CN114216991A - Method for detecting genotoxic impurities in rifampicin - Google Patents
Method for detecting genotoxic impurities in rifampicin Download PDFInfo
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- CN114216991A CN114216991A CN202111602354.XA CN202111602354A CN114216991A CN 114216991 A CN114216991 A CN 114216991A CN 202111602354 A CN202111602354 A CN 202111602354A CN 114216991 A CN114216991 A CN 114216991A
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Abstract
The invention relates to the technical field of drug analysis, and particularly discloses a method for detecting genotoxic impurities in rifampicin. The detection method comprises the steps of preparing a blank solution, a reference solution and a test solution, and detecting by adopting a headspace sampling-gas chromatography-mass spectrometry combined method. The detection method provided by the invention realizes quantitative and qualitative analysis of formaldehyde in the rifampicin raw material medicine, has strong specificity, excellent repeatability, accuracy, sensitivity and linear relation, and has lower detection limit, wherein the detection limit of the formaldehyde is 0.026 mu g/mL, and the quantitative limit is 0.077 mu g/mL, thereby meeting the detection requirement of the formaldehyde in the rifampicin raw material medicine.
Description
Technical Field
The invention relates to the technical field of drug analysis, in particular to a method for detecting genotoxic impurities in rifampicin.
Background
Rifampicin (Rifampicin) has a molecular formula of C43H58N4O12The compound is a rifamycin semi-synthetic broad-spectrum antibacterial drug, is red or dark red crystalline powder and is insoluble in water. Antituberculous drugs have antibacterial activity against a variety of pathogenic microorganisms, and also have therapeutic effects against gram-positive or gram-negative bacteria, viruses, and the like.
Rifampin was first synthesized in 1959 in the laboratory in italy and was first marketed in 1968 in italy by the company cenoft in the form of capsules, tablets and syrups. The rifampicin imitation drugs have been on the market for many years in China, and with the change of relevant drug regulations, it is a necessary trend to require all imitation drugs to reach the quality and curative effect consistent with the original research, which brings great challenges to the domestic imitation pharmaceutical enterprises while ensuring the effectiveness and quality of the drugs. As for impurities, the requirements of pharmaceuticals are more stringent than those of the original research.
Residual genotoxic impurity formaldehyde may be generated in the process of generating rifampicin bulk drug by the existing process, and no relevant literature reports about a detection method of formaldehyde in rifampicin at present. Therefore, the development of a method for detecting formaldehyde in rifampicin is of great significance for improving the medication safety and the quality control of rifampicin.
Disclosure of Invention
In view of this, the application provides a method for detecting genotoxic impurities in rifampicin, which has strong specificity, excellent repeatability, accuracy and sensitivity, and can meet the detection requirement of genotoxic impurity formaldehyde in rifampicin bulk drugs.
In order to achieve the purpose of the invention, the embodiment of the invention adopts the following technical scheme:
a method for detecting genotoxic impurities in rifampicin, comprising at least the following steps:
step one, preparing a blank solution, a reference solution and a test solution:
blank solution: putting p-benzenesulfonic acid and ethanol in a headspace bottle, sealing by a gland, and preparing a blank solution by using a solvent;
preparing a reference substance solution: putting a formaldehyde aqueous solution, p-toluenesulfonic acid and ethanol into a headspace bottle, and preparing a reference substance solution by using a solvent;
preparing a test solution: putting p-toluenesulfonic acid, a rifampicin sample and ethanol into a headspace bottle, sealing by a gland, and preparing a test solution by using a solvent;
step two, detecting the blank solution, the reference solution and the test solution by adopting a headspace sampling-gas chromatography-mass spectrometry combined method, wherein the gas chromatography conditions are as follows:
DB-WAX capillary chromatographic column is adopted, and the injection port temperature is as follows: 135-145 ℃; the split ratio is as follows: 17: 1-23: 1; flow rate: 0.9mL/min to 1.1 mL/min; temperature programmed heating of furnace temperature: the initial temperature is 33-37 ℃, the temperature is kept for 4-6 min, the temperature is raised to 200-210 ℃ at the speed of 28-32 ℃/min, and the temperature is kept for 4-6 min;
the conditions of the mass spectrum are as follows:
the quantitative ions of the formaldehyde were 31m/z, 59m/z and 103m/z using EI ion source, single quadrupole.
Compared with the prior art, the detection method for the genotoxic impurities in the rifampicin has the following advantages:
the method adopts a pre-column derivation method, uses p-toluenesulfonic acid as a catalyst, generates diethanol formal from ethanol and formaldehyde, has complete reaction, stable product and higher correspondence to a mass spectrum detector, and therefore, the method adopts a headspace sampling-gas chromatography-mass spectrum combined method to realize quantitative and qualitative analysis of formaldehyde in rifampicin bulk drugs, has strong specificity and excellent repeatability, accuracy, sensitivity and linear relation, and has a lower detection limit, wherein the detection limit of the formaldehyde is 0.026 mu g/mL, the quantitative limit is 0.077 mu g/mL, and the detection requirement of the formaldehyde in the rifampicin bulk drugs is met.
The invention adopts the combination of headspace sampling, gas chromatography and mass spectrometry to detect the formaldehyde in the rifampicin, has reliable and controllable whole operation process, is suitable for practical application and popularization, and has wide application prospect.
Optionally, the injection port temperature is: 140 ℃; the split ratio is as follows: 20: 1; flow rate: 1.0 mL/min; temperature programmed heating of furnace temperature: the initial temperature was 35 deg.C, held for 5min, and the temperature was raised to 205 deg.C at a rate of 30 deg.C/min, held for 5 min.
The preferable detection condition is favorable for separating the impurity derivative diethanol formal from other components, and the peak type is better and the separation degree is higher.
The temperature rise program of the furnace temperature is an important factor influencing the performance of the gas chromatographic column, and the initial temperature and the final temperature determine the elution capacity of each component, so the initial temperature, the temperature rise rate and the final temperature are preferably selected in the application, the impurity derivative diethanol formal can be rapidly separated from the rifampicin bulk drug, and the impurity derivative diethanol formal and the rifampicin bulk drug have excellent separation degrees.
Optionally, the gas chromatography conditions further comprise: helium as carrier gas, 1.0mL of sample, flame ionization detector, and 245-255 deg.C of detector temperature.
Optionally, the conditions of the mass spectrum further include: the ion source temperature is 230 ℃, the quadrupole rod temperature is 150 ℃, and the transmission line temperature is 270-290 ℃.
Further optionally, the transmission line temperature is 280 ℃.
Under the preferable mass spectrum condition, the impurity derivative diethanol formal has higher response value, so that the separation result is more accurate.
Optionally, the headspace sampling conditions are as follows: the temperature of the sampling needle is 105-115 ℃, the gas phase circulation time is 26-30 min, and the sample injection time is 0.4-0.6 min; the temperature of the transmission line is 110-130 ℃, and the equilibrium pressure is 13-16 psi.
Further optionally, the conditions of headspace sampling are as follows: the temperature of the sampling needle is 110 ℃, the gas phase circulation time is 28min, and the sample introduction time is 0.5 min; the transfer line temperature was 120 ℃ and the equilibrium pressure was 15 psi.
Optionally, the headspace sampling conditions are as follows: the bottle temperature of the headspace bottle is 45-55 ℃, the heat preservation time is 30-40 min, and the pressurization time is 0.4-0.6 min.
Further, optionally, the conditions of headspace sample injection are as follows: the bottle temperature of the headspace bottle is 50 ℃, the heat preservation time is 35min, and the pressurization time is 0.5 min.
The temperature and the heat preservation time of the optimized headspace bottle are favorable for complete reaction between formaldehyde and ethanol, and the accuracy of a detection result is ensured.
The solvent is dimethyl sulfoxide.
The preferable solvent has no interference on the detection of the formaldehyde, and the accuracy of the detection result is ensured.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained according to these drawings without creative efforts.
FIG. 1 is a gas chromatogram of a blank solution provided in example 1 of the present invention;
FIG. 2 is a gas chromatogram of a control solution provided in example 1 of the present invention;
FIG. 3 is a gas chromatogram of a test solution provided in example 1 of the present invention;
FIG. 4 is a gas chromatogram of a 100% sample spiking solution provided in example 1 of the present invention;
fig. 5 is a linear relationship diagram provided in embodiment 4 of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1
The embodiment of the application provides a method for detecting genotoxic impurities in rifampicin, which comprises the following steps:
step one, preparing a blank solution, a reference solution, a test solution and a 100% test sample adding standard solution:
blank solution: weighing 200mg of p-benzenesulfonic acid, placing the p-benzenesulfonic acid in a 20mL headspace bottle, adding 1mL of dimethyl sulfoxide to dissolve the p-benzenesulfonic acid, transferring 50 mu L of ethanol, and sealing by a gland to obtain a blank solution;
control stock solution 1: adding 5mL of dimethyl sulfoxide into a 50mL volumetric flask, taking 420.31mg of formaldehyde aqueous solution with the concentration of 38.25 wt% into the volumetric flask, diluting the solution to a scale by using the dimethyl sulfoxide, and shaking up; precisely transferring the solution into a volumetric flask with the volume of 1.0-50 mL, diluting the solution to a scale with DMSO, and shaking up to obtain a reference substance stock solution 1;
control stock solution 2: precisely transferring 0.5mL of reference stock solution into a volumetric flask of 1-25 mL, diluting with DMSO to a scale, and shaking up to obtain reference stock solution 2;
control solution: weighing 200mg of p-toluenesulfonic acid, placing the p-toluenesulfonic acid into a 20mL headspace bottle, adding 1mL of reference stock solution 2, transferring 50uL of ethanol, and sealing by a gland to obtain a reference solution;
test solution: weighing 200mg of p-toluenesulfonic acid and 500mg of rifampicin samples, precisely weighing the samples into a 20mL headspace bottle, adding 1.0mL of methyl sulfoxide for dissolution, transferring 50 mu L of ethanol, and sealing by a gland to obtain a test solution;
adding standard solution to 100% of test sample: weighing 200mg of p-toluenesulfonic acid, then weighing 500mg of rifampicin sample, precisely weighing the rifampicin sample into a 20mL headspace bottle, adding 1.0mL of reference substance stock solution 2, transferring 50uL of ethanol, and sealing by a gland to obtain a 100% sample standard solution;
and step two, detecting the blank solution, the reference solution, the test solution and the 100% test sample added standard solution by adopting a headspace sampling-gas chromatography-mass spectrometry combined method, and recording spectrograms, wherein the chromatograms are respectively shown in figure 1, figure 2, figure 3 and figure 4.
Wherein the gas chromatography conditions are as follows:
using a DB-WAX capillary chromatographic column, wherein the injection port temperature is as follows: 140 ℃; the split ratio is as follows: 20: 1; flow rate: 1.0 mL/min; temperature programmed heating of furnace temperature: the initial temperature is 35 ℃, the temperature is kept for 5min, the temperature is increased to 205 ℃ at the speed of 30 ℃/min, and the temperature is kept for 5 min; a flame ionization detector with a temperature of 250 ℃,
the conditions of the mass spectrum are as follows:
adopting an EI ion source, wherein the temperature of the ion source is 230 ℃, the temperature of a single quadrupole is 150 ℃, the temperature of a transmission line is 280 ℃, and the mass ion SIM of formaldehyde: 31/59/103;
the headspace sampling conditions are as follows: the temperature of the headspace bottle is 50 ℃, the heat preservation time is 35min, and the pressurization time is 0.5 min; the temperature of the sampling needle is 110 ℃, the gas phase circulation time is 28min, and the sample introduction time is 0.5 min; the transfer line temperature was 120 ℃ and the equilibrium pressure was 15 psi.
As can be seen from FIGS. 2 to 4, the chromatographic peaks of the ethanol formal appear in the reference solution, the test solution and the 100% test solution added with the standard solution at a retention time of 5.2 min.
As can be seen from fig. 1 to 4, each component in the blank solution has no significant interference with each analyte in the reference solution; the minimum value of the separation degree of each peak of the substance to be detected and the adjacent chromatographic peak in the test solution, the reference solution and the 100 percent test substance added standard solution is 22.32 and is far more than 1.5; the retention time of each peak of the substance to be detected in the test solution or the 100 percent test solution added standard solution is consistent with that of the reference solution; therefore, the detection method provided by the application has good specificity.
Example 2 repeatability
Taking the same batch of samples, preparing 6 parts of 100% sample solution for adding a standard, and detecting by adopting a headspace sampling-gas chromatography-mass spectrometry combined method, wherein the specific conditions of headspace sampling, gas chromatography and mass spectrometry are described in example 1, and the results are shown in table 1.
The preparation method of the 100% sample labeling solution comprises the following steps: firstly weighing 200mg of p-toluenesulfonic acid, then weighing about 500mg of rifampicin sample, precisely weighing the rifampicin sample into a 20mL headspace bottle, adding 1.0mL of the reference stock solution 2 prepared in the example 1, then transferring 50uL of ethanol, and sealing through a gland to obtain a 100% sample standard solution.
TABLE 1 repeatability results
As can be seen from Table 1, the RSD value of the content of the diethanol formal to be detected in 6 analysis repetitive solutions is 1.9% (< 15.0%), which indicates that the detection method provided by the application has good repeatability.
Example 3 limits of quantitation and detection
And (3) quantifying limited mother liquor: transferring 0.15mL of the reference stock solution prepared in example 1 to a volumetric flask of 25mL, adding DMSO to dilute to a scale, and shaking up to obtain a quantitative limit mother solution.
Quantitative limiting solution: weighing 200mg of p-toluenesulfonic acid, precisely weighing the p-toluenesulfonic acid into a 20mL headspace bottle, adding 1.0mL of the quantitative limiting mother liquor, transferring 50uL of ethanol, and sealing by a gland to obtain quantitative limiting solution, and preparing three parts in parallel.
Detection limit mother liquor: transferring 1mL of the reference stock solution prepared in example 1 to a 10mL volumetric flask, adding DMSO to dilute to a scale, and shaking up to obtain the detection limit mother solution.
Detection limiting solution: weighing 200mg of p-toluenesulfonic acid, precisely weighing the p-toluenesulfonic acid into a 20mL headspace bottle, adding 1.0mL of the detection limit mother solution, transferring 50uL of ethanol, and sealing by a gland to obtain a detection limit solution, and preparing three parts in parallel.
The quantitative limiting solution and the detection limiting solution are detected by adopting a headspace sampling-gas chromatography-mass spectrometry combined method, 3 needles are continuously sampled respectively, the specific conditions of headspace sampling, gas chromatography and mass spectrometry are described in example 1, and the results are shown in tables 2 and 3 below.
TABLE 2 quantitative limit results
As can be seen from Table 2, the minimum value of the signal-to-noise ratio of the analyte for 3 times of the quantitative limiting solution is 34.3 (not less than 10); the RSD value of the peak area of the diethanol formal is 12.3 percent (less than or equal to 15.0 percent); therefore, the quantitative limit of the detection method provided by the application meets the detection requirement.
TABLE 3 detection Limit results
As can be seen from Table 3, the minimum value of the signal-to-noise ratio of the analyte for 3 times of the detection limit solution is 13.6 (not less than 3); therefore, the detection limit of the detection method provided by the application meets the detection requirement.
Example 4 Linear relationship
Linear solution: firstly, 200mg of p-toluenesulfonic acid is precisely weighed and placed in a 20mL headspace bottle, and 10 parts are prepared in parallel; different volumes of the control stock solution 1 prepared in example 1 were transferred to the headspace bottle as shown in Table 4 below, and 50uL of ethanol was added to prepare 2 parts of each concentration.
TABLE 4
| Concentration level | LOQ％ | 50％ | 80％ | 100％ | 150％ |
| Reference stock solution 1(mL) | 0.15 | 0.25 | 0.4 | 0.5 | 0.75 |
The prepared linear solution is detected by adopting a headspace sampling-gas chromatography-mass spectrometry combined method, each concentration is sampled for 2 times, wherein the specific conditions of headspace sampling, gas chromatography and mass spectrometry are as described in example 1, and a spectrogram is recorded. The results are shown in Table 5 and the linear relationship chart is shown in FIG. 5, wherein the concentration of the diethanol formal (. mu.g/mL) is used as the abscissa and the peak area is used as the ordinate, a standard curve is drawn, and a regression equation is calculated. From the results, it can be seen that the linear correlation coefficient r of the diethanol formal is not less than 0.990, the peak area ratio of the absolute value of the Y-axis intercept to the 100% limit concentration is 1.2% (. ltoreq.10.0%), and the diethanol formal has a good linear relationship in the concentration range of 0.386. mu.g/mL to 1.929. mu.g/mL.
TABLE 5 Linear results
Example 5 accuracy
Accuracy solution: comprises 50% of standard adding solution, 100% of standard adding solution and 150% of standard adding solution
50% spiked solution: 200mg of p-toluenesulfonic acid and about 500mg of rifampicin sample are weighed precisely and placed in a 20mL headspace bottle, 0.25mL of the reference stock solution 1 prepared in example 1 is transferred into the headspace bottle, and 50 μ L of ethanol is added to prepare 3 parts in parallel.
100% spiked solution: 200mg of p-toluenesulfonic acid and about 500mg of rifampicin sample are weighed precisely and placed in a 20mL headspace bottle, 0.5mL of the control stock solution 1 prepared in example 1 is transferred into the headspace bottle, and 50 μ L of ethanol is added to prepare 3 parts in parallel.
150% spiking solution: 200mg of p-toluenesulfonic acid and about 500mg of rifampicin sample are weighed precisely and placed in a 20mL headspace bottle, 0.75mL of the reference stock solution 1 prepared in example 1 is transferred into the headspace bottle, and 50 μ L of ethanol is added to prepare 3 parts in parallel.
The prepared accurate solution is detected by adopting a headspace sampling-gas chromatography-mass spectrometry combined method, each concentration is subjected to sampling for 3 times, wherein the specific conditions of headspace sampling, gas chromatography and mass spectrometry are as described in example 1, a spectrogram is recorded, and the recovery rate result is shown in table 6.
The recovery was calculated according to the following formula:
recovery (%). measured/theoretical X100%
TABLE 6 accuracy results
As can be seen from Table 6, the individual values of the recovery of the diethanol formal at 50%, 100%, 150% of the limiting concentrations ranged from 72.7% to 106.8% (all ranged from 70.0% to 130.0%); thus, the accuracy of the detection method provided by the parent is good.
EXAMPLE 6 durability
The control solution prepared in example 1 was tested by headspace sampling-gas chromatography-mass spectrometry, and the specific conditions of headspace sampling, gas chromatography and mass spectrometry were fine-tuned.
The conditions of the gas chromatography were fine-tuned, both the headspace injection and mass spectrometry conditions were the same as in example 1:
fine adjustment I, sample inlet temperature adjustment: the inlet temperature was 135 ℃ and other conditions of the gas chromatography were the same as those of example 1.
Fine adjustment II, sample inlet temperature adjustment: the inlet temperature was 145 ℃ and other conditions of the gas chromatography were the same as those of example 1.
Fine adjustment, adjustment of the flow splitting ratio: the split ratio was 17:1 and other conditions of the gas chromatograph were the same as in example 1.
Fine adjustment, adjustment of the flow splitting ratio: the split ratio was 23:1 and other conditions of the gas chromatograph were the same as in example 1.
Fifthly, fine adjustment and flow rate adjustment: the flow rate was 0.9mL/min, and other conditions of gas chromatography were the same as those of example 1.
Fine adjustment six, flow rate adjustment: the flow rate was 1.1mL/min, and other conditions of gas chromatography were the same as those of example 1.
Seventhly, fine adjustment and temperature rise program adjustment: the program of the temperature raising program is as follows:
the starting temperature was 33 ℃ and held for 6min, the temperature was raised to 200 ℃ at a rate of 28 ℃/min and held for 6min, and the other conditions of the gas chromatograph were the same as those of example 1.
Eighthly, adjusting a temperature rise program: the program of the temperature raising program is as follows:
the starting temperature was 37 ℃ and held for 4min, the temperature was raised to 210 ℃ at a rate of 32 ℃/min and held for 4min, and other conditions of the gas chromatograph were the same as those of example 1.
The mass spectrometry conditions were fine-tuned, and the conditions for headspace injection and gas chromatography were the same as in example 1:
fine adjustment nine, transmission line temperature: the temperature of the transmission line was 270 ℃ and the other conditions of the mass spectrum were the same as in example 1;
fine adjustment ten, transmission line temperature: the temperature of the transmission line was 290 ℃ and the other conditions of the mass spectrum were the same as in example 1;
the headspace injection temperature was adjusted and the conditions for gas chromatography and mass spectrometry were the same as in example 1:
eleven fine adjustment, the bottle temperature of the headspace bottle is 45 ℃, the heat preservation time is 40min, the pressurization time is 0.4min, and other conditions of headspace sample injection are the same as those of the embodiment 1;
fine adjustment twelve, the bottle temperature of the headspace bottle is 55 ℃, the heat preservation time is 35min, the pressurization time is 0.6min, and other conditions of headspace sample injection are the same as those of the embodiment 1;
thirteen fine adjustment, the temperature of the sampling needle is 105 ℃, the cycle time is 30min, the sample injection time is 0.6min, and other conditions of headspace sample injection are the same as those of the embodiment 1;
fourteen fine adjustments are made, the temperature of a sampling needle is 115 ℃, the cycle time is 26min, the sample introduction time is 0.4min, and other conditions of headspace sample introduction are the same as those of the embodiment 1;
fifteen fine adjustment, the temperature of the transmission line is 110 ℃, the equilibrium pressure is 16psi, and other conditions of headspace sample injection are the same as those of the embodiment 1;
sixteen fine adjustments, 130 ℃ transmission line temperature, 13psi equilibrium pressure, and the other conditions for headspace injection were the same as in example 1.
The results of headspace sample injection-gas chromatography-mass spectrometry combined detection after fine tuning one to sixteen were substantially the same as those of example 1, and the resolution of chromatographic peaks was good, thus demonstrating that the fine tuning of chromatographic conditions, mass spectrometry conditions or headspace sample injection conditions had no effect on the detection of the genotoxic impurity formaldehyde in rifampicin.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents or improvements made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (10)
1. A method for detecting genotoxic impurities in rifampicin, characterized in that: the detection method at least comprises the following steps:
step one, preparing a blank solution, a reference solution and a test solution:
blank solution: putting p-benzenesulfonic acid and ethanol in a headspace bottle, sealing by a gland, and preparing a blank solution by using a solvent;
preparing a reference substance solution: putting a formaldehyde aqueous solution, p-toluenesulfonic acid and ethanol into a headspace bottle, and preparing a reference substance solution by using a solvent;
preparing a test solution: putting p-toluenesulfonic acid, a rifampicin sample and ethanol into a headspace bottle, sealing by a gland, and preparing a test solution by using a solvent;
step two, detecting the blank solution, the reference solution and the test solution by adopting a headspace sampling-gas chromatography-mass spectrometry combined method, wherein the gas chromatography conditions are as follows:
DB-WAX capillary chromatographic column is adopted, and the injection port temperature is as follows: 135-145 ℃; the split ratio is as follows: 17: 1-23: 1; flow rate: 0.9mL/min to 1.1 mL/min; temperature programmed heating of furnace temperature: the initial temperature is 33-37 ℃, the temperature is kept for 4-6 min, the temperature is raised to 200-210 ℃ at the speed of 28-32 ℃/min, and the temperature is kept for 4-6 min;
the conditions of the mass spectrum are as follows:
the quantitative ions of the formaldehyde were 31m/z, 59m/z and 103m/z using EI ion source, single quadrupole.
2. The method of detecting a genotoxic impurity in rifampicin according to claim 1, characterized by: the sample inlet temperature is as follows: 140 ℃; the split ratio is as follows: 20: 1; flow rate: 1.0 mL/min; temperature programmed heating of furnace temperature: the initial temperature was 35 deg.C, held for 5min, and the temperature was raised to 205 deg.C at a rate of 30 deg.C/min, held for 5 min.
3. The method of detecting a genotoxic impurity in rifampicin according to claim 1, characterized by: the conditions of the gas chromatograph further include: helium as carrier gas, 1.0mL of sample, flame ionization detector, and 245-255 deg.C of detector temperature.
4. The method of detecting a genotoxic impurity in rifampicin according to claim 1, characterized by: the conditions of the mass spectrum further include: the ion source temperature is 230 ℃, the quadrupole rod temperature is 150 ℃, and the transmission line temperature is 270-290 ℃.
5. The method of detecting a genotoxic impurity in rifampicin according to claim 4, characterized by: the transmission line temperature was 280 ℃.
6. The method of detecting a genotoxic impurity in rifampicin according to claim 1, characterized by: the headspace sampling conditions are as follows: the temperature of the sampling needle is 105-115 ℃, the gas phase circulation time is 26-30 min, and the sample injection time is 0.4-0.6 min; the temperature of the transmission line is 110-130 ℃, and the equilibrium pressure is 13-16 psi.
7. The method of detecting a genotoxic impurity in rifampicin according to claim 6, characterized by: the headspace sampling conditions are as follows: the temperature of the sampling needle is 110 ℃, the gas phase circulation time is 28min, and the sample introduction time is 0.5 min; the transfer line temperature was 120 ℃ and the equilibrium pressure was 15 psi.
8. The method of detecting a genotoxic impurity in rifampicin according to claim 1, characterized by: the headspace sampling conditions are as follows: the bottle temperature of the headspace bottle is 45-55 ℃, the heat preservation time is 30-40 min, and the pressurization time is 0.4-0.6 min.
9. The method of detecting a genotoxic impurity in rifampicin according to claim 8, characterized by: the headspace sampling conditions are as follows: the bottle temperature of the headspace bottle is 50 ℃, the heat preservation time is 35min, and the pressurization time is 0.5 min.
10. The method of detecting genotoxic impurities in rifampicin according to any one of claims 1 to 9, characterized by: the solvent is dimethyl sulfoxide.
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