A general analysis method for 10 halogenated alkanes in pharmaceuticals

Abstract

The application discloses a general analysis and detection method for 10 kinds of halogenated alkanes in medicines, which comprises sample solution, preparation of control solution, gas chromatography mass spectrometry analysis and detection, qualitative and quantitative analysis, and steps are as follows: step 1: acid is added in an organic solvent to prepare a diluent, and the control solution and the sample solution are prepared; step 2: the solution in step 1 is injected into a gas chromatography mass spectrometer through headspace sampling for detection; and step 3: qualitative and quantitative analysis. The application develops a general analysis method for 10 kinds of common halogenated alkanes, can simultaneously detect the 10 kinds of common halogenated alkanes in a later project process, the detection sensitivity can reach 1 ppm, the stability of the halogenated alkanes can be more than 48 hours, the method has strong universality, and can be widely applied to different kinds of raw materials, intermediates and API samples.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical analysis and detection technology, and relates to the analysis of genotoxic impurities such as haloalkanes in active pharmaceutical ingredients. In particular, it relates to a highly sensitive analytical detection method for haloalkanes using GCMS, specifically a general analytical detection method for 10 haloalkanes in a drug. Background Technology

[0002] The analysis and control of genotoxic impurities such as haloalkanes is an important part of drug quality control. Traditional methods for detecting haloalkanes mainly rely on gas chromatography, which has insufficient sensitivity and exhibits significant instability issues, particularly tert-butyl chloride and carbon tetrachloride, whose stability degrades rapidly within hours. Furthermore, later-stage tests typically require analytical methods with a stability requirement of 24-48 hours. Summary of the Invention

[0003] The technical problem to be solved by this invention is to provide a general analytical detection method for 10 common haloalkanes in later-stage project processes, and to solve the stability problem, so as to meet the needs of later-stage projects.

[0004] To solve the above-mentioned technical problems, the specific technical solution adopted by the present invention is as follows:

[0005] A universal analytical method for detecting 10 haloalkanes in a pharmaceutical product, characterized by comprising the preparation of sample solution and reference solution, gas chromatography-mass spectrometry analysis, qualitative and quantitative analysis, with the specific steps as follows:

[0006] Step 1: Prepare a diluent by adding acid to an organic solvent, and then prepare the reference solution and sample solution;

[0007] Step 2: The solution from Step 1 is injected into a gas chromatograph-mass spectrometer via headspace injection for detection;

[0008] Step 3: Qualitative and quantitative analysis.

[0009] The 10 haloalkanes are chloromethane, chloroethane, 2-chloropropane, 1,1-dichloroethylene, tert-butane 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-imidazolinone, 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 non-volatile acids.

[0012] In some specific implementations, the sample solutions in steps 1 and 2 are selected from one or more of the following: control samples, raw material samples, intermediate samples, and product samples.

[0013] In some specific implementations, the sample solution and reference solution in step 1 are prepared by weighing and adding the diluent, then mixing to a final volume; step 1 also includes a limit of quantitation solution, which is obtained by diluting the reference solution.

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

[0015] In some specific implementations, the parameters of the headspace sampler for the gas chromatograph-mass spectrometer in step 2 are as follows: headspace furnace temperature 80℃; sample quantitative loop temperature 130℃; transfer line temperature 150℃; sample vial equilibration time 20 min; sample vial pressure 17 psi; headspace sample vial loading volume 1.0 mL; headspace vial size 20 mL.

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

[0017]

[0018]

[0019] In the formula:

[0020] A Res.Sol. Peak areas representing specific solvents observed in the sample solution;

[0021] W SPL This represents the mass of the sample, expressed in mg.

[0022] V SPL The volume of the sample solution diluted is expressed in mL.

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

[0024] The formulas for calculating the response factor RF of each specific solvent in reference solution-1 and reference solution-2 are as follows:

[0025]

[0026] In the formula:

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

[0028] W STD This represents the mass of each specific solvent in the reference solution, expressed in mg.

[0029] V STD This 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] This invention develops a universal analytical method for 10 common haloalkanes, which can simultaneously detect 10 common haloalkanes in later-stage project processes. The detection sensitivity can reach 1 ppm, and the stability of haloalkanes can be maintained for more than 48 hours. The method is highly versatile and can be widely applied to different types of raw materials, intermediates and API samples.

[0032] The following will further explain the concept and technical effects of the present invention in conjunction with the accompanying drawings, so as to fully understand the purpose, features and effects of the present invention. Attached Figure Description

[0033] Figure 1 This is a GCMS TIC plot of ten haloalkanes.

[0034] Figure 2 This is a linear graph of chloromethane.

[0035] Figure 3 This is a linear graph of chloroethane.

[0036] Figure 4 This is a linear graph of 2-chloropropane.

[0037] Figure 5 This is a linear graph of 1,1-dichloroethylene.

[0038] Figure 6 The linear graph of tert-butane chloride.

[0039] Figure 7 This is a linear graph of 2-bromopropane.

[0040] Figure 8 This is a linear graph of 1,1,1-trichloroethane.

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

[0042] Figure 10 This is a linear graph of 1,2-dichloroethane.

[0043] Figure 11 This is a linear graph of 1,2-dibromoethane. Detailed Implementation

[0044] To make the technical means, inventive features, objectives, and effects of the invention readily understandable, the invention is further illustrated below with reference to specific figures. However, the invention is not limited to the embodiments described below.

[0045] It should be noted that the accompanying drawings are only used to complement the content disclosed in this specification, so as to enable those skilled in the art to understand and read them, and are not intended to limit the conditions under which the present invention can be implemented. Therefore, they have no substantial technical significance. Any adjustments that do not involve creative innovation, without affecting the effects and objectives that the present invention can produce, should still fall within the scope of the technical content disclosed in the present invention.

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

[0047] This invention incorporates a non-volatile acid into the headspace method diluent, ensuring a consistently acidic environment regardless of sample pH, thus stabilizing the haloalkanes. Instrumentally, this invention utilizes headspace GCMS technology to establish a universal analytical method for detecting 10 common haloalkanes in later stages of clinical processes. This technology is applicable to the GCMS determination of 10 haloalkanes—chloromethane, chloroethane, 2-chloropropane, 1,1-dichloroethylene, tert-butane, 2-bromopropane, 1,1,1-trichloroethane, carbon tetrachloride, 1,2-dichloroethane, and 1,2-dibromoethane—in raw materials, intermediates, and finished products in clinical trials. The corresponding ranges and retention times are listed in Table 1 below.

[0048] Table 1. Detection range and retention time for 10 chloroalkanes

[0049] Chloromethane 0.05-5 4.5 chloroethane 0.05-5 6.0 2-Chloropropane 0.05-5 7.9 1,1-Dichloroethylene 0.05-5 8.4 tert-butane 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 this invention are shown in Table 2 below:

[0051] Table 2 Main Reagents

[0052]

[0053]

[0054]

[0055] Example 1: Chromatographic and mass spectrometric conditions, headspace sampler parameters

[0056] The chromatographic and mass spectrometry conditions, headspace sampler parameters, etc. are set 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 are established based on the Agilent system. When using other instruments, some non-critical parameters may be adjusted (e.g., solvent delay, MS on, MS off, etc.); the cycle time may be extended as needed (if applicable).

[0063] Table 5 MS Headspace Sampler Parameters

[0064]

[0065] Example 2 Solution Preparation

[0066] The preparation of reference stock solution 1, reference stock solution 2, reference solution, limit of quantitation solution, and sample solution are shown in Tables 6-8 below.

[0067] Table 6 Preparation of Reference Stock Solution 1 and Reference Stock Solution 2

[0068]

[0069]

[0070] Table 7 Preparation of Reference Solution 1, Reference Solution 2, and Limit of Quantitation Solution

[0071]

[0072] Table 8 Sample Solution Preparation

[0073]

[0074] Example 3: Sample Injection Analysis and Detection

[0075] The system adaptability requirements are as shown in Table 9:

[0076] Table 9 System Adaptability Requirements

[0077]

[0078]

[0079] Example 4: Calculation of Detection Results

[0080] Calculate the response factor (RF) of each specific solvent in STD-1 and STD-2 solutions using the following formula:

[0081]

[0082] in:

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

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

[0085] V STD This represents the dilution volume (mL) of the standard solution.

[0086] Calculate the content of each specific residual solvent observed in the chromatogram using one of the following two formulas:

[0087]

[0088]

[0089] in:

[0090] A Res.Sol. Peak areas representing specific solvents observed in the sample solution;

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

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

[0093] This represents the average response factor of each specific solvent in the standard solution.

[0094] Example 5 Analysis of Detection Results

[0095] For each specific solvent with a LOQ of at least 1: the average result should be reported as the final result. However, if a result is below the LOQ, the maximum result should be reported as the final result.

[0096] For each specific solvent less than LOQ, the result will be reported as " <LOQ”。

[0097] If any result exceeds the quality standard limit, an OOS investigation will be conducted.

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

[0099] Table 10 LOQ for each specific solvent

[0100]

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

[0102] Table 11 Experimental Results

[0103]

[0104]

[0105]

[0106] This demonstrates that the detection sensitivity for all 10 haloalkanes can reach 1 ppm, and the haloalkanes exhibit good stability, lasting for more than 48 hours, indicating strong versatility of the method.

[0107] The preferred embodiments of the present invention have been described in detail above. It should be understood that those skilled in the art can make numerous modifications and variations based on the concept of the present invention without creative effort. Therefore, all technical solutions that can be obtained by those skilled in the art based on the concept of the present invention through logical analysis, reasoning, or limited experimentation on the basis of existing technology should be within the scope of protection defined by the claims.

Claims

1. A universal analytical method for detecting 10 haloalkanes in a pharmaceutical product, characterized in that, The process includes sample solution and reference solution preparation, gas chromatography-mass spectrometry analysis, qualitative and quantitative analysis, and the specific steps are as follows: Step 1: Prepare a diluent by adding acid to an organic solvent, and then prepare the reference solution and sample solution; Step 2: The solution from Step 1 is injected into a gas chromatograph-mass spectrometer via headspace injection for detection; Step 3: Qualitative and quantitative analysis; The 10 haloalkanes were also detected as chloromethane, chloroethane, 2-chloropropane, 1,1-dichloroethylene, tert-butane chloride, 2-bromopropane, 1,1,1-trichloroethane, carbon tetrachloride, 1,2-dichloroethane, and 1,2-dibromoethane. The chromatographic conditions for step 2 are as follows: Column: Agilent DB-624, 60 m × 0.32 mm, ID 1.8 µm, PN:123-1364; Injector temperature: 240℃; Column temperature program: 40℃ for 10 min, 10℃ / min to 70℃ for 5 min, 50℃ / min to 250℃ for 8 min; Flow rate: 3 mL / min; Split ratio: 5:

1. The mass spectrometry conditions for step 2 are as follows: Ion source: EI; Ion source temperature: 230℃; Quadrupole temperature: 150℃; Acquisition mode: SIM. The formulas for qualitative and quantitative analysis in step 3 are selected from one of the following calculation formulas: 1） 2） In the formula: A Res. Sol. Peak areas representing specific solvents observed in the sample solution; W SPL This represents the mass of the sample, expressed in mg. V SPL The volume of the sample solution diluted is expressed in mL. This represents the average response factor of each specified solvent in the reference solution; The formulas for calculating the response factor RF of each specific solvent in reference solution-1 and reference solution-2 are as follows: In the formula: A STD Peak areas representing specific solvents in the reference solution; W STD This represents the mass of each specific solvent in the reference solution, expressed in mg. V STD This represents the dilution volume of the reference solution, in mL.

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-imidazolinone, 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 non-volatile acids.

4. The method according to claim 1, characterized in that, The preparation method of the sample solution and reference solution in step 1 is as follows: after weighing, add the diluent and mix well to make up to the required volume; step 1 also includes a limit of quantitation solution, which is obtained by diluting the reference solution.

5. The method according to claim 1, characterized in that, The parameters of the headspace sampler for the gas chromatograph-mass spectrometer in step 2 are as follows: headspace furnace temperature 80℃; sample quantitative loop temperature 130℃; transfer line temperature 150℃; sample vial equilibration time 20 min; sample vial pressure 17 psi; headspace sample vial loading volume 1.0 mL; headspace vial size 20 mL.

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