General analysis and detection method for 10 haloalkanes in medicine

Abstract

The invention discloses a general analysis and detection method for 10 haloalkanes in a medicine, which comprises preparation of a sample solution and a reference substance solution, gas chromatography-mass spectrometry analysis and detection, and qualitative and quantitative analysis, and comprises the following steps: step 1, adding an acid into an organic solvent to prepare a diluent, and preparing the reference substance solution and the sample solution; 2, injecting the solution in the step 1 into a gas chromatography-mass spectrometer through headspace sample injection for detection; and 3, qualitative and quantitative analysis. According to the method, general analysis method development is carried out on 10 common halogenated alkanes, the 10 common halogenated alkanes in the later project process can be detected at the same time, the detection sensitivity can reach 1 ppm, the stability of the halogenated alkanes reaches 48 h or above, and the method is high in universality and can be widely applied to different types of raw materials, intermediates and API samples.

Description

Technical Field

[0001] The present invention belongs to the technical field of drug analysis and detection, and relates to the analysis of genotoxic impurities of halogenated alkanes in raw materials, and in particular relates to a method for detecting halogenated alkanes by GCMS and a high-sensitivity analysis and detection method, and specifically relates to a universal analysis and detection method for 10 halogenated alkanes in a drug. Background Art

[0002] The analysis and control of genotoxic impurities of halogenated alkanes is an important part of drug quality control. The traditional method for detecting halogenated alkanes is mainly gas chromatography, which has low sensitivity and the stability of halogenated alkanes is a prominent problem, especially tert-butyl chloride and carbon tetrachloride, which have poor stability. The reference substance decreases rapidly within a few hours, while the stability requirement of the analytical method for later projects is generally 24-48 hours. Summary of the invention

[0003] The technical problem to be solved by the present invention is to provide a universal analysis and detection method for 10 halogenated alkanes commonly used in later project processes, and to solve the stability problem to meet the needs of later projects.

[0004] In order to solve the above technical problems, the specific technical solutions adopted by the present invention are as follows:

[0005] A universal analytical detection method for 10 halogenated alkanes in medicines, characterized by comprising sample solution, reference solution preparation, gas chromatography-mass spectrometry analysis and detection, qualitative and quantitative analysis, and the specific steps are as follows:

[0006] Step 1: Add acid to an organic solvent to prepare a diluent, and prepare a reference solution and a sample solution;

[0007] Step 2: injecting the solution in step 1 into a gas chromatograph-mass spectrometer for detection by headspace injection;

[0008] Step 3: Qualitative and quantitative analysis.

[0009] The 10 halogenated alkanes are methyl chloride, ethyl chloride, 2-chloropropane, 1,1-dichloroethylene, tert-butyl chloride, 2-bromopropane, 1,1,1-trichloroethane, carbon tetrachloride, 1,2-dichloroethane, and 1,2-dibromoethane.

[0010] In some specific embodiments, the organic solvent in the diluent in step 1 is selected from one or more of N,N-dimethylacetamide, N-methylpyrrolidone, dimethyl sulfoxide, 1,3-dimethyl-2-imidazolidinone, and N,N-dimethylformamide.

[0011] In some specific embodiments, the acid in the diluent in step 1 is selected from one or more of phosphoric acid, sulfuric acid, p-toluenesulfonic acid or other less volatile acids.

[0012] In some specific embodiments, the sample solution in step 1 and step 2 is selected from one or more of in-process control samples, raw material samples, intermediate samples, and product samples.

[0013] In some specific embodiments, the sample solution and the reference solution in step 1 are prepared by adding the diluent after weighing, and mixing to a fixed volume; step 1 also includes a quantitative limit solution, which is obtained by diluting the reference solution.

[0014] In some specific embodiments, the injection operation in step 2 is to inject diluent, quantitation limit solution (LOQ), reference solution, sample solution, and spiked sample solution in sequence. In some specific embodiments, the chromatographic conditions of step 2 are: the chromatographic column is Agilent DB-624, 60m×0.32mm, ID 1.8μm, PN:123-1364; the injection port temperature is 240°C; the chromatographic column temperature program is 40°C for 10min, the temperature program is 10°C / min to 70°C and then maintained for 5min, the temperature program is 50°C / min to 250°C and then maintained for 8min; the flow rate is 3mL / min; the split ratio is 5:1; the mass spectrometry conditions of step 2 are: the ion source is EI; the ion source temperature is 230°C; the quadrupole temperature is 150°C; the acquisition mode is SIM.

[0015] In some specific embodiments, the parameters of the headspace sampler equipped with the gas chromatograph-mass spectrometer in step 2 are: headspace furnace temperature 80°C; sample quantitative loop temperature 130°C; transfer line temperature 150°C; sample bottle equilibration time 20min; sample bottle pressure 17psi; headspace sample bottle loading volume 1.0mL; headspace bottle specification 20mL.

[0016] In some specific embodiments, the formula for qualitative and quantitative analysis in step 3 is selected from one of the following calculation formulas:

[0017]

[0018]

[0019] Where:

[0020] A Res.Sol. represents the peak area of ​​each specific solvent observed in the sample solution;

[0021] W SPL Represents the mass of the sample in mg;

[0022] V SPL Represents the dilution volume of the sample solution, in mL;

[0023] It represents the average response factor of each specified solvent in the reference solution;

[0024] The calculation formula of the response factor RF of each specific solvent in reference solution-1 and reference solution-2 is as follows:

[0025]

[0026] Where:

[0027] A STD Represents the peak area of ​​each specific solvent in the reference solution;

[0028] W STD Represents the mass of each specific solvent in the reference solution, in mg;

[0029] V STD Represents the dilution volume of the reference solution in mL.

[0030] Compared with the prior art, the present invention has the following beneficial effects:

[0031] The present invention develops a universal analytical method for 10 common halogenated alkanes, which can simultaneously detect the 10 common halogenated alkanes in the later project process, with a detection sensitivity of up to 1 ppm, and makes the halogenated alkanes stable for more than 48 hours. The method has strong versatility and can be widely applied to different types of raw materials, intermediates and API samples.

[0032] The concept of the present invention and the technical effects produced will be further described below in conjunction with the accompanying drawings to fully understand the purpose, characteristics and effects of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 These are the GCMS TIC diagrams of ten halogenated alkanes.

[0034] Figure 2 is a linear graph for methyl chloride.

[0035] Figure 3 is a linear graph for ethyl chloride.

[0036] Figure 4 It is a linear graph for 2-chloropropane.

[0037] Figure 5 is the linear graph of 1,1-dichloroethylene

[0038] Figure 6 is the linear graph of tert-butyl chloride

[0039] Figure 7 is the linear graph of 2-bromopropane

[0040] Figure 8 is the linear graph of 1,1,1-trichloroethane

[0041] Fig. 9 is a linear graph of carbon tetrachloride

[0042] Fig.10 is the linear graph of 1,2-dichloroethane

[0043] Fig.11 is the linear graph of 1,2-dibromoethane DETAILED DESCRIPTION

[0044] In order to make the technical means, creative features, objectives and effects of the invention easier to understand, the invention is further described below with reference to specific diagrams. However, the invention is not limited to the following implementation cases.

[0045] It should be noted that the drawings in this specification are only used to match the contents disclosed in the specification so as to facilitate understanding and reading by persons familiar with this technology, and are not used to limit the conditions under which the present invention can be implemented. Therefore, they have no substantive technical significance. Any adjustments without creative effort, without affecting the effects and purposes that can be achieved by the present invention, should still fall within the scope of the technical contents disclosed by the present invention.

[0046] All kinds of reagents, antibodies, and consumables involved in the examples are commercially available products, and all commercial products can be used.

[0047] The present invention adds a low-volatile acid to the diluent of the headspace method, so that no matter what the acidity or alkalinity of the sample is, the acid in the diluent is sufficient to maintain an acidic environment to help stabilize the halogenated alkanes. In terms of instruments, the present invention uses headspace GCMS technology to establish a universal analytical method for detecting 10 common halogenated alkanes in late-stage project processes. This technology is applicable to the GCMS content determination of 10 halogenated alkanes in raw materials, in-process controls, intermediates and products of clinical projects: methyl chloride, ethyl chloride, 2-chloropropane, 1,1-dichloroethylene, tert-butyl chloride, 2-bromopropane, 1,1,1-trichloroethane, carbon tetrachloride, 1,2-dichloroethane, and 1,2-dibromoethane. The corresponding ranges and retention times are listed in Table 1 below:

[0048] Table 1 Detection range and retention time of 10 chlorinated alkanes

[0049] name Working range (μg / mL) Retention time (min) Methyl chloride 0.05-5 4.5 Ethyl chloride 0.05-5 6.0 2-Chloropropane 0.05-5 7.9 1,1-Dichloroethylene 0.05-5 8.4 tert-Butyl chloride 0.05-5 10.1 2-Bromopropane 0.05-5 12.2 1,1,1-Trichloroethane 0.05-5 15.5 Carbon tetrachloride 0.05-5 16.0 1,2-Dichloroethane 0.05-5 16.8 1,2-Dibromoethane 0.05-5 21.5

[0050] The main reagents of the present invention are shown in Table 2 below:

[0051] Table 2 Main reagents

[0052]

[0053]

[0054]

[0055] Example 1 Chromatographic, mass spectrometric conditions, headspace injector parameters

[0056] Set the chromatographic, mass spectrometric conditions, headspace sampler parameters, etc. as shown in Tables 3, 4, and 5 below:

[0057] Table 3 Chromatographic conditions

[0058]

[0059] Table 4 Mass spectrometry conditions

[0060]

[0061]

[0062] Note: The above conditions were established based on the Agilent system. When using other instruments, some non-critical parameters will be adjusted (e.g., solvent delay, MS on, MS off, etc.); the cycle time (if any) may be extended based on actual conditions.

[0063] Table 5 MS headspace sampler parameters

[0064]

[0065] Example 2 Solution Preparation

[0066] The preparations of reference substance stock solution 1, reference substance stock solution 2, reference substance solution, quantitative limit solution, and sample solution are shown in Tables 6 to 8 below.

[0067] Table 6 Preparation of reference substance stock solution 1 and reference substance stock solution 2

[0068]

[0069]

[0070] Table 7 Preparation of reference solution 1, reference solution 2 and quantitative limit solution

[0071]

[0072] Table 8 Sample solution preparation

[0073]

[0074] Example 3 Sample injection analysis and detection

[0075] System adaptability requirements are as follows:

[0076] Table 9 System adaptability requirements

[0077]

[0078]

[0079] Example 4 Calculation of test results

[0080] The response factor (RF) for each specific solvent in STD-1 and STD-2 solutions was calculated according to the following formula:

[0081]

[0082] in:

[0083] A STD represents the peak area of ​​each specific solvent in the standard solution (the average peak area of ​​the first 6 injections using STD-1 solution);

[0084] W STD Represents the weight of each specified solvent in the standard solution (mg);

[0085] V STD Represents the diluted volume of the standard solution (mL).

[0086] The amount of each specific residual solvent observed in the chromatogram was calculated according to one of the following two formulas:

[0087]

[0088]

[0089] in:

[0090] A Res.Sol. represents the peak area of ​​each specific solvent observed in the sample solution;

[0091] W SPL Represents the weight of the sample (mg);

[0092] V SPL represents the dilution volume of the sample solution (mL);

[0093] Represents the average response factor for each specific solvent in the standard solution.

[0094] Example 5 Analysis and Test Results

[0095] For each specific solvent not below LOQ: the average result should be reported as the final result. However, if one test result is below LOQ, the maximum result should be reported as the final result.

[0096] For each specific solvent below LOQ, report the result as “ <LOQ”。

[0097] If a result is outside the specification limit, an OOS investigation is conducted.

[0098] The LOQ of each specific solvent in the sample is shown in Table 10 below:

[0099] Table 10 LOQ of each specific solvent

[0100]

[0101] The test results are shown in Table 11 below:

[0102] Table 11 Experimental results

[0103]

[0104]

[0105]

[0106] It can be seen that the detection sensitivity of the 10 halogenated alkanes can reach 1 ppm, and the halogenated alkanes have good stability, reaching more than 48 hours, and the method has strong versatility.

[0107] The preferred specific embodiments of the present invention are described in detail above. It should be understood that ordinary technicians in the field can make many modifications and changes based on the concept of the present invention without creative work. Therefore, all technical solutions that can be obtained by technicians in the technical field based on the concept of the present invention through logical analysis, reasoning or limited experiments on the basis of the prior art should be within the scope of protection determined by the claims.

Claims

1. A universal analytical method for detecting 10 halogenated alkanes in medicines, characterized in that: Including sample solution, reference solution preparation, gas chromatography-mass spectrometry analysis and detection, qualitative and quantitative analysis, the specific steps are as follows: Step 1: Add acid to an organic solvent to prepare a diluent, and prepare a reference solution and a sample solution; Step 2: injecting the solution in step 1 into a gas chromatograph mass spectrometer through headspace injection for detection; Step 3: Qualitative and quantitative analysis. The 10 halogenated alkanes are methyl chloride, ethyl chloride, 2-chloropropane, 1,1-dichloroethylene, tert-butyl chloride, 2-bromopropane, 1,1,1-trichloroethane, carbon tetrachloride, 1,2-dichloroethane, and 1,2-dibromoethane.

2. The method according to claim 1, characterized in that The organic solvent in the diluent in step 1 is selected from one or more of N,N-dimethylacetamide, N-methylpyrrolidone, dimethyl sulfoxide, 1,3-dimethyl-2-imidazolidinone, and N,N-dimethylformamide.

3. The method according to claim 1, characterized in that The acid in the diluent in step 1 is selected from one or more of phosphoric acid, sulfuric acid, p-toluenesulfonic acid or other less volatile acids.

4. The method according to claim 1, characterized in that: The sample solution and the reference solution in step 1 are prepared by adding the diluent after weighing, and mixing to a fixed volume; step 1 also includes a quantitative limit solution, which is obtained by diluting the reference solution.

5. The method according to claim 1, characterized in that The chromatographic conditions of step 2 are as follows: the chromatographic column is Agilent DB-624, 60m×0.32mm, ID 1.8μm, PN:123-1364; the injection port temperature is 240°C; the chromatographic column temperature program is 40°C for 10 min, the temperature program is 10°C / min to 70°C and then maintained for 5 min, the temperature program is 50°C / min to 250°C and then maintained for 8 min; the flow rate is 3mL / min; the split ratio is 5:1; the mass spectrometry conditions of step 2 are as follows: the ion source is EI; the ion source temperature is 230°C; the quadrupole temperature is 150°C; the acquisition mode is SIM.

6. The method according to claim 1, characterized in that The parameters of the headspace sampler equipped with the gas chromatograph-mass spectrometer in step 2 are: headspace furnace temperature 80°C; sample quantitative loop temperature 130°C; transfer line temperature 150°C; sample bottle equilibration time 20min; sample bottle pressure 17psi; headspace sample bottle loading volume 1.0mL; headspace bottle specification 20mL.

7. The method according to claim 1, characterized in that The formula for qualitative and quantitative analysis in step 3 is selected from one of the following calculation formulas: 1) 2) Where: A Res.Sol. represents the peak area of ​​each specific solvent observed in the sample solution; W SPL Represents the mass of the sample in mg; V SPL Represents the dilution volume of the sample solution, in mL; RF represents the average response factor of each specified solvent in the reference solution; The calculation formula of the response factor RF of each specific solvent in reference solution-1 and reference solution-2 is as follows: Where: A STD Represents the peak area of ​​each specific solvent in the reference solution; W STD Represents the mass of each specific solvent in the reference solution, in mg; V STD Represents the dilution volume of the reference solution in mL.

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