Detection method of related substances in sacubitril valsartan sodium bulk drug and quality control standard of related substances in sacubitril valsartan sodium bulk drug

Abstract

The invention relates to a detection method of sacubitril valsartan sodium related substances and a quality control standard of the sacubitril valsartan sodium related substances, and belongs to the technical field of pharmaceutical analysis. In order to accurately reflect the amount of related substances in sacubitril-valsartan sodium, a reasonable basis can be provided for formulating a quality standard, so that the product quality can be better controlled and mastered, and the safety and effectiveness of clinical medication are improved. The invention provides a method for detecting related substances in sacubitril valsartan sodium and a quality control standard of the related substances in sacubitril valsartan sodium. The detection method has high accuracy and precision and strong durability, and is suitable for industrial inspection and quality control of related substances in sacubitril valsartan sodium.

Description

Technical Field

[0001] The invention belongs to the technical field of drug analysis and relates to a detection method for related substances in a sacubitril-valsartan sodium raw material drug and a quality control standard thereof. Background Art

[0002] Sacubitril-valsartan sodium is an angiotensin receptor neprilysin inhibitor (ARNI) developed by Novartis. It releases sacubitril and valsartan simultaneously in the body to exert its effects, and its therapeutic effect is much higher than the simple combination of valsartan and sacubitril. On the one hand, it inhibits neprilysin (neutral endopeptidase; NEP) through the active metabolite of sacubitril, LBQ657, to achieve the effects of sodium excretion, diuresis, vasodilation and heart protection. On the other hand, it blocks the type 1 receptor (AT1) of angiotensin II through valsartan to achieve vasodilation and reduce the heart load. This drug is mainly used to treat chronic heart failure and hypertension.

[0003] Sacubitril / valsartan sodium is the first supramolecular complex that can regulate different pharmacological targets. Its mechanism of action in vivo is unique in the medical field. Through the dual mechanism and synergistic effect of these two components, it can effectively treat chronic heart failure with reduced ejection fraction. Its effectiveness and safety have been confirmed by large-scale clinical studies such as PARADIGM-HF.

[0004] The structural formula of sacubitril valsartan sodium is as follows:

[0005]

[0006] The raw materials of sacubitril-valsartan sodium may lead to the formation of impurities during the synthesis, production and storage, and introduce the following 17 impurities:

[0007]

[0008]

[0009] Among the above impurities, FSVS-SM3-IMP-F is the chiral isomer of valsartan, and FSVS-IMP-A, FSVS-IMP-B, and FSVS-IMP-C are the chiral isomers of sacubitril.

[0010] FSVS-SM1 is the starting material for producing sacubitril, FSVS-II is the remaining intermediate residue, FSVS-SM3-IMP-A is the intermediate for synthesizing FSVS-SM3, FSVS-SM3-IMP-E is a process impurity generated by the reaction of FSVS-SM3-IMP-A with butyryl chloride contained in valeryl chloride, FSVS-IV-IMP-C and FSVS-IV-IMP-F are the impurity transfer reactions of FSVS-SM1-IMP-E and may also be derived from the hydrolysis reaction of sacubitril, FSVS-IV-IMP-B is generated by the impurity transfer reaction of FSVS-II-IMP-A, FSVS-SM1-IMP-D is an impurity generated by the cyclization reaction of FSVS-SM1 and may also come from the degradation of sacubitril / valsartan at high temperature. FSVS-SM3-IMP-C is an intermediate in the synthesis of FSVS-SM3. FSVS-IV-IMP-A is generated by the impurity transfer reaction of FSVS-SM1-IMP-D. FSVS-IV-IMP-D is generated by the impurity transfer reaction of FSVS-SM1-IMP-D. FSVS-SM3-IMP-B is an intermediate in the synthesis of FSVS-SM3. FSVS-SM3-IMP-D is a process impurity in the synthesis of FSVS-SM3.

[0011] In the complex preparation process, sacubitril valsartan sodium will produce a variety of impurities with similar structures, which is difficult to separate. In addition, Chinese patent application CN112611813A discloses a method for testing genotoxic impurities of sacubitril valsartan sodium starting materials. Although the method can sensitively, accurately and precisely measure the genotoxic impurities, it is impossible to simultaneously measure the content of the effective ingredient of sacubitril valsartan sodium, and only the genotoxic impurities can be measured. Other related substances contained in sacubitril valsartan sodium cannot be measured, and the limitations are relatively large in practical applications. Chinese patent application CN116953122A discloses a method for detecting impurities in sacubitril valsartan sodium. This method uses a trifluoroacetic acid system to detect 10 impurities in sacubitril valsartan sodium, but the 10 impurities studied are impurities that may be produced under its corresponding preparation process, and trifluoroacetic acid itself has a certain ion pair effect, which has a certain degree of influence on the chromatographic column and the chromatographic system. In order to improve the purity of sacubitril-valsartan sodium products and control the quality of sacubitril-valsartan sodium products, it is urgently necessary to provide an analytical method with a simple system, high sensitivity, high precision and good specificity, which can detect sacubitril, valsartan and various sacubitril-valsartan sodium impurities under the same conditions, so as to effectively control the quality of sacubitril-valsartan sodium raw materials in subsequent processes. Summary of the invention

[0012] In order to accurately reflect the situation of related substances in sacubitril valsartan sodium raw materials, a reasonable basis can be provided for the formulation of quality standards, so as to better control and master product quality, thereby improving the safety and effectiveness of products. The present invention provides a detection method for related substances in sacubitril valsartan sodium raw materials and a quality control standard thereof. The detection method has high accuracy and precision, strong durability, and is suitable for industrial use in the inspection and quality control of related substances in sacubitril valsartan sodium raw materials.

[0013] The present invention achieves the above object through the following technical solutions:

[0014] The method for detecting related substances in sacubitril-valsartan sodium raw material comprises the following steps:

[0015] A method for detecting related substances in sacubitril-valsartan sodium, using high performance liquid chromatography for detection, wherein the

[0016] The conditions for HPLC were:

[0017] Chromatographic column: octadecylsilane bonded silica gel, detection wavelength: 245nm~255nm, flow rate: 0.8ml / min~1.2ml / min, column temperature: 25℃~35℃, mobile phase A: 0.01mol / L ammonium dihydrogen phosphate aqueous solution (add 1.3ml~1.7ml glacial acetic acid to every 1000ml), mobile phase B: methanol-acetonitrile (85:15~95:5), detection is carried out under the following gradient elution conditions.

[0018]

[0019] The technology of the present invention can simultaneously detect the active ingredients valsartan and sacubitril in sacubitril-valsartan sodium, as well as 13 impurity compounds with similar structures under the above-mentioned specific and identical chromatographic conditions, and can completely separate and accurately quantify various process impurities and degradation impurities in sacubitril-valsartan sodium. The technology has a fast detection speed, does not involve special detection equipment, chromatographic columns or reagents, has a wide range of applications, and can better control the quality of sacubitril-valsartan sodium raw materials.

[0020] Furthermore, the chromatographic column is Waters XBridge Shield RP18 or Phenomenex GeminiC18.

[0021] Preferably, the specifications of the chromatographic column are 4.6 mm×250 mm, 5 μm; or other specifications with equivalent energy efficiency.

[0022] Furthermore, in the detection method, the test sample, the reference sample or the control is prepared into a solution with acetonitrile-water (1:1) and then detected.

[0023] Furthermore, the detection method simultaneously detects valsartan, sacubitril and 13 impurities, and the structures of the 13 impurities are shown in Table 1.

[0024] Table 1: Impurity structure table

[0025]

[0026]

[0027] 1) Preparation of system suitability solution: Take appropriate amounts of this product and FSVS-IV-IMP-C, add acetonitrile-water (1:1) to dissolve and quantitatively dilute to prepare a mixed solution containing approximately 1 mg of sacubitril-valsartan sodium and approximately 3 μg of FSVS-IV-IMP-C per 1 ml.

[0028] 2) Preparation of test solution: Take an appropriate amount of the product, weigh accurately, add acetonitrile-water (1:1) to dissolve and quantitatively dilute to make a solution containing about 1 mg per 1 ml.

[0029] 3) Preparation of control solution: Accurately measure an appropriate amount of the test solution and quantitatively dilute it with acetonitrile-water (1:1) to make a solution containing approximately 1 μg per 1 ml.

[0030] 4) Chromatographic conditions: octadecylsilane bonded silica gel was used as filler; mobile phase A was 0.01 mol / L aqueous ammonium dihydrogen phosphate solution (1.5 ml of glacial acetic acid was added to every 1000 ml), and mobile phase B was methanol-acetonitrile (9:1); the flow rate was 1.0 ml per minute, and linear gradient elution was performed; the column temperature was 30°C; the detection wavelength was 250 nm; and the injection volume was 20 μl.

[0031] 5) System suitability requirements: In the system suitability solution spectrum, the order of peak elution of each component is FSVS-IV-IMP-C, valsartan, and sacubitril, and the separation between each peak should meet the requirements.

[0032] 6) Determination of related substances in sacubitril-valsartan sodium raw material: Accurately measure the test solution and the reference solution, inject them into the liquid chromatograph respectively, and record the chromatogram. If there are impurity peaks in the chromatogram of the test solution, calculate according to the main component self-reference method with the correction factor.

[0033] The above-mentioned method for detecting related substances in sacubitril-valsartan sodium raw material medicine specifically comprises the following steps:

[0034] 1) Preparation of system suitability solution: Take 3 mg of FSVS-IV-IMP-C reference substance, place it in a 50 ml volumetric flask, dissolve it with acetonitrile-water (1:1) and dilute it to the mark. Accurately measure 1 ml and place it in a 20 ml volumetric flask to which 23 mg of sacubitril-valsartan trisodium reference substance has been added, add acetonitrile-water (1:1) to dissolve it and dilute it to the mark. Shake well to prepare the system suitability solution.

[0035] 2) Preparation of test solution: Take 20 mg of the product, weigh accurately, place in a 20 ml volumetric flask, add acetonitrile-water (1:1) to dissolve and quantitatively dilute to the scale, shake well, and use as the test solution.

[0036] 3) Preparation of control solution: Accurately measure 0.1 ml of the test solution, place it in a 100 ml volumetric flask, add acetonitrile-water (1:1) to dilute to the scale, shake well, and use as the control solution.

[0037] 4) Chromatographic conditions: Use octadecylsilane bonded silica gel as filler (Waters XBridge Shield RP18, 4.6 mm × 250 mm, 5 μm or chromatographic column with equivalent performance); mobile phase A is 0.01 mol / L ammonium dihydrogen phosphate aqueous solution (1.5 ml of glacial acetic acid is added to every 1000 ml), and mobile phase B is methanol-acetonitrile (9:1); the flow rate is 1.0 ml per minute, and linear gradient elution is performed according to the table below; the column temperature is 30°C; the detection wavelength is 250 nm; and the injection volume is 20 μl.

[0038]

[0039] 5) System suitability requirements: In the system suitability solution spectrum, the order of peak elution of each component is FSVS-IV-IMP-C, valsartan, and sacubitril, and the separation between each peak should meet the requirements.

[0040] 6) Determination of related substances in sacubitril-valsartan sodium raw material: Accurately measure 20 μl of the test solution and the control solution, respectively, inject them into the liquid chromatograph, and record the chromatogram. If there are impurity peaks in the chromatogram of the test solution, calculate them according to the main component self-control method with the correction factor.

[0041] The present invention also provides a quality control standard for related substances in a sacubitril-valsartan sodium bulk drug, wherein the related substances are determined by the above-mentioned detection method. When the above-mentioned detection method is used for determination, the limits of the related substances are as follows: the peak area of ​​FSVS-IV-IMP-C (FSVS-IV-IMP-F co-eluted therewith) multiplied by 0.4506 shall not exceed 0.3% by the self-control method based on the peak area of ​​sacubitril, the peak area of ​​other single impurities multiplied by 0.4770 shall not exceed 0.10% by the self-control method based on the peak area of ​​valsartan, except for the FSVS-IV-IMP-C peak, the sum of all impurities shall not exceed 0.3%, and the total amount of each impurity shall not exceed 0.6%. Other impurity peaks less than 0.01% can be ignored.

[0042] Experimental Example 1: Selection of Detection Wavelength

[0043] FSVS-SM1, FSVS-SM1-IMP-D, FSVS-II, FSVS-SM3-IMP-A, FSVS-SM3-IMP-B, FSVS-SM3-IMP-C, FSVS-SM3-IMP-D, FSVS-SM3-IMP-E, FSVS-SM3-IMP-F, FSVS-IV-IMP-A, FSVS-IV-IMP-B, FSVS-IV-IMP-C, FSVS-IV-IMP-D, FSVS-IV-IMP-F, FSVS-IMP-A, FSVS-IMP-B and FSVS-IMP-C were respectively prepared into appropriate concentrations with acetonitrile-water (1:1) and scanned with a PAD detector in the wavelength range of 400nm to 200nm. Each component had a large absorption at a wavelength of 250nm, so the wavelength was finally set to 250nm.

[0044] Experimental Example 2: Selection of Chromatographic Conditions

[0045] Table 2: Selection of chromatographic conditions for determination of related substances in sacubitril-valsartan sodium API

[0046]

[0047]

[0048]

[0049] The experimental results show that, based on the exploration and optimization process of the above-mentioned chromatographic conditions, condition 6 was determined as the chromatographic condition for the detection of related substances in sacubitril valsartan sodium raw materials. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] The attached figure is a spectrum of the mixed solution of each component detected in the sacubitril-valsartan sodium raw material. The peak order is FSVS-SM3-IMP-A, FSVS-II, FSVS-SM3-IMP-E, FSVS-IV-IMP-C, main peak 1 valsartan, FSVS-IV-IMP-B, FSVS-SM1-IMP-D, FSVS-SM3-IMP-C, main peak 2 sacubitril, FSVS-IMP-B / C, FSVS-IV-IMP-A, FSVS-IV-IMP-D, FSVS-SM3-IMP-B, FSVS-SM1, FSVS-SM3-IMP-D, among which FSVS-IMP-B / C is controlled under the isomer inspection item and is not quantified under the related substances. DETAILED DESCRIPTION

[0051] The present invention is further described in detail by the following examples, including but not limited to the following examples.

[0052] Embodiment 1: Methodological investigation of the method for detecting related substances in the sacubitril-valsartan sodium bulk drug of the present invention

[0053] 1.1 Method specificity test

[0054] Take appropriate amounts of FSVS-SM1, FSVS-SM1-IMP-D, FSVS-II, FSVS-SM3-IMP-A, FSVS-SM3-IMP-B, FSVS-SM3-IMP-C, FSVS-SM3-IMP-D, FSVS-SM3-IMP-E, FSVS-SM3-IMP-F, FSVS-IV-IMP-A, FSVS-IV-IMP-B, FSVS-IV-IMP-C, FSVS-IV-IMP-D, FSVS-IMP-A, FSVS-IMP-B, and FSVS-IMP-C, add solvent to dissolve and dilute to appropriate concentrations, as positioning solutions for each component;

[0055] System suitability solution: Take appropriate amounts of sacubitril / valsartan sodium, FSVS-SM1, FSVS-SM1-IMP-D, FSVS-II, FSVS-SM3-IMP-A, FSVS-SM3-IMP-B, FSVS-SM3-IMP-C, FSVS-SM3-IMP-D, FSVS-SM3-IMP-E, FSVS-SM3-IMP-F, FSVS-IV-IMP-A, FSVS-IV-IMP-B, FSVS-IV-IMP-C, FSVS-IV-IMP-D, FSVS-IMP-A, FSVS-IMP-B, and FSVS-IMP-C, add solvent to dissolve them, and quantitatively dilute to make a 1 ml solution. l contains about 1 mg of sacubitril-valsartan sodium, and a mixed solution of about 1 μg each of FSVS-SM1, FSVS-SM1-IMP-D, FSVS-II, FSVS-SM3-IMP-A, FSVS-SM3-IMP-B, FSVS-SM3-IMP-C, FSVS-SM3-IMP-D, FSVS-SM3-IMP-E, FSVS-SM3-IMP-F, FSVS-IV-IMP-A, FSVS-IV-IMP-B, FSVS-IV-IMP-C, FSVS-IV-IMP-D, FSVS-IMP-A, FSVS-IMP-B, and FSVS-IMP-C is used as the system suitability solution;

[0056] Accurately measure 20 μl of each component positioning solution and system suitability solution, inject them into the liquid chromatograph, and record the chromatogram. The results are shown in Table 3.

[0057] Table 3: System suitability results table

[0058]

[0059] Conclusion: In the system suitability results table 1, the separation degree between FSVS-IMP-B / C and the main peak 2 Sacubitril is 1.403, and the minimum separation degree between the other components is 1.593. Since FSVS-IMP-B / C and the main peak 2 Sacubitril are diastereomers, FSVS-IMP-A and the main peak 2 Sacubitril are enantiomers, so the two overlap; FSVS-SM3-IMP-F and the main peak 1 valsartan are enantiomers, so the two also overlap; therefore, the four impurities are investigated under normal phase conditions, and the peak separation degree of FSVS-IMP-B / C and Sacubitril is greater than 1.2, and the minimum separation degree of the other impurities is greater than 1.5, which meets the requirements. This shows that the method has good specificity.

[0060] 1.2 Forced degradation test

[0061] Table 4: Summary of forced degradation methods

[0062]

[0063] According to the chromatographic conditions of the above-mentioned method for detecting related substances in sacubitril-valsartan sodium raw materials, 20 μl of blank solvent, undamaged sample solution and the above-mentioned damaged solution were accurately measured and injected into the liquid chromatograph for detection with a PDA detector. The results are shown in Tables 5 and 6.

[0064] Table 5: Forced degradation test results table-1

[0065]

[0066]

[0067] Table 6: Forced degradation test results table-2

[0068]

[0069] Conclusion: According to the damage of this product under various conditions, the results are summarized as follows:

[0070] The blank solvent has no interference with the determination of related substances, and the blank solution has no interference with the detection. New degradation products are detected under strong degradation conditions such as alkali, high temperature, and oxidation. Each degradation product can be completely separated from the main peak and does not interfere with the detection of other known impurities. After peak purity inspection, the main peak purity similarity under each condition is less than the purity threshold, indicating that the peak purity is qualified, verifying the good specificity of the chromatographic conditions. The material balance under the above-mentioned destruction conditions was calculated by the external standard method with the undestroyed content as 100%, and the material conservation further verified the good specificity of the chromatographic conditions.

[0071] 1.3 Limit of detection and limit of quantification test

[0072] Take appropriate amounts of valsartan reference substance, sacubitril calcium reference substance, FSVS-SM1, FSVS-SM1-IMP-D, FSVS-II, FSVS-SM3-IMP-A, FSVS-SM3-IMP-B, FSVS-SM3-IMP-C, FSVS-SM3-IMP-D, FSVS-SM3-IMP-E, FSVS-IV-IMP-A, FSVS-IV-IMP-B, FSVS-IV-IMP-C, FSVS-IV-IMP-F, and FSVS-IV-IMP-D, add solvent to dissolve and dilute step by step to appropriate concentrations. Under the above chromatographic conditions, the concentration at a signal-to-noise ratio of about 3:1 is used as the detection limit, and the concentration at a signal-to-noise ratio of about 10:1 is used as the quantification limit. The test results are shown in Table 7.

[0073] Table 7: Detection limit and quantification limit results

[0074]

[0075] Conclusion: The proposed concentration of the test solution is 1 mg / ml, the detection limit concentration is equivalent to less than 0.0057% of the main component percentage, and the quantification limit concentration is equivalent to less than 0.0189% of the main component percentage; therefore, each impurity can be accurately quantified above 0.0189%, and this method has high sensitivity.

[0076] 1.4 Linearity and range

[0077] Take appropriate amount of valsartan reference substance, sacubitril calcium reference substance, FSVS-SM1, FSVS-SM1-IMP-D, FSVS-II, FSVS-SM3-IMP-E, and FSVS-IV-IMP-C reference substances, add solvent to dissolve and dilute step by step to a series of appropriate concentrations, as linear test solutions 1-1 to 5. Take appropriate amount of FSVS-IV-IMP-F reference substance, add solvent to dissolve and dilute step by step to a series of appropriate concentrations, as linear test solutions 2-1 to 5. Accurately measure 20 μl of each, inject into liquid chromatograph respectively, record chromatograms, and the test results are shown in Tables 8 to 15.

[0078] Table 8: FSVS-II linearity test results

[0079]

[0080]

[0081] Table 9: FSVS-SM3-IMP-E linear test results

[0082]

[0083] Table 10: FSVS-IV-IMP-C linearity test results

[0084]

[0085] Table 11: FSVS-IV-IMP-F linearity test results

[0086]

[0087] Table 12: Valsartan linearity test results

[0088]

[0089] Table 13: FSVS-SM1-IMP-D linearity test results

[0090]

[0091] Table 14: Shakuba curve test results

[0092]

[0093] Table 15: FSVS-SM1 linearity test results

[0094]

[0095]

[0096] Conclusion: The components have good linear relationship within the above range.

[0097] 1.5 Correction Factor

[0098] According to the provisions of Part 4 of the 2020 edition of the Chinese Pharmacopoeia, the correction factor formula can be defined as:

[0099]

[0100] According to the requirements for the calculation of the correction factor, the correction factor should be verified by multi-point calculation, that is, "A / C" is defined as the slope of the curve. Referring to the linear test and regression linear equation of the above-mentioned sacubitril valsartan sodium raw material, the slope of the peak area of ​​each chromatographic peak to the concentration is obtained within a given concentration range and compared with the slope of the sacubitril valsartan sodium raw material. The calculation formula is as follows:

[0101]

[0102] Where: Slope: Impurity: The slope of the linear curve of impurity peak area versus concentration

[0103] Slope Sacubitril-Valsartan Sodium API: Slope of the linear curve of peak area versus concentration of Sacubitril-Valsartan Sodium API

[0104] (Note: FSVS-SM3-IMP-E is calculated using the slope of valsartan, and the other impurities are calculated using the slope of sacubitril.)

[0105] The calculation of the correction factor refers to the above linear investigation, and the slope of the linear equation is used to calculate the correction factor. The linear concentration range and linear equation for calculating the correction factor are summarized in Table 16.

[0106] Table 16: Correction factor results table

[0107] Element Linear equations Slope Correction Factor FSVS-II y=80826.7772x-563.0061 80826.7772 0.77 FSVS-SM3-IMP-E y=37430.1994x-111.0785 37430.1994 0.99 FSVS-IV-IMP-C y=64551.3773x-911.7353 64551.3773 0.97 FSVS-IV-IMP-F y=67442.2755x+1033.2972 67442.2755 0.92 Valsartan y=37065.1000x+253.0070 37065.1000 / FSVS-SM1-IMP-D y=101572.8651x-879.8380 101572.8651 0.61 Shakubaqu y=62383.9854x+401.9576 62383.9854 / FSVS-SM1 y=66476.0041x+806.6441 66476.0041 0.94

[0108] Conclusion: The correction factors of impurities FSVS-SM3-IMP-E, FSVS-IV-IMP-C, FSVS-IV-IMP-F, and FSVS-SM1 are all between 0.9 and 1.1, so no correction is required. The correction factors of FSVS-II and FSVS-SM1-IMP-D are not between 0.9 and 1.1, but between 0.2 and 3.0, so they can be corrected using the correction factors. Therefore, the quantitative method can adopt the principal component self-control method with the correction factor added.

[0109] 1.6 Accuracy test

[0110] Reference substance stock solution ①: Take appropriate amount of FSVS-SM1, FSVS-SM1-IMP-D, FSVS-II, FSVS-SM3-IMP-E, FSVS-IV-IMP-C, and FSVS-IV-IMP-F, accurately weigh them, add solvent to dissolve them, and quantitatively dilute them to make a mixed solution containing 10 μg of each component per 1 ml, shake well, and use it as reference substance stock solution ①;

[0111] Reference substance stock solution ②: Take an appropriate amount of FSVS-IV-IMP-F, weigh accurately, add solvent to dissolve and quantitatively dilute to make a solution containing 10 μg of FSVS-IV-IMP-F per 1 ml, shake well, and use it as reference substance stock solution ②;

[0112] Reference substance solution 1: Take 2 ml of reference substance stock solution ① and dilute it to 20 ml with solvent as reference substance solution 1.

[0113] Reference substance solution 2: Take 2 ml of reference substance stock solution ② and dilute it to 20 ml with solvent as reference substance solution 2.

[0114] Test solution: Take about 20 mg of this product (batch L201907001), place in a 20 ml volumetric flask, and add 1 ml, 2 ml, and 3 ml of reference stock solution ① respectively, dilute to the mark with solvent, shake well, and use as the test solution. Prepare 3 copies of each concentration in parallel.

[0115] Test solution: Take about 20 mg of this product (L201907001 batch), place in a 20 ml volumetric flask, and add 1 ml, 2 ml, and 3 ml of reference substance stock solution ② respectively, dilute to the scale with solvent, shake well, and use as the test solution. Prepare 3 copies of each concentration in parallel.

[0116] Control solution: accurately measure the test solution and quantitatively dilute it with solvent to make a solution containing 1 μg of the main component per 1 ml, shake well, and use it as the control solution;

[0117] Under the above chromatographic conditions, accurately measure 20 μl of reference solution 1, reference solution 2, control solution and test solution, respectively, and inject them into the liquid chromatograph. Record the chromatograms, and calculate the recovery of each impurity by the external standard method and the principal component self-reference method with correction factor. The test results are shown in Tables 17 to 22.

[0118] Table 17: FSVS-II recovery test results

[0119]

[0120]

[0121] Table 18: FSVS-SM3-IMP-E recovery test results

[0122]

[0123] Table 19: FSVS-IV-IMP-C recovery test results

[0124]

[0125]

[0126] Table 20: FSVS-IV-IMP-F recovery test results

[0127]

[0128] Table 21: FSVS-SM1-IMP-D recovery test results

[0129]

[0130]

[0131] Table 22: FSVS-SM1 recovery test results

[0132]

[0133] Conclusion: The results of the external standard method and the main component self-control method with correction factors showed that the average recovery rate of each impurity was between 90% and 108%, indicating that the accuracy of this method is good. It also shows that the main component self-control method with correction factors can be accurately quantified.

[0134] 1.7 Precision test

[0135] 1.7.1 Injection precision test

[0136] Reference solution: Take appropriate amount of valsartan reference substance, sacubitril calcium reference substance, FSVS-SM1, FSVS-SM1-IMP-D, FSVS-II, FSVS-SM3-IMP-E, FSVS-IV-IMP-C, accurately weigh, add acetonitrile-water (1:1) to dissolve and dilute to make a mixed solution containing 1 μg of each component per 1 ml, shake well, as the reference solution, accurately measure 20 μl respectively and inject into the liquid chromatograph 6 times continuously, and record the chromatogram. The test results are shown in Tables 23 and 24.

[0137] Table 23: Peak area results of injection precision test

[0138]

[0139] Table 24: Injection precision test retention time results

[0140]

[0141] Conclusion: The test results show that the injection precision is good.

[0142]

[0143]

[0144]

[0145]

[0146]

[0147]

[0148]

[0149]

[0150]

[0151]

[0152]

[0153]

[0154] Example 2 The present invention discloses a method for detecting related substances in a sacubitril-valsartan sodium bulk drug and a quality inspection method thereof. A method for detecting related substances in a sacubitril-valsartan sodium bulk drug specifically comprises the following steps:

[0155] 1) Preparation of system suitability solution: Take 3 mg of FSVS-IV-IMP-C reference substance, place it in a 50 ml volumetric flask, dissolve it with acetonitrile-water (1:1) and dilute it to the mark. Accurately measure 1 ml and place it in a 20 ml volumetric flask to which 23 mg of sacubitril-valsartan trisodium reference substance has been added, add acetonitrile-water (1:1) to dissolve it and dilute it to the mark. Shake well to prepare the system suitability solution.

[0156] 2) Preparation of test solution: Take 20 mg of the product, weigh accurately, place in a 20 ml volumetric flask, add acetonitrile-water (1:1) to dissolve and quantitatively dilute to the scale, shake well, and use as the test solution.

[0157] 3) Preparation of control solution: Accurately measure 0.1 ml of the test solution, place it in a 100 ml volumetric flask, add acetonitrile-water (1:1) to dilute to the scale, shake well, and use as the control solution.

[0158] 4) Chromatographic conditions: Use octadecylsilane bonded silica gel as filler (Waters XBridge Shield RP18, 4.6 mm × 250 mm, 5 μm or chromatographic column with equivalent performance); mobile phase A is 0.01 mol / L ammonium dihydrogen phosphate aqueous solution (1.5 ml of glacial acetic acid is added to every 1000 ml), and mobile phase B is methanol-acetonitrile (9:1); the flow rate is 1.0 ml per minute, and linear gradient elution is performed according to the table below; the column temperature is 30°C; the detection wavelength is 250 nm; and the injection volume is 20 μl.

[0159]

[0160] 5) System suitability requirements: In the system suitability solution spectrum, the order of peak elution of each component is FSVS-IV-IMP-C, valsartan, and sacubitril, and the separation between the peaks should meet the requirements.

[0161] 6) Determination of related substances in sacubitril-valsartan sodium raw material: Accurately measure 20 μl of the test solution and the control solution, respectively, inject them into the liquid chromatograph, and record the chromatogram. If there are impurity peaks in the chromatogram of the test solution, calculate them according to the main component self-control method with the correction factor.

[0162] The present invention also provides a quality control standard for related substances in a sacubitril-valsartan sodium bulk drug, wherein the related substances are determined by the above-mentioned detection method. When the above-mentioned detection method is used for determination, the peak area of ​​FSVS-IV-IMP-C (FSVS-IV-IMP-F co-eluted therewith) multiplied by 0.4506 shall not exceed 0.3% by the self-control method based on the peak area of ​​sacubitril, and the peak area of ​​other single impurities multiplied by 0.4770 shall not exceed 0.10% by the self-control method based on the peak area of ​​valsartan, except for the FSVS-IV-IMP-C peak, the sum of all impurities shall not exceed 0.3%, and the total amount of each impurity shall not exceed 0.6%. Other impurity peaks less than 0.01% can be ignored.

[0163] The results of the inspection of related substances in multiple batches of samples and stability samples using the above detection method are shown in Tables 37 to 40.

[0164] Table 37: Test results of related substances in sacubitril-valsartan sodium API

[0165]

[0166] Table 38: Results of related substance inspection in samples of sacubitril-valsartan sodium API influencing factors

[0167]

[0168]

[0169] Table 39: Results of related substances testing in samples of sacubitril-valsartan sodium API accelerated stability test

[0170]

[0171]

[0172] Table 40: Results of related substances test in samples of sacubitril-valsartan sodium API long-term stability test

[0173]

[0174]

[0175]

[0176] Conclusion: The related substances in multiple batches of samples of this product are all below the limit of 30%, and no significant changes were observed in the stability study. In order to better control the quality of the product and ensure the safety of medication, the related substances are included in the final quality standards. The limits of related substances are as follows: the peak area of ​​FSVS-IV-IMP-C (FSVS-IV-IMP-F co-eluted with it) multiplied by 0.4506 shall not exceed 0.3% by the self-control method based on the peak area of ​​sacubitril; the peak area of ​​other individual impurities multiplied by 0.4770 shall not exceed 0.10% by the self-control method based on the peak area of ​​valsartan; except for the FSVS-IV-IMP-C peak, the sum of all impurities shall not exceed 0.3%, and the total amount of each impurity shall not exceed 0.6%.

Claims

1. A method for detecting related substances of sacubitril-valsartan sodium and its quality control standard, characterized in that The steps include: 1) Preparation of system suitability solution: Take appropriate amount of this product and FSVS-IV-IMP-C, add acetonitrile-water (1:1) to dissolve and quantitatively dilute to make a mixed solution containing about 1 mg of sacubitril-valsartan sodium and about 3 μg of FSVS-IV-IMP-C per 1 ml; 2) Preparation of test solution: Take an appropriate amount of the product, weigh accurately, add acetonitrile-water (1:1) to dissolve and quantitatively dilute to make a solution containing about 1 mg per 1 ml; 3) Preparation of control solution: Accurately measure an appropriate amount of the test solution and quantitatively dilute it with acetonitrile-water (1:1) to make a solution containing approximately 1 μg per 1 ml; 4) Chromatographic conditions: octadecylsilane bonded silica gel was used as filler; mobile phase A was 0.01 mol / L ammonium dihydrogen phosphate aqueous solution (1.5 ml of glacial acetic acid was added to every 1000 ml), and mobile phase B was methanol-acetonitrile (9:1); the flow rate was 1.0 ml per minute, and linear gradient elution was performed; the column temperature was 30°C; the detection wavelength was 250 nm; the injection volume was 20 μl; 5) System suitability requirements: In the system suitability solution spectrum, the peak order of each component is FSVS-IV-IMP-C, valsartan, and sacubitril, and the separation between each peak should meet the requirements; 6) Determination of related substances in sacubitril-valsartan sodium raw material: Accurately measure the test solution and the reference solution, inject them into the liquid chromatograph respectively, and record the chromatogram. If there are impurity peaks in the chromatogram of the test solution, calculate according to the main component self-reference method with the correction factor.

2. The method for detecting related substances of sacubitril-valsartan sodium as claimed in claim 1, further characterized by comprising the following steps: 1) Preparation of system suitability solution: Take 3 mg of FSVS-IV-IMP-C reference substance, place it in a 50 ml volumetric flask, dissolve it with acetonitrile-water (1:1) and dilute it to the mark. Accurately measure 1 ml and place it in a 20 ml volumetric flask with 23 mg of sacubitril-valsartan trisodium reference substance, add acetonitrile-water (1:1) to dissolve it and dilute it to the mark. Shake well to prepare the system suitability solution. 2) Preparation of test solution: Take 20 mg of the product, weigh accurately, place in a 20 ml volumetric flask, add acetonitrile-water (1:1) to dissolve and quantitatively dilute to the scale, shake well, and use as the test solution; 3) Preparation of control solution: Accurately measure 0.1 ml of the test solution, place it in a 100 ml volumetric flask, add acetonitrile-water (1:1) to dilute to the mark, shake well, and use as the control solution; 4) Chromatographic conditions: Use octadecylsilane bonded silica gel as filler (Waters XBridge Shield RP18, 4.6mm×250mm, 5μm or chromatographic column with equivalent performance); mobile phase A is 0.01mol / L ammonium dihydrogen phosphate aqueous solution (add 1.5ml of glacial acetic acid to every 1000ml), mobile phase B is methanol-acetonitrile (9:1); flow rate is 1.0ml per minute, linear gradient elution is performed according to the table below; column temperature is 30℃; detection wavelength is 250nm; injection volume is 20μl; 5) System suitability requirements: In the system suitability solution spectrum, the peak order of each component is FSVS-IV-IMP-C, valsartan, and sacubitril, and the separation between each peak should meet the requirements; 6) Determination of related substances in sacubitril-valsartan sodium raw material: Accurately measure 20 μl of the test solution and the reference solution, respectively, and inject them into the liquid chromatograph to record the chromatogram. If there are impurity peaks in the chromatogram of the test solution, calculate them according to the main component self-reference method with the addition of correction factors.

3. A quality control standard for related substances of sacubitril-valsartan sodium, characterized in that: The method for detecting sacubitril-valsartan sodium related substances adopts the detection method described in claim 1 or 2.

4. The quality control standard according to claim 3, characterized in that: When the above detection method is used for determination, the limits of relevant substances are as follows: the peak area of ​​FSVS-IV-IMP-C (FSVS-IV-IMP-F co-elutes with it) multiplied by 0.4506 shall not exceed 0.3% after calculation based on the peak area of ​​sacubitril according to the self-control method; the peak area of ​​other individual impurities multiplied by 0.4770 shall not exceed 0.10% after calculation based on the peak area of ​​valsartan according to the self-control method; the sum of all impurities except the FSVS-IV-IMP-C peak shall not exceed 0.3%, and the total amount of each impurity shall not exceed 0.6%.

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