Method for determining contents of seven genotoxic impurities in epichlorohydrin by gas chromatography-mass spectrometry

Abstract

The invention discloses a method for determining the content of seven genotoxic impurities in epichlorohydrin through gas chromatography-mass spectrometry, and belongs to the technical field of pharmaceutical analysis. According to the method, a quartz capillary chromatographic column taking polyethylene glycol as a stationary phase is adopted, split-flow sample injection is performed, and the genotoxic impurities in an epoxy chloropropane sample are detected by using a gas chromatography-mass spectrometry method; the genetic toxicity impurities are glycidyl, 1, 3-dichloro-2-propyl alcohol, 2, 3-dichloro-1-propyl alcohol, 3-chloro-1, 2-propylene glycol, 2-chloro-1, 3-propylene glycol, 2-chloro-2-propylene-1-alcohol, and 1, 2, 3-trichloropropane. The invention further discloses a preparation method of the compound. According to the present invention, the seven genotoxic impurities in the epoxy chloropropane can be rapidly, effectively, accurately and reliably separated and detected, and the genotoxic impurities can be controlled within the low detection range through the mass spectrometer, such that the quality of the epoxy chloropropane can be easily controlled, the product quality of the hexyl aminolevulinate hydrochloride can be easily improved, and the medication safety of the patient can be easily improved.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical analysis, and particularly relates to a method for determining the contents of seven genotoxic impurities in epichlorohydrin by gas chromatography-mass spectrometry Background Art

[0002] Epichlorohydrin, chemically named 3-chloro-1,2-epoxypropane, with the molecular formula C3H5ClO and molecular weight 92.52, has the following chemical structural formula:

[0003]

[0004] Epichlorohydrin is the starting material for the synthesis of the raw material drug hexyl aminolevulinate hydrochloride. Epichlorohydrin itself is a genotoxic impurity, and genotoxic impurities will be generated during its synthesis process. The main genotoxic impurities are glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, 1,2,3-trichloropropane. Once these impurities are introduced into the subsequent reaction for synthesizing hexyl aminolevulinate hydrochloride, they will have an important impact on the raw material drug and preparation of hexyl aminolevulinate hydrochloride. Therefore, it is urgent to establish an analytical method and conduct quality control for the seven genotoxic impurities in epichlorohydrin.

[0005] Chinese Patent No. CN117074545A discloses a method for simultaneously determining five impurities in epichlorohydrin by gas chromatography. The injection method is to directly inject epichlorohydrin into the gas chromatograph, with the diluent being dimethyl sulfoxide, the carrier gas being nitrogen, the carrier gas flow rate being 0.5 mL / min to 3 mL / min, the initial column temperature being 40 - 80 °C, holding for 5 min, then increasing the temperature to 220 °C at a rate of 5 °C / min and holding for 2 min, the injection port temperature being 225 °C, and the detector temperature being 250 °C. This method only detects five related substance impurities in epichlorohydrin, and its response is poor (the detection concentration of the quantification limit is relatively high), which cannot meet the requirements of the low control limit of genotoxic impurities. Summary of the Invention

[0006] The technical problem solved by the present invention is to provide a gas chromatography-mass spectrometry detection method that can simultaneously detect seven genotoxic impurities, has high sensitivity, good resolution, and low interference from the product matrix, and can be widely used for determining genotoxic impurities in epichlorohydrin, fully meeting the qualitative and quantitative analysis of genotoxic impurities in the product and effectively controlling the product quality.

[0007] To solve the above technical problems, the technical solution adopted by the present invention is as follows:

[0008] Method for determining the contents of seven genotoxic impurities in epichlorohydrin by gas chromatography-mass spectrometry. A fused silica capillary column with polyethylene glycol as the stationary phase is used, and split injection is adopted. The genotoxic impurities in the epichlorohydrin sample are detected by gas chromatography-mass spectrometry. The genotoxic impurities are glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane.

[0009] Preferably, the split ratio of the split injection is 10:1.

[0010] Preferably, in the gas chromatography-mass spectrometry method, the inlet temperature is 200 °C.

[0011] Preferably, in the gas chromatography-mass spectrometry method, the carrier gas is helium, and the carrier gas flow rate is 1 mL / min.

[0012] Preferably, in the gas chromatography-mass spectrometry method, the detector ion source is an electron impact source, the ion source temperature is 230 °C, the mass spectrometry transfer interface temperature is 260 °C, the mass spectrometry monitoring mode is selected ion monitoring, and the detected characteristic ion mass-to-charge ratios are 43, 44.1, 57, 61, 62, 75, and 79.

[0013] Preferably, in the gas chromatography-mass spectrometry method, the column temperature is programmed. The first stage: starting temperature: 30 - 50 °C, running time: 3 - 7 min; the second stage: running temperature: 110 - 120 °C, running time: 2 - 3 min; the third stage: running temperature: 250 - 260 °C, running time: 1 - 2 min; the heating rate from the first stage to the second stage is 5 - 6 °C / min; the heating rate from the second stage to the third stage is 15 - 18 °C / min.

[0014] Preferably, in the gas chromatography-mass spectrometry method, the injection volume is 1 μL.

[0015] Preferably, the detection of the genotoxic impurities in the epichlorohydrin sample includes the following steps:

[0016] 1) Prepare a test solution and a reference solution containing genotoxic impurities;

[0017] 2) Detect the test solution and the reference solution by gas chromatography-mass spectrometry, and record the peak area of the corresponding genotoxic impurity in the test solution, denoted as A 样 , and the peak area of the genotoxic impurity in the reference solution, denoted as A 对 ;

[0018] 3) Calculate the content w / w of the genotoxic impurity in epichlorohydrin according to the following formula;

[0019] Content w / w = (C 对 *A 样 ) / (A 对 *C 样 );

[0020] In the formula: C 对 is the concentration of the reference substance solution; C 样 is the concentration of the test sample solution; A 对 is the peak area of the corresponding genotoxic impurity in the reference substance solution; A 样 is the peak area of the corresponding genotoxic impurity in the test sample solution.

[0021] Preferably, the preparation process of the reference substance solution containing the genotoxic impurity is as follows: Take appropriate amounts of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane, dissolve and dilute with a solvent to make a mixed solution containing 2.5 μg of each genotoxic impurity per 1 mL, mix well, and obtain the reference substance solution.

[0022] Preferably, the solvent is methanol.

[0023] Beneficial effects: Compared with the prior art, the present invention has the following advantages:

[0024] 1) The gas chromatography-mass spectrometry method of the present invention can quickly, effectively, accurately, and reliably separate and detect seven genotoxic impurities in epichlorohydrin, the starting material of aminolevulinic acid hexyl ester hydrochloride, and the genotoxic impurities can be controlled within a lower detection range through a mass spectrometer, which is beneficial to controlling the quality of epichlorohydrin, improving the product quality of aminolevulinic acid hexyl ester hydrochloride, and improving the drug safety of patients;

[0025] 2) Compared with the existing Chinese invention patent of CN117074545A, the present invention can strictly control multiple genotoxic impurities in epichlorohydrin, accurately locate the genotoxic impurities by using the gas chromatography-mass spectrometry method, and control them at a lower detection concentration. Description of the Drawings

[0026] Figure 1 is the chromatogram of the reference substance solution detected in Example 1 of the present invention. Among them, a is the chromatogram of glycidol, b is the chromatogram of 1,3-dichloro-2-propanol, c is the chromatogram of 2,3-dichloro-1-propanol and 2-chloro-1,3-propanediol, d is the chromatogram of 3-chloro-1,2-propanediol, e is the chromatogram of 2-chloro-2-propen-1-ol, and f is the chromatogram of 1,2,3-trichloropropane;

[0027] Figure 2 For the test solution spectrum detected in Example 1 of the present invention, where a is the chromatogram of glycidol, b is the chromatogram of 1,3-dichloro-2-propanol, c is the chromatogram of 2,3-dichloro-1-propanol, d is the chromatogram of 3-chloro-1,2-propanediol, e is the chromatogram of 2-chloro-2-propen-1-ol, and f is the chromatogram of 1,2,3-trichloropropane;

[0028] Figure 3 It is a standard curve graph, where 2a is the standard curve of glycidol, 2b is the standard curve graph of 1,3-dichloro-2-propanol, 2c is the standard curve of 2,3-dichloro-1-propanol, 2d is the standard curve of 3-chloro-1,2-propanediol, 2e is the standard curve of 2-chloro-1,3-propanediol, 2f is the standard curve of 2-chloro-2-propen-1-ol, and 2g is the standard curve of 1,2,3-trichloropropane. Detailed implementation manners

[0029] The following further clarifies the present invention in combination with specific embodiments. The embodiments are implemented on the premise of the technical solution of the present invention. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. For those conditions not specified in the embodiments, they are carried out according to conventional conditions or the conditions recommended by the manufacturer. For the reagents or instruments whose manufacturers are not specified, they are all conventional products that can be obtained through commercial purchase.

[0030] In the following embodiments, the glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, and 1,2,3-trichloropropane selected are all purchased from Aladdin Reagent (Shanghai) Co., Ltd. and are of analytical purity. 3-chloro-1,2-propanediol and 2-chloro-1,3-propanediol are both purchased from Shanghai Macklin Biochemical Co., Ltd. and are of analytical purity. 2-chloro-2-propen-1-ol is purchased from Merck Chemical Technology (Shanghai) Co., Ltd. and is of analytical purity. Epichlorohydrin is purchased from Jiangsu Ruixiang Chemical Co., Ltd.

[0031] Example 1

[0032] A method for determining the contents of seven genotoxic impurities in epichlorohydrin by gas chromatography-mass spectrometry, comprising the following steps:

[0033] (1) Prepare a reference solution and a test solution:

[0034] Preparation of the reference solution: Appropriate amounts of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane are taken and dissolved and diluted with methanol to prepare a mixed solution containing 2.5 μg of each genotoxic impurity per 1 mL as the reference solution;

[0035] Preparation of the test solution: Take 500 mg of epichlorohydrin and place it in a 10 mL volumetric flask. Dissolve it with methanol and dilute to the mark to obtain the test solution.

[0036] (2) Set the detection conditions for gas chromatography - mass spectrometry:

[0037] Chromatographic column: A fused - silica capillary column with polyethylene glycol as the stationary phase (MEGA - WAX Plus, 30 m × 0.25 mm, 0.25 μm);

[0038] Injector temperature: 200 °C;

[0039] Injection mode: Split injection, split ratio is 10:1;

[0040] Carrier gas: Helium, carrier gas flow rate: 1.0 mL / min;

[0041] Injection volume is 1 μL;

[0042] The detector ion source is an electron impact source (EI), the ion source temperature is 230 °C, the mass spectrometry transfer interface temperature is 260 °C, the mass spectrometry monitoring mode is selected ion monitoring (SIM), and the detection characteristic ion mass - to - charge ratios (m / z) are 43, 44.1, 57, 61, 62, 75, 79;

[0043] The column temperature is programmed as follows: First stage: Initial temperature: 30 - 50 °C, running time: 3 - 7 min; Second stage: Running temperature: 110 - 120 °C, running time: 2 - 3 min; Third stage: Running temperature: 250 - 260 °C, running time: 1 - 2 min; The heating rate from the first stage to the second stage is 5 - 6 °C / min; The heating rate from the second stage to the third stage is 15 - 18 °C / min;

[0044] (3) Take 1 μL of the reference solution and inject it into the gas chromatography - mass spectrometry instrument. Adjust the detection sensitivity to make the peak height reach the full scale of the peak height of the reference solution. Then accurately measure 1 μL of the test solution and inject it into the gas chromatography - mass spectrometry instrument, and record the chromatogram. The results are as Figure 1 shown.

[0045] It can be seen from Figure 1 that under the chromatographic conditions of the present invention, there is no interference for each genotoxic impurity under its corresponding characteristic ion, and there is no interference from the blank solvent.

[0046] It can be seen from Figure 2It can be seen that among the impurities in epichlorohydrin, except for glycidol and 2-chloro-1,3-propanediol, the other 5 genotoxic impurities were all detected; the genotoxic impurities in the chromatogram of the test solution did not interfere with each other, and there were no unknown impurities in the test solution interfering with the detection at the corresponding mass-to-charge ratios of each genotoxic impurity.

[0047] Example 2

[0048] Method validation was carried out on the detection method of Example 1, and the validation was carried out from several aspects such as system suitability, limit of quantitation, limit of detection, linear relationship, precision, accuracy, and solution stability.

[0049] 1. System suitability

[0050] Positioning solution: Weigh accurately appropriate amounts of glycidol (SM2-1), 1,3-dichloro-2-propanol (SM2-2), 2,3-dichloro-1-propanol (SM2-3), 3-chloro-1,2-propanediol (SM2-4), 2-chloro-1,3-propanediol (SM2-7), 2-chloro-2-propen-1-ol (SM2-8), 1,2,3-trichloropropane (SM2-9) reference substances and acetic acid (SM2-5), glycerol (SM2-6) reference substances, and dilute with methanol to prepare a solution containing 50 μg of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, 1,2,3-trichloropropane, acetic acid, and glycerol in each 1 mL as the positioning solution for each impurity.

[0051] System suitability solution: Weigh accurately 500 mg of epichlorohydrin and place it in a 10 mL volumetric flask containing a small amount of methanol. Add an appropriate amount of each positioning solution and dilute with methanol to prepare a solution containing 50 mg of epichlorohydrin and 5 μg of each impurity in each 1 mL as the system suitability solution.

[0052] Test solution: Weigh accurately 500 mg of epichlorohydrin and place it in a 10 mL volumetric flask containing a small amount of methanol. Dilute with methanol to the mark to prepare a solution containing 50 mg of epichlorohydrin in each 1 mL.

[0053] Respectively take 1 μL of blank solvent, test solution, system suitability solution, and positioning solution for each impurity, inject them into the gas chromatography-mass spectrometry instrument, record the chromatogram, and examine the separation of each impurity. The results are shown in Table 1.

[0054] Table 1 Results of system suitability

[0055]

[0056]

[0057] As can be seen from Table 1, under the characteristic ions corresponding to each impurity, the blank solvent and epichlorohydrin do not interfere with the detection of impurities, there is no interference among the impurities, and the system suitability of the method of the present invention meets the detection requirements.

[0058] 2. Quantitation Limit and Detection Limit

[0059] Accurately weigh appropriate amounts of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane reference substances, and dilute with methanol to prepare a mixed solution containing 2.5 μg of each of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane per 1 mL. Use the stepwise dilution method to measure its quantitation limit (S / N≥10) and detection limit (S / N≥3), and the results are shown in Table 2.

[0060] Table 2 Results of Quantitation Limit and Detection Limit

[0061]

[0062] As can be seen from Table 2, under the chromatographic conditions of the present invention, the quantitation limit and detection limit meet the inspection requirements.

[0063] 3. Linear Relationship

[0064] Accurately weigh appropriate amounts of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane reference substances, and dilute with methanol to prepare a mixed solution containing 10 μg of each of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane per 1 mL as a 400% linear mother liquor (200 ppm). Prepare linear solutions according to Table 3.

[0065] Table 3 Preparation of Linear Solutions

[0066]

[0067]

[0068] Precisely measure 1 μL of each of the above solutions and inject them into a gas chromatography-mass spectrometry instrument. Inject 2 needles for each sample, record the chromatogram, measure the peak area, and perform linear regression with the peak area A as the ordinate and the concentration C as the abscissa. The results are shown in Table 4 below.

[0069] Table 4 Results of the linear relationship test

[0070]

[0071]

[0072]

[0073] As can be seen from Table 4, glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane showed good linearity (r > 0.99) with their peak areas within their respective concentration ranges.

[0074] 4. Injection precision

[0075] Accurately weigh appropriate amounts of reference substances of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane, and dilute them with methanol to prepare a mixed solution containing 2.5 μg of each of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane per 1 mL. Accurately measure 1 μL and inject it into the gas chromatography-mass spectrometry instrument, and inject samples continuously for 6 times, and record the peak areas. The results are shown in Table 5.

[0076] Table 5 Results of injection precision

[0077]

[0078] As can be seen from Table 5, when injecting samples continuously for 6 times, the RSD of the target peak area ≤ 10%, and the injection precision is good.

[0079] 5. Solution stability

[0080] Conduct solution stability tests on the reference substance solution and the test solution respectively.

[0081] 1) Stability of the reference substance solution

[0082] Accurately weigh appropriate amounts of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane reference substances, and dilute them with methanol to prepare a mixed solution containing 2.5 μg of each of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane per 1 mL. Inject the solution at different times to examine its within-day stability. The results are shown in Table 6.

[0083] Table 6 Results of the stability of the impurity reference substance solution

[0084]

[0085]

[0086] As can be seen from Table 6, each impurity reference substance is stable in the solvent within 12.5 hours (RSD ≤ 10%).

[0087] 2) Stability of the test solution

[0088] Accurately weigh an appropriate amount of epichlorohydrin, dissolve it in methanol and dilute to prepare a solution containing 50 mg of epichlorohydrin per 1 mL as the test solution. Inject the solution at different times to examine the within-day stability of its impurities (calculated by peak area). The results are shown in Table 7.

[0089] Table 7 Results of the stability of the test solution

[0090]

[0091] As can be seen from Table 7, the test solution is stable within 2.5 hours (RSD ≤ 15%).

[0092] 6. Repeatability

[0093] Accurately weigh appropriate amounts of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane reference substances, and dilute them with methanol to prepare a mixed solution containing 2.5 μg of each of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane per 1 mL as the reference solution. Accurately weigh an appropriate amount of epichlorohydrin, dissolve it in methanol and dilute to prepare a solution containing 50 mg per 1 mL as the test solution. The test solution was repeatedly prepared in 6 portions for determination. The results are shown in Table 8.

[0094] Table 8 Repeatability Results

[0095]

[0096]

[0097] As can be seen from Table 8, the absolute value of the difference between the impurity contents of the 6 repeatability samples and the mean value is ≤ 10% of the limit, indicating good repeatability.

[0098] 7. Accuracy Test (Spiking Recovery Rate)

[0099] Preparation of reference solution: Weigh accurately appropriate amounts of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane reference substances, and dilute with methanol to prepare a mixed solution containing 2.5 μg of each of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane per 1 mL.

[0100] Preparation of 200% recovery reserve solution: Weigh accurately appropriate amounts of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane reference substances, and dilute with methanol to prepare a mixed solution containing 5 μg of each of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane per 1 mL.

[0101] Preparation of 100% recovery reserve solution: Weigh accurately appropriate amounts of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane reference substances, and dilute with methanol to prepare a mixed solution containing 2.5 μg of each of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane per 1 mL.

[0102] Preparation of stock solution with 50% recovery rate: Weigh accurately appropriate amounts of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane reference substances, and dilute them with methanol to prepare a mixed solution containing 1.25 μg of each of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, and 1,2,3-trichloropropane in 1 mL.

[0103] Accurately weigh 500 mg of epichlorohydrin into nine dry 10-mL volumetric flasks. Divide them into three groups of three. Before injection, add low, medium, and high recovery rate stock solutions at 50%, 100%, and 200% respectively, and make up to the mark with solvent to obtain the corresponding spiked test solutions.

[0104] Precisely measure 1 μL of each of the above samples and inject them into a gas chromatography-mass spectrometry (GC-MS) instrument. Record the peak areas of each genotoxic impurity, and calculate their recovery rates and relative standard deviations (RSDs). The results are shown in Table 9.

[0105] Table 9 Results of recovery rates

[0106]

[0107]

[0108]

[0109]

[0110] As can be seen from Table 9, the recovery rates and average recovery rates of each genotoxic impurity are within the range of 80% - 115%, and the RSD ≤ 10%, indicating good recovery rates.

[0111] 8. Intermediate precision

[0112] The content of genotoxic impurities in the same batch of samples was examined by different operators at different times according to the repeatability test method. The results are shown in Table 10.

[0113] Table 10 Results of intermediate precision test

[0114]

[0115] As can be seen from Table 10, there are no obvious changes in the detection results of the genotoxic impurity content of this product, indicating good intermediate precision of this method.

[0116] 9. Robustness

[0117] The durability of the analytical method for genetic toxic impurities of epichlorohydrin was verified mainly from three aspects: different initial temperatures, different flow rates, and different inlet temperatures. Experiments were conducted according to the conditions in Table 11, and the results are shown in Table 12.

[0118] Table 11 Durability Conditions

[0119] parameter specified parameter range of variation inlet temperature 200℃ ±5℃ flow rate 1.0 mL / min flow rate ± 0.05 mL / min initial temperature 40℃ ±5℃

[0120] Table 12 Durability Results

[0121]

[0122] As can be seen from Table 12, when there are slight changes in the initial temperature, flow rate, and inlet temperature, the detection results of genetic toxic impurities of epichlorohydrin show no obvious changes (the absolute value of the impurity content compared with the original conditions ≤ 20% of the limit), indicating that the durability of this chromatographic condition is good.

[0123] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A method for determining the contents of seven genotoxic impurities in epichlorohydrin by gas chromatography-mass spectrometry, characterized in that, A quartz capillary chromatographic column with polyethylene glycol as the stationary phase is used, and split injection is adopted. The genotoxic impurities in epichlorohydrin samples are detected by gas chromatography-mass spectrometry; the genotoxic impurities are glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, 1,2,3-trichloropropane.

2. The method for determining the contents of seven genotoxic impurities in epichlorohydrin by gas chromatography-mass spectrometry according to claim 1, wherein The split ratio of the split injection is 10:

1.

3. The method for determining the contents of seven genotoxic impurities in epichlorohydrin by gas chromatography-mass spectrometry according to claim 1, characterized in that, In the gas chromatography-mass spectrometry method, the inlet temperature is 200 °C.

4. The method for determining the contents of seven genotoxic impurities in epichlorohydrin by gas chromatography-mass spectrometry according to claim 1, characterized in that, In the gas chromatography-mass spectrometry method, the carrier gas is helium, and the carrier gas flow rate is 1 mL / min.

5. The method for determining the contents of seven genotoxic impurities in epichlorohydrin by gas chromatography-mass spectrometry according to claim 1, characterized in that, In the gas chromatography-mass spectrometry method, the detector ion source is an electron impact source, the ion source temperature is 230 °C, the mass spectrometry transfer interface temperature is 260 °C, the mass spectrometry monitoring mode is selected ion scanning, and the detected characteristic ion mass-to-charge ratios are 43, 44.1, 57, 61, 62, 75, 79.

6. The method for determining the contents of seven genotoxic impurities in epichlorohydrin by gas chromatography-mass spectrometry according to claim 1, wherein In the gas chromatography-mass spectrometry method, the column temperature is programmed heating. The first stage: starting temperature: 30 - 50 °C, running time: 3 - 7 min; the second stage: running temperature: 110 - 120 °C, running time: 2 - 3 min; the third stage: running temperature: 250 - 260 °C, running time: 1 - 2 min; the heating rate from the first stage to the second stage is 5 - 6 °C / min; the heating rate from the second stage to the third stage is 15 - 18 °C / min.

7. The method for determining the contents of seven genotoxic impurities in epichlorohydrin by gas chromatography-mass spectrometry according to claim 1, characterized in that, In the gas chromatography-mass spectrometry method, the injection volume is 1 μL.

8. The method for determining the contents of seven genotoxic impurities in epichlorohydrin by gas chromatography-mass spectrometry according to claim 1, wherein, The detection of genotoxic impurities in epichlorohydrin samples includes the following steps: 1) Prepare a test solution and a reference solution containing genotoxic impurities; 2) Detect the test solution and the reference solution by gas chromatography-mass spectrometry, record the peak area of the corresponding genotoxic impurity in the test solution, denoted as A 样 , and the peak area of the genotoxic impurity in the reference solution, denoted as A 对 ; 3) Calculate the content w / w of genotoxic impurities in epichlorohydrin according to the following formula; Content w / w = (C 对 * A 样 ) / (A 对 * C 样 ); Where: C 对 is the concentration of the reference solution; C 样 is the concentration of the test solution; A 对 is the peak area of the genotoxic impurity corresponding to the reference solution; A 样 is the peak area of the genotoxic impurity corresponding to the test solution.

9. The method for determining the contents of seven genotoxic impurities in epichlorohydrin by gas chromatography-mass spectrometry according to claim 8, characterized in that, The preparation process of the reference solution containing genotoxic impurities is as follows: Appropriate amounts of glycidol, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol, 2-chloro-1,3-propanediol, 2-chloro-2-propen-1-ol, 1,2,3-trichloropropane are taken, dissolved in a solvent and diluted to make a mixed solution containing 2.5 μg of each genotoxic impurity per 1 mL, and mixed evenly to obtain the reference solution.

10. The method for determining the contents of seven genotoxic impurities in epichlorohydrin by gas chromatography-mass spectrometry according to claim 9, characterized in that, The solvent is methanol.

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