Analysis method for detecting multiple residual solvents in medicine

Abstract

The invention belongs to the field of analytical chemistry, and particularly relates to an analysis method for detecting multiple residual solvents in a medicine preparation process. According to the method, 21 organic solvents including methanol, tetrahydrofuran, ethanol, cyclohexane, diethyl ether, isopropyl acetate, acetone, n-heptane, isopropanol, n-butyl alcohol, acetonitrile, dioxane, dichloromethane, pyridine, tert-butyl alcohol, toluene, tert-butyl methyl ether, N, N-dimethylformamide, n-hexane, dimethyl sulfoxide and ethyl acetate are simultaneously detected by adopting a gas chromatographic method. The method is not influenced by drug types, can be applied to content detection of various drug residual solvents, is simple to operate and wide in application, and saves drug detection time and detection cost.

Description

Technical Field

[0001] The invention belongs to the field of analytical chemistry, and in particular relates to a method for detecting multiple residual solvents in medicines by using gas chromatography. Background Art

[0002] Residual solvents in drugs refer to organic volatile compounds that are used in the production of raw materials, excipients and preparations but are not completely removed during the process. In the synthesis process of raw materials, the selection of appropriate solvents can increase the yield or determine the properties of the drug, such as crystal form, purity, dissolution rate, etc. In biological drugs, the selection of appropriate solvents increases the solubility of materials in production, such as ADC, and the selection of appropriate solvents to dissolve the intermediate toxin linker. Therefore, organic solvents are indispensable and very critical substances in drug synthesis reactions. When the level of residual solvents contained in drugs is higher than the safety value, it will cause harm to the human body or the environment. Therefore, it is very necessary for drugs to require the study and effective control of organic solvents and reaction materials used in their process when registering and declaring drugs. The control of residual solvents has received more and more attention.

[0003] The International Council for Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) harmonized guidelines require that residual solvents in the final drug product must be controlled within the specified limits. Currently, the methods for determining residual solvents in drugs include loss on drying, spectrophotometry, and gas chromatography. Loss on drying is a method of determining the content of residual solvents by the loss of mass of the sample during heating. Spectrophotometry can use the reaction of specific solvents and specific chemical reagents to determine the residual solvents in drugs. In the prior art, headspace gas chromatography is mainly used to determine the residual amount of organic solvents in drugs. Since the method of gas chromatography analysis to detect organic solvent residues is specific, the detection method can only detect solvent residues in a targeted manner, and cannot simultaneously detect organic solvents used in the drug synthesis process. Due to the wide variety of organic solvents, it is necessary to provide an analytical method that is not affected by the type of drug, can be applied to a variety of drugs, and can simultaneously detect the content of multiple organic solvents during drug development. Summary of the invention

[0004] The purpose of the present invention is to provide an analytical method for simultaneously detecting multiple residual solvents in a drug, thereby achieving effective control of the drug quality and ensuring the safety and controllability of subsequent drug quality.

[0005] To achieve the above object, the present invention adopts the following technical solution:

[0006] A gas chromatography analysis method for simultaneously detecting multiple residual solvents in a drug, wherein the gas chromatography conditions of the analysis method include:

[0007] Inlet temperature: 200-250℃;

[0008] Column box heating program: initial column temperature is 30-40°C, holding time is 6-10min, preferably holding time is 8-10min, more preferably holding time is 6min, 8min or 10min, heating to 220°C at a rate of 15-25°C per minute, holding time after heating is 3-7min, preferably holding time is 5-7min, more preferably holding time is 3min, 5min or 7min.

[0009] In certain embodiments of the present invention, the injection port temperature is preferably 200°C, 230°C or 250°C.

[0010] In certain embodiments of the present invention, the initial temperature in the column oven temperature program is preferably 30°C, 35°C or 40°C.

[0011] In certain embodiments of the present invention, the rate of heating to 220° C. in the temperature heating program is preferably 15° C. / min, 20° C. / min or 25° C. / min.

[0012] In certain embodiments of the present invention, the analytical method reference substance or test substance needs to be diluted with a diluent before injection.

[0013] In certain embodiments of the present invention, the diluent is N-methylpyrrolidone.

[0014] In certain embodiments of the present invention, the detector used in the analysis method is a hydrogen flame ionization detector.

[0015] In certain embodiments of the present invention, the chromatographic column selected for the analytical method is a medium-polarity gas chromatography capillary column or a non-polar gas chromatography capillary column.

[0016] In certain embodiments of the present invention, the medium polarity gas chromatography capillary column is a capillary chromatography column using 6% cyanopropylphenyl-94% dimethylpolysiloxane as a stationary phase. Preferably, the medium polarity gas chromatography capillary column is a VF624 capillary chromatography column.

[0017] In certain embodiments of the present invention, the non-polar gas chromatography capillary column is a capillary chromatography column using 100% dimethyl polysiloxane as a stationary liquid. Preferably, the non-polar gas chromatography capillary column is an HP-1 capillary chromatography column.

[0018] In some embodiments of the present invention, the detection temperature of the detector is greater than 200°C.

[0019] In certain embodiments of the present invention, the detection temperature of the detector is preferably 200-300°C, more preferably 250-300°C.

[0020] In certain embodiments of the present invention, the detection temperature of the detector is preferably 200°C, 250°C or 300°C.

[0021] In certain embodiments of the present invention, the carrier gas of the analytical method is an inert gas. Preferably, the carrier gas is nitrogen.

[0022] In certain embodiments of the present invention, the carrier gas flow rate in the gas chromatography is 2.0-3.0 ml / min.

[0023] In certain embodiments of the present invention, the carrier gas flow rate in the gas chromatograph is preferably 2.0 ml / min or 3.0 ml / min.

[0024] In certain embodiments of the present invention, the analysis method uses split injection with a split ratio of 1-10:1, preferably a split ratio of 5-10:1, and more preferably a split ratio of 1:1, 5:1 or 10:1.

[0025] In certain embodiments of the present invention, the headspace equilibrium temperature of the analytical method is greater than 70°C, preferably the headspace equilibrium temperature is 70-90°C, and more preferably the headspace equilibrium temperature is 70°C or 90°C.

[0026] In certain embodiments of the present invention, the plurality of residual solvents comprises two or more of methanol, tetrahydrofuran, ethanol, cyclohexane, diethyl ether, isopropyl acetate, acetone, n-heptane, isopropanol, n-butanol, acetonitrile, dioxane, dichloromethane, pyridine, tert-butanol, toluene, tert-butyl methyl ether, N,N-dimethylformamide (DMF), n-hexane, dimethyl sulfoxide (DMSO), and ethyl acetate.

[0027] Preferably, the plurality of residual solvents comprises three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, eleven or more, twelve or more, thirteen or more, fourteen or more, fifteen or more, sixteen or more, seventeen or more, eighteen or more, nineteen or more, or twenty or more of the group consisting of methanol, tetrahydrofuran, ethanol, cyclohexane, ether, isopropyl acetate, acetone, n-heptane, isopropanol, n-butanol, acetonitrile, dioxane, dichloromethane, pyridine, tert-butyl alcohol, toluene, tert-butyl methyl ether, N,N-dimethylformamide, n-hexane, dimethyl sulfoxide, and ethyl acetate.

[0028] In certain embodiments of the present invention, the plurality of residual solvents are one or more combinations of methanol, tetrahydrofuran, ethanol, cyclohexane, ether, isopropyl acetate, acetone, n-heptane, isopropanol, n-butanol, acetonitrile, dioxane, dichloromethane, pyridine, tert-butanol, toluene, tert-butyl methyl ether, N,N-dimethylformamide, n-hexane, dimethyl sulfoxide and ethyl acetate.

[0029] In certain embodiments of the present invention, the analytical method can be used to detect residual solvents in a drug, wherein the drug is a biological targeted drug or a chemical targeted drug, preferably, the biological targeted drug is one or more of a monoclonal antibody, a biclonal antibody, and an antibody-drug conjugate (ADC).

[0030] The antibody-drug conjugate (ADC) described in the present invention comprises Ab or an antigen-binding Ab fragment and a drug conjugated to the Ab or the antigen-binding Ab fragment, wherein the drug is an anticancer drug, an antiproliferative drug, a cytotoxic drug, an antiangiogenic drug, an antithrombotic drug, an apoptotic drug, an immunostimulatory drug, an antimicrobial drug, an antibiotic drug, an antiviral drug, an anti-inflammatory drug, an antifibrotic drug, an immunosuppressive drug, a steroid, a bronchodilator, a beta-blocker, a matrix metalloproteinase inhibitor, an ADAM12 inhibitor, an ADAM12 signaling inhibitor, an enzyme, a hormone, a neurotransmitter, a toxin, a radioisotope, a compound, a small molecule, a small molecule inhibitor, a protein, a peptide, a vector, a plasmid, a viral particle, a nanoparticle, a DNA molecule, an RNA molecule, siRNA, shRNA, microRNA, an oligonucleotide or an imaging drug.

[0031] The beneficial effects of the present invention are:

[0032] 1. The prior art requires the development of multiple analytical methods to detect multiple residual solvents in drugs, or different analytical methods need to be developed for different drugs to achieve the purpose of detecting residual solvents of different drugs. The present invention uses a gas chromatography analysis method to achieve the purpose of effectively separating and analyzing up to 21 solvents at the same time, and the analysis method of the present invention has good precision, good separation, high sensitivity, and good linear correlation.

[0033] 2. The analytical method of the present invention is not affected by the type of drug. As long as the solvent used in the drug synthesis process is included in the range of 21 solvents defined in the present invention, whether it is one or more, the analytical method of the present invention can be used to confirm at one time whether the drug has relevant solvent residues.

[0034] 3. The analytical method of the present invention is simple to operate, the drug detection time is short, and the detection cost is low. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 The gas chromatogram of the system suitability of Example 1;

[0036] Figure 2 The gas chromatogram of Example 2 for investigating the temperature program holding time condition 1;

[0037] FIG. 3A to FIG. 3DThe gas chromatograms of Example 2 for the temperature program holding time conditions 2-5;

[0038] Figure 4 The gas chromatogram of the chromatographic column HP-1 was investigated for Example 3;

[0039] FIG. 5A to FIG. 5D The gas chromatogram of the initial temperature and the heating rate of Example 4 is shown;

[0040] FIG. 6A to FIG. 6D The gas chromatogram of Example 5 for investigating the injection port temperature and the detector temperature;

[0041] Figure 7 The gas chromatogram of the headspace equilibrium temperature of Example 5 is investigated;

[0042] Fig. 8A and Figure 8B This is a gas chromatogram of the split ratio of Example 6;

[0043] Fig.9A and Fig. 9B The gas chromatogram of the carrier gas flow rate investigated in Example 6;

[0044] FIG. 10A to FIG. 10C They are gas chromatograms of blank solvent, reference solution 1, and test solution of compound of formula I batch 01 of Example 7, respectively;

[0045] FIG. 11A to FIG. 11C They are gas chromatograms of reference solution 2 of Example 8, test solution batch 01 and batch 02 of compound of formula II, respectively. DETAILED DESCRIPTION

[0046] The gas phase detection and analysis method involved in the present invention is further described in detail below in the form of embodiments, but this should not be understood as the scope of the above subject matter of the present invention being limited to the following embodiments. All technologies realized based on the above content of the present invention belong to the scope of the present invention.

[0047] Example 1

[0048] 1. Test instruments and reagents

[0049] Table 1. Instruments

[0050] Serial number name supplier model 1 Gas Chromatograph Agilent 7890B 2 Headspace Sampler Agilent 7697A 3 Electronic balance Sartorius BSA224S-CW 4 Chromatographic columns Agilent VF624ms 60m×320μm, 1.80μm

[0051] 2. Chromatographic conditions

[0052] Chromatographic column: capillary column with 6% cyanopropylphenyl-94% dimethylpolysiloxane as stationary phase

[0053] Sample dilution solvent: N-methylpyrrolidone

[0054] Inlet temperature: 230°C

[0055] Detector: Hydrogen flame ionization detector

[0056] Carrier gas: Nitrogen

[0057] Carrier gas flow rate: 2.0ml / min

[0058] Heating program: Initial column temperature is 35°C, maintained for 8 min, then increased to 220°C at a rate of 20°C per minute, maintained for 5 min

[0059] Detector temperature: 300°C

[0060] Split ratio: 10:1

[0061] Headspace bottle equilibrium temperature: 90℃, equilibrium for 20min

[0062] Quantitative loop temperature: 100°C

[0063] Transfer line temperature: 110°C

[0064] 3. Solution preparation

[0065] 3.1 Preparation of reference stock solution

[0066] Take a 100ml volumetric flask, weigh in 20ml N-methylpyrrolidone, and weigh in 0.3041g methanol, 0.5001g ethanol, 0.5180g ether, 0.5071g acetone, 0.4996g isopropanol, 0.0417g acetonitrile, 0.065g dichloromethane, 0.5076g tert-butyl alcohol, 0.0297g n-hexane, 0.5080g ethyl acetate, 0.0760g tetrahydrofuran, 0.3824g cyclohexane, 0.5028g isopropyl acetate, 0.5004g n-heptane, 0.5103g n-butanol, 0.0422g dioxane, 0.0209g pyridine, 0.0914g toluene, 0.0908g DMF, 0.5455g DMSO and 0.5033 g of tert-butyl methyl ether are dissolved, and N-methylpyrrolidone is added to the scale. Shake well and use this as the reference stock solution.

[0067] 3.2 Preparation of reference solution

[0068] Accurately measure 1 ml of the reference stock solution, place it in a 10 ml volumetric flask, add N-methylpyrrolidone to dilute to the scale, and shake well; accurately transfer 1 ml, place it in an empty bottle, and cap it as the reference solution.

[0069] 4. System suitability test

[0070] The reference solution was used as the system suitability test solution. The solution was sampled in the headspace and measured according to the above chromatographic conditions, and the chromatogram was recorded. Under the above chromatographic conditions, each component can be well separated, and the blank solvent has no interference. The retention time of each component is shown in Table 2, and the chromatogram is shown in Figure 1 .

[0071] Table 2. Retention time of each component

[0072] Serial number Solvents Retention time (min) Serial number Solvents Retention time (min) 1 Methanol 6.57 12 Tetrahydrofuran 13.01 2 Ethanol 8.85 13 Cyclohexane 13.38 3 Ether 9.15 14 Isopropyl acetate 13.67 4 acetone 9.86 15 n-Heptane 13.92 5 Isopropyl alcohol 10.16 16 n-Butanol 14.18 6 Acetonitrile 10.48 17 Dioxane 14.65 7 Dichloromethane 10.81 18 Pyridine 15.42 8 Tert-Butanol 10.97 19 Toluene 15.51 9 Tert-Butyl Methyl Ether 11.23 20 N,N-Dimethylformamide 16.38 10 n-Hexane 11.67 21 Dimethyl sulfoxide 17.30 11 Ethyl acetate 12.71

[0073] 5. Verification Process

[0074] 5.1 Exclusive

[0075] The chromatogram separation obtained from the system suitability test is shown in Table 3 below:

[0076] Table 3. Separation of each component

[0077]

[0078] 5.2 Limit of quantitation, limit of detection

[0079] 5.2.1 Experimental process

[0080] Take appropriate amounts of methanol, tetrahydrofuran, ethanol, cyclohexane, ether, isopropyl acetate, acetone, n-heptane, isopropanol, n-butanol, acetonitrile, dioxane, dichloromethane, pyridine, tert-butanol, toluene, tert-butyl methyl ether, N,N-dimethylformamide, n-hexane, dimethyl sulfoxide and ethyl acetate respectively, weigh accurately, dissolve with N-methylpyrrolidone to prepare a solution of appropriate concentration, and determine under the above-mentioned chromatographic conditions. The quantification limit is calculated based on the standard of S / N not less than 10 and concentration not more than 30% of the respective limit concentration. The detection limit is calculated based on the standard of S / N not less than 3 and concentration not more than 10% of the respective limit concentration.

[0081] 5.2.2 Results

[0082] The results of the limit of quantification and detection are shown in Table 4.

[0083] Table 4. Summary of quantitation limits and detection limits of each component

[0084]

[0085]

[0086] 5.3 Linear

[0087] 5.3.1 Preparation of linear solution

[0088] Transfer 2.5 ml of "3.1 Reference substance stock solution" into a 25 ml volumetric flask, add N-methylpyrrolidone to the mark, shake well, transfer 1000 μl into 5 20 ml headspace bottles respectively, and seal with crimp caps as reference substance solution.

[0089] The reference solution was diluted step by step to different concentrations; each solution was injected and analyzed from low concentration to high concentration according to the above chromatographic conditions.

[0090] 5.3.2 Linear Results

[0091] The linearity and range of each component were statistically analyzed and the results are shown in Table 5.

[0092] Table 5 Summary of linearity and range of each component

[0093]

[0094]

[0095] The linear relationship between the residual solvent concentration and the peak area was good in the range from the limit of quantification to 200% of the limit concentration.

[0096] Through the above verification process and verification results, it is proved that the chromatographic conditions can accurately and effectively detect 21 solvents. The method is simple, highly sensitive, precise and accurate.

[0097] Example 2

[0098] This example investigates the effect of the temperature program maintenance time on the gas chromatography analysis results. The different temperature programs set in the experiment are shown in Table 6. The other experimental conditions are the same as those in Example 1. The injected sample is the 3.2 reference solution in Example 1. The experimental results are shown in Tables 6 and Figure 2 , FIG. 3A to FIG. 3D .

[0099] Table 6. Results of optimization of the heating program maintenance time

[0100]

[0101] From the experimental results of the temperature program holding time in Table 6, it can be seen that when the initial temperature holding time is 5 minutes, it is impossible to achieve complete separation of all 21 solvents. However, when the initial temperature holding time is set to 6-10 minutes and the temperature is raised to 220°C and maintained for 3-7 minutes, chromatographic detection results that meet the analysis requirements can be obtained.

[0102] Example 3

[0103] This example investigates the effect of the chromatographic column on the gas chromatography analysis results. The chromatographic column used in the experiment is different from that in Example 1 as shown in Table 7. The other experimental conditions are the same as in Example 1. The injected sample is the 3.2 reference solution in Example 1. The experimental results are shown in Tables 7 and Figure 4 .

[0104] Table 7. Column selection results

[0105] Serial number Chromatographic columns Experimental Results 1 HP-1(30m×0.53mm×3μm) Separation of 21 solvents with a minimum resolution of 1.21

[0106] From the chromatographic column investigation experimental results in Table 7, it can be seen that the HP-1 chromatographic column can obtain chromatographic detection results that meet the analysis requirements.

[0107] Example 4

[0108] This example investigates the effects of initial temperature and heating rate on gas chromatography analysis results. The different initial temperatures and heating rates set in the experiment are shown in Table 8. The other experimental conditions are the same as those in Example 1. The injected sample is the 3.2 reference solution in Example 1. The experimental results are shown in Tables 8 and FIG. 5A to FIG. 5D .

[0109] Table 8. Results of optimization of initial temperature and heating rate

[0110] Serial number Experimental conditions Experimental Results 1 The initial temperature is 30℃( Figure 5A ) Separation of 21 solvents with a minimum resolution of 1.38 2 The initial temperature is 40℃( Figure 5B ) Separation of 21 solvents with a minimum resolution of 1.50 3 The heating rate is 15℃ / min( Figure 5C ) Separation of 21 solvents with a minimum resolution of 1.29 4 The heating rate is 25℃ / min( Figure 5D ) Separation of 21 solvents with a minimum resolution of 1.44

[0111] From the initial temperature and heating rate optimization experimental results in Table 8, it can be seen that when the initial temperature is 30-40°C and the heating rate is 15-25°C / min, chromatographic detection results that meet the analysis requirements can be obtained.

[0112] Example 5

[0113] This example investigates the effects of the injection port temperature, detector temperature and headspace bottle equilibrium temperature on the gas chromatography analysis results. The different injection port temperatures, detector temperatures and headspace bottle equilibrium temperatures set in the experiment are shown in Table 9. The other experimental conditions are the same as those in Example 1. The injection sample is the 3.2 reference solution in Example 1. The experimental results are shown in Tables 9 and Figures 6A to 6D , Figure 7 .

[0114] Table 9. Results of optimization of injection port temperature, detector temperature, and headspace vial equilibrium temperature

[0115] Serial number Experimental conditions Experimental Results Condition 1 The injection port temperature was 200°C ( Fig. 6A ) Separation of 21 solvents with a minimum resolution of 1.44 Condition 2 The injection port temperature was 250°C ( Figure 6B ) Separation of 21 solvents with a minimum resolution of 1.47 Condition 3 The detector temperature is 200°C ( Figure 6C ) Separation of 21 solvents with a minimum resolution of 1.46 Condition 4 The detector temperature is 250°C ( Fig.6D ) Separation of 21 solvents with a minimum resolution of 1.45 Condition 5 The headspace equilibrium temperature is 70°C ( Figure 7 ) Separation of 21 solvents with a minimum resolution of 1.39

[0116] From the experimental results of the injection port temperature, detector temperature and headspace bottle equilibrium temperature in Table 9, it can be seen that when the injection port temperature is 200-250°C, the detector temperature is 200-300°C and the headspace bottle equilibrium temperature is 70-90°C, chromatographic detection results that meet the analysis requirements can be obtained.

[0117] Example 6

[0118] This example investigates the effect of split ratio and carrier gas flow rate on the gas chromatography analysis results. The different split ratios and carrier gas flow rates set in the experiment are shown in Table 10. The other experimental conditions are the same as those in Example 1. The injected sample is the 3.2 reference solution in Example 1. The experimental results are shown in Tables 10 and Fig. 8A , Figure 8B , Fig.9A and Fig. 9B .

[0119] Table 10. Optimization results of split ratio and carrier gas flow rate

[0120]

[0121] From the experimental results of split ratio and carrier gas flow rate in Table 10, it can be seen that when the split ratio is 1:1-10:1, chromatographic detection results that meet the analysis requirements can be obtained. When the carrier gas flow rate is 1.0ml / min, the baseline rises, which interferes with the solvent detection at 9.388min and cannot realize the solvent content detection. However, when the carrier gas flow rate is 2.0-3.0ml / min, chromatographic detection results that meet the analysis requirements can be obtained.

[0122] Example 7

[0123] This example examines various residual solvents in the drug portion of the ADC (compound of Formula I).

[0124] The compound of formula I is the compound of Example 15 in patent publication number WO2018113652, which is mainly used to treat chronic hepatitis B. Five solvents, methanol, ethyl acetate, acetonitrile, dichloromethane and toluene, are used in the production process of the compound.

[0125]

[0126] 1. Chromatographic conditions

[0127] The chromatographic conditions were the same as in Example 1.

[0128] 2. Solution preparation

[0129] Preparation of test solution: Take about 0.1 g of the ADC drug part (compound of formula I, batch 01), accurately weigh it, place it in an empty bottle, accurately add 1 ml of N-methylpyrrolidone, seal it, and use it as the test solution.

[0130] Preparation of reference solution 1: Take appropriate amounts of methanol, acetonitrile, dichloromethane, ethyl acetate and toluene, weigh accurately, and dilute with N-methylpyrrolidone to make a mixed solution containing about 0.3 mg of methanol, 0.041 mg of acetonitrile, 0.06 mg of dichloromethane, 0.5 mg of ethyl acetate and 0.089 mg of toluene per 1 ml. Accurately measure 1 ml, place in a 20 ml headspace bottle, and seal.

[0131] 3. Determination method:

[0132] Accurately measure the blank solution, reference solution 1 and test solution, inject them into the headspace respectively, and record the chromatograms.

[0133] Calculation formula: Calculated by peak area using the external standard method.

[0134]

[0135] Where: W std1 is the sample amount of the solvent in the reference solution 1, mg;

[0136] A spl is the peak area of ​​the solvent in the test solution;

[0137] DF spl is the dilution factor of the test solution;

[0138] W spl is the sample weight of the test product, mg;

[0139] AHr std1 is the average peak area of ​​the solvent in reference solution 1 for 5 consecutive measurements;

[0140] DF std1 It is the dilution multiple of reference solution 1.

[0141] 4. Measurement results

[0142] The results were calculated by external standard method using peak area. See Table 11 for the chromatograms. See Figure 10 for the chromatograms of blank solvent. Fig. 10A , Chromatography of reference solution 1 Fig. 10B , Chromatography of the test solution Formula I compound batch 01 Fig. 10C .

[0143] Table 11 Determination results of residual solvents in compounds of formula I

[0144]

[0145] Example 8

[0146] This example examines various residual solvents in the drug portion of the ADC (compound of Formula II).

[0147] The compound of formula II is the compound of Example 5 in patent publication number WO2019184744, a white to yellow solid, mainly used for the prevention and treatment of thromboembolic diseases. In the production process of the compound, 12 solvents including methanol, ethanol, isopropanol, acetonitrile, dichloromethane, tert-butyl methyl ether, ethyl acetate, tetrahydrofuran, n-heptane, 1,4-dioxane, toluene, and N,N-dimethylformamide are used.

[0148]

[0149] 1. Chromatographic conditions

[0150] The chromatographic conditions were the same as in Example 1.

[0151] 2. Solution preparation

[0152] Preparation of test solution: Take about 0.1 g of the ADC drug part (compound of formula II), accurately weigh it, place it in an empty bottle, accurately add 1 ml of N-methylpyrrolidone, seal it, and use it as the test solution.

[0153] Compounds of formula II from two batches 01 and 02 were prepared into test solutions according to the above method, affixed with batch labels, and then tested.

[0154] Preparation of reference solution 2: Take appropriate amounts of methanol, ethanol, isopropanol, acetonitrile, dichloromethane, tert-butyl methyl ether, ethyl acetate, tetrahydrofuran, n-heptane, 1,4-dioxane, toluene, and N,N-dimethylformamide, weigh accurately, and dilute with N-methylpyrrolidone to make a mixed solution containing about 300μg of methanol, 500μg of ethanol, 500μg of isopropanol, 41μg of acetonitrile, 60μg of dichloromethane, 500μg of tert-butyl methyl ether, 500μg of ethyl acetate, 72μg of tetrahydrofuran, 500μg of n-heptane, 38μg of 1,4-dioxane, 89μg of toluene, and 88μg of N,N-dimethylformamide per 1ml. Accurately measure 1ml, place in an empty bottle, and seal it as the reference solution.

[0155] 3. Determination method:

[0156] The determination method is the same as that in Example 7.

[0157] 4. Measurement results

[0158] The results were calculated by external standard method using peak area. See Table 12 for the chromatograms. See Figure 11 for the chromatograms of reference solution 2. Fig.11A , Chromatogram of the test solution Formula II compound batch 01 Fig. 11B , Chromatogram of Batch 02 Fig. 11C .

[0159] Table 12. Results of residual solvent determination of compound of formula II

[0160]

Claims

1. A gas chromatography analysis method for simultaneously detecting multiple residual solvents in a drug, characterized in that: The gas chromatography conditions were as follows: Inlet temperature: 200-250°C; preferably 230°C; Column box heating program: initial column temperature is 30-40°C, preferably 35°C, maintained for 6-10 min, preferably 8-10 min, heated to 220°C at a rate of 15-25°C per minute, preferably heated to 220°C at a rate of 15-20°C per minute, maintained for 3-7 min, preferably 5-7 min.

2. The analysis method according to claim 1, characterized in that The sample to be tested needs to be diluted with a diluent before injection, and the diluent is N-methylpyrrolidone.

3. The analysis method according to any one of claims 1 or 2, characterized in that The chromatographic column used in the analytical method is a medium polarity or non-polar capillary chromatographic column.

4. The analysis method according to claim 3, characterized in that The medium polarity capillary chromatographic column is a capillary chromatographic column using 6% cyanopropylphenyl-94% dimethylpolysiloxane as a stationary liquid, preferably a VF624 capillary chromatographic column.

5. The analysis method according to claim 3, characterized in that The non-polar capillary chromatographic column is a capillary chromatographic column with 100% dimethyl polysiloxane as the stationary liquid, preferably an HP-1 capillary chromatographic column.

6. The analysis method according to any one of claims 1 to 5, characterized in that The carrier gas flow rate in the gas chromatograph is 2.0-3.0 ml / min.

7. The analysis method according to any one of claims 1 to 6, characterized in that The injection mode of the analysis method is split injection, and the split ratio is 1-10:1, preferably the split ratio is 5-10:

1.

8. The analysis method according to any one of claims 1 to 7, characterized in that The headspace equilibrium temperature of the analytical method is greater than 70°C, preferably 70-90°C.

9. The analysis method according to any one of claims 1 to 8, characterized in that The analytical method can be used to detect a variety of residual solvents, which are one or more combinations of methanol, tetrahydrofuran, ethanol, cyclohexane, ether, isopropyl acetate, acetone, n-heptane, isopropanol, n-butanol, acetonitrile, dioxane, dichloromethane, pyridine, tert-butanol, toluene, tert-butyl methyl ether, N,N-dimethylformamide, n-hexane, dimethyl sulfoxide and ethyl acetate.

10. The analysis method according to any one of claims 1 to 9, characterized in that: The drug is a biological targeted drug or a chemical targeted drug; preferably, the biological targeted drug is one or more of a monoclonal antibody, a biclonal antibody, and an antibody-drug conjugate (ADC).

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