A quality control method for new antiviral drug intermediates

Abstract

The present invention relates to the field of compound detection technology, and in particular to a quality control method for an intermediate of an antiviral new drug, comprising the following steps: taking the intermediate sample and dissolving it in an organic solvent to obtain an intermediate solution; taking the intermediate solution and detecting impurities by high performance liquid chromatography. The quality control method for the intermediate of an antiviral new drug provided by the present invention can effectively separate the intermediate of the antiviral new drug from adjacent impurities and various known impurities, accurately determine the impurity content, and has a high degree of separation, effectively solving the problem of the difficulty in separating and detecting the intermediate of the antiviral new drug and its related substances, thereby ensuring the quality control of the intermediate of the antiviral new drug, and further providing a strong guarantee for the quality control of the final product antiviral new drug.

Description

Technical Field

[0001] The invention relates to the technical field of compound detection, and in particular to a quality control method for a new antiviral drug intermediate. Background Art

[0002] WXSH0208 is a Class 1 chemical small molecule RNA polymerase PA subunit inhibitor, currently in Phase III clinical trials, and is indicated for the treatment of influenza A and B. The chemical structure of WXSH0208-3, an intermediate of the new antiviral drug WXSH0208, is as follows:

[0003]

[0004] Chemical name: (7 aS ,16 R )-10-(Benzyloxy)-2,3-difluoro-8-methyl-16-phenyl-6,7,7 a ,8-Tetrahydro-16 H -Benzo[ b ]pyrido[1',2':1,6][1,2,4]triazine[2,3- e ][1,5]Oxazolidin-9,11-dione.

[0005] WXSH0208-3 intermediates may include various related substances as impurities (including 16 impurities, see Table 1 Summary of Impurities in Related Substances) from starting materials, intermediates, process impurities, by-products, degradation impurities, etc. In order to ensure the quality of WXSH0208-3 intermediates, it is necessary to conduct qualitative and quantitative analysis of its related substances.

[0006] Table 1 Summary of impurities in new antiviral drug intermediates

[0007]

[0008]

[0009] Summary of the invention

[0010] The present invention aims to solve at least one of the technical problems existing in the related art. To this end, the object of the present invention is to provide a quality control method for an intermediate of a new antiviral drug.

[0011] In order to achieve this purpose, the technical solution adopted by the present invention is:

[0012] A quality control method for a new antiviral drug intermediate, wherein the chemical structure of the intermediate is as follows:

[0013] ;

[0014] The steps include:

[0015] S100, dissolving the intermediate sample in an organic solvent to obtain the intermediate solution;

[0016] S200, taking the intermediate solution and detecting impurities by high performance liquid chromatography;

[0017] The impurities include at least one of the following compounds:

[0018] , , , , , , , , , , , , , , and .

[0019] Furthermore, the high performance liquid chromatography is a reverse phase high performance liquid chromatography.

[0020] Furthermore, the chromatographic conditions of the reversed-phase high performance liquid chromatography are as follows:

[0021] Chromatographic column: C18 column, specification: 4.6×150mm, 3.5μm;

[0022] Mobile phase: 0.1% phosphoric acid aqueous solution as mobile phase A, methanol as mobile phase B, gradient elution;

[0023] Detection wavelength: 220nm;

[0024] Flow rate: 0.7 ml / min ~0.9 ml / min;

[0025] Column temperature: 38℃~42℃.

[0026] Furthermore, the process of gradient elution is as follows: it is constant within 0min to 15min, with the volume proportion of mobile phase A in the eluent maintained at 70%, and the volume proportion of mobile phase B maintained at 30%; then it changes at a constant speed to the 40th minute, with the volume proportion of mobile phase A at 25%, and the volume proportion of mobile phase B at 75%; then it changes at a constant speed to the 45th minute, and is constant within 45min to 50min, with the volume proportion of mobile phase A maintained at 10%, and the volume proportion of mobile phase B maintained at 90%; then it changes at a constant speed to the 50.1min, and is constant within 50.1min to 60min, with the volume proportion of mobile phase A maintained at 70%, and the volume proportion of mobile phase B maintained at 30%.

[0027] Furthermore, the process of gradient elution is as follows: it is constant within 0min to 5min, with the volume proportion of mobile phase A of the eluent maintained at 70%, and the volume proportion of mobile phase B maintained at 30%; then it changes at a constant speed to the 25th minute, with the volume proportion of mobile phase A being 50%, and the volume proportion of mobile phase B being 50%; then it changes at a constant speed to the 55th minute, with the volume proportion of mobile phase A being 40%, and the volume proportion of mobile phase B being 60%; then it changes at a constant speed to the 70th minute, with the volume proportion of mobile phase A being 10%, and the volume proportion of mobile phase B being 90%; then it changes at a constant speed to the 70.1th minute, and is constant from 70.1min to 80min, with the volume proportion of mobile phase A being maintained at 70%, and the volume proportion of mobile phase B being maintained at 30%.

[0028] Furthermore, the process of gradient elution is as follows: it is constant within 0min~5min, the volume proportion of mobile phase A is maintained at 70%, and the volume proportion of mobile phase B is maintained at 30%; then it changes at a constant speed to the 25th minute, the volume proportion of mobile phase A is 50%, and the volume proportion of mobile phase B is 50%; then it changes at a constant speed to the 55th minute, the volume proportion of mobile phase A is 40%, and the volume proportion of mobile phase B is 60%; then it changes at a constant speed to the 70th minute, and is constant from 70min to 80min, the volume proportion of mobile phase A is maintained at 10%, and the volume proportion of mobile phase B is maintained at 90%; then it changes at a constant speed to the 80.1min, and is constant from 80.1min to 90min, the volume proportion of mobile phase A is 70%, and the volume proportion of mobile phase B is 30%.

[0029] Furthermore, the flow rate is 0.8 ml / min.

[0030] Furthermore, the column temperature is 40°C.

[0031] Furthermore, in S100, the organic solvent is dichloromethane.

[0032] Furthermore, a ghost peak capture column is added, and the ghost peak capture column is used to eliminate interference from solvent and gradient change peaks.

[0033] The above one or more technical solutions in the embodiments of the present invention have at least one of the following technical effects:

[0034] The quality control method for the new antiviral drug intermediate provided by the present invention can effectively separate the new antiviral drug intermediate from adjacent impurities and various known impurities, can accurately measure the impurity content, and has a high degree of separation, effectively solving the problem of difficulty in separating and detecting the new antiviral drug intermediate and its related substances, thereby ensuring the controllable quality of the new antiviral drug intermediate.

[0035] The quality control method for the new antiviral drug intermediate provided by the present invention has high sensitivity and good accuracy, can more comprehensively perform accurate quantitative detection of various impurities in the sample, formulate reasonable control limits, and provide a strong guarantee for the quality control of the final product, the new antiviral drug WXSH0208.

[0036] Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0038] Figure 1 The chromatogram is a mixed solution of the antiviral new drug intermediate and five related substances in Example 1 of the present invention.

[0039] Figure 2 The chromatogram is a mixed solution of the antiviral new drug intermediate and five related substances in Example 2 of the present invention.

[0040] Figure 3 The chromatogram is a mixed solution of the new antiviral drug intermediate and 16 related substances in Example 3 of the present invention.

[0041] Figure 4 The chromatogram is a mixed solution of the antiviral new drug intermediate and 14 related substances in Example 4 of the present invention.

[0042] Figure 5 The chromatogram is a mixed solution of the new antiviral drug intermediate and 16 related substances in Example 5 of the present invention.

[0043] Figure 6 This is a chromatogram of the impurity detection limits of related substances in Example 6 of the present invention.

[0044] Figure 7 The chromatogram is the quantitative limit of impurities of related substances in Example 6 of the present invention.

[0045] Figure 8 This is the chromatogram of the 100% horizontal loading solution in Example 6 of the present invention.

[0046] Among them, WXSH0208-3 represents the new antiviral drug intermediate. DETAILED DESCRIPTION

[0047] In order to make the purpose, technical scheme and advantages of the present invention clearer, the technical scheme of the present invention will be clearly and completely described below in conjunction with the drawings in the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in the field without creative work are within the scope of protection of the present invention. The following embodiments are used to illustrate the present invention, but cannot be used to limit the scope of the present invention.

[0048] In the following examples, the experimental methods used are conventional methods unless otherwise specified, and the materials, reagents, etc. used are all available from commercial sources unless otherwise specified.

[0049] In the following examples and comparative examples, the drugs, reagents and instruments used are as follows:

[0050] Instruments and equipment: High performance liquid chromatograph (Vanquish-DAD, ThermoFisher); electronic balance (XPR205, Mettler-Toledo Instrument Shanghai Co., Ltd.), electronic balance (MSX (SG EA), Sartorius);

[0051] Chromatographic column: XBridge C18, chromatographic column (4.6×150mm, 3.5μm) (Waters); WatersXBridge Shield RP18 chromatographic column (4.6×150mm, 3.5μm) (Waters); Ghost-Sniper Column (4.0×30mm), Mr. Chromatography;

[0052] Reagents: methanol (chromatographic grade, batch number 240102, Concord); acetonitrile (chromatographic grade, batch number 240109, Concord); phosphoric acid (chromatographic grade, batch number 20240110, Komiou); water (ultrapure water, homemade);

[0053] WXSH0208-BP33a(Z-1) (Batch number: E040641-030-04, source: Duchuang (Chongqing) Pharmaceutical Technology Co., Ltd.);

[0054] WXSH0208-BP104(Z-2) (Batch No.: E040639-019-09, Source: Duchuang (Chongqing) Pharmaceutical Technology Co., Ltd.);

[0055] WXSH0208-BP27(Z-3) (Batch number: WS-ES18228-492-P1, source: Duchuang (Chongqing) Pharmaceutical Technology Co., Ltd.);

[0056] WXSH0208-BP12(Z-4) (Batch number: E040639-048-01, source: Duchuang (Chongqing) Pharmaceutical Technology Co., Ltd.);

[0057] WXSH0208-4(Z-5) (Batch number: WS-QC00003270-07-K-202312001, source: Duchuang (Chongqing) Pharmaceutical Technology Co., Ltd.);

[0058] WXSH0208-SM1 (Z-6) (Batch number: WS-WXSH0208-SM1-240119-1, source: Tianjin Chenxin Pharmaceutical Research Co., Ltd.);

[0059] TPPO(Z-7) (Batch No.: 972DRF4R, Source: Anaiji Chemical);

[0060] WXSH0208-BP108b(Z-8) (Batch number: E040639-052-04, source: Duchuang (Chongqing) Pharmaceutical Technology Co., Ltd.);

[0061] WXSH0208-BP30(Z-9) (Batch number: WS-WXSH0208-BP30-240511-1, source: Tianjin Chenxin Pharmaceutical Research Co., Ltd.);

[0062] WXSH0208-208-107a(Z-10) (Batch No.: E040638-093-01, Source: Duchuang (Chongqing) Pharmaceutical Technology Co., Ltd.);

[0063] WXSH0208-BP106b(Z-11) (Batch No.: E040639-056-01, Source: Duchuang (Chongqing) Pharmaceutical Technology Co., Ltd.);

[0064] WXSH0208-1(Z-12) (Batch number: WS-WXSH0208-1-52220737011-1AA, source: Tianjin Chenxin Pharmaceutical Research Co., Ltd.);

[0065] WXSH0208-1A(Z-13) (Batch number: WS-QC00003270-05-K-202311002, source: Duchuang (Chongqing) Pharmaceutical Technology Co., Ltd.);

[0066] WXSH0208-BP5(Z-14) (Batch No.: E040638-104-01, Source: Duchuang (Chongqing) Pharmaceutical Technology Co., Ltd.);

[0067] WXSH0208-BP24a(Z-15) (Batch No.: ES18228-494-P1, Source: Shanghai WuXi AppTec Pharmaceutical Development Co., Ltd.);

[0068] WXSH0208-BP24b(Z-16) (Batch No.: ES18228-494-P2, Source: Shanghai WuXi AppTec Pharmaceutical Development Co., Ltd.);

[0069] Sample of new antiviral drug intermediate (abbreviated as WXSH0208-3) (batch number: E040583-132-01, source: Du Chuang (Chongqing) Pharmaceutical Technology Co., Ltd.).

[0070] Example 1

[0071] The equipment and conditions are as follows:

[0072] Chromatographic column: RP18 chromatographic column, specification: 4.6×150mm, 3.5μm;

[0073] Mobile phase: 0.1% phosphoric acid aqueous solution as mobile phase A, methanol as mobile phase B;

[0074] The gradient elution process is:

[0075]

[0076] Flow rate: 0.8 ml / min;

[0077] The column temperature was 40°C;

[0078] The detection wavelength is 220nm;

[0079] The injection volume was 6 μl.

[0080] Diluent: water-methanol (volume ratio 20:80)

[0081] Operation steps: Weigh about 20 mg of WXSH0208-3 sample into a 10 ml volumetric flask, add 1 ml of dichloromethane (DCM) to dissolve it, add 2 ml of methanol, add impurities Z-2, Z-4, Z-5, Z-7, and Z-11 respectively at 0.2% of the test sample concentration, then add diluent to dilute to the scale, mix well, and you have it.

[0082] Test results: chromatogram of mixed solution, such as Figure 1As shown, this method was used to detect related substances of WXSH0208-3. Z-7 and Z-2 did not achieve baseline separation, with a separation degree of 1.84. Z-11 was inseparable from its previous impurities, with a separation degree of 1.31. Therefore, an attempt was made to replace the chromatographic column.

[0083] Example 2

[0084] The equipment and conditions are as follows:

[0085] Chromatographic column: C18 column, specification: 4.6×150mm, 3.5μm;

[0086] Mobile phase: 0.1% phosphoric acid aqueous solution as mobile phase A, methanol as mobile phase B;

[0087] The gradient elution process is:

[0088]

[0089] The flow rate is 0.8 ml / min;

[0090] The column temperature was 40°C;

[0091] The detection wavelength is 220nm;

[0092] The injection volume was 6 μl.

[0093] Diluent: water-methanol (volume ratio 20:80)

[0094] Operation steps: Weigh about 20 mg of WXSH0208-3 sample into a 10 ml volumetric flask, add 1 ml of dichloromethane to dissolve it, add 2 ml of methanol, add impurities Z-2, Z-4, Z-5, Z-7, and Z-11 respectively at 0.2% of the test sample concentration, then add diluent to dilute to the scale, mix well, and you have it.

[0095] Test results: chromatogram of mixed solution, such as Figure 2 As shown, this method was used to detect related substances of WXSH0208-3, and the separation effect of various impurities was good.

[0096] Example 3

[0097] The equipment and conditions are as follows:

[0098] Chromatographic column: C18 column, specification: 4.6×150mm, 3.5μm;

[0099] Mobile phase: 0.1% phosphoric acid aqueous solution as mobile phase A, methanol as mobile phase B;

[0100] The gradient elution process is:

[0101]

[0102] The flow rate is 0.8 ml / min;

[0103] The column temperature was 40°C;

[0104] The detection wavelength is 220nm;

[0105] The injection volume was 6 μl.

[0106] Diluent: water-methanol (volume ratio 20:80)

[0107] Operation steps: Weigh about 20 mg of the test sample and put it into a 10 ml volumetric flask, add 1 ml of dichloromethane to dissolve it, add 2 ml of methanol to dilute it, and then add appropriate amounts of impurities Z-1, Z-2, Z-3, Z-4, Z-5, Z-6, Z-7, Z-8, Z-9, Z-10, Z-11, Z-12, Z-13, Z-14, Z-15, and Z-16 respectively, then add 80% methanol to dilute to the scale and shake well; the solution is ready.

[0108] Among them, each impurity was added at 0.2% of the test sample concentration.

[0109] Test results: chromatogram of mixed solution, as shown Figure 3 As shown, this method was used to detect related substances of WXSH0208-3, and the main peak was not separated from the impurities before and after it, and the gradient was further optimized.

[0110] Example 4

[0111] The equipment and conditions are as follows:

[0112] Chromatographic column: C18 column, specification: 4.6×150mm, 3.5μm;

[0113] Mobile phase: 0.1% phosphoric acid aqueous solution as mobile phase A, methanol as mobile phase B;

[0114] The gradient elution process is:

[0115]

[0116] The flow rate is 0.8 ml / min;

[0117] The column temperature was 40°C;

[0118] The detection wavelength is 220nm;

[0119] The injection volume was 6 μl.

[0120] Diluent: water-methanol (volume ratio 20:80)

[0121] Operation steps: Weigh about 20 mg of the test sample and place it in a 10 ml volumetric flask, add 1 ml of dichloromethane to dissolve it, add 2 ml of methanol to dilute it, and then add appropriate amounts of impurities Z-1, Z-2, Z-3, Z-4, Z-5, Z-7, Z-9, Z-10, Z-11, Z-12, Z-13, Z-14, Z-15, and Z-16 respectively, then add 80% methanol to dilute to the scale, and shake well; the solution is ready.

[0122] Among them, each impurity was added at 0.2% of the test sample concentration.

[0123] Test results: chromatogram of mixed solution, such as Figure 4 As shown, this method is used to detect related substances of WXSH0208-3, and the separation effect of various impurities is good. In order to elute all small polar impurities in the sample, the elution time at 90% methanol can be extended.

[0124] Example 5

[0125] 1. Instruments and conditions

[0126] Chromatographic column: C18 column, specification: 4.6×150mm, 3.5μm;

[0127] Mobile phase: 0.1% phosphoric acid aqueous solution as mobile phase A, methanol as mobile phase B;

[0128] The gradient elution process is:

[0129]

[0130] Flow rate: 0.8 ml / min;

[0131] The column temperature was 40°C;

[0132] The detection wavelength is 220nm;

[0133] The injection volume was 10 μl.

[0134] Diluent: water-methanol (volume ratio 20:80)

[0135] Steps:

[0136] Test solution: weigh about 20 mg of the test sample and place it in a 10 ml volumetric flask, add 1 ml of dichloromethane to dissolve it, add 2 ml of methanol to dilute it, then add 80% methanol to dilute to the mark, and shake well; the solution is ready.

[0137] Mixed solution: weigh about 20 mg of the test sample and place it in a 10 ml volumetric flask, add 1 ml of dichloromethane to dissolve it, add 2 ml of methanol to dilute it, and then add appropriate amounts of impurities Z-1, Z-2, Z-3, Z-4, Z-5, Z-6, Z-7, Z-8, Z-9, Z-10, Z-11, Z-12, Z-13, Z-14, Z-15, and Z-16, then add 80% methanol to dilute to the scale and shake well; the solution is ready.

[0138] Among them, Z-9 was added at 0.1%, and the remaining impurities were added at 0.2% of the test sample concentration.

[0139] Mixed solution chromatogram, such as Figure 5 As shown, the method was used to detect related substances of WXSH0208-3, and impurities were eluted at 77.342 min, with good separation between the impurities.

[0140] (B) Examination of the durability range of method parameters.

[0141] The durability range of this method was investigated from the perspective of the initial ratio of the mobile phase to the organic phase, column temperature, flow rate and other conditions, as shown in the following table:

[0142]

[0143] Column temperature: range: 38°C ~ 42°C, initial proportion of organic phase: range: 28% ~ 32%.

[0144] The column temperatures of 38°C, 40°C and 42°C were examined in turn, and the initial proportion of the organic phase was 28%, 30% and 32%. The higher the column temperature, the higher the initial proportion of the organic phase, the earlier the peak time, and the smaller the separation between impurities, but the separation can meet the detection requirements.

[0145] Flow rate range: 0.7ml / min~0.9ml / min.

[0146] The flow rate conditions of 0.7ml / min, 0.8ml / min and 0.9ml / min were examined in turn. Under the same conditions, the higher the flow rate, the earlier the peak time. Under the three flow rate conditions, the separation between each impurity and sample can meet the requirements. The flow rate of 0.8ml / min with better separation and appropriate peak time is preferred.

[0147] Example 6

[0148] (I) Investigation of limit of quantification and limit of detection.

[0149] Chromatographic conditions: the same as those used in Example 5.

[0150] Operation steps: Accurately weigh appropriate amount of each impurity reference substance, accurately weigh, dissolve in methanol and dilute to scale to obtain each impurity reference substance solution; dilute step by step to prepare each impurity quantitative limit and detection limit solution, inject into liquid chromatograph, record chromatogram, and detect the spectrum as shown in Figure 6 , Figure 7 shown.

[0151] The quantitative limits and detection limits of each impurity are shown in the following table:

[0152]

[0153] This method was used to detect related substances in WXSH0208-3. Based on the test solution concentration of 2 mg / ml, the detection limits were all less than 0.010%, with high sensitivity, which met the detection requirements.

[0154] (B) Recovery rate investigation

[0155] Chromatographic conditions: the same as the optimal method conditions determined in Example 5.

[0156] Operation steps: Investigate the recovery rates at the 50% level, 100% level and 150% level (add approximately 50% limit, 100% limit and 150% limit of each impurity to the test solution, and calculate the recovery rate of each impurity).

[0157] Reference substance solution: Weigh the appropriate amount of each impurity reference substance accurately, weigh accurately, dissolve and dilute with diluent to make a solution containing about 10μg of WXSH0208-BP30 (Z-9) and 20μg of other impurities per ml as the mixed reference substance stock solution. Then accurately measure 2ml of the mixed reference substance stock solution, place it in a 10ml volumetric flask, add 1ml of dichloromethane, and then add diluent to dilute to the scale as the reference substance solution.

[0158] Unsampled test solution: Accurately weigh about 20 mg of WXSH0208-3 sample, place it in a 10 ml volumetric flask, add 1 ml of dichloromethane to dissolve it, add 2 ml of methanol to mix, and then add diluent to the scale to obtain the sample solution.

[0159] 50% horizontal loading solution: accurately weigh about 20 mg of WXSH0208-3 sample, place it in a 10 ml volumetric flask, add 1 ml of dichloromethane to dissolve it, add 2 ml of methanol to mix, accurately measure 1 ml of the mixed reference stock solution into the flask, add diluent to the scale, as the 50% horizontal loading solution.

[0160] 100% horizontal loading solution: accurately weigh about 20 mg of WXSH0208-3 sample, place it in a 10 ml volumetric flask, add 1 ml of dichloromethane to dissolve it, add 2 ml of methanol to mix, accurately measure 2 ml of the mixed reference stock solution into the flask, add diluent to the scale, as the 100% horizontal loading solution.

[0161] 150% horizontal loading solution: accurately weigh about 20 mg of WXSH0208-3 sample, place it in a 10 ml volumetric flask, add 1 ml of dichloromethane to dissolve it, add 2 ml of methanol to mix, accurately measure 3 ml of the mixed reference stock solution into the flask, add diluent to the scale, as the 150% horizontal loading solution.

[0162] Respectively measure the reference solution, test solution and sample solution, inject them into the liquid chromatograph, record the chromatogram and calculate the recovery rate.

[0163] 100% horizontal sample solution detection spectrum, such as Figure 8 shown.

[0164] The recovery results at each level are shown in the following table:

[0165]

[0166]

[0167] It can be seen from the above table that the 16 impurities are in the range of 50% to 150%, and the average recovery rates are all between 85% and 110%, which meets the requirements, proving that the quality control method for a new antiviral drug intermediate provided by the present invention has good accuracy.

[0168] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A quality control method for a new antiviral drug intermediate, characterized in that: The chemical structural formula of the intermediate is as follows: ； The steps include: S100, dissolving an intermediate sample in an organic solvent to obtain an intermediate solution; S200, taking the intermediate solution and detecting impurities by high performance liquid chromatography; The impurities include at least one of the following compounds: , , , , , , , , , , , , , , and ; The high performance liquid chromatography method is a reversed phase high performance liquid chromatography method, and the chromatographic conditions of the reversed phase high performance liquid chromatography method are as follows: Chromatographic column: C18 column, specification: 4.6×150mm, 3.5μm; Mobile phase: 0.1% phosphoric acid aqueous solution as mobile phase A, methanol as mobile phase B, gradient elution; Detection wavelength: 220nm; Flow rate: 0.7 ml / min ~0.9 ml / min; Column temperature: 38℃~42℃.

2. The quality control method of the antiviral new drug intermediate according to claim 1, characterized in that: The process of gradient elution is as follows: it is constant from 0min to 15min, with the volume proportion of mobile phase A in the eluent maintained at 70%, and the volume proportion of mobile phase B maintained at 30%; then it changes at a constant speed to the 40th minute, with the volume proportion of mobile phase A at 25%, and the volume proportion of mobile phase B at 75%; then it changes at a constant speed to the 45th minute, and is constant from 45min to 50min, with the volume proportion of mobile phase A maintained at 10%, and the volume proportion of mobile phase B maintained at 90%; then it changes at a constant speed to the 50.1th minute, and is constant from 50.1min to 60min, with the volume proportion of mobile phase A maintained at 70%, and the volume proportion of mobile phase B maintained at 30%.

3. The quality control method of the antiviral new drug intermediate according to claim 1, characterized in that: The process of gradient elution is as follows: it is constant from 0min to 5min, with the volume proportion of mobile phase A of the eluent maintained at 70%, and the volume proportion of mobile phase B maintained at 30%; then it changes at a constant speed to the 25th min, with the volume proportion of mobile phase A being 50%, and the volume proportion of mobile phase B being 50%; then it changes at a constant speed to the 55th min, with the volume proportion of mobile phase A being 40%, and the volume proportion of mobile phase B being 60%; then it changes at a constant speed to the 70th min, with the volume proportion of mobile phase A being 10%, and the volume proportion of mobile phase B being 90%; then it changes at a constant speed to the 70.1min, and is constant from 70.1min to 80min, with the volume proportion of mobile phase A being maintained at 70%, and the volume proportion of mobile phase B being maintained at 30%.

4. The quality control method of the antiviral new drug intermediate according to claim 1, characterized in that: The process of gradient elution is as follows: it is constant from 0min to 5min, with the volume proportion of mobile phase A maintained at 70%, and the volume proportion of mobile phase B maintained at 30%; then it changes at a constant speed to the 25th min, with the volume proportion of mobile phase A at 50%, and the volume proportion of mobile phase B at 50%; then it changes at a constant speed to the 55th min, with the volume proportion of mobile phase A at 40%, and the volume proportion of mobile phase B at 60%; then it changes at a constant speed to the 70th min, and is constant from 70min to 80min, with the volume proportion of mobile phase A maintained at 10%, and the volume proportion of mobile phase B maintained at 90%; then it changes at a constant speed to the 80.1min, and is constant from 80.1min to 90min, with the volume proportion of mobile phase A at 70%, and the volume proportion of mobile phase B at 30%.

5. The quality control method for the new antiviral drug intermediate according to any one of claims 1 to 4, characterized in that: The flow rate is 0.8 ml / min.

6. The quality control method for the intermediate of the new antiviral drug according to any one of claims 1 to 4, characterized in that: The column temperature was 40°C.

7. The quality control method of the antiviral new drug intermediate according to claim 1, characterized in that: In S100, the organic solvent is dichloromethane.

8. The quality control method for the intermediate of the new antiviral drug according to claim 1, characterized in that: A ghost peak trapping column is added, and the ghost peak trapping column is used to eliminate the interference of solvent and gradient change peaks.

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