CN112326860B - Method for simultaneously detecting genotoxic impurities of 1, 4-dichlorobutane and 1-chloro-4-hydroxybutane in bulk drug or preparation - Google Patents

Abstract

The invention relates to the technical field of analysis and detection, in particular to a method for simultaneously detecting genetic toxicity impurities of 1, 4-dichlorobutane and 1-chloro-4-hydroxybutane in a bulk drug or a preparation. The method for simultaneously detecting the genetic toxicity impurities of the 1, 4-dichlorobutane and the 1-chloro-4-hydroxybutane in the bulk drugs or the preparations adopts gas chromatography-mass spectrometry combined detection, EI ion sources and top air phase analysis, and uses methyl tert-butyl ether as an internal standard substance to calculate the content of the impurities in the samples by the internal standard method. The invention solves the problem that the detection limit of the existing genotoxic impurities can not be reached, simultaneously fills the vacancy of the method for simultaneously detecting the genotoxic impurities of 1, 4-dichlorobutane and 1-chloro-4-hydroxybutane in the bulk drugs or preparations, and has the advantages of reaching ppm of the detection limit, improving the sensitivity, good repeatability and high response.

Description

Method for simultaneously detecting genetic toxicity impurities of 1, 4-dichlorobutane and 1-chloro-4-hydroxybutane in bulk drug or preparation

Technical Field

The invention relates to the technical field of analysis and detection, in particular to a method for simultaneously detecting genetic toxicity impurities of 1, 4-dichlorobutane and 1-chloro-4-hydroxybutane in a bulk drug or a preparation.

Background

Genotoxic Substances (GTIs) are substances which cause DNA mutations, chromosomal breaks or DNA recombinations and may also lead to the development of cancer. Genotoxic impurities are mainly derived from starting materials, intermediates, reagents and reaction byproducts in the synthesis process of bulk drugs. In addition, drugs may also degrade during synthesis, storage or formulation to produce genotoxic impurities. The European Medicines Agency (EMA) in 2007 discovered that a partial batch of nelfinavir mesylate produced by rochon pharmacy contains a high dose of the genotoxic impurity ethyl methanesulfonate; detecting that the valsartan used for hypertension produced by 7-month-China-sea pharmacy in 2018 contains trace genetic toxicity impurity N, N-dimethyl Nitrosamine (NDMA); the inclusion of NDMA was also detected in valsartan tablets produced by Torrent pharmaceuticals in india 8.8.month of the year. The medicines are recalled after the genetic toxicity impurities exceed the standard after being listed on the market. The research field of genotoxic impurities is relatively blank, three toxinomen events not only bring potential safety hazards and health problems to patients, but also bring immeasurable economic losses to pharmaceutical enterprises, and the whole pharmaceutical industry is knocked to sound a safe production alarm clock, so that more and more attention is paid in recent years.

At present, only a method for detecting 1-chloro-4-hydroxybutane, a method for detecting 1, 4-dichlorobutane and a method for simultaneously detecting two genetic toxic impurities are available; and because the limit requirement of the genotoxic impurities is extremely low, most enterprises do not have corresponding matched equipment research, related data are less, and no direct method can directly detect the impurities, so that the impurities cannot be accurately quantified.

Disclosure of Invention

The invention aims to provide a method for simultaneously detecting 1, 4-dichlorobutane and 1-chloro-4-hydroxybutane genotoxic impurities in a bulk drug or a preparation, solves the problem that the detection limit of the genotoxic impurities cannot be reached at present, fills the gap of the method for simultaneously detecting the 1, 4-dichlorobutane and 1-chloro-4-hydroxybutane genotoxic impurities in the bulk drug or the preparation, improves the sensitivity, has good repeatability and high response, and the detection limit reaches ppm.

The method for simultaneously detecting the genetic toxicity impurities of the 1, 4-dichlorobutane and the 1-chloro-4-hydroxybutane in the bulk drugs or the preparations comprises the steps of detecting by a gas chromatography-mass spectrometry combined method, adopting an EI ion source, carrying out top air phase analysis, taking methyl tert-butyl ether as an internal standard substance, and calculating the content of the impurities in a sample by the internal standard method.

Specifically, the method for simultaneously detecting genotoxic impurities of 1, 4-dichlorobutane and 1-chloro-4-hydroxybutane in the bulk drug or the preparation comprises the following steps:

(1) Preparing a diluent: mixing purified water and an organic solvent according to a volume ratio of 1;

(2) Preparing a blank solution: taking the diluent as a blank solution;

(3) Preparing an internal standard solution: respectively preparing methyl tert-butyl ether solutions with different concentrations as internal standard solutions by taking the diluent as a solvent and the methyl tert-butyl ether as the internal standard substances;

(4) Preparation of standard solutions: respectively preparing 1, 4-dichlorobutane solution, 1-chloro-4-hydroxybutane solution, 1, 4-dichlorobutane solution and 1-chloro-4-hydroxybutane mixed solution with different concentrations as standard solutions by using the diluent as a solvent and using the 1, 4-dichlorobutane and the 1-chloro-4-hydroxybutane as standard substances;

(5) Preparation of sample test solutions: taking the diluent as a solvent, preparing a sample solution with the concentration of 1g/mL, adding 0.2mL of the sample solution with the concentration of 1g/mL, 0.2mL of an internal standard solution with the concentration of 1 mu g/mL of methyl tert-butyl ether and 1.6mL of the diluent into a 20mL headspace bottle, and shaking up to obtain a sample test solution;

(6) And (3) detection: and (3) introducing a sample test solution into a gas chromatography-mass spectrometer, recording a spectrogram, and calculating the content of impurities in the sample by an internal standard method.

The organic solvent in the diluent is one of N-butyl alcohol, N-dimethylformamide, N-dimethylacetamide and petroleum ether.

The organic solvent and methyl tert-butyl ether were both of AR grade purity.

The selection of the internal standard substance and the organic solvent is very important, and the scanning sections of the organic solvent, the internal standard substance and the object to be detected must be free of interference, so that the detection result is ensured to be accurate.

Preferably, the internal standard solution, the standard solution and the sample solution to be tested can be prepared as follows:

internal standard solution: weighing 20mg of methyl tert-butyl ether, precisely weighing, placing in a 20mL volumetric flask, dissolving with a diluent, fixing the volume to a scale, and shaking up; and precisely taking 10 mu L of the solution, placing the solution into a 10mL volumetric flask, dissolving the solution by using a diluent, fixing the volume to a scale, and shaking up to obtain an internal standard solution with the concentration of the methyl tert-butyl ether of 1 mu g/mL.

Standard solution: respectively weighing 20mg of 1, 4-dichlorobutane standard substance and 20mg of 1-chloro-4-hydroxybutane standard substance, accurately weighing, respectively placing in 10mL volumetric flasks, dissolving with a diluent, fixing the volume to a scale, and shaking up; respectively taking 10 mu L of 2mg/mL 1, 4-dichlorobutane and 10 mu L of 2mg/mL 1-chloro-4-hydroxybutane, precisely adding 980 mu L of diluent, and shaking up to prepare a mixed standard solution of 20 mu g/mL; precisely adding 975 mu L of diluent into 25 mu L of the mixed standard solution, and shaking up to obtain 500ng/mL of mixed standard solution; precisely taking 1.0mL of the standard solution, placing the standard solution in a 10mL volumetric flask, precisely sucking 1.0mL of 1 mu g/mL internal standard solution, placing the internal standard solution in the volumetric flask, adding diluent to a constant volume to scale, and shaking up to obtain the standard solution with the concentration of 1, 4-dichlorobutane of 50ng/mL and the concentration of 1-chloro-4-hydroxybutane of 50ng/mL, wherein the standard solution contains 100ng/mL of the internal standard substance.

Sample solution to be tested: weighing 2g of sample, precisely weighing, placing in a 2mL volumetric flask, dissolving with diluent, fixing the volume to a scale, and shaking up; precisely taking 0.2mL of the solution, placing the solution in a 20mL headspace bottle, precisely adding 0.2mL of 1 microgram/mL methyl tert-butyl ether internal standard solution, precisely adding 1.6mL of diluent, and shaking up to obtain a sample test solution with the sample concentration of 0.1g/mL, wherein the internal standard solution is 100ng/mL; three sample test solutions were prepared in the same manner.

The gas phase conditions were:

a chromatographic column: a quartz capillary column with the specification of 30m multiplied by 0.25mm and 1.4 mu m;

temperature rising procedure: keeping the temperature at 45 ℃ for 2min, heating to 170 ℃ at 10 ℃/min, and then heating to 250 ℃ at 50 ℃/min and keeping the temperature for 1min;

sample inlet temperature: 220 to 240 ℃;

flow rate: 0.9-1.1mL/min, adopting a split-flow mode for sample injection, wherein the split-flow ratio is 5;

injecting sample in headspace, and balancing for 30min.

Preferably, the gas phase conditions are:

and (3) chromatographic column: a quartz capillary column with the specification of 30m multiplied by 0.25mm and 1.4 mu m;

temperature rising procedure: maintaining at 45 deg.C for 2min, heating to 170 deg.C at 10 deg.C/min, heating to 250 deg.C at 50 deg.C/min, and maintaining for 1min;

sample inlet temperature: 230 ℃;

flow rate: 1.0mL/min, adopting a split-flow mode for sample injection, wherein the split-flow ratio is 5;

and (4) headspace sampling, and balancing for 30min.

The mass spectrum conditions are as follows:

adopting an EI source;

the scanning mode is as follows: SIM scanning;

scanning segment 1 for 2.5min, wherein the name of an internal standard substance is methyl tert-butyl ether, and the quantitative ion is m/z 73.0;

scanning segment 2 for 4.9min, wherein the name of the target object is 1-chloro-4-hydroxybutane, and the quantitative ion is m/z 72.1;

scanning section 3 for 7min, wherein the name of the target object is 1, 4-dichlorobutane, and the quantitative ion is m/z 55.1;

the solvent is delayed for 1-3min.

The chromatographic column, the temperature raising program and the scanning section have great influence on the detection result, and the target peaks of the object to be detected and the internal standard substance cannot be completely separated after adjustment, so that the detection result is interfered.

Compared with the prior art, the invention has the following beneficial effects:

the invention solves the problem that the detection limit of the prior genotoxic impurity can not be reached, simultaneously fills the gap of the method for simultaneously detecting the genotoxic impurity of 1, 4-dichlorobutane and 1-chloro-4-hydroxybutane in the raw material medicine or the preparation, and has the advantages of reaching the detection limit to ppm, improving the sensitivity, good repeatability and high response.

Drawings

FIG. 1 is a typical gas phase spectrum of a blank solution of example 2;

FIG. 2 is a typical gas phase spectrum of a 10. Mu.g/mL 1, 4-dichlorobutane standard solution of example 2;

FIG. 3 is a typical spectrum of mass spectrum of 10. Mu.g/mL 1, 4-dichlorobutane standard solution of example 2;

FIG. 4 is a gas phase representative spectrum of a 10 μ g/mL 1-chloro-4-hydroxybutane standard solution of example 2;

FIG. 5 is a typical spectrum of mass spectrum of 10. Mu.g/mL 1-chloro-4-hydroxybutane standard solution of example 2;

FIG. 6 is a typical gas phase spectrum of a 10. Mu.g/mL internal standard solution of methyl t-butyl ether of example 2;

FIG. 7 is a typical spectrum of mass spectrum of an internal standard solution of methyl t-butyl ether of example 2 at 10. Mu.g/mL;

FIG. 8 is a typical gas phase diagram of a mixed standard solution of 1, 4-dichlorobutane and 1-chloro-4-hydroxybutane (containing 10. Mu.g/mL of an internal standard solution) of example 2;

in fig. 8, a:1, 4-dichlorobutane; b: 1-chloro-4-hydroxybutane; c: an internal standard substance;

FIG. 9 is a typical spectrum of the gas phase of 1, 4-dichlorobutane in a 0.5g/mL sample solution of example 2;

FIG. 10 is a typical spectrum of mass spectrum of 1-chloro-4-hydroxybutane in a 0.5g/mL sample solution of example 2;

FIG. 11 is a typical spectrum of mass spectrum of MTBE in 0.5g/mL sample solution of example 2;

FIG. 12 is a quantitative limit representative map of 1, 4-dichlorobutane of example 3;

FIG. 13 is a typical spectrum of the detection limit of 1, 4-dichlorobutane of example 3;

FIG. 14 is a quantitative limit representative spectrum of 1-chloro-4-hydroxybutane of example 3;

FIG. 15 is a typical spectrum of detection limit of 1-chloro-4-hydroxybutane of example 3;

FIG. 16 is a typical spectrum of the 1 st parallel sample of the sample of example 7 (batch No. 1).

Detailed Description

The present invention is further described below with reference to examples.

The raw materials used in the examples were as follows:

1, 4-dichlorobutane, manufacturer: TCI;

1-chloro-4-hydroxybutane, manufacturer: a Mecang;

n, N-Dimethylformamide (DMF), grade AR, manufacturer: chemical agents of the national drug group, ltd;

methyl tert-butyl ether, AR grade, manufacturer: chemical agents of the national drug group, ltd.

The diluents used in the examples were:

pure water and N, N-dimethylformamide were mixed in a volume ratio of 1.

The apparatus used in the examples is as follows:

a triple quadrupole gas chromatograph-mass spectrometer was used. The mass spectrum is an Agilent Technologies 7000 triple quadrupole; the gas phase is Agilent Technologies 7890A; the headspace sampler is Agilent Technologies 7697A.

The detection conditions in the examples are as follows:

gas phase conditions: the chromatographic column is a VF-624MS quartz capillary column (30 m × 0.25mm,1.4 μm); temperature rising procedure: maintaining at 45 deg.C for 2min, heating to 170 deg.C at 10 deg.C/min, heating to 250 deg.C at 50 deg.C/min, and maintaining for 1min; sample inlet temperature: 230 ℃; the flow rate is 1.0mL/min; adopting a split-flow mode for sample injection, wherein the split-flow ratio is 5; injecting sample in headspace, and balancing for 30min.

Mass spectrum conditions: adopting an EI ion source; the scanning mode is as follows: SIM scanning; scanning segment 1 for 2.5min, wherein the name of an internal standard substance is methyl tert-butyl ether, and the quantitative ion is m/z 73.0; scanning segment 2 for 4.9min, wherein the name of the target object is 1-chloro-4-hydroxybutane, and the quantitative ion is m/z 72.1; scanning for 3min, and obtaining a target object with the name of 1, 4-dichlorobutane; the solvent delay was 2.5min.

Example 1

Verifying the applicability of the system:

the method is verified by repeatedly measuring RSD (standard deviation) values of peak areas of the 1, 4-dichlorobutane and the 1-chloro-4-hydroxybutane in standard solutions (the concentrations of the 1, 4-dichlorobutane and the 1-chloro-4-hydroxybutane are both 50ng/mL and the concentration of the methyl tert-butyl ether is 100 ng/mL) and the ratio of peak areas of internal standard solutions (the concentration of the methyl tert-butyl ether is 100 ng/mL) and RSD values of retention time of each target for 6 times, and the RSD values of the relative standard deviation RSD values of the peak areas of the target and the ratio of the peak areas of the internal standard solutions and the RSD values of the retention time of the target in chromatograms obtained for 6 times are required to meet acceptable standards (the RSD values of the peak areas of the target and the internal standard peak areas are not more than 15%, and the RSD values of the retention time of the target are not more than 2 blank), and the solution has no interference with peaks. The test results are shown in tables 1-2.

TABLE 1, 4-Dichlorobutane System suitability measurement results

TABLE 2 results of measuring the applicability of 1-chloro-4-hydroxybutane system

Example 2:

verifying the specificity of the system:

50. Mu.g/mL of a DMF-water solution as a blank solution, 10. Mu.g/mL of a 1, 4-dichlorobutane standard solution, 10. Mu.g/mL of a 1-chloro-4-hydroxybutane standard solution, 10. Mu.g/mL of a methyl t-butyl ether internal standard solution, 10. Mu.g/mL of a mixed standard solution (1, 4-dichlorobutane and 1-chloro-4-hydroxybutane each having a concentration of 5. Mu.g/mL, containing 10. Mu.g/mL of methyl t-butyl ether), 0.5g/mL of a sample solution were each introduced.

A typical spectrum of the blank solution is shown in figure 1.

A typical spectrum of a 10. Mu.g/mL 1, 4-dichlorobutane standard solution is shown in FIGS. 2-3.

A typical spectrum of a 10. Mu.g/mL 1-chloro-4-hydroxybutane standard solution is shown in FIGS. 4-5.

Typical spectra of internal standard solutions of 10. Mu.g/mL methyl tert-butyl ether are shown in FIGS. 6-7.

A typical spectrum of a mixed standard solution of 1, 4-dichlorobutane and 1-chloro-4-hydroxybutane (containing 10. Mu.g/mL of the internal standard solution) is shown in FIG. 8.

Typical patterns for the 0.5g/mL sample solution are shown in FIGS. 9-11.

As can be seen from FIGS. 3-11, the interference peaks are found in all the spectra, which proves that the specificity of the system is good.

Example 3

And (3) verifying the sensitivity of the system:

diluting a mixed standard solution of 1, 4-dichlorobutane and 1-chloro-4-hydroxybutane step by using a diluent to obtain a sample with a signal-to-noise ratio (S/N) of 10 as a quantitative limit; the sample concentration with a signal-to-noise ratio (S/N) of 3 was the detection limit.

Typical spectra of the quantitative and detection limits of 1, 4-dichlorobutane are shown in FIGS. 12-13.

Typical spectra of the quantitative limit and the detection limit of 1-chloro-4-hydroxybutane are shown in FIGS. 14-15.

As can be seen from FIGS. 12 to 15, the limit of quantitation of 1, 4-dichlorobutane was 3ng/mL (S/N: 10.5) and the limit of detection was 1ng/mL (S/N: 5.5); the quantitative limit of 1-chloro-4-hydroxybutane was 0.5ng/mL (S/N: 12.2), and the detection limit was 0.3ng/mL (S/N: 5.7).

Example 4

Verify linearity and range of the system:

taking 5 concentration points within the range of 20-500 ng/mL, taking the concentration of a target as a horizontal coordinate, taking the ratio of the peak area of the target to the peak area of an internal standard as a vertical coordinate, and drawing a curve, wherein the value of a linear regression coefficient r of the curve is not less than 0.99.

The linearity results for 1, 4-dichlorobutane and 1-chloro-4-hydroxybutane are shown in tables 3-4.

TABLE 3 linear results for 1, 4-dichlorobutane

TABLE 4 Linear results for 1-chloro-4-hydroxybutane

Example 4

Precision of the verification system:

(1) Repeatability of

The method is verified to have good precision by preparing 6 sample limit test solutions, injecting 1 needle into each solution, calculating the RSD value of the peak area of the target object and the peak area of the internal standard peak of the 6-needle spectrogram and requiring that the RSD of 6-time result measurement is not more than 15%. The test results are shown in tables 5 to 6.

TABLE 5, 4-Dichlorobutane repeatability results

TABLE 6 repeatability results for 1-chloro-4-hydroxybutane

(2) Intermediate precision

In order to examine the influence of random variation factors on precision, different analysts respectively prepare 6 mixed standard solutions of 1, 4-dichlorobutane and 1-chloro-4-hydroxybutane to perform detection, and the detection needs to be performed on different dates. The RSD of the peak area ratio of 12 times of total sample injection of the precision result and the repeatability result is required to be not more than 15 percent to confirm good intermediate precision. The test results are shown in tables 7 to 8.

TABLE 7 intermediate precision results for 1, 4-dichlorobutane

TABLE 8 intermediate precision results for 1-chloro-4-hydroxybutane

Example 5

And (3) verifying the sample adding recovery rate of the system:

the method is confirmed to have good sample recovery by adding the mixed standard solution to the test solution (batch No. 1, sample in example 7) at the limit of 80%, 100% and 120%, and determining the recovery rate of the standard sample added at different concentrations (actually measured amount-target content in sample solution)/added amount x 100%, wherein the recovery rate is required to be between 80% and 120%. The test results are shown in tables 9 to 10.

TABLE 9 results of sample recovery of 4-dichlorobutane

TABLE 10 results of sample recovery for 1-chloro-4-hydroxybutane

Example 6

And (3) verifying the stability of the system:

according to the change of the time of the mixed standard solution of 50ng/mL of 1, 4-dichlorobutane and 1-chloro-4-hydroxybutane (containing 100ng/mL of the internal standard substance of the methyl tert-butyl ether) and the sample of 0.1g/mL (the sample in the batch No. 1 and the example 7) added with the mixed standard solution of 50ng/mL (containing 100ng/mL of the internal standard substance of the methyl tert-butyl ether), under the natural environment, after the mixed standard solution and the sample added standard solution in the headspace bottle are placed for 0, 3.5, 6, 11, 13.5 and 24 hours, the peak area is compared with 0 hour, the recovery rate is required to be between 80 and 120 percent, and the method provides a basis for detecting the placing time of the mixed standard solution and the sample added standard solution. The test results are shown in tables 11-12.

TABLE 11, 4-Dichlorobutane stability results

TABLE 12 stability results for 1-chloro-4-hydroxybutane

Example 6

And (3) verifying the durability of the system:

and evaluating the degree of the measurement result which is not influenced when the measurement condition parameters have slight changes by changing different flow rates and different injection port temperatures. It is desirable that the tailing factor of 1, 4-dichlorobutane and 1-chloro-4-hydroxybutane be no greater than 2.0 for each of the varying conditions. The chromatographic condition variation parameters are shown in Table 13. The test results are shown in table 14.

TABLE 13 chromatographic Condition variation parameters

Chromatographic parameters	Specified value	Range of variation
			Flow rate of flow	1.0mL/min	0.9 and 1.1mL/min
Temperature at sample inlet	230℃	220 and 240 ℃ C

TABLE 14 test results for variations in various Condition parameters

Example 7

The detection method of the invention is adopted to detect the contents of the genetic toxicity impurities of 1, 4-dichlorobutane and 1-chloro-4-hydroxybutane in a certain medicine injection.

(1) And (3) setting the limit: the risk of a mutagenic impurity at 1.5. Mu.g/day per human intake is considered negligible according to the ICH M7 requirements. Referring to the original preparation specification of the injection, the maximum daily intake of the sample per day can be calculated according to the weight of 60kg by calculation, and the limit of each genotoxic impurity in the sample is preliminarily determined to be not more than 0.0000466%.

The test solutions of the drug injections of the batches 1, 2 and 3 are prepared according to 0.1g/mL, and 3 parallel test solutions are prepared for each batch of samples and are analyzed by an instrument.

Preparation of test solution: weighing 2g of sample, precisely weighing, placing in a 2mL volumetric flask, dissolving with diluent, fixing the volume to a scale, and shaking up; precisely taking 0.2mL of the solution, placing the solution in a 20mL headspace bottle, precisely adding 0.2mL of 1 mu g/mL methyl tert-butyl ether internal standard solution, precisely adding 1.6mL of diluent, and shaking up. Three test solutions (sample concentration: 0.1g/mL, containing internal standard 100 ng/mL) were prepared in the same manner.

The test results are shown in table 15.

TABLE 15 determination of impurity content in samples

Claims (3)

1. A method for simultaneously detecting genetic toxicity impurities of 1, 4-dichlorobutane and 1-chloro-4-hydroxybutane in bulk drugs or preparations is characterized in that: detecting by a gas chromatography-mass spectrometry combined method, adopting an EI ion source, carrying out top air phase analysis, taking methyl tert-butyl ether as an internal standard substance, and calculating the content of impurities in the sample by the internal standard method;

the method comprises the following steps:

(1) Preparing a diluent: mixing purified water and an organic solvent according to a volume ratio of 1;

(2) Preparing a blank solution: taking the diluent as a blank solution;

(3) Preparing an internal standard solution: respectively preparing methyl tert-butyl ether solutions with different concentrations as internal standard solutions by taking the diluent as a solvent and the methyl tert-butyl ether as the internal standard substance;

(4) Preparation of standard solutions: respectively preparing 1, 4-dichlorobutane solution, 1-chloro-4-hydroxybutane solution, 1, 4-dichlorobutane solution and 1-chloro-4-hydroxybutane mixed solution with different concentrations as standard solutions by using the diluent as a solvent and using the 1, 4-dichlorobutane and the 1-chloro-4-hydroxybutane as standard substances;

(5) Preparation of sample test solutions: taking the diluent as a solvent, preparing a sample solution with the concentration of 1g/mL, adding 0.2mL of the sample solution with the concentration of 1g/mL, 0.2mL of an internal standard solution with the concentration of 1 mu g/mL of methyl tert-butyl ether and 1.6mL of the diluent into a 20mL headspace bottle, and shaking up to obtain a sample test solution;

(6) And (3) detection: sample test solution is injected into a gas chromatography-mass spectrometer, a spectrogram is recorded, and the content of impurities in the sample is calculated by an internal standard method;

the organic solvent in the diluent is one of N-butyl alcohol, N-dimethylformamide, N-dimethylacetamide and petroleum ether;

the gas phase conditions were:

a chromatographic column: a quartz capillary column with the specification of 30m multiplied by 0.25mm and 1.4 mu m;

temperature rising procedure: keeping the temperature at 45 ℃ for 2min, heating to 170 ℃ at 10 ℃/min, and then heating to 250 ℃ at 50 ℃/min and keeping the temperature for 1min;

sample inlet temperature: 220 to 240 ℃;

flow rate: 0.9-1.1mL/min, adopting a split-flow mode for sample injection, wherein the split-flow ratio is 5;

introducing sample in headspace, and balancing for 30min;

the mass spectrum conditions are as follows:

adopting an EI source;

the scanning mode comprises the following steps: SIM scanning;

scanning segment 1 for 2.5min, wherein the name of an internal standard substance is methyl tert-butyl ether, and the quantitative ion is m/z 73.0;

scanning segment 2 for 4.9min, wherein the name of the target object is 1-chloro-4-hydroxybutane, and the quantitative ion is m/z 72.1;

scanning for a period of 3min, wherein the time is 7min, the name of a target object is 1, 4-dichlorobutane, and the quantitative ion is m/z 55.1;

the solvent is delayed for 1-3min.

2. The method for simultaneously detecting genotoxic impurities of 1, 4-dichlorobutane and 1-chloro-4-hydroxybutane in a bulk drug substance or a preparation according to claim 1, wherein the genotoxic impurities comprise: the organic solvent and methyl tert-butyl ether were both of AR grade purity.

3. The method for simultaneously detecting genotoxic impurities of 1, 4-dichlorobutane and 1-chloro-4-hydroxybutane in a bulk drug substance or a preparation according to claim 1, wherein the genotoxic impurities comprise: the gas phase conditions were:

a chromatographic column: a quartz capillary column with the specification of 30m multiplied by 0.25mm and 1.4 mu m;

temperature rising procedure: keeping the temperature at 45 ℃ for 2min, heating to 170 ℃ at 10 ℃/min, and then heating to 250 ℃ at 50 ℃/min and keeping the temperature for 1min;

sample inlet temperature: 230 ℃;

flow rate: 1.0mL/min, adopting a split-flow mode for sample injection, wherein the split-flow ratio is 5;

injecting sample in headspace, and balancing for 30min.

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