Method for detecting 2-bromoethanol and ethylene glycol in imidafenacin starting material 1, 2-dibromoethane

Abstract

The invention discloses a method for detecting 2-bromoethanol and ethylene glycol in an imidafenacin starting material 1, 2-dibromoethane, and belongs to the field of chemical medicine analysis. According to the present invention, the gas chromatography is adopted to detect and separate the impurities such as 2-bromoethanol and ethylene glycol in the main component 1, 2-dibromoethane, the method has characteristics of high specificity, strong sensitivity and good durability, and compared with the complex research means of the conventional potential genetic toxicity impurity, the analysis method of the present invention has characteristics of rapidness, simpleness, effectiveness and reliability, and can be used for the industrial production. 2-bromoethanol and ethylene glycol in the imidafenacin starting material 1, 2-dibromoethane can be effectively detected, so that the quality of the imidafenacin bulk drug is ensured.

Description

Technical Field

[0001] The invention belongs to the technical field of chemical drug analysis, and in particular relates to a method for detecting 2-bromoethanol and ethylene glycol in 1,2-dibromoethane, a starting material of imidafenacin. Background Art

[0002] Imidafenacin is a new therapeutic agent for overactive bladder with antimycin activity, used to treat urothelial mediator release and detrusor overactivity caused by cerebral infarction. It is a new diphenylbutyramide anticholinergic drug jointly developed by Ono Pharmaceutical Co., Ltd. and Kyorin Pharmaceutical. It has high bladder selectivity and is used to treat urinary urgency, urinary frequency and urinary incontinence caused by overactive bladder. It was launched in Japan in June 2007.

[0003] The existing synthesis process of imidafenacin requires the starting material 1,2-dibromoethane. The synthesis process of the starting material 1,2-dibromoethane may produce by-products ethylene glycol and 2-bromoethanol, among which 2-bromoethanol has a genotoxic impurity warning structure with a relatively low limit.

[0004] At present, there are almost no reports on the relevant research on the impurity control of 1,2-dibromoethane in the literature or patents at home and abroad. Therefore, how to reduce the by-products (ethylene glycol and 2-bromoethanol) produced by the synthesis of the starting material 1,2-dibromoethane in the process of preparing synthetic imidafenacin has become a problem that needs to be solved urgently. Summary of the invention

[0005] The purpose of the present invention is to provide a method for detecting 2-bromoethanol and ethylene glycol in 1,2-dibromoethane, a starting material of imidafenacin, and to reduce the risk of impurities being introduced into imidafenacin raw materials by controlling the impurities, thereby ensuring the quality of the imidafenacin raw materials.

[0006] In order to solve the above problems, the technical solution adopted by the present invention is as follows:

[0007] A method for detecting 2-bromoethanol and ethylene glycol in 1,2-dibromoethane, a starting material of imidafenacin, uses gas chromatography to detect and separate the impurities 2-bromoethanol and ethylene glycol in the main component 1,2-dibromoethane; wherein the gas phase detection conditions are as follows:

[0008] Chromatographic column: A capillary column with 6% cyanopropylphenyl and 94% dimethylpolysiloxane as stationary liquid;

[0009] Carrier gas: high-purity nitrogen, flow rate 4.62~4.82mL / min;

[0010] Detector temperature: 220~240℃;

[0011] Inlet temperature: 190~210℃;

[0012] Detector: FID detector;

[0013] Split ratio: 3:1~1:1;

[0014] Injection volume: 1 μL;

[0015] Solvent: methanol;

[0016] Heating program: starting temperature is 75-85°C, maintained for 4 minutes, heated to 180°C at a rate of 10°C per minute, maintained for 5 minutes.

[0017] Furthermore, the specification of the chromatographic column is 30m×0.530mm, and the film thickness is 3.00μm.

[0018] Furthermore, the detector temperature is 230°C.

[0019] Furthermore, the injection port temperature is 200°C.

[0020] Furthermore, the carrier gas flow rate is 4.72 mL / min.

[0021] Furthermore, the split ratio is 2:1.

[0022] Furthermore, the starting temperature of the temperature rising program is 80°C.

[0023] Further, any of the above methods for detecting 2-bromoethanol and ethylene glycol in 1,2-dibromoethane, the starting material of imidafenacin, comprises the following steps:

[0024] (1) Take 1,2-dibromoethane, add methanol to dissolve and dilute to make a solution containing 10 mg of 1,2-dibromoethane per 1 mL, which is used as the test solution;

[0025] (2) Take 2-bromoethanol and ethylene glycol, add methanol to dissolve and dilute to prepare a solution containing 10 μg of 2-bromoethanol and 10 μg of ethylene glycol per 1 mL, which is used as the reference solution;

[0026] Take 1,2-dibromoethane and place it in a volumetric bottle, add 2-bromoethanol fixation solution and ethylene glycol fixation solution, then add solvent to dissolve and dilute to the scale, shake well, and you will get a mixed solution of reference substance and test substance;

[0027] (3) Inject the blank solvent, test solution, reference solution, reference and test solution, and positioning solution into the gas chromatograph respectively and record the chromatogram.

[0028] Furthermore, the volume of each solution injection was 1 μL.

[0029] Furthermore, the separation degree between the 1,2-dibromoethane peak and adjacent peaks in the test solution is greater than 1.5.

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

[0031] 1) The present invention uses gas chromatography to effectively detect the impurities 2-bromoethanol and ethylene glycol in the main component 1,2-dibromoethane, and the method has high specificity, strong sensitivity and good durability. Compared with the more complicated research methods of conventional potential genotoxic impurities, the analysis method of the present invention is fast, simple, effective and reliable, thereby ensuring the quality of the imidafenacin raw material.

[0032] 2) The present invention has no obvious difference in the content determination results of residual solvents when the chromatographic parameters change slightly under different injection port temperatures, different detector temperatures, different column starting temperatures, different carrier gas flow rates, different split ratios, and chromatographic columns of the same model and manufacturer with different numbers, and has good durability. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 is a chromatogram of a mixed solution of 1,2-dibromoethane, 2-bromoethanol and ethylene glycol in Example 1;

[0034] Figure 2 The chromatogram of the test solution in Example 1;

[0035] Figure 3 is the linear relationship diagram of 2-bromoethanol;

[0036] Figure 4 This is the linear relationship diagram for ethylene glycol. DETAILED DESCRIPTION

[0037] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are described in detail below in conjunction with specific embodiments.

[0038] Unless otherwise specified, the materials and equipment used in the following examples are commercially available products.

[0039] According to the synthesis route of 1,2-dibromoethane, the starting material of imidafenacin, and the impurity analysis, it can be determined that the by-products are ethylene glycol and 2-bromoethanol, among which 2-bromoethanol is a halogenated alkane compound and a potential hereditary impurity. The structure and generation route of the impurity are as follows:

[0040]

[0041] The gas phase detection conditions of the present invention are as follows:

[0042] Chromatographic column: a capillary column with 6% cyanopropylphenyl and 94% dimethylpolysiloxane as stationary phase (or similar polarity) (DB-624, 30m×0.530mm, 3.00μm);

[0043] Carrier gas: high-purity nitrogen, flow rate 4.62~4.82mL / min;

[0044] Detector temperature: 220~240℃

[0045] Inlet temperature: 190~210℃

[0046] Detector: FID detector

[0047] Split ratio: 3:1~1:1

[0048] Injection volume: 1 μL

[0049] Solvent: Methanol

[0050] Heating program: starting temperature is 75-85°C, maintained for 4 minutes, heated to 180°C at a rate of 10°C per minute, maintained for 5 minutes.

[0051] Example 1

[0052] A method for detecting 2-bromoethanol and ethylene glycol in 1,2-dibromoethane, a starting material of imidafenacin, comprises the following steps:

[0053] (1) Preparation of the test sample solution: Take an appropriate amount of 1,2-dibromoethane, add methanol to dissolve and dilute to make a solution containing 10 mg of 1,2-dibromoethane per 1 mL, which is used as the test sample solution;

[0054] (2) Preparation of reference solution: Accurately weigh appropriate amounts of 2-bromoethanol and ethylene glycol, add methanol to dissolve and dilute to prepare a solution containing 10 μg of 2-bromoethanol and 10 μg of ethylene glycol per 1 mL, which serves as the reference solution;

[0055] Reference substance + test substance mixed solution: Accurately weigh 100 mg of 1,2-dibromoethane and place it in a 10 mL volumetric flask. Accurately measure 1.0 mL each of 2-bromoethanol fixing solution and ethylene glycol fixing solution and place them in a volumetric flask. Add solvent to dissolve and dilute to the scale. Shake well to obtain.

[0056] (3) Detection: Using a gas chromatograph, accurately measure 1 μL of blank solvent, test solution, reference solution, reference + test solution mixture, and positioning solution and inject them into the gas chromatograph to record the chromatogram; the results are shown in Table 1 and Figure 1 , Figure 2 shown.

[0057] Table 1 Retention time and separation results of the corresponding peaks of each solution

[0058]

[0059] Result analysis: The retention time of ethylene glycol peak and 2-bromoethanol peak in the reference solution was consistent with the retention time of the corresponding peaks in the reference + test solution and the positioning solution; the blank solvent had no interference with the detection of each component; the separation degree between the 1,2-dibromoethane peak and the adjacent peaks in the test solution was greater than 1.5.

[0060] Example 2 Method Verification

[0061] The detection method disclosed in Example 1 was verified from the aspects of quantitative limit, detection limit, linear range, injection precision, repeatability, intermediate precision, accuracy, solution stability and durability, as follows:

[0062] 1. Limit of quantification and detection limit

[0063] Accurately measure the reference solution, dilute the solution and inject it according to the content detection chromatographic conditions, record the chromatogram, take the signal-to-noise ratio S / N≈10 as the quantitative limit, and take the signal-to-noise ratio S / N≈3 as the detection limit. The results are shown in Table 2 below.

[0064] Table 2 Quantitation limit and detection limit test results

[0065]

[0066] 2. Linear range

[0067] Take appropriate amount of 2-bromoethanol and ethylene glycol reference substances respectively, prepare linear series solutions step by step, and perform gas chromatography detection to obtain the linear relationship of 2-bromoethanol, as shown in Table 3 and Figure 3 As shown; the linear relationship of ethylene glycol is shown in Table 4 and Figure 4 shown.

[0068] Table 3 2-bromoethanol linearity test results

[0069]

[0070] Table 4 Ethylene glycol linear test results

[0071]

[0072] 3. Injection precision

[0073] The reference solution was injected 6 times continuously, 1 μL each time, into the gas chromatograph, and the chromatogram was recorded. The results are shown in Table 5 below.

[0074] Table 5 Results of 6 consecutive injections of reference solution

[0075]

[0076] Result analysis: In the chromatogram of the reference solution, the sample was injected 6 times, and the RSD of the peak areas of ethylene glycol and 2-bromoethanol were both <2.0%, and the RSD of the retention time were both <2.0%, which met the requirements and the injection precision was good.

[0077] 4. Repeatability test

[0078] Accurately measure 1 μL of the reference solution and the test solution, inject them into the gas chromatograph, measure them in parallel 6 times, record the chromatograms, and the results are shown in Table 6 below.

[0079] Table 6 Comparison of the test results of the reference solution and the test solution repeated 6 times

[0080] name Ethylene glycol content (%) 2-Bromoethanol content (%) Repeatability 1 ND 0.044 Repeatability 2 ND 0.043 Repeatability 3 ND 0.043 Repeatability 4 ND 0.043 Repeatability 5 ND 0.044 Repeatability 6 ND 0.043 average value N / A 0.043 RSD% N / A 0.94

[0081] Result analysis: Ethylene glycol was not detected in any of the 6 samples, and the RSD of 2-bromoethanol detection content was less than 10.0%, which met the requirements and had good repeatability.

[0082] 5. Intermediate precision test

[0083] Replace different test personnel, use different instruments at different time periods, and prepare blank solvent, reference solvent, and test solution according to the above method; accurately measure 1 μL of each solution and inject it into the gas chromatograph respectively, repeat the experiment 6 times, record the chromatogram, and the results are shown in Table 7 below.

[0084] Table 7 Intermediate precision test results

[0085] name Ethylene glycol content (%) 2-Bromoethanol content (%) Repeatability 1 ND 0.044 Repeatability 2 ND 0.043 Repeatability 3 ND 0.043 Repeatability 4 ND 0.043 Repeatability 5 ND 0.044 Repeatability 6 ND 0.043 Intermediate precision 1 ND 0.039 Intermediate precision 2 ND 0.037 Intermediate precision 3 ND 0.040 Intermediate precision 4 ND 0.041 Intermediate precision 5 ND 0.039 Intermediate precision 6 ND 0.039 average value N / A 0.041 RSD% N / A 5.63

[0086] Results showed that 12 samples were statistically analyzed for repeatability and intermediate precision, and ethylene glycol was detected in all samples. The RSD of 2-bromoethanol detection content was less than 10.0%, which met the requirements and had good precision.

[0087] 6. Accuracy test

[0088] Quantitatively add three concentrations (80%, 100%, 120%) of each known component to the test solution, calculate the content of the known components in the test solution, deduct the content of each component obtained after the known component content in the test solution is the measured value, and divide the measured value by the added value to obtain the recovery rate of each component. The test results are shown in Tables 8 and 9 below.

[0089] Table 8 Ethylene glycol accuracy test results

[0090]

[0091]

[0092] Table 9 2-bromoethanol accuracy test results

[0093]

[0094] 7. Solution stability

[0095] Take the test solution and the reference solution, place them at room temperature, measure the stability according to the above method, and record the chromatogram. The results are shown in Table 10 below.

[0096] Table 10 Solution stability test results

[0097]

[0098]

[0099] Result analysis: At room temperature, the test solution is stable within 51 hours, and the mixed control solution is stable within 51 hours.

[0100] 8. Durability test

[0101] The changes of the injection port temperature, detector temperature, carrier gas flow rate, chromatographic column, split ratio, and column starting temperature on the system were tested respectively to verify the durability of the chromatographic conditions, as shown in Table 11 below.

[0102] Table 11 Durability test results

[0103]

[0104] The results showed that slight changes in chromatographic parameters, such as different injection port temperatures, different detector temperatures, different column starting temperatures, different carrier gas flow rates, different split ratios, and different numbered chromatographic columns of the same model and manufacturer, had no significant difference in the determination results of the residual solvent content, and the durability was good.

[0105] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for detecting 2-bromoethanol and ethylene glycol in 1,2-dibromoethane, a starting material of imidafenacin, characterized in that: Gas chromatography was used to detect the impurities 2-bromoethanol and ethylene glycol in the main component 1,2-dibromoethane; wherein the gas phase detection conditions were as follows: Chromatographic column: A capillary column with 6% cyanopropylphenyl and 94% dimethylpolysiloxane as stationary liquid; Carrier gas: high-purity nitrogen, flow rate 4.62~4.82mL / min; Detector temperature: 220~240℃; Inlet temperature: 190~210℃; Detector: FID detector; Split ratio: 3:1~1:1; Injection volume: 1 μL; Solvent: methanol; Heating program: starting temperature is 75-85°C, maintained for 4 minutes, heated to 180°C at a rate of 10°C per minute, maintained for 5 minutes.

2. The method for detecting 2-bromoethanol and ethylene glycol in 1,2-dibromoethane, the starting material of imidafenacin, according to claim 1, characterized in that: The specification of the chromatographic column is 30m×0.530mm, and the film thickness is 3.00μm.

3. The method for detecting 2-bromoethanol and ethylene glycol in 1,2-dibromoethane, the starting material of imidafenacin, according to claim 1, characterized in that: The detector temperature was 230°C.

4. The method for detecting 2-bromoethanol and ethylene glycol in 1,2-dibromoethane, the starting material of imidafenacin according to claim 1, characterized in that: The injection port temperature is 200°C.

5. The method for detecting 2-bromoethanol and ethylene glycol in 1,2-dibromoethane, the starting material of imidafenacin according to claim 1, characterized in that: The carrier gas flow rate is 4.72 mL / min.

6. The method for detecting 2-bromoethanol and ethylene glycol in 1,2-dibromoethane, the starting material of imidafenacin according to claim 1, characterized in that: The split ratio is 2:

1.

7. The method for detecting 2-bromoethanol and ethylene glycol in 1,2-dibromoethane, the starting material of imidafenacin according to claim 1, characterized in that: The starting temperature of the heating program was 80°C.

8. The method for detecting 2-bromoethanol and ethylene glycol in 1,2-dibromoethane, a starting material of imidafenacin, according to any one of claims 1 to 7, characterized in that: Here are the steps: (1) Take 1,2-dibromoethane, add methanol to dissolve and dilute to make a solution containing 10 mg of 1,2-dibromoethane per 1 mL, which is used as the test solution; (2) Take 2-bromoethanol and ethylene glycol, add methanol to dissolve and dilute to prepare a solution containing 10 μg of 2-bromoethanol and 10 μg of ethylene glycol per 1 mL, which is used as the reference solution; Take 1,2-dibromoethane and place it in a volumetric bottle, add 2-bromoethanol fixation solution and ethylene glycol fixation solution, then add solvent to dissolve and dilute to the scale, shake well, and you will get a mixed solution of reference substance and test substance; (3) Inject the blank solvent, test solution, reference solution, reference and test solution, and positioning solution into the gas chromatograph respectively and record the chromatogram.

9. The method for detecting 2-bromoethanol and ethylene glycol in 1,2-dibromoethane, the starting material of imidafenacin according to claim 8, characterized in that: The injection volume of each solution was 1 μL.

10. The method for detecting 2-bromoethanol and ethylene glycol in 1,2-dibromoethane, a starting material of imidafenacin according to claim 8, characterized in that: The separation degree between the 1,2-dibromoethane peak and the adjacent peaks in the test solution is greater than 1.5.

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