Method for detecting sulfonate impurities in medicine and application

Abstract

The invention discloses a method for detecting sulfonate impurities in a medicine and application. The method comprises the following step: detecting a test solution by adopting a gas chromatographic method, wherein the test solution is a solution containing a medicine to be detected; the gas chromatography conditions are as follows: the sample introduction flow rate is 0.8-2.8 mL / min; the sample injection split ratio is (15-30): 1; the temperature of a sample inlet is 180-200 DEG C; the temperature of a detector is 220-300 DEG C; the temperature rise mode is gradient temperature rise; the initial temperature is 30-35 DEG C; the detection method disclosed by the invention can be used for simultaneously detecting four genotoxic impurities including methyl methanesulfonate, ethyl methanesulfonate, isopropyl methanesulfonate and propyl methanesulfonate in the medicine, and has the advantages of good separation effect, high sensitivity, no blank interference, easiness in popularization of detection equipment, low cost, high analysis speed and the like.

Description

Technical Field

[0001] The invention belongs to the technical field of drug analysis, and particularly relates to a detection method and application of sulfonate impurities in drugs. Background Art

[0002] Rasagiline mesylate is a first-line drug for the early treatment of Parkinson's disease (PD), and it also has an improving effect on patients whose efficacy of long-term use of dopa preparations has declined. Its structural formula is:

[0003]

[0004] The chemical name is (R)-N-2-propynyl-1-hydroindaneamine mesylate. Rasagiline mesylate is prone to form mesylate compounds during the synthesis process. Mesylate compounds, as a type of genotoxic impurities, can act on the purine group of DNA molecules to generate alkylated purines, destroy the spatial conformation and stability of the DNA double helix, and induce gene mutations and cancer in the body. Due to its potential destructiveness to DNA, it needs to be controlled both in raw materials and preparations.

[0005] At present, the method of direct injection of gas chromatograph after extraction provided by the supplier of raw materials is difficult to avoid the problems such as the pollution of the raw material itself to the instrument and the poor separation effect of the main component and each impurity in the sample. In recent years, although there are more literature reports on the detection method of mesylate substances at home and abroad, methyl mesylate, ethyl mesylate, propyl mesylate and isopropyl mesylate in rasagiline mesylate are detected simultaneously by gas chromatography (GC), and there is no report in the prior art. For the mesylate impurities with lower detection limit, domestic and foreign literature usually chooses to detect by mass spectrometry, and these methods are all higher to equipment price and equipment maintenance cost. Summary of the invention

[0006] The present invention provides a method and application of detecting sulfonate impurities in a drug to solve the problem that the detection method in the prior art cannot simultaneously detect the mesylate impurities in the drug (e.g., methyl mesylate, ethyl mesylate, propyl mesylate and isopropyl mesylate, four genotoxic impurities in rasagiline mesylate). The detection method of the present invention can simultaneously detect four genotoxic impurities in the drug, namely, methyl mesylate, ethyl mesylate, isopropyl mesylate and propyl mesylate, and has the advantages of good separation effect, strong specificity, high sensitivity, no blank interference, low detection cost and fast analysis speed.

[0007] The present invention adopts a sodium thiosulfate derivatization method as a pretreatment method, reacts a mesylate with sodium iodide in acetonitrile to obtain a corresponding iodinated alkane product, and can achieve qualitative and quantitative detection of four mesylate genotoxic impurities in rasagiline mesylate while simultaneously determining the genotoxic impurities in rasagiline mesylate, and can be used for the detection of mesylate genotoxic impurities in rasagiline mesylate bulk drug and its preparation. (The chemical structural formulas and names of the four genotoxic impurities are shown in Table 1)

[0008] Table 1

[0009]

[0010] The present invention solves the above technical problems through the following technical solutions.

[0011] The present invention provides a method for detecting sulfonate impurities in a drug, which comprises the following steps: using gas chromatography to detect a test solution, and obtaining:

[0012] Wherein, the test solution is a solution containing the drug to be tested;

[0013] The methanesulfonate impurities include methyl methanesulfonate, ethyl methanesulfonate, isopropyl methanesulfonate, and propyl methanesulfonate;

[0014] In the gas chromatography method, the gas chromatography conditions include: injection flow rate of 0.8-2.8mL / min; injection split ratio of (15-30):1; injection port temperature of 180-200°C; detector temperature of 220-300°C; heating method of gradient heating; initial temperature of 30-35°C.

[0015] In the present invention, the injection flow rate is preferably 0.8-2.5 mL / min, such as 1.0 mL / min, 1.5 mL / min, 1.8 mL / min, 2.0 mL / min or 2.5 mL / min. In the present invention, in the gas chromatography, the operation mode is generally constant flow.

[0016] In the present invention, the injection split ratio can be 15: 1, 20: 1, 25: 1 or 30: 1. Those skilled in the art are aware that the injection split ratio refers to the ratio of the flow rate of the sample entering the chromatographic column through the split injector to the flow rate discharged through the splitter under the condition that the injected sample is completely gasified and fully mixed with the carrier gas.

[0017] In the present invention, the injection port temperature may be 180°C, 190°C or 200°C.

[0018] In the present invention, the detector temperature is preferably 220-290°C, such as 230°C, 240°C, 260°C or 280°C.

[0019] In the present invention, preferably, the process of the gradient heating is shown in the following table:

[0020] Initial temperature(℃) Heating rate (℃ / min) Final temperature(℃) Retention time (min) 30-35 0 30-35 0-5 30-35 5-10 100-120 0-5 100-120 5-30 220-250 3-10

[0021] The data in the above table are explained as follows:

[0022] The initial temperature is 30-35°C; during 0-5min, the temperature increases from 30-35°C at a heating rate of 5-10°C / min; the temperature is increased to 100-120°C; at 100-120°C, it is retained for 0-5min; from 100-120°C, the temperature is increased at a heating rate of 5-30°C / min; the temperature is increased to 220-250°C; at 220-250°C, it is retained for 3-10min.

[0023] In some preferred embodiments of the present invention, the process of the gradient heating is as follows:

[0024] The initial temperature is 30°C; from 0 to 5 minutes, the temperature is 30°C; the temperature increases from 30°C at a heating rate of 5-10°C / min; the temperature is increased to 100°C; at 100°C, it is retained for 0-5 minutes; from 100°C, the temperature is increased at a heating rate of 5-30°C / min; the temperature is increased to 220°C; at 220°C, it is retained for 3-10 minutes.

[0025] In some preferred embodiments of the present invention, the process of the gradient heating is as follows:

[0026] The initial temperature is 35°C; from 0 to 5 minutes, the temperature is 35°C; the temperature increases from 35°C at a heating rate of 5-10°C / min; the temperature is increased to 100°C; at 100°C, it is retained for 0-5 minutes; from 100°C, the temperature is increased at a heating rate of 5-30°C / min; the temperature is increased to 220°C; at 220°C, it is retained for 3-10 minutes.

[0027] In some preferred embodiments of the present invention, the process of the gradient heating is as follows:

[0028] The initial temperature is 35°C; from 0 to 5 minutes, the temperature is 35°C; the temperature increases from 35°C at a heating rate of 5-10°C / min; the temperature is increased to 120°C; at 120°C, it is retained for 0-5 minutes; from 120°C, the temperature is increased at a heating rate of 5-30°C / min; the temperature is increased to 220°C; at 220°C, it is retained for 3-10 minutes.

[0029] In some preferred embodiments of the present invention, the process of the gradient heating is as follows:

[0030] The initial temperature is 30°C; from 0 to 5 minutes, the temperature is 30°C; the temperature increases from 30°C at a heating rate of 5-10°C / min; the temperature is increased to 100°C; at 100°C, it is retained for 0-5 minutes; from 100°C, the temperature is increased at a heating rate of 5-30°C / min; the temperature is increased to 250°C; at 250°C, it is retained for 3-10 minutes.

[0031] In the present invention, in the gas chromatography method, the carrier gas is preferably nitrogen or helium.

[0032] In the present invention, in the gas chromatography, the injection method is headspace injection. The headspace injection is generally performed using a headspace injector. The headspace injector can be a conventional headspace injector in the art, and the specification of the quantitative loop is preferably 1 mL.

[0033] The headspace temperature of the headspace sampler may be a conventional temperature in the art, preferably 50-80°C, such as 50°C, 60°C or 70°C.

[0034] The temperature of the quantitative loop of the head space sampler may be a conventional temperature in the art, preferably 50-80°C, such as 60°C or 70°C.

[0035] The transmission line temperature of the head space sampler may be a conventional temperature in the art, preferably 60-100°C, such as 70°C or 80°C.

[0036] Among them, the headspace temperature, quantitative loop temperature and transmission line temperature of the headspace injection can be conventional temperatures in the art, and the temperatures are increased by 10°C to 20°C in sequence, for example, the headspace temperature is 60°C to 80°C, the quantitative loop temperature is the headspace temperature plus 10°C to 20°C, and the transmission line temperature is the quantitative loop temperature plus 10°C to 20°C.

[0037] Preferably, the headspace temperature, the quantitative loop temperature and the transfer line temperature of the headspace sampler are increased by 10-20°C in sequence. That is, the quantitative loop temperature is the headspace temperature plus 10-20°C, and the transfer line temperature is the quantitative loop temperature plus 10-20°C. For example, the headspace temperature, the quantitative loop temperature and the transfer line temperature of the headspace sampler are 50°C, 60°C and 70°C in sequence; or, the headspace temperature, the quantitative loop temperature and the transfer line temperature of the headspace sampler are 60°C, 70°C and 80°C in sequence.

[0038] In the present invention, in the gas chromatography method, the stationary phase is preferably 6% cyanopropyl-94% dimethylsiloxane, 5% phenyl-95% dimethylsiloxane or polyethylene glycol.

[0039] In the present invention, in the gas chromatography, the length of the chromatographic column can be 30-75 mm, preferably 30 mm, 50 mm or 75 mm, and more preferably 30 mm.

[0040] In the present invention, in the gas chromatography method, the inner diameter of the chromatographic column can be 250-530 μm, preferably 250 μm, 320 μm or 530 μm, and more preferably 250 μm or 530 μm.

[0041] In the present invention, in the gas chromatography method, the coating thickness of the chromatographic column can be 0.25-2.0 μm, preferably 0.25 μm, 1.0 μm, 1.4 μm, 1.8 μm or 2.0 μm, more preferably 1.0 μm or 1.4 μm.

[0042] In the present invention, in the gas chromatography method, preferably, the chromatographic column is a chromatographic column produced by Agilent, model HP-INNOWAX, specification 30mm*530μm, 1.0μm; or a chromatographic column produced by Agilent, model DB-624, specification 30mm*250μm, 1.4μm.

[0043] In the present invention, the drug to be tested may be rasagiline mesylate. The rasagiline mesylate may be a conventional rasagiline mesylate bulk drug and its preparation in the art, such as the rasagiline mesylate bulk drug purchased from Amino Chemicals Ltd.

[0044] In the present invention, the test solution may generally further include a derivatization solution and an organic solvent.

[0045] The derivatization solution may include sodium iodide, sodium thiosulfate and water. The preparation method of the derivatization solution may generally include the following steps: mixing sodium iodide, sodium thiosulfate and water. The sodium thiosulfate may be sodium thiosulfate pentahydrate or anhydrous sodium thiosulfate. The water is generally purified water.

[0046] The mass ratio of the sodium iodide to the sodium thiosulfate may be conventional in the art, preferably 2000:1.

[0047] The organic solvent may be a conventional organic solvent in the art, preferably acetonitrile.

[0048] Wherein, in the test solution, the mass volume ratio of the drug to be tested and the derivatization solution can be conventional in the art, preferably 100:1 mg / L.

[0049] Wherein, in the test solution, the volume ratio of the derivatization solution to the organic solvent can be a conventional volume ratio in the art, preferably 1:1.

[0050] In the present invention, the detection method may also include calculating the content according to the external standard method; detecting the test solution according to the gas chromatography method to obtain the peak area Ax of the sulfonate impurity in the test solution; detecting the reference solution according to the gas chromatography method to obtain the peak area Ar of the sulfonate impurity in the reference solution, and the concentration Cr of the sulfonate impurity in the reference solution is known; according to the external standard method formula Cx=Cr*Ax / Ar, the concentration Cx of the sulfonate impurity in the test solution is calculated.

[0051] The reference solution may include a derivatization solution and a reference stock solution. The preparation method of the reference solution may generally include the following steps: mixing the derivatization solution and the reference stock solution. The obtained reference solution is generally sealed in a headspace bottle for standby use.

[0052] The derivatization solution can be as described above.

[0053] The reference substance stock solution may include methyl methanesulfonate, ethyl methanesulfonate, isopropyl methanesulfonate, propyl methanesulfonate and a diluent.

[0054] The diluent may be a conventional organic solvent in the art, preferably acetonitrile.

[0055] The concentration of the genotoxic impurities (methyl methanesulfonate, ethyl methanesulfonate, isopropyl methanesulfonate and propyl methanesulfonate) of each component in the reference substance stock solution can be a conventional concentration in the art, preferably 0.01 mg / mL.

[0056] In the reference substance solution, the volume ratio of the derivatization solution to the reference substance stock solution can be a conventional volume ratio in the art, preferably 1:1.

[0057] The present invention also provides an application of the above-mentioned method for detecting sulfonate impurities in a drug in detecting genotoxic impurities in a rasagiline mesylate raw material or a preparation thereof.

[0058] On the basis of being in accordance with the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0059] The reagents and raw materials used in the present invention are commercially available.

[0060] Compared with the existing analysis methods, the beneficial effects of the present invention are:

[0061] The invention establishes a method for simultaneously detecting four genotoxic impurities, namely methyl methanesulfonate, ethyl methanesulfonate, isopropyl methanesulfonate and propyl methanesulfonate, in a drug, which has the advantages of good separation effect, strong specificity, high sensitivity, no blank interference, low detection cost and fast analysis speed.

[0062] Through methodological verification, the specificity, linearity, range, repeatability, detection limit, quantitation limit and accuracy of the detection method of the present invention all meet the requirements of the guidance principles for validation of analytical methods for drug quality standards in the appendix of Part IV of the 2015 edition of the Chinese Pharmacopoeia. The present invention has important practical significance in the quality control of the synthesis and preparation process of rasagiline mesylate. BRIEF DESCRIPTION OF THE DRAWINGS

[0063] Figure 1 This is a reference spectrum of the genotoxicity analysis method of rasagiline mesylate-system suitability solution in Example 1.

[0064] Figure 2 This is a reference spectrum of the genotoxicity analysis method of rasagiline mesylate-system suitability solution in Example 2.

[0065] Figure 3 This is a reference spectrum of the genotoxicity analysis method of rasagiline mesylate-test solution in Example 2.

[0066] Figure 4 This is a reference spectrum of the genotoxicity analysis method of rasagiline mesylate in Comparative Example 1 - system suitability solution.

[0067] Figure 5 This is a partial enlarged view of the reference spectrum of the genotoxicity analysis method of rasagiline mesylate-system suitability solution in Comparative Example 1.

[0068] Figure 6 This is a reference spectrum of the genotoxicity analysis method of rasagiline mesylate in Comparative Example 2 - system suitability solution.

[0069] Figure 7 This is a reference spectrum of the genotoxicity analysis method of rasagiline mesylate in Comparative Example 3 - system suitability solution. DETAILED DESCRIPTION

[0070] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0071] Unless otherwise specified, the reagents used in the following examples and comparative examples can be obtained and used from conventional commercial sources.

[0072] In the following examples and comparative examples, the gas chromatograph used is an Agilent 7890B series gas chromatograph.

[0073] Example 1

[0074] (1) Chromatographic conditions:

[0075] Stationary phase: polyethylene glycol (manufacturer Agilent, model HP-INNOWAX, 30mm*530μm, 1.0μm)

[0076] The carrier gas is helium;

[0077] The operation mode was constant flow, and the flow rate was 0.8 mL / min;

[0078] The injection mode was split injection with a split ratio of 30:1;

[0079] Inlet temperature 200°C, detector temperature 240°C;

[0080] The initial temperature was 30 °C, and the temperature was increased gradually;

[0081] The heating program is as follows:

[0082]

[0083] The injection method was headspace injection, the headspace temperature was 50°C, the quantitative loop temperature was 60°C, and the transfer line temperature was 70°C; injection volume: 1000 μL.

[0084] (2) Preparation of derivatization solution:

[0085] Weigh 60 g of sodium iodide and 30 mg of sodium thiosulfate pentahydrate, add 50 mL of purified water, mix well, and obtain the product.

[0086] (3) Preparation of blank solution:

[0087] Accurately pipette 1 mL of the derivatization solution and 1 mL of acetonitrile into a 20 mL headspace bottle and seal it.

[0088] (4) Preparation of reference solution:

[0089] Accurately weigh methyl methanesulfonate, ethyl methanesulfonate, isopropyl methanesulfonate and propyl methanesulfonate respectively, use acetonitrile as diluent, and prepare reference substance stock solutions containing 0.01 mg / mL of genotoxic impurities of each component.

[0090] Accurately pipette 1 mL of the derivatization solution and 1 mL of the reference stock solution into a 20 mL headspace bottle and seal it.

[0091] (5) Preparation of test solution:

[0092] Weigh 100 mg of rasagiline mesylate raw material (commercially available from Amino Chemicals Ltd.) and place it in a 20 mL headspace bottle. Accurately pipette 1 mL of the derivatization solution and 1 mL of acetonitrile and seal the bottle to obtain the product.

[0093] (6) System suitability solution:

[0094] Take 100 mg of rasagiline mesylate raw material and place it in a 20 mL headspace bottle, accurately weigh it, add 1 mL of derivatization solution and 1 mL of reference substance stock solution, seal it, and shake it well to obtain the product.

[0095] The system suitability solution can be used to prove that the blank solution and the peak positions of each impurity do not affect each other.

[0096] (7) Determination

[0097] Accurately measure 1000 μL of the blank solution, reference solution, test solution and system suitability solution mentioned above respectively, and inject them into the gas chromatograph for determination and calculation.

[0098] (8) Acceptance limit

[0099] The blank solution chromatogram has no interference at the main peak position; the separation degree between the main peak and the adjacent chromatographic peaks on the system suitability solution chromatogram should be no less than 1.5. The tailing factor of each peak should be less than 2.0; the RSD of the main peak area in the reference solution for 6 consecutive measurements is ≤10.0%; the RSD of the main peak retention time is ≤1.0%.

[0100] The separation effects of the four gene impurities measured in Example 1 are shown in Table 2.

[0101] Table 2

[0102]

[0103] Note: The peak order of the system suitability solution is methyl methanesulfonate, ethyl methanesulfonate, isopropyl methanesulfonate, propyl methanesulfonate and solvent peak (acetonitrile). Figure 1 The separation degree is the separation degree between two adjacent peaks. The data specifically refers to the separation degree with the latter peak. In the test solution, methyl methanesulfonate, ethyl methanesulfonate, isopropyl methanesulfonate and propyl methanesulfonate were not detected.

[0104] Example 2

[0105] (1) Chromatographic conditions:

[0106] Stationary phase: 6% cyanopropyl-94% dimethylsiloxane; (Manufacturer Agilent, Model DB-624, 30mm*250μm, 1.4μm)

[0107] The carrier gas is helium;

[0108] The operation mode was constant flow, and the flow rate was 1.0 mL / min;

[0109] The injection mode was split injection with a split ratio of 15:1;

[0110] Inlet temperature 200°C, detector temperature 240°C;

[0111] The initial temperature was 35 °C, and the temperature was increased gradually;

[0112] The heating program is as follows:

[0113]

[0114] The injection method was headspace injection, the headspace temperature was 60°C, the quantitative loop temperature was 70°C, and the transfer line temperature was 80°C; injection volume: 1000 μL.

[0115] (2) Preparation of derivatization solution:

[0116] Weigh 60 g of sodium iodide and 30 mg of sodium thiosulfate pentahydrate, add 50 mL of purified water, mix well, and obtain the product.

[0117] (3) Preparation of blank solution:

[0118] Accurately pipette 1 mL of the derivatization solution and 1 mL of acetonitrile into a 20 mL headspace bottle and seal it.

[0119] (4) Preparation of reference solution:

[0120] Methyl methanesulfonate, ethyl methanesulfonate, isopropyl methanesulfonate and propyl methanesulfonate were accurately weighed respectively, and acetonitrile was used as a diluent to prepare a reference stock solution containing 0.01 mg / mL of the genotoxic impurity of each component.

[0121] Accurately pipette 1 mL of the derivatization solution and 1 mL of the reference stock solution into a 20 mL headspace bottle and seal it.

[0122] (5) Preparation of test solution:

[0123] Weigh 100 mg of rasagiline raw material and place it in a 20 mL headspace bottle. Accurately pipette 1 mL of derivatization solution and 1 mL of acetonitrile and seal the bottle to obtain the product.

[0124] (6) System suitability solution:

[0125] Take 100 mg of rasagiline mesylate raw material and place it in a 20 mL headspace bottle, accurately weigh it, add 1 mL of derivatization solution and 1 mL of reference substance stock solution, seal it, and shake it well to obtain the product.

[0126] (7) Determination

[0127] Accurately measure 1000 μL of the blank solution, reference solution, test solution and system suitability solution mentioned above respectively, and inject them into the gas chromatograph for determination and calculation.

[0128] (8) Acceptance limit

[0129] The blank solution chromatogram has no interference at the main peak position; the separation of each chromatographic peak on the specific solution chromatogram should be no less than 1.5. The tailing factor of each peak should be less than 2.0; the RSD of the main peak area in the reference solution for 6 consecutive measurements is ≤10.0%; the RSD of the main peak retention time is ≤1.0%.

[0130] The methodological investigation is as follows:

[0131] (1) Specificity and system adaptability

[0132] The separation effects of the four gene impurities measured in Example 2 are shown in Table 3.

[0133] Table 3

[0134]

[0135]

[0136] Note: The peak order of the system suitability solution is methyl methanesulfonate, solvent peak (acetonitrile), ethyl methanesulfonate, isopropyl methanesulfonate and propyl methanesulfonate. Figure 2 The separation degree is the separation degree between two adjacent peaks. The data specifically refers to the separation degree with the latter peak. In the test solution, methyl methanesulfonate, ethyl methanesulfonate, isopropyl methanesulfonate and propyl methanesulfonate were not detected. For details, see Figure 3 .

[0137] The blank solution chromatogram has no interference at each peak position; the specific solution chromatogram (such as Figure 2 As shown in the figure, the separation of each chromatographic peak is greater than 1.5, and the tailing factor of each peak is greater than 2.0; the RSD of the main peak area in the reference solution for 6 consecutive times is ≤10.0%; and the RSD of the main peak retention time is ≤1.0%. It can be seen that the specificity and system adaptability of Example 2 meet the methodological requirements.

[0138] (2) Linear

[0139] Weigh the reference substances of methyl methanesulfonate, ethyl methanesulfonate, isopropyl methanesulfonate and propyl methanesulfonate respectively, use acetonitrile as diluent, and take the concentration of the reference substance stock solution (0.01 mg / mL) as 100% as the basis, respectively configure into 15%, 40%, 80%, 100%, 120% and 160% series standard solutions. Take 1 mL of each linear standard solution and place it in a 20 mL headspace bottle, accurately weigh it, add 1 mL of derivatization solution, seal it and mix it. Inject the sample according to the proposed chromatographic conditions, and perform linear regression on the mass concentration (X, mg / mL) of the corresponding solvent with the peak area (Y). It is found that each impurity has a good linear relationship in the concentration range of 0.8-8.0 μg / mL, and the correlation coefficient is greater than 0.999.

[0140] The linear regression equation is as follows:

[0141] The regression equation and correlation coefficient of methyl methanesulfonate were: y = 2.3466x + 0.0945, 0.9984;

[0142] The regression equation and residual sum of squares for ethyl methanesulfonate were: y = 3.8667x-0.0711, 0.9995;

[0143] The regression equation and residual sum of squares for isopropyl methanesulfonate were: y = 4.3840x + 0.1105, 0.9995;

[0144] The regression equation and residual sum of squares of propyl methanesulfonate are: y=3.7899x+0.1979, 0.9991 respectively.

[0145] The residual sums of squares for methyl methanesulfonate, ethyl methanesulfonate, isopropyl methanesulfonate, and propyl methanesulfonate are 0.5445, 0.6047, 0.8458, and 0.8496, respectively.

[0146] The slope intercepts of methyl methanesulfonate, ethyl methanesulfonate, isopropyl methanesulfonate and propyl methanesulfonate are 0.84%, 0.33%, 0.45% and 1.03%, respectively.

[0147] (3) Precision

[0148] 100 mg of rasagiline mesylate raw material was placed in a 20 mL headspace bottle, accurately weighed, 1 mL of derivatization solution and 1 mL of reference substance stock solution were added, sealed, shaken, and used as a repeatable solution. Six replicates were prepared in parallel. The RSD of the measured results of each genotoxic impurity was less than 3%.

[0149] (4) Sensitivity

[0150] Take 1mL of the reference stock solution and dilute it with acetonitrile to a 20mL volumetric flask as the sensitivity stock solution (0.0005mg / mL). Take a 20mL headspace bottle, add 1mL of the derivatization solution and 1mL of the sensitivity stock solution, seal it, shake it well, and use it as the sensitivity solution. The signal-to-noise ratio of each component is greater than 3.

[0151] (5) Accuracy

[0152] Weigh methyl methanesulfonate, ethyl methanesulfonate, isopropyl methanesulfonate and propyl methanesulfonate reference substances respectively, use acetonitrile as diluent, and take the concentration of reference substance stock solution (0.01mg / mL) as 100% benchmark, respectively configure into 40%, 100% and 120% three concentration gradients, as accuracy stock solution. Take 100mg of rasagiline mesylate raw material and place it in a 20mL headspace bottle, accurately weigh, add 1mL of derivatization solution and 1mL of each concentration gradient accuracy stock solution, seal, shake well, as accuracy solution, prepare 3 parts in parallel for each concentration. According to the proposed chromatographic conditions, the sample injection is measured, and the results show that the single recovery rate and average recovery rate of each genotoxic impurity content are all in the range of 80.0-120.0%.

[0153] As described above, the test method of Example 2 was validated by methodology for specificity, linearity, precision, sensitivity and accuracy, and the validation results all met the requirements of the Guiding Principles for Validation of Analytical Methods for Drug Quality Standards in Appendix IV of the 2015 Edition of the Chinese Pharmacopoeia.

[0154] Example 3

[0155] The difference between Example 3 and Example 1 is that the temperature is raised according to the following procedure:

[0156]

[0157] Example 4

[0158] The difference between Example 4 and Example 2 is that the temperature is raised according to the following procedure:

[0159]

[0160] The test results show that the separation effect, specificity, linearity, precision, sensitivity and accuracy of Example 3 are equivalent to those of Example 1; the separation effect, specificity, linearity, precision, sensitivity and accuracy of Example 4 are equivalent to those of Example 2.

[0161] Example 5

[0162] The only difference between Example 5 and Example 1 is that the carrier gas is nitrogen, the flow rate is 1.0 mL / min, and the split ratio is 25:1.

[0163] The separation effects of the four gene impurities measured in Example 5 are shown in Table 4.

[0164] Table 4

[0165]

[0166] Note: The peak order of the system suitability solution is methyl methanesulfonate, solvent peak (acetonitrile), ethyl methanesulfonate, isopropyl methanesulfonate and propyl methanesulfonate. The resolution is the separation degree between two adjacent peaks, and the data specifically refers to the resolution with the latter peak. In the test solution, methyl methanesulfonate, ethyl methanesulfonate, isopropyl methanesulfonate and propyl methanesulfonate were not detected.

[0167] It can be seen from Table 4 that the separation effect of Example 5 is equivalent to that of Example 1.

[0168] Comparative Example 1

[0169] The difference between Comparative Example 1 and Example 2 is that the injection split ratio is 3:1. The results show that the retention time of the methyl methanesulfonate peak and the solvent peak (acetonitrile peak) are close and difficult to separate (the separation degree is less than 1.5, see Figure 4 and Figure 5 ), resulting in the inability to separate the four genetic impurities simultaneously.

[0170] Comparative Example 2

[0171] The only difference between Comparative Example 2 and Example 1 is that the injection flow rate is 3 mL / min. The results show that the retention time of the methyl methanesulfonate peak and the solvent peak (acetonitrile peak) are close and difficult to separate (the separation degree is less than 1.5, see Figure 6 ), resulting in the inability to separate the four genetic impurities simultaneously.

[0172] Comparative Example 3

[0173] The only difference between Comparative Example 3 and Example 2 is that the injection flow rate is 0.5 mL / min. The results show that the retention time of the methyl methanesulfonate peak is close to that of the solvent peak (acetonitrile peak), and it is difficult to separate (the separation degree is less than 1.5, see Figure 7 ), resulting in the inability to separate the four genetic impurities simultaneously.

Claims

1. A method for detecting mesylate impurities in a drug, characterized in that: The method comprises the following steps: using gas chromatography to detect the test solution, and obtaining; Wherein, the test solution is a solution containing the drug to be tested; The methanesulfonate impurities include methyl methanesulfonate, ethyl methanesulfonate, isopropyl methanesulfonate, and propyl methanesulfonate; In the gas chromatography method, the gas chromatography conditions include: an injection flow rate of 0.8-2.8 mL / min; an injection split ratio of (15-30):1; an injection port temperature of 180-200°C; a detector temperature of 220-300°C; a temperature increase method of gradient temperature increase; and an initial temperature of 30-35°C.

2. The detection method according to claim 1, characterized in that It satisfies one or more of the following conditions ad; a. The injection flow rate is 0.8-2.5 mL / min, for example, 1.0 mL / min, 1.5 mL / min, 1.8 mL / min, 2.0 mL / min or 2.5 mL / min; b. The injection split ratio is 15:1, 20:1, 25:1 or 30:1; c. The injection port temperature is 180°C, 190°C or 200°C; d. The detector temperature is 220-290°C, such as 230°C, 240°C, 260°C or 280°C.

3. The detection method according to claim 1, characterized in that The process of the gradient heating is as follows: The initial temperature is 30-35°C; during 0-5min, the temperature increases from 30-35°C at a heating rate of 5-10°C / min; the temperature is increased to 100-120°C; at 100-120°C, it is retained for 0-5min; from 100-120°C, the temperature is increased at a heating rate of 5-30°C / min; the temperature is increased to 220-250°C; at 220-250°C, it is retained for 3-10min.

4. The detection method according to claim 3, characterized in that The process of the gradient heating is as follows: The initial temperature is 30°C; from 0 to 5 minutes, the temperature is 30°C; the temperature is increased from 30°C at a heating rate of 5-10°C / min; the temperature is increased to 100°C; at 100°C, it is retained for 0-5 minutes; from 100°C, the temperature is increased at a heating rate of 5-30°C / min; the temperature is increased to 220°C; at 220°C, it is retained for 3-10 minutes; Alternatively, the initial temperature is 35°C; from 0 to 5 minutes, the temperature is 35°C; the temperature is increased from 35°C at a heating rate of 5 to 10°C / min; the temperature is increased to 100°C; at 100°C, it is retained for 0 to 5 minutes; from 100°C, the temperature is increased at a heating rate of 5 to 30°C / min; the temperature is increased to 220°C; at 220°C, it is retained for 3 to 10 minutes; Alternatively, the initial temperature is 35°C; from 0 to 5 minutes, the temperature is 35°C; the temperature is increased from 35°C at a heating rate of 5 to 10°C / min; the temperature is increased to 120°C; at 120°C, it is retained for 0 to 5 minutes; from 120°C, the temperature is increased at a heating rate of 5 to 30°C / min; the temperature is increased to 220°C; at 220°C, it is retained for 3 to 10 minutes; Alternatively, the initial temperature is 30°C; from 0 to 5 minutes, the temperature is 30°C; the temperature increases from 30°C at a heating rate of 5 to 10°C / min; the temperature is increased to 100°C; at 100°C, it is retained for 0 to 5 minutes; from 100°C, the temperature is increased at a heating rate of 5 to 30°C / min; the temperature is increased to 250°C; at 250°C, it is retained for 3 to 10 minutes.

5. The detection method according to any one of claims 1 to 4, characterized in that: It satisfies one or more of the following conditions ac: a. In the gas chromatography, the carrier gas used is nitrogen or helium; b. In the gas chromatography method, the injection method is headspace injection; c. In the gas chromatography method, the stationary phase is 6% cyanopropyl-94% dimethylsiloxane, 5% phenyl-95% dimethylsiloxane or polyethylene glycol, preferably 6% cyanopropyl-94% dimethylsiloxane.

6. The detection method according to claim 5, characterized in that Using a headspace sampler to perform the headspace injection; The specification of the quantitative loop in the headspace sampler is preferably 1 mL; The headspace temperature of the headspace sampler is preferably 50-80°C, such as 50°C, 60°C or 70°C; The temperature of the quantitative loop of the head space sampler is preferably 50-80°C, such as 60°C or 70°C; The transfer line temperature of the head space sampler is preferably 60-100°C, such as 70°C or 80°C; Preferably, the headspace temperature, the quantitative loop temperature and the transfer line temperature of the headspace sampler are increased by 10-20°C in sequence; for example, the headspace temperature, the quantitative loop temperature and the transfer line temperature of the headspace sampler are 50°C, 60°C and 70°C in sequence; or, the headspace temperature, the quantitative loop temperature and the transfer line temperature of the headspace sampler are 60°C, 70°C and 80°C in sequence.

7. The detection method according to any one of claims 1 to 4, characterized in that: It satisfies one or more of the following conditions ac: a. In the gas chromatography, the length of the chromatographic column is 30-75 mm, preferably 30 mm, 50 mm or 75 mm; b. In the gas chromatography method, the inner diameter of the chromatographic column is 250-530 μm, preferably 250 μm, 320 μm or 530 μm; c. In the gas chromatography method, the coating thickness of the chromatographic column is 0.25-2.0 μm, preferably 0.25 μm, 1.0 μm, 1.4 μm, 1.8 μm or 2.0 μm; In the gas chromatography, the chromatographic column is preferably a chromatographic column produced by Agilent, model HP-INNOWAX, with specifications of 30mm*530μm, 1.0μm, or a chromatographic column produced by Agilent, model DB-624, with specifications of 30mm*250μm, 1.4μm.

8. The detection method according to claim 1, characterized in that It satisfies one or more of the following conditions ab: a. The drug to be tested is rasagiline mesylate; b. The test solution further comprises a derivatization solution and an organic solvent; the derivatization solution preferably comprises sodium iodide, sodium thiosulfate and water; c. The detection method further comprises calculating the content according to an external standard method; the external standard method comprises detecting the reference solution by the gas chromatography method; Wherein, the reference substance solution preferably includes the derivatization solution and a reference substance stock solution; the reference substance stock solution preferably includes methyl methanesulfonate, ethyl methanesulfonate, isopropyl methanesulfonate, propyl methanesulfonate and a diluent.

9. The detection method according to claim 8, characterized in that It satisfies one or more of the following conditions ag: a. In the test solution, the mass volume ratio of the drug to be tested and the derivatization solution is 100:1 mg / L; b. In the test solution, the volume ratio of the derivatization solution to the organic solvent is 1:1; c. In the derivatization solution, the mass ratio of the sodium iodide to the sodium thiosulfate is 2000:1; d. In the test solution, the organic solvent is acetonitrile; e. In the reference substance solution, the volume ratio of the derivatization solution to the reference substance stock solution is 1:1; f. In the reference substance stock solution, the diluent is acetonitrile; g. In the reference substance stock solution, the concentrations of methyl methanesulfonate, ethyl methanesulfonate, isopropyl methanesulfonate and propyl methanesulfonate are all 0.01 mg / mL.

10. Use of the detection method according to any one of claims 1 to 9 in detecting genotoxic impurities in rasagiline mesylate raw materials or preparations thereof.

Images