium sucrose octasulfate as claimed in claim 7, characterized in that: The flow rate was 1.0 mL / min.

9. The method for detecting related substances in potassium sucrose octasulfate according to claim 1, characterized in that: The injection volume was 50 μL.

10. The method for detecting related substances in potassium sucrose octasulfate according to claim 1, characterized in that: The preparation of the test solution specifically includes the following steps: taking a sucrose potassium octasulfate sample, adding a mobile phase to dissolve and dilute it to obtain a test solution with a concentration of 50 mg / mL; The preparation of the control solution specifically includes the following steps: taking the test solution, diluting it with the mobile phase, and obtaining a control solution with a concentration of 0.05 mg / mL.

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