Method for detecting triphenylphosphine oxide in sacubitril valsartan sodium starting material

Abstract

The method for detecting the triphenylphosphine oxide in the sacubitril valsartan sodium starting material is established by adopting a high performance liquid chromatography method. Through verification, the detection method has the advantages of high sensitivity, strong specificity, high accuracy, durability and reliability, and can realize effective control of triphenylphosphine oxide in sacubitril-valsartan sodium, thereby ensuring the medicine quality of sacubitril-valsartan sodium and the effectiveness and safety of patient medication.

Description

Technical Field

[0001] The invention relates to the technical field of drug analysis, and in particular to an analysis and detection method for triphenylphosphine oxide. Background Art

[0002] Triphenylphosphine oxide (see the following formula II) is an organic, weakly acidic substance, irritating, and toxic. After entering the human body, triphenylphosphine oxide will have an irritating effect on the central nervous system and respiratory system, and in severe cases will cause fatal damage. At the same time, it is also corrosive and will cause damage to the skin, eyes, respiratory tract, etc., so it needs to be strictly controlled in human use.

[0003]

[0004] Triphenylphosphine oxide is an impurity introduced in the preparation process of sacubitril-valsartan sodium starting material. The chemical name of the starting material is {(R,E)-5-(biphenyl-4-yl)-4-[(tert-butoxycarbonyl)amino]-2-methylpentenoic acid} (see the following formula I for details)

[0005]

[0006] Since triphenylphosphine oxide may remain in the final product sacubitril valsartan sodium, in order to reduce the occurrence of side effects of medication in patients and ensure the safety of medication, its limit should be strictly controlled and sufficient quality research should be carried out. Currently, there is no literature reporting its detection method, and a reliable, accurate and durable detection method needs to be established to achieve safe and effective monitoring. Summary of the invention

[0007] The purpose of the present invention is to provide a method for detecting triphenylphosphine oxide, so as to achieve effective monitoring of triphenylphosphine oxide in sacubitril-valsartan sodium starting materials, so as to reduce the occurrence of side effects of medication in patients, ensure the safety of medication, and thus jointly maintain human life and health.

[0008] The technical solution of the present invention is as follows:

[0009] A method for detecting triphenylphosphine oxide in a sacubitril-valsartan sodium starting material, wherein the structural formula of the sacubitril-valsartan sodium starting material is formula (I), and the structural formula of triphenylphosphine oxide is formula (II), characterized in that: high performance liquid chromatography is used for detection, and the chromatographic conditions are: the chromatographic column stationary phase uses octadecylsilane bonded silica gel as a filler; the mobile phase consists of a mobile phase A and a mobile phase B, the mobile phase A is an ammonium acetate solution, and the mobile phase B is an organic phase.

[0010]

[0011] Furthermore, the chromatographic conditions at least meet at least one of the following conditions (1) to (9):

[0012] (1) Chromatographic column: 4.6 mm × 150 mm, 3.5 μm, preferably Waters C18 column, 4.6 mm × 150 mm, 3.5 μm; more preferably Waters Sunfire C18 column, 4.6 mm × 150 mm, 3.5 μm;

[0013] (2) Mobile phase A is 10-30 mmol / L ammonium acetate solution; preferably 10-30 mmol / L ammonium acetate solution, the pH value is adjusted to 5.7±0.5; more preferably 10-30 mmol / L ammonium acetate solution, the pH value is adjusted to 5.7±0.1 with glacial acetic acid;

[0014] (3) mobile phase B is acetonitrile;

[0015] (4) Mobile phase ratio: the volume ratio of mobile phase A to mobile phase B is 58-62:42-38;

[0016] (5) Flow rate: 0.8 ml / min to 1.2 ml / min;

[0017] (6) Column temperature: 30-40°C;

[0018] (7) Injection volume: 10-30 μl; preferably, the injection volume is 20 μl;

[0019] (8) Detector: UV detector;

[0020] (9) Detection wavelength: 199~220nm.

[0021] Furthermore, the elution method is isocratic elution.

[0022] Furthermore, the running time is 25 min.

[0023] Further, the solution is prepared as follows:

[0024] Test solution: Take an appropriate amount of sacubitril / valsartan sodium starting material and add solvent to dissolve and dilute to a solution of a certain concentration;

[0025] Reference solution: Take an appropriate amount of triphenylphosphine oxide reference substance and add solvent to dissolve and dilute to a solution of a certain concentration;

[0026] System suitability solution: Take an appropriate amount of sacubitril / valsartan sodium starting material, add an appropriate amount of triphenylphosphine oxide reference substance, and add solvent to dissolve and dilute to a solution of a certain concentration.

[0027] Further, the solution is prepared as follows:

[0028] Test solution: Take about 100 mg of sacubitril-valsartan sodium starting material, weigh accurately, place in a 10 ml volumetric flask, add appropriate amount of solvent, sonicate to dissolve and dilute to scale, shake well;

[0029] Reference solution: take about 4 mg of triphenylphosphine oxide reference substance, accurately weigh it, put it in a 100 ml volumetric flask, add solvent to dissolve and dilute to the scale, shake well, as the stock solution; accurately measure 5 ml of the stock solution, put it in a 50 ml volumetric flask, dilute to the scale with solvent, and shake well;

[0030] System suitability solution: Take about 100 mg of sacubitril / valsartan sodium starting material, accurately weigh it, put it in a 10 ml volumetric flask, accurately measure 1 ml of stock solution and add it, add an appropriate amount of solvent, sonicate to dissolve and dilute to the scale, and shake well.

[0031] Furthermore, the solvent is selected from at least one of mobile phase, methanol, chloroform, acetic acid, tetrahydrofuran, dimethyl sulfoxide and ethanol; preferably, the solvent is ethanol.

[0032] Furthermore, in the system suitability solution, the elution order is triphenylphosphine oxide and sacubitril-valsartan sodium starting material, and the separation degree between the two is greater than 2.0.

[0033] Furthermore, the content of triphenylphosphine oxide in the test solution, calculated by external standard method, does not exceed 0.04%; preferably does not exceed 0.036%.

[0034] Furthermore, in the method, the mean signal-to-noise ratio of the quantitative limit solution (n=6) is not less than 10, preferably 13.6; in the detection limit solution, the mean signal-to-noise ratio is not less than 3, preferably 6.3.

[0035] Furthermore, the method can quantitatively detect triphenylphosphine oxide in a concentration range of 1.0236 μg / ml to 8.2078 μg / ml.

[0036] The present invention also provides an application of the above-mentioned detection method in detecting triphenylphosphine oxide in sacubitril-valsartan sodium starting materials.

[0037] Unless otherwise specified, when the present invention relates to the percentage between liquids, the percentage is volume / volume percentage; when the present invention relates to the percentage between liquids and solids, the percentage is volume (ml) / weight (g) percentage; when the present invention relates to the percentage between solids and liquids, the percentage is weight (g) / volume (ml) percentage; the rest are weight / weight percentages.

[0038] Compared with the prior art, the present invention has the following beneficial technical effects:

[0039] 1) A method for detecting triphenylphosphine oxide in sacubitril-valsartan sodium starting materials was established by high performance liquid chromatography. This method has high sensitivity, strong specificity, high accuracy and durability, and can effectively monitor triphenylphosphine oxide in sacubitril-valsartan sodium starting materials.

[0040] 2) Considering the safety of the experimenters, the simplicity of the experiment and the operability of the HPLC method, anhydrous ethanol, which is relatively cheap and safe, is preferably used as the solvent.

[0041] 3) The mobile phase A of this method is 10-30 mmol / L ammonium acetate solution. By adjusting the pH value, the service life of the chromatographic column can be better protected.

[0042] 4) Change the time-consuming and difficult column equilibration gradient conditions to a simpler and time-saving isocratic method.

[0043] 5) The control of triphenylphosphine oxide has been achieved from the starting material stage of sacubitril valsartan sodium, making it have higher industrial value and the cost loss is lower than that of control research at the intermediate or finished product stage. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] Figure 1 It is the chromatogram of the blank solution of Example 1.

[0045] Figure 2 This is the chromatogram of the system suitability solution of Example 1.

[0046] Figure 3 It is the chromatogram of the reference substance solution of Example 1.

[0047] Figure 4 The chromatogram of the test solution of Example 1 is shown in FIG. DETAILED DESCRIPTION

[0048] In order to make the purpose, technical scheme and advantages of the present invention more clearly understood, the specific implementation methods are described in the form of embodiments below, and the above contents of the present invention are further described in detail. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following embodiments. In the absence of conflict, the embodiments of the present invention and the features in the embodiments can be combined with each other at will.

[0049] Example 1

[0050] High performance liquid chromatography

[0051] Blank solution: anhydrous ethanol.

[0052] Test solution: Take about 100 mg of sacubitril-valsartan sodium starting material (hereinafter referred to as starting material), accurately weigh it, put it in a 10 ml volumetric flask, add an appropriate amount of anhydrous ethanol, ultrasonically dissolve it and dilute it to the scale, and shake it well.

[0053] Reference solution: Take about 4 mg of triphenylphosphine oxide reference substance, accurately weigh it, place it in a 100 ml volumetric flask, add anhydrous ethanol to dissolve and dilute to the scale, shake well, and use it as the stock solution; accurately measure 5 ml of the stock solution, place it in a 50 ml volumetric flask, dilute to the scale with anhydrous ethanol, and shake well.

[0054] System suitability solution: Take about 100 mg of the starting material, weigh it accurately, put it in a 10 ml volumetric flask, accurately measure 1 ml of the stock solution and add it, add an appropriate amount of anhydrous ethanol, sonicate to dissolve and dilute to the scale.

[0055] The chromatographic conditions were as follows: octadecylsilane bonded silica gel was used as the filler (Waters Sunfire C18, 4.6 mm×150 mm, 3.5 μm); 30 mmol / L ammonium acetate solution (pH adjusted to 5.7±0.1 with glacial acetic acid)-acetonitrile (60:40) was used as the mobile phase, the column temperature was 35°C; the flow rate was 1.0 ml per minute; the detection wavelength was 210 nm; the running time was 25 min; and the injection volume was 20 μl.

[0056] System suitability requirements: In the system suitability solution, the peak order is triphenylphosphine oxide and starting material, and the resolution between the two should be greater than 2.0.

[0057] Determination method: Take 20 μl of each of the above solutions and inject them into the liquid chromatograph to record the chromatogram (see the attached figure for relevant spectra). Figures 1 to 4 ).

[0058] Above Figure 1 is the chromatogram of the blank solution, indicating that the solvent has no interference with the detection of triphenylphosphine oxide and the starting material; Figure 2 This is the chromatogram of the system suitability solution, which shows that triphenylphosphine oxide peaks at about 10 minutes and the starting material peaks at about 21 minutes, and the resolution between the two is 4.8, which can achieve baseline separation; Figure 3 This is the chromatogram of the reference solution, indicating that triphenylphosphine oxide peaks at about 10 minutes; Figure 4 This is the chromatogram of the test solution, which shows that the starting material peaks at about 21 minutes and triphenylphosphine oxide is not detected.

[0059] If a triphenylphosphine oxide peak is detected in the chromatogram of the limit test solution, it should not exceed 0.036% when calculated by the external standard method.

[0060] Example 2

[0061] Specificity and instrument precision: examine the interference and separation between substances, as well as the stability of the instrument.

[0062] 1. Solution preparation:

[0063] ①Solvent: anhydrous ethanol.

[0064] ②Blank solution: anhydrous ethanol.

[0065] ③Location Solution 1~⑥Location Solution 4: Take about 1.0 mg of impurity 1, impurity 2, impurity 3 and impurity 4 reference substances, accurately weigh, place in different 100 ml volumetric flasks, add solvent to dissolve and dilute to the scale, shake well, and use as location solution 1~location solution 4.

[0066] Note: The reason for adding positioning solutions 1 to 4 to the specificity investigation is that impurities 1 to 4 exist in the starting materials, and they are studied and strictly controlled in the relevant substances under the inspection items. In order to prevent the impurities from interfering with the triphenylphosphine oxide method under the detection conditions of triphenylphosphine oxide, and to make the method more specific, the impurities are positioned under this method (to ensure that each impurity is completely peaked, positioning solutions 1 to 4 are run for 60 minutes).

[0067] ⑦Triphenylphosphine oxide positioning solution: Take about 4 mg of triphenylphosphine oxide reference substance, accurately weigh it, put it in a 100 ml volumetric flask, add solvent to dissolve and dilute to the scale, shake well, and obtain.

[0068] ⑧System suitability solution: Take about 100 mg of the starting material, accurately weigh it, put it in a 10 ml volumetric flask, dissolve it with an appropriate amount of solvent by ultrasonication, accurately measure 1 ml of triphenylphosphine oxide positioning solution and add it, add solvent to dilute to the scale, shake well, and obtain.

[0069] ⑨ Reference solution: Accurately measure 5 ml of triphenylphosphine oxide fixing solution, place it in a 50 ml volumetric flask, dilute to the scale with solvent, shake well, and obtain.

[0070] ⑩ Test solution: Take about 100 mg of the starting material, weigh it accurately, put it in a 10 ml volumetric flask, add solvent and ultrasonically dissolve it and dilute it to the scale, shake well, and you have it.

[0071] 2. Determination method

[0072] Take the above solutions, inject them into liquid chromatograph respectively, and record the chromatogram.

[0073] 3. The results are shown in Tables 1 and 2

[0074] Table 1 Specificity verification results

[0075]

[0076] Table 2 Instrument precision verification results

[0077]

[0078] 4. Conclusion

[0079] The solvent and impurities did not interfere with the determination of the impurity triphenylphosphine oxide; in the system suitability solution, the peak separation between the impurity triphenylphosphine oxide and the starting material was 4.830, which was greater than 2.0; the peak area RSD value of the reference solution after 6 consecutive injections was 4.3%, which was less than 5.0%.

[0080] This method has strong specificity and high instrument precision, which proves that the instrument system is stable and can ensure data reliability.

[0081] Example 3

[0082] Solution stability: Investigate the solution stability of system suitability solution, reference solution, test solution and mixed solution at room temperature.

[0083] 1. Solution preparation

[0084] System suitability solution: Continue to use the specific properties

[0085] Reference solution: continue to use the specificity item

[0086] Test solution: continue to use the specificity item

[0087] 2. Determination method

[0088] Take the above solutions, inject them into liquid chromatograph respectively, and record the chromatogram.

[0089] 3. The results are shown in Table 3

[0090] Table 3 Solution stability verification results

[0091]

[0092]

[0093] 4. Conclusion

[0094] Under room temperature conditions, 0 hours to 73 hours: in the system suitability solution, the separation between the triphenylphosphine oxide peak and the starting material peak was 4.803, greater than 2.0; the peak area RSD of triphenylphosphine oxide in the reference solution was 3.8%; the peak area of ​​triphenylphosphine oxide in the system suitability solution was 3.8%; both were not greater than 5.0%; triphenylphosphine oxide was not detected in the test solution.

[0095] The reference and test solutions of this method are stable within 73 hours at room temperature, indicating that the solvent is appropriately selected and is not prone to volatilization and degradation of components. The solution stability time can support the experiment within 73 hours, thereby ensuring the stability and reliability of the data.

[0096] Example 4

[0097] Linearity, limit of quantitation and limit of detection: Investigate the sensitivity of the method and whether the response value is linearly related to the concentration within the set concentration range.

[0098] 1. Solution preparation

[0099] ①Solvent / blank solution: anhydrous ethanol.

[0100] ②Quantitative limit solution: Accurately measure 2.5 ml of the reference solution under the specificity item, place it in a 10 ml volumetric flask, dilute it to the mark with solvent, shake well, and prepare 6 portions in parallel.

[0101] ③ Detection limit solution: Accurately measure 3 ml of the quantitative limit solution, place it in a 10 ml volumetric flask, dilute to the scale with solvent, shake well, and obtain.

[0102] ④ Positioning solution: Take the triphenylphosphine oxide positioning solution under the specificity item.

[0103] ⑤25% linear solution: take the quantitative limit solution.

[0104] ⑥50% linear solution: Accurately measure 0.5 ml of the positioning solution, place it in a 10 ml volumetric flask, dilute it to the scale with solvent, shake well, and you have it.

[0105] ⑦80% linear solution: Accurately measure 0.8 ml of the positioning solution, place it in a 10 ml volumetric flask, dilute it to the scale with solvent, shake well, and you have it.

[0106] ⑧100% linear solution: Accurately measure 1 ml of the positioning solution, place it in a 10 ml volumetric flask, dilute it to the scale with solvent, shake well, and you have it.

[0107] ⑨150% linear solution: Accurately measure 1.5 ml of the positioning solution, place it in a 10 ml volumetric flask, dilute it to the scale with solvent, shake well, and you are done.

[0108] ⑩200% linear solution: Accurately measure 2 ml of the positioning solution, place it in a 10 ml volumetric flask, dilute it to the scale with solvent, shake well, and you are done.

[0109] 2. Determination method

[0110] Take the above solutions, inject them into liquid chromatograph respectively, and record the chromatogram.

[0111] 3. The results are shown in Tables 4 and 5.

[0112] Table 4 Quantitation limit and detection limit verification results

[0113]

[0114] Table 5 Linearity verification results

[0115]

[0116] 4. Conclusion

[0117] In the 6 quantitative limit solutions, the average signal-to-noise ratio of triphenylphosphine oxide was 13.6, not less than 10, and the RSD value of the peak area was 7.49%, less than 8.0%; in the detection limit solution, the signal-to-noise ratio of triphenylphosphine oxide was 6.3, not less than 3; in the linear solution, the peak area and concentration of triphenylphosphine oxide were linear in the concentration range of 1.0236μg / ml to 8.2078μg / ml, the linear equation was y=73618.9321x+22830.4854, the correlation coefficient r was 0.9995, greater than 0.999, and the intercept / Y100% was 7.20, less than 25%.

[0118] This method has high sensitivity, and the peak area and concentration have good linearity within the concentration range of quantitative limit to 200% limit; the triphenylphosphine oxide reference solution is 4ug / ml, and this method can quantitatively detect triphenylphosphine oxide within the concentration range of 1.0236μg / ml to 8.2078μg / ml, proving that the reference concentration is appropriately selected and can be accurately and effectively detected, thereby not underestimating the content of impurities.

[0119] Example 5

[0120] Accuracy and repeatability: examine whether the repeated test results of the sample are consistent; whether the test is accurate within different concentration ranges.

[0121] 1. Solution preparation

[0122] ①Solvent / blank solution: anhydrous ethanol.

[0123] ② System suitability solution: select under the specificity item.

[0124] ③ Reference substance stock solution: Take about 4 mg of triphenylphosphine oxide reference substance, weigh accurately, place in a 100 ml volumetric flask, add solvent to dissolve and dilute to the scale, shake well, and prepare 2 portions in parallel.

[0125] ④ Reference substance solution: Accurately measure 5.0 mg of reference substance stock solution, place in a 50 ml volumetric flask, add solvent to dissolve and dilute to the scale, shake well, and prepare 2 portions in parallel.

[0126] ⑤ Test solution: Take about 100 mg of the starting material, weigh accurately, place in a 10 ml volumetric flask, add solvent and ultrasonically dissolve and dilute to the scale, shake well, and prepare 3 portions in parallel.

[0127] ⑥25% recovery rate solution: Take 100 mg of the starting material, weigh it accurately, put it in a 10 ml volumetric bottle, add an appropriate amount of solvent and sonicate to dissolve it, accurately measure 2.5 ml of the reference solution, add it, dilute it to the scale with solvent, shake well, and you have it. Prepare 3 copies in parallel.

[0128] ⑦100% recovery solution: Take 100 mg of the starting material, weigh it accurately, put it in a 10 ml volumetric bottle, add an appropriate amount of solvent and sonicate to dissolve it, accurately measure 1.0 ml of the reference stock solution and add it, dilute it to the scale with solvent, shake well, and you have it. Prepare 3 copies in parallel.

[0129] ⑧200% recovery rate solution: Take 100 mg of the starting material, weigh it accurately, put it in a 10 ml volumetric bottle, add an appropriate amount of solvent and sonicate to dissolve it, accurately measure 2.0 ml of the reference stock solution, add it, dilute it to the scale with solvent, shake well, and you have it. Prepare 3 copies in parallel.

[0130] 2. Determination method

[0131] Take the above solutions, inject them into liquid chromatograph respectively, and record the chromatogram.

[0132] 3. The results are shown in Tables 6 and 7.

[0133] Table 6 Accuracy and repeatability verification results 1

[0134] name Detection amount% Mean % Test solution 1 Not detected / Test solution 2 Not detected / Test solution 3 Not detected /

[0135] Table 7 Accuracy and repeatability verification results 2

[0136]

[0137] 4. Conclusion

[0138] In the system suitability solution, the peak separation between triphenylphosphine oxide and the starting material was 4.795, which was greater than 2.0; in the test solution, triphenylphosphine oxide was not detected; in the three 100% recovery solutions, the RSD value of the impurity triphenylphosphine oxide detected was 0.71%, which did not exceed 4.0%. Within the concentration range of 25% (LOQ) to 200% limit, the average recovery of the nine samples was 105.5%, which should be within the range of 85.0% to 110.0%; the RSD value was 1.8%, which did not exceed 10.0%.

[0139] This method has high accuracy and can ensure the accuracy and repeatability of the test results.

[0140] Example 6

[0141] Precision: Under certain conditions, the same batch of samples is tested multiple times to examine the degree of similarity between the test results.

[0142] 1. Repeatability

[0143] 2. Intermediate precision (changing different instruments)

[0144] 1) Solution preparation

[0145] ①Solvent / blank solution: anhydrous ethanol.

[0146] ② Reference substance stock solution: Take about 4 mg of triphenylphosphine oxide reference substance, weigh accurately, place in a 100 ml volumetric flask, add solvent to dissolve and dilute to the scale, shake well, and obtain.

[0147] ③ System suitability solution: Take about 100 mg of the starting material, weigh it accurately, put it in a 10 ml volumetric flask, dissolve it by ultrasonication with an appropriate amount of solvent, accurately measure 1.0 ml of the reference substance stock solution, add it, dilute it to the scale with solvent, shake well, and you have it.

[0148] ④ Reference substance solution: Take 5 ml of reference substance stock solution, weigh accurately, place in a 50 ml volumetric flask, add solvent to dissolve and dilute to the scale, shake well, and prepare 2 portions in parallel.

[0149] ⑤ Test solution: Take about 100 mg of the starting material, weigh accurately, place in a 10 ml volumetric flask, add solvent and ultrasonically dissolve and dilute to the scale, shake well, and prepare 3 copies in parallel.

[0150] ⑥100% spiked solution: Take about 100 mg of the starting material, weigh accurately, place in a 10 ml volumetric flask, add an appropriate amount of solvent and sonicate to dissolve, add 1.0 ml of the reference stock solution, dilute to the mark with solvent, shake well, and prepare 3 portions in parallel.

[0151] 2) Determination method

[0152] Take the above solutions, inject them into liquid chromatograph respectively, and record the chromatogram.

[0153] 3) The results are shown in Tables 8 and 9.

[0154] Table 8 Precision-Verification Results-1

[0155]

[0156] Table 9 Precision-Verification Results 2

[0157]

[0158] 4) Conclusion

[0159] In the 12 test solutions with repeatability and intermediate precision, triphenylphosphine oxide was not detected; in the 6 100% repeatability spiked solutions, the RSD value of triphenylphosphine oxide detected was 1.02%, not exceeding 4.0%; in the 12 spiked solutions with repeatability and intermediate precision, the RSD value of triphenylphosphine oxide detected was 2.59%, not exceeding 8.0%.

[0160] The same batch of samples were tested multiple times, and the test results were all within the qualified range. The results show that this method has good repeatability and precision. It can be used to provide technical support for routine inspections and meet the requirements of system suitability tests, ensuring the repeatability and precision of the method.

[0161] Example 7

[0162] According to the method described in Example 1, multiple batches of samples were tested, and the results showed that none of the samples were detected, indicating that the test results of this method were consistent. See Table 10 below for details.

[0163] Table 10 Multiple batch test results

[0164] batch number Triphenylphosphine oxide content % First batch of samples Not detected The second batch of samples Not detected The third batch of samples Not detected The fourth batch of samples Not detected The fifth batch of samples Not detected The sixth batch of samples Not detected

[0165] Example 8

[0166] Robustness: Examines how well the method tolerates small changes in chromatographic conditions.

[0167] 1. Solution preparation

[0168] ①Solvent / blank solution: anhydrous ethanol.

[0169] ② System suitability solution: select under the specificity item.

[0170] ③ Reference solution: Take reference solution 1 under the accuracy item.

[0171] ④ Test solution: Take test solution 1 under the accuracy item.

[0172] 2. Determination method

[0173] Take the above solutions, inject them into liquid chromatograph respectively, and record the chromatogram.

[0174] 3. The results are shown in Table 11.

[0175] Table 11 Durability test data results

[0176]

[0177] The bold words in the table represent the chromatographic conditions for normal detection.

[0178] 4. Conclusion

[0179] Under different chromatographic conditions: in the system suitability solution, the separation between triphenylphosphine oxide and the starting material peak was greater than 2.0, and the RSD value of the detected amount of triphenylphosphine oxide should not exceed 5.0%; triphenylphosphine oxide was not detected in the test solution.

[0180] When the chromatographic conditions of this method are slightly changed (flow rate ±0.2ml / min, column temperature ±5℃, mobile phase A ratio ±2%, pH ±0.2 and different instruments), the determination is not affected compared with normal conditions. This method has a high tolerance. It can be used to provide technical support for routine inspections and meet the requirements of system suitability tests to ensure the reliability of the method.

Claims

1. A method for detecting triphenylphosphine oxide in a sacubitril-valsartan sodium starting material, wherein: The structural formula of the sacubitril-valsartan sodium starting material is formula (I), and the structural formula of triphenylphosphine oxide is formula (II). The method is characterized in that: high performance liquid chromatography is used for detection, and the chromatographic conditions are: the chromatographic column uses octadecylsilane bonded silica gel as a filler; the mobile phase consists of a mobile phase A and a mobile phase B, the mobile phase A is an ammonium acetate solution, and the mobile phase B is an organic phase.

2. The detection method according to claim 1, characterized in that: The chromatographic conditions at least meet at least one of the following conditions (1) to (9): (1) Chromatographic column: 4.6 mm × 150 mm, 3.5 μm, preferably Waters C18 column, 4.6 mm × 150 mm, 3.5 μm; more preferably Waters Sunfire C18 column, 4.6 mm × 150 mm, 3.5 μm; (2) Mobile phase A is 10-30 mmol / L ammonium acetate solution; preferably 10-30 mmol / L ammonium acetate solution, the pH value is adjusted to 5.7±0.5; more preferably 10-30 mmol / L ammonium acetate solution, the pH value is adjusted to 5.7±0.1 with glacial acetic acid; (3) Mobile phase B is acetonitrile; (4) Mobile phase ratio: the volume ratio of mobile phase A to mobile phase B is 58-62:42-38; (5) Flow rate: 0.8 ml / min to 1.2 ml / min; (6) Column temperature: 30-40°C; (7) Injection volume: 10-30 μl; (8) Detector: UV detector; (9) Detection wavelength: 199-220 nm.

3. The detection method according to any one of claims 1 to 2, characterized in that: Solution preparation: Test solution: Take an appropriate amount of sacubitril / valsartan sodium starting material and add solvent to dissolve and dilute to a solution of a certain concentration; Reference solution: Take an appropriate amount of triphenylphosphine oxide reference substance and add solvent to dissolve and dilute to a solution of a certain concentration; System suitability solution: Take an appropriate amount of sacubitril / valsartan sodium starting material, add an appropriate amount of triphenylphosphine oxide reference substance, and add solvent to dissolve and dilute to a solution of a certain concentration; Preferably, Solution preparation: Test solution: Take about 100 mg of sacubitril-valsartan sodium starting material, weigh accurately, place in a 10 ml volumetric flask, add appropriate amount of solvent, sonicate to dissolve and dilute to scale, shake well; Reference solution: take about 4 mg of triphenylphosphine oxide reference substance, accurately weigh it, put it in a 100 ml volumetric flask, add solvent to dissolve and dilute to the scale, shake well, as the stock solution; accurately measure 5 ml of the stock solution, put it in a 50 ml volumetric flask, dilute to the scale with solvent, and shake well; System suitability solution: Take about 100 mg of sacubitril / valsartan sodium starting material, accurately weigh it, put it in a 10 ml volumetric flask, accurately measure 1 ml of stock solution and add it, add an appropriate amount of solvent, sonicate to dissolve and dilute to the scale, and shake well.

4. The detection method according to claim 3, characterized in that: The solvent is selected from at least one of mobile phase, methanol, chloroform, acetic acid, tetrahydrofuran, dimethyl sulfoxide and ethanol; Preferably, the solvent is ethanol.

5. The detection method according to any one of claims 1 to 4, characterized in that: In the system suitability solution, the elution order was triphenylphosphine oxide and sacubitril-valsartan sodium starting material, and the resolution between the two was greater than 2.

0.

6. The detection method according to any one of claims 1 to 5, characterized in that: The content of triphenylphosphine oxide in the test solution, calculated by external standard method, does not exceed 0.04%; preferably does not exceed 0.036%.

7. The detection method according to any one of claims 1 to 6, characterized in that: In the method, the mean signal-to-noise ratio of the quantitative limit solution (n=6) is not less than 10, preferably 13.6; in the detection limit solution, the mean signal-to-noise ratio is not less than 3, preferably 6.

3.

8. The detection method according to any one of claims 1 to 7, characterized in that: The method can quantitatively detect the concentration range of triphenylphosphine oxide in the range of 1.0236 μg / ml to 8.2078 μg / ml.

9. Use of the detection method according to any one of claims 1 to 8 in detecting triphenylphosphine oxide in sacubitril-valsartan sodium starting materials.

Images