Method for detecting polymer impurities in potassium sodium dehydroandroan drographolide succinate and application of method

Abstract

The invention relates to a method for detecting polymer impurities in potassium sodium dehydroandroan drographolide succinate and application of the method, and the detection method comprises the step of detecting a sample by adopting a liquid chromatography-mass spectrometry method to obtain the content of the polymer impurities. The detection method provided by the invention can realize qualitative and quantitative analysis of polymer impurities in potassium sodium dehydroandroan drographolide succinate, has the advantages of high impurity separation degree, good specificity, high precision, good repeatability and high sensitivity, is accurate and reliable, and can be widely applied to quality detection of potassium sodium dehydroandroan drographolide succinate.

Description

Technical Field

[0001] The present invention relates to the technical field of drug detection, and in particular to a detection method for polymer impurities in niacinamide and application thereof. Background Art

[0002] The andrographolide derivative injections on the domestic market mainly include Chuanhuning, Yanhuning, Lianbizhi (andrographolide sulfite), and Xiyanping.

[0003] As a compound, "Yanhuning" has the chemical name: 14-dehydroxy-11,12-didehydroandrographolide-3,19-disuccinic acid half ester potassium sodium salt, and the molecular formula is: C 28 H 34 KNaO 10 ·H2O (molecular weight is 610.68). As a derivative of andrographolide, Yanhuning can inhibit the early increase of capillary permeability, inhibit the exudation of inflammatory substances and edema, can specifically excite the function of the pituitary-adrenal cortex axis, increase the biosynthesis of ACTH in the anterior pituitary, and promote the release of ACTH; in vitro experiments have shown that Yanhuning also has the effect of killing adenovirus, influenza virus, respiratory virus and other viruses. Yanhuning injection has good antibacterial, antiviral, heat-clearing and detoxifying effects, and is mainly used in the treatment of viral upper respiratory tract infections and viral pneumonia.

[0004] Andrographolide is a very unstable compound. In aqueous solution, the lactone ring is easily hydrolyzed, or even easily polymerized at different levels at a certain temperature. The resulting products can cause serious adverse reactions.

[0005] During the storage of yanhuning, polymer impurities are easily generated due to degradation. This type of impurity is different from other impurities (such as hydrolysis and oxidation impurities). Due to its small polarity, it is difficult to detect with conventional detection methods and is easy to miss, thus affecting the safety and effectiveness of the final product. There is no report on the detection of polymer impurities in yanhuning in the prior art. Summary of the invention

[0006] In order to solve the above technical problems, the present invention provides a method for detecting polymer impurities in yanhuning, and also provides the application of the detection method in the quality detection of yanhuning.

[0007] The detection method provided by the present invention adopts liquid chromatography-mass spectrometry, wherein the liquid chromatography achieves a good separation effect, and the mass spectrometry performs qualitative analysis on polymer impurities. Thus, the detection method of the present invention can realize the qualitative and quantitative analysis of polymer impurities in yanhuning, and has the advantages of high impurity separation, good specificity, high precision, good repeatability and high sensitivity. The method is accurate and reliable, and can be widely used in the quality detection of yanhuning.

[0008] The present invention adopts the following technical solutions:

[0009] In a first aspect, the present invention provides a method for detecting polymer impurities in indomethacin, the detection method comprising: detecting a sample using a liquid chromatography-mass spectrometry method to obtain the content of polymer impurities in indomethacin.

[0010] In the present invention, the polymer impurities include but are not limited to impurities generated by oxidation, degradation, polymerization, etc. of the raw material of yanhuning during storage, and impurities generated by the above reactions in the aqueous solution of yanhuning. The detection method provided by the present invention uses liquid chromatography-mass spectrometry, liquid chromatography achieves a good separation effect, and mass spectrometry performs qualitative analysis on polymer impurities. The detection method is accurate and reliable, has the advantages of good specificity, high precision, good repeatability and high sensitivity, and can be widely used in the quality detection of yanhuning.

[0011] Preferably, the polymer impurities in the yanhuning include any one or both of the following structures:

[0012]

[0013]

[0014] In the present invention, liquid chromatography-mass spectrometry is used to determine the structural information of the polymer.

[0015] Preferably, the mobile phase used in the liquid chromatography includes mobile phase A and mobile phase B, the mobile phase A is ammonium acetate buffer, and the mobile phase B is any one or both of the group consisting of acetonitrile and methanol, preferably acetonitrile.

[0016] Preferably, the pH value of the mobile phase A is 2.0-3.0, for example, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, etc., preferably 2.30-2.70, for example, 2.30, 2.31, 2.32, 2.33, 2.34, 2.35, 2.36, 2.37, 2.38, 2.39, 2.40, 2.41, 2.42, 2.43, 2.44, 2.45, 2.46, 2.47, 2.48, 2.49, 2.50, 2.51, 2.52, 2.53, 2.54, 2.55, 2.56, 2.57, 2.58, 2.59, 2.60, 2.61, 2.62, 2.63, 2.64, 2.65 41, 2.42, 2.43, 2.44, 2.45, 2.46, 2.47, 2.48, 2.49, 2.50, 2.51, 2.52, 2.53, 2.54, 2.55, 2.56, 2.57, 2.58, 2.59, 2.60, 2.61, 2.62, 2.63, 2.64, 2.65, 2.66, 2.67, 2.68, 2.69, 2.70, etc.

[0017] In the present invention, when the pH value of mobile phase A is too low or too high, the impurity peaks in the liquid phase spectrum overlap and the peak shape is poor; when the pH is within the preferred range, the separation effect of polymer impurities is better.

[0018] Preferably, the liquid chromatography adopts a gradient elution mode, and the procedure of the gradient elution is:

[0019] (1) from 0 to 8 minutes, wherein the volume proportion of mobile phase A is 15-25%, for example, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, etc., and the volume proportion of mobile phase B is 75-85%, for example, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, etc.;

[0020] (2) when the mobile phase A changes at a uniform rate to 30 minutes, the volume proportion of mobile phase A is 45-55%, for example, it can be 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, etc., and the volume proportion of mobile phase B is 45-55%, for example, it can be 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, etc.;

[0021] (3) when the mobile phase A changes at a uniform rate to 40 minutes, the volume proportion of mobile phase A is 70-85%, for example, it can be 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, etc., and the volume proportion of mobile phase B is 15-30%, for example, it can be 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, etc.;

[0022] (4) when the speed changes uniformly to the 51st minute, the volume proportion of mobile phase A is 15-25%, for example, it can be 15%, 16%, 18%, 20%, 22%, 24%, 25%, etc., and the volume proportion of mobile phase B is 75-85%, for example, it can be 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, etc.;

[0023] (5) From the 51st minute to the 60th minute, the proportion of mobile phase remained unchanged.

[0024] Preferably, the change rate of mobile phase A in the above step (3) in the gradient elution program is 2.3-3.5% / min, for example, it can be 2.3% / min, 2.4% / min, 2.5% / min, 2.6% / min, 2.7% / min, 2.8% / min, 2.9% / min, 3.0% / min, 3.1% / min, 3.2% / min, 3.3% / min, 3.4% / min, 3.5% / min, etc., preferably 2.5-3.0% / min.

[0025] In the present invention, when the change rate of mobile phase A in the above step (3) is too high or too low, the separation degree of polymer impurities in the liquid phase spectrum is poor; when the change rate is within the preferred range of the present invention, the separation degree is good.

[0026] Preferably, the chromatographic column used in the liquid chromatography is a silanol-active octadecylsilane bonded chromatographic column.

[0027] Preferably, the particle size of the chromatographic column used in the liquid chromatography is 1.5-5μm, for example, it can be 1.7μm, 2.1μm, 2.5μm, 3μm, 3.5μm, 4μm, 4.6μm, 5μm, etc., the column length is 100-300mm, for example, it can be 100mm, 150mm, 200mm, 250mm, 300mm, etc., and the inner diameter of the column is 2-4.6mm, for example, it can be 2.1mm, 2.5mm, 3mm, 3.5mm, 4mm, 4.6mm, etc.

[0028] In the present invention, when the silanol-active octadecylsilane bonded chromatographic column is used, such as CAPCELL PAKC18 MGII silica gel chromatographic column, 3 μm, 150 mm*4 mm, the peak shape of the polymer is better and the separation degree is higher.

[0029] Preferably, the column temperature of the liquid chromatography column is 30-40°C, for example, it can be 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, etc., preferably 30-35°C.

[0030] Preferably, the injection flow rate of the liquid chromatography is 0.4-1.0 mL / min, for example, it can be 0.4 mL / min, 0.5 mL / min, 0.6 mL / min, 0.7 mL / min, 0.8 mL / min, 0.9 mL / min, 1.0 mL / min, etc., preferably 0.6-0.8 mL / min.

[0031] In the present invention, when either the column temperature or the injection flow rate is too high or too low, the separation degree of the obtained liquid phase spectrum is poor. When the column temperature and the injection flow rate are within the preferred range of the present invention, the separation effect is further improved.

[0032] Preferably, the ultraviolet detection wavelength of the liquid chromatography is 245-255 nm, for example, it can be 245 nm, 247 nm, 249 nm, 251 nm, 253 nm, 255 nm, etc.

[0033] Preferably, the mass analyzer of the mass spectrometer is a triple quadrupole mass spectrometer.

[0034] Preferably, the mass spectrometer is an ESI mass spectrometer.

[0035] Preferably, the ion source of the mass spectrometer is set to ESI positive ion mode.

[0036] Preferably, the sheath gas pressure of the mass spectrometer is 35-45arb, for example, it can be 35arb, 36arb, 38arb, 40arb, 42arb, 44arb, 45arb, etc.

[0037] In the present invention, when the sheath gas pressure of the mass spectrometer is too high or too low, the detection sensitivity is reduced. When the pressure is within the range provided by the present invention, the sensitivity is higher.

[0038] Preferably, the auxiliary gas pressure of the mass spectrometer is 8-12arb, for example, 8arb, 9arb, 10arb, 11arb, 12arb, etc.

[0039] In a second aspect, the present invention provides application of the method for detecting polymer impurities in indomethacin according to the first aspect in quality detection of indomethacin.

[0040] Compared with the prior art, the present invention has at least the following beneficial effects:

[0041] (1) The liquid chromatography-mass spectrometry detection method provided by the present invention can simultaneously qualitatively and quantitatively determine two types of polymer impurities in inflammatory oxaline (for example, impurity 1 and impurity 2 in the present invention). The method is interference-free, has good specificity, high impurity separation, high precision, good durability, and is suitable for application in quality control. The method is accurate and reliable.

[0042] (2) The detection method provided by the present invention has high sensitivity. For example, based on inflammatory oxadiazine, the detection sensitivity quantitative limit of each polymer can reach 0.03 μg / mL, and the detection limit can be as low as 0.01 μg / mL. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1This is the liquid phase spectrum of the Inertsil C18 silica gel chromatographic column in Example 1, wherein the retention time of polymer 1 is 14.92 minutes, and the retention time of polymer 2 is 15.75 minutes.

[0044] Figure 2 This is the liquid phase spectrum of the CAPCELL PAK C18 MGII silica gel chromatography column in Example 1, wherein the retention time of polymer 1 is 13.63 minutes, and the retention time of polymer 2 is 14.58 minutes.

[0045] Figure 3 This is a liquid phase spectrum of the mobile phase ratio of group 1 in Example 2, wherein the retention time of polymer 1 is 13.76 minutes, and the peak of polymer 2 is cracked.

[0046] Figure 4 This is the liquid phase spectrum when the pH value of mobile phase A in Example 3 is 2.3. The retention time of polymer 1 is 14.95 minutes, and the peak of polymer 2 overlaps with the peak of polymer 1.

[0047] Figure 5 This is the liquid phase spectrum in Example 4 when the column temperature is 40° C. The retention time of polymer 1 is 15.01 minutes, and the peak of polymer 2 partially overlaps with the peak of polymer 1.

[0048] Figure 6 This is the mass spectrometry qualitative spectrum of polymer 1 in the specificity test of Example 7.

[0049] Figure 7 This is the qualitative mass spectrometry spectrum of polymer 2 in the specificity test of Example 7. DETAILED DESCRIPTION

[0050] The technical solution of the present invention is further described below in conjunction with the accompanying drawings and through specific implementation methods. However, the following embodiments are only simple examples of the present invention and do not represent or limit the scope of protection of the present invention. The scope of protection of the present invention shall be subject to the claims.

[0051] Herein, when not specifically indicated, the term "indigoferin" may refer to any of any dosage forms of indigoferin raw materials, indigoferin preparations, indigoferin injections, indigoferin injections, indigoferin for injection, etc., containing any of the aqueous solutions of "indigoferin" as a compound. In many cases, the indigoferin raw materials may be in any physical form such as powder, granules, or blocks. Those skilled in the art will easily know the specific meaning of the term "indigoferin" herein based on the overall meaning of the context, and will not cause deviations in understanding.

[0052] Example 1 Determination of chromatographic column

[0053] This example compares the types of chromatographic columns used in liquid chromatography, specifically including:

[0054] (1) Solution preparation

[0055] Test solution: Take the raw material of yanhuning that has been stored at 60℃ for 30 days, weigh it accurately, dissolve it in water and quantitatively dilute it to make a solution with a concentration of about 1.0 mg / mL as the test solution;

[0056] (2) Chromatographic conditions

[0057] Mobile phase A: Ammonium acetate buffer: Add appropriate amount of ammonium acetate to water to prepare 0.5% (w / v) ammonium acetate solution, and adjust the pH to 2.5 with acetic acid;

[0058] Mobile phase B: anhydrous acetonitrile (i.e., acetonitrile mass fraction is above 99.8%);

[0059] The flow rate was 0.6 mL / min.

[0060] Detection wavelength: 251nm;

[0061] Column temperature: 35°C;

[0062] Mobile phase elution gradient:

[0063] Time (min) Mobile phase B (vt%) Mobile phase A (vt%) 0 80 20 8 80 20 30 50 50 40 23 77 51 80 20 60 80 20

[0064] (3) Experimental steps and conclusions

[0065] The chromatographic column models used and the test results are shown in the following table:

[0066]

[0067]

[0068] Conclusion: Using column 2, the polymer peak shape is significantly better than that obtained using column 1.

[0069] Example 2 Determination of chromatographic mobile phase

[0070] This example compares different mobile phases in the gradient elution of high performance liquid chromatography, specifically including:

[0071] (1) Based on the detection conditions of Example 1, chromatographic column 2 was used, and the type of mobile phase was changed according to the conditions described below, while other conditions remained unchanged:

[0072] Group 1: same mobile phase as in Example 1;

[0073] Group 2: The mobile phase differed from that in Example 1 only in that mobile phase A was replaced with a 0.05% potassium dihydrogen phosphate aqueous solution (pH value was adjusted to 2.5 with phosphoric acid);

[0074] Group 3: The mobile phase is different from that in Example 1 only in that the mobile phase B is replaced with anhydrous methanol (i.e., the methanol mass fraction is greater than 99.9%);

[0075] (2) Experimental steps and conclusions

[0076] The corresponding test results using the mobile phases of Groups 1 to 3 are shown in the following table:

[0077]

[0078] Conclusion: The peak separation is better when using ammonium acetate buffer-acetonitrile as the mobile phase system.

[0079] Example 3 Determination of Chromatographic Mobile Phase Ratio

[0080] This example compares the mobile phase ratios in the gradient elution of high performance liquid chromatography, specifically including:

[0081] (1) Based on the detection conditions of Example 1, chromatographic column 2 was used, and the proportion of the mobile phase was changed according to the conditions described below, while other conditions remained unchanged:

[0082] Group 1: The difference between the mobile phase elution gradient and that in Example 1 is that at the 40th minute, the volume proportion of mobile phase A is 73%, and the volume proportion of mobile phase B is 27%, that is, within 30-40 minutes, the change rate of mobile phase A is 2.3% / min;

[0083] Group 2: The difference between the mobile phase elution gradient and that in Example 1 is that at the 40th minute, the volume proportion of mobile phase A is 75%, and the volume proportion of mobile phase B is 25%, that is, within the 30th to 40th minute, the change rate of mobile phase A is 2.5% / min;

[0084] Group 3: The mobile phase elution gradient is the same as that in Example 1;

[0085] Group 4: The difference between the mobile phase elution gradient and that in Example 1 is that at the 40th minute, the volume proportion of mobile phase A is 80%, and the volume proportion of mobile phase B is 20%, that is, within the 30th to 40th minute, the change rate of mobile phase A is 3% / min;

[0086] Group 5: The difference between the mobile phase elution gradient and that in Example 1 is that at the 40th minute, the volume proportion of mobile phase A is 85%, and the volume proportion of mobile phase B is 15%, that is, within 30-40 minutes, the change rate of mobile phase A is 3.5% / min;

[0087] Group 6: The difference from the mobile phase elution gradient in Example 1 is that, at 0-8 minutes and 51-60 minutes, the volume proportion of mobile phase A is 25%, and the volume proportion of mobile phase B is 75%; at 30 minutes, the volume proportion of mobile phase A is 45%, and the volume proportion of mobile phase B is 55%; at 40 minutes, the volume proportion of mobile phase A is 70%, and the volume proportion of mobile phase B is 30%;

[0088] Group 7: The only difference from the mobile phase elution gradient in Example 1 is that, at 0-8 minutes and 51-60 minutes, the volume of mobile phase A accounts for 15%, and the volume of mobile phase B accounts for 85%; at the 30th minute, the volume of mobile phase A accounts for 55%, and the volume of mobile phase B accounts for 45%; at the 40th minute, the volume of mobile phase A accounts for 85%, and the volume of mobile phase B accounts for 15%.

[0089] (2) Experimental steps and conclusions

[0090] The test results under the gradient elution program of different groups are shown in the following table:

[0091]

[0092] Conclusion: When the change rate of mobile phase A between 30 and 40 minutes was between 2.5 and 3.0, the separation was good, and the optimal change rate was 2.7.

[0093] Example 4 Determination of pH value of chromatographic mobile phase A

[0094] This example compares the pH value of mobile phase A of liquid chromatography, specifically including:

[0095] (1) Based on Example 1, chromatographic column 2 was used, and the pH value of mobile phase A was changed to 2.0, 2.3, 2.5, 2.7 and 3.0, respectively, while other conditions remained unchanged;

[0096] (2) Experimental steps and conclusions

[0097] The detection results of different pH values ​​are shown in the following table:

[0098]

[0099]

[0100] Conclusion: When the pH of mobile phase A is between 2.3-2.7, the polymer peak shape is good.

[0101] Example 5 Determination of chromatographic column temperature range

[0102] This example compares the column temperature of a chromatographic column in a high performance liquid chromatography, specifically including:

[0103] (1) Based on Example 1, chromatographic column 2 was used, and the column temperature of the chromatographic column was changed to 30° C., 35° C. and 40° C., respectively, while other conditions remained unchanged;

[0104] (2) Experimental steps and conclusions

[0105] The detection effects of different column temperatures are shown in the following table:

[0106]

[0107] Conclusion: When the column temperature is between 30-35℃, the separation of polymer peaks is good.

[0108] Example 6 Determination of Chromatographic Condition Flow Rate

[0109] This example compares the injection flow rate in high performance liquid phase detection, specifically including:

[0110] (1) Based on Example 1, chromatographic column 2 was used, and the injection flow rate was changed to 0.4 mL / min, 0.6 mL / min, 0.8 mL / min and 1.0 mL / min, respectively, while other conditions remained unchanged;

[0111] (2) Experimental steps and conclusions

[0112] The test results of different injection flow rates are shown in the following table:

[0113]

[0114]

[0115] Conclusion: When the flow rate is between 0.6-0.8mL / min, the system applicability (peak shape, separation and sensitivity, etc.) of the chromatographic method is good, and the optimal flow rate is 0.6mL / min.

[0116] Example 7 Determination of Sheath Gas Pressure in Mass Spectrometry Detection

[0117] This example compares the corresponding detection sensitivities of different sheath gas pressures in mass spectrometry detection, specifically including:

[0118] (1) Based on Example 1, chromatographic column 2 was used to perform detection according to the chromatographic conditions provided in Example 1. The mass spectrometry conditions were as follows:

[0119] Ion source: ESI+;

[0120] Spray voltage: 3500V(+);

[0121] Sheath gas pressure: select 30arb, 35arb, 40arb, 45arb respectively;

[0122] Auxiliary gas pressure: 10arb;

[0123] Evaporation temperature: 350℃;

[0124] Ion transfer tube temperature: 350°C;

[0125] Scan mode: FulMS-ddMS2;

[0126] Ful MS: Scan range is m / z 200-3000, resolution is 70000;

[0127] MS2: Resolution is 17500;

[0128] Step NCE: 20, 40, 60;

[0129] TopN:3.

[0130] (2) Experimental steps and conclusions

[0131] The detection sensitivity results under different sheath gas pressures are shown below:

[0132]

[0133] Conclusion: When the sheath gas pressure is between 35-45arb, the mass spectrometer response sensitivity is good.

[0134] Example 8 Specificity Test

[0135] (1) Chromatographic conditions

[0136] Mobile phase A: ammonium acetate buffer: prepare a 0.5% (w / v) ammonium acetate solution with ammonium acetate and adjust the pH to 2.5 with acetic acid;

[0137] Mobile phase B: anhydrous acetonitrile;

[0138] Flow rate: 0.6 mL / min;

[0139] Chromatographic column: CAPCELL PAK C18 MGII silica gel chromatographic column (3μm, 150mm*4mm);

[0140] Detection wavelength: 251nm;

[0141] Column temperature: 35°C;

[0142] Gradient elution program:

[0143]

[0144]

[0145] (2) Mass spectrometry conditions

[0146] Ion source: ESI+;

[0147] Spray voltage: 3500V(+);

[0148] Sheath gas pressure: select 40arb respectively;

[0149] Auxiliary gas pressure: 10arb;

[0150] Evaporation temperature: 350℃;

[0151] Ion transfer tube temperature: 350°C;

[0152] Scan mode: FulMS-ddMS2;

[0153] Ful MS: Scan range is m / z 200-3000, resolution is 70000;

[0154] MS2: Resolution is 17500;

[0155] Step NCE: 20, 40, 60;

[0156] TopN:3.

[0157] (3) Test method

[0158] Blank solvent: mobile phase A: mobile phase B = (80:20, V / V);

[0159] System suitability solution: Take the raw material of indomethacin (high temperature 60°C, 30 days), accurately weigh it, add water to dissolve it and quantitatively dilute it to make a solution with a concentration of about 1.0 mg / mL as the system suitability solution.

[0160] The test results are shown in the following table:

[0161] project Condition Blank solvent No interference System Suitability Solutions The polymer peak is well separated from adjacent peaks and does not contain other impurity peaks.

[0162] Conclusion: The blank solvent does not interfere with the sample determination, the polymer peak components are single, and the separation between the polymer peaks is good, indicating that the detection method of the present invention has good specificity. The qualitative mass spectra corresponding to polymer 1 and polymer 2 are Figure 6 and Figure 7 .

[0163] Example 9 Sensitivity Test

[0164] Test method:

[0165] Blank solvent: mobile phase A: mobile phase B = (80:20, V / V);

[0166] Sensitivity test solution: accurately weigh the raw material of yanhuning (high temperature 60°C, 30 days), dissolve it in water and dilute it quantitatively, and measure it when it is diluted to a signal / noise ratio (S / N) = 10:1 and a signal / noise ratio (S / N) = 3:1, which is the quantitative limit and detection limit of the method of the present invention;

[0167] The chromatographic and mass spectrometry conditions were the same as those in Example 7.

[0168] The test results are shown in the following table:

[0169] name Limit of quantification (μg / mL) Detection limit (μg / mL) Yanhuning 0.03 0.01

[0170] Conclusion: Calculated based on inflammatory oxadiazine, the sensitivity of each polymer has a quantitative limit of 0.03μg / mL and a detection limit of 0.01μg / mL.

[0171] Example 10 Repeatability Test

[0172] Test method:

[0173] Blank solvent: mobile phase A: mobile phase B = (80:20, V / V);

[0174] Repeatability test solution: Take the raw material of yanhuning (high temperature 60℃, 30 days) and accurately weigh it, add water to dissolve it and quantitatively dilute it to make a solution with a concentration of about 1.0 mg / mL as the repeatability test solution. Prepare six parts in parallel, that is, polymer 1 and polymer 2 are prepared to prepare test samples-1 to test samples-6 shown in the table below;

[0175] The chromatographic and mass spectrometry conditions were the same as those in Example 7.

[0176] The test results are shown in the following table:

[0177] name Polymer 1 content (%) Polymer 2 content (%) Test sample-1 0.18 0.02 Test sample-2 0.20 0.03 Test sample-3 0.19 0.02 Test sample-4 0.19 0.03 Test sample-5 0.18 0.02 Test sample-6 0.19 0.02

[0178] Conclusion: The detection data between different groups of polymer 1 and polymer 2 are stable, indicating that the method of the present invention has good repeatability.

[0179] Example 11 Stability Test

[0180] Test method:

[0181] Blank solvent: mobile phase A: mobile phase B = (80:20, V / V);

[0182] Stability test solution: accurately weigh the raw material of yanhuning, dissolve it in blank solvent and quantitatively dilute it to make a solution with a concentration of about 1.0 mg / mL as the stability test solution;

[0183] The chromatographic and mass spectrometry conditions were the same as those in Example 7.

[0184] The test results are shown in the following table:

[0185]

[0186]

[0187] Conclusion: The results show that the test solution is unstable and needs to be prepared extemporaneously before use.

[0188] The above embodiments only represent specific implementation methods of the present invention, but the protection scope of the present invention is not limited thereto. Those skilled in the art should understand that any changes or substitutions made by those skilled in the art based on the technical solutions disclosed in the present invention fall within the protection scope and disclosure scope of the present invention.

Claims

1. A method for detecting polymer impurities in yanhuning, characterized in that: The detection method comprises: using a liquid chromatography-mass spectrometry method to detect a sample to obtain the content of polymer impurities in the yanhuning.

2. The detection method according to claim 1, characterized in that: The polymer impurities in the yanhuning include any one or both of the following structures:

3. The detection method according to claim 1 or 2, characterized in that: The mobile phase used in the liquid chromatography includes mobile phase A and mobile phase B, wherein the mobile phase A is ammonium acetate buffer, and the mobile phase B is any one or both of the group consisting of acetonitrile and methanol, preferably acetonitrile; Preferably, the pH value of the mobile phase A is 2.0-3.0, preferably 2.3-2.

7.

4. The detection method according to claim 3, characterized in that: The liquid chromatography adopts a gradient elution method, and the procedure of the gradient elution is: from 0 to 8 minutes, the volume of mobile phase A accounts for 15-25%, and the volume of mobile phase B accounts for 75-85%; then the volume changes at a constant speed to the 30th minute, the volume of mobile phase A accounts for 45-55%, and the volume of mobile phase B accounts for 45-55%; then the volume changes at a constant speed to the 40th minute, the volume of mobile phase A accounts for 70-85%, and the volume of mobile phase B accounts for 15-30%; then the volume changes at a constant speed to the 51st minute, the volume of mobile phase A accounts for 15-25%, and the volume of mobile phase B accounts for 75-85%; from 51st to 60th minute, the mobile phase proportion remains constant; Preferably, within 30-40 minutes, the change rate of mobile phase A is 2.3-3.5% / min, preferably 2.5-3.0% / min.

5. The detection method according to any one of claims 1 to 4, characterized in that: The chromatographic column used in the liquid chromatography is a silanol-active octadecylsilane bonded chromatographic column; Preferably, the particle size of the chromatographic column used in the liquid chromatography is 1.5-5 μm, the column length is 100-300 mm, and the column inner diameter is 2-4.6 mm.

6. The detection method according to any one of claims 1 to 5, characterized in that: The column temperature of the liquid chromatography column is 30-40°C, preferably 30-35°C; Preferably, the injection flow rate of the liquid chromatography is 0.4-1.0 mL / min, preferably 0.6-0.8 mL / min.

7. The detection method according to any one of claims 1 to 6, characterized in that: The ultraviolet detection wavelength of the liquid chromatography is 245-255 nm.

8. The detection method according to any one of claims 1 to 7, characterized in that: The mass analyzer of the mass spectrometer is a triple quadrupole mass spectrometer; Preferably, the mass spectrometer is an ESI mass spectrometer; Preferably, the ion source of the mass spectrometer is set to ESI positive ion mode.

9. The detection method according to any one of claims 1 to 8, characterized in that: The sheath gas pressure of the mass spectrometer is 35-45arb; Preferably, the auxiliary gas pressure of the mass spectrometer is 8-12 arb.

10. Use of the method for detecting polymer impurities in indomethacin according to any one of claims 1 to 9 in the quality detection of indomethacin.

Images