Method for analyzing content of 2-glycol ether and unknown impurities in bilastine starting material 2-ethoxychloroethane

Abstract

The invention discloses a method for analyzing the content of 2-glycol ether and unknown impurities in bilastine starting material 2-ethoxychloroethane, and belongs to the technical field of chemical analysis. The method adopts a gas chromatographic method to measure the content of 2-glycol ether and unknown impurities in 2-ethoxychloroethane, and comprises the following steps: firstly, preparing a test solution and a reference solution for later use; and then setting gas chromatography detection conditions, and detecting 2-glycol ether and unknown impurities in 2-ethoxychloroethane by using a capillary column with 6% of hydrogen propyl phenyl-94% of dimethyl polysiloxane as a stationary liquid. The method disclosed by the invention is high in specificity, strong in sensitivity and good in durability; the analysis method provided by the invention is rapid, simple, effective and reliable, and can effectively determine the content of 2-glycol ether and unknown impurities in 2-ethoxychloroethane, thereby ensuring the quality of the bilastine bulk drug.

Description

Technical Field

[0001] The invention belongs to the technical field of chemical analysis, and particularly relates to an analysis method for the content of 2-ethylene glycol ethyl ether and unknown impurities in 2-ethoxyethyl chloride, a starting material of bilastine. Background Art

[0002] Bilastin is a non-sedating, long-acting antihistamine that selectively antagonizes peripheral H1 receptors, has no affinity for muscarinic receptors, and has low affinity for other receptors. It is suitable for the treatment of allergic rhinitis and urticaria, including adults and adolescents aged 12 and above. Developed by the Spanish company FAES FARMA, it was approved for marketing by the European Union in August 2010 and is available in 10 European countries including the United Kingdom, Spain, Germany, Denmark, Bulgaria, and Sweden. In April 2006, the FDA approved it for clinical trials and it is currently in Phase III clinical trials. On June 26, 2023, Menarini, Italy's largest biopharmaceutical group, announced that its second-generation non-sedating, long-acting antihistamine Liminon (generic name: Bilastin Tablets) was officially approved for marketing by the National Medical Products Administration (NMPA).

[0003] According to the synthetic process route provided by the manufacturer, 2-ethoxyethyl chloride is obtained by using 2-ethylene glycol ethyl ether as raw material, undergoing chlorination reaction with thionyl chloride and undergoing a distillation process.

[0004] At present, there are almost no reports on related research on the impurity control of 2-ethoxyethyl chloride in domestic and foreign literature or patents. The present invention aims to provide an analytical method for the unknown impurity content of 2-ethylene glycol ethyl ether in 2-ethoxyethyl chloride, so as to control the impurities and reduce the risk of the impurities being brought into the bilastine raw material drug, thereby ensuring the quality of the bilastine raw material drug. Summary of the invention

[0005] The technical problem solved by the invention is to provide an analysis method for the content of 2-ethylene glycol ethyl ether and unknown impurities in 2-ethoxychloroethane, a starting material of bilastine. The method is rapid, simple, effective, has high specificity, strong sensitivity and good durability.

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

[0007] A method for analyzing the content of 2-ethylene glycol ethyl ether and unknown impurities in 2-ethoxychloroethane, a starting material of bilastine, adopts gas chromatography and comprises the following steps:

[0008] 1) Prepare the test solution and reference solution and set aside;

[0009] 2) Set gas chromatography detection conditions;

[0010] 3) Take the test solution and the reference solution respectively, inject them into the gas chromatograph, and calculate the content of 2-ethylene glycol ethyl ether and unknown impurities in 2-ethoxychloroethane.

[0011] The chromatographic column is a capillary column with 6% hydropropylphenyl-94% dimethylpolysiloxane as the stationary liquid, a length of 30 m, an inner diameter of 0.530 mm, and a filler particle size of 3.0 μm.

[0012] The carrier gas is high-purity nitrogen, the detector temperature is 250°C, the injection port temperature is 230°C, and the detector is a FID detector.

[0013] The split ratio is 4:1.

[0014] The injection volume was 1 μL.

[0015] The flow rate is 2.0 ml / min

[0016] The temperature rise program is as follows: the initial temperature is 60°C, maintained for 5 minutes, then raised to 75°C at a rate of 1°C per minute, maintained for 5 minutes, then raised to 250°C at a rate of 50°C per minute, maintained for 6 minutes.

[0017] The test sample and reference sample were prepared using ethanol as solvent.

[0018] Preparation of the test solution: Accurately weigh 200 mg of 2-ethoxyethyl chloride, place in a 100 mL volumetric flask, add ethanol to dissolve and dilute to the scale, shake well, and obtain.

[0019] Preparation of the reference solution: accurately weigh 100 mg of 2-ethylene glycol ethyl ether reference substance, place it in a 100 mL volumetric flask, add ethanol to dissolve and dilute to the scale, shake well, then accurately measure 1.0 ml, place it in a 10 ml volumetric flask, add ethanol to dilute to the scale, shake well, and obtain.

[0020] Compared with the prior art, the present invention has the following advantages:

[0021] The invention provides an analysis method for the content of 2-ethylene glycol ethyl ether and unknown impurities in 2-ethoxyethyl chloride. The impurity 2-ethylene glycol ethyl ether and the unknown impurities are effectively separated from the main component 2-ethoxyethyl chloride by gas chromatography. The method has high specificity, strong sensitivity and good durability. The analysis method provided by the invention is fast, simple, effective and reliable, and can effectively measure the content of 2-ethylene glycol ethyl ether and the unknown impurities in 2-ethoxyethyl chloride, thereby ensuring the quality of bilastine raw material medicine. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 It is the chromatogram of the positioning solution of 2-ethylene glycol ethyl ether of the present invention;

[0023] Figure 2 It is the chromatogram of the test solution of the present invention;

[0024] Figure 3 It is a chromatogram of the mixed reference substance solution of the present invention;

[0025] Figure 4 is the quantitative limit solution chromatogram of the present invention;

[0026] Figure 5 is the detection limit solution chromatogram of the present invention;

[0027] Figure 6 It is a linear curve diagram of 2-ethylene glycol ethyl ether of the present invention;

[0028] Figure 7 It is the linear curve diagram of 2-ethoxyethyl chloride of the present invention. DETAILED DESCRIPTION

[0029] The present invention is further illustrated below in conjunction with specific examples. The examples are implemented based on the technical solutions of the present invention. It should be understood that these examples are only used to illustrate the present invention and are not used to limit the scope of the present invention.

[0030] The instrument parameters and sample information in the following examples are shown in Tables 1-3.

[0031] Table 1 Instruments and reagents

[0032]

[0033] Table 2 Reference substance information

[0034]

[0035] Table 3 Sample information

[0036]

[0037] According to the synthetic process provided by the manufacturer, 2-ethoxychloroethane is obtained by using 2-ethylene glycol ethyl ether as raw material, reacting with thionyl chloride and undergoing distillation. No special metal elements or other organic solvents are used in the reaction process. The reaction path is as follows:

[0038] .

[0039] Example 1

[0040] A method for analyzing the content of 2-ethylene glycol ethyl ether and unknown impurities in 2-ethoxyethyl chloride, a starting material of bilastine, is determined by gas chromatography and specifically comprises the following steps:

[0041] 1) Prepare test solution and reference solution:

[0042] Preparation of test solution: accurately weigh 200 mg of 2-ethoxyethyl chloride, place in a 100 mL volumetric flask, add ethanol to dissolve and dilute to the scale, shake well, and obtain;

[0043] Preparation of reference solution: Accurately weigh 100 mg of 2-ethylene glycol ether reference substance, place it in a 100 mL volumetric flask, add ethanol to dissolve and dilute to the scale, shake well, then accurately measure 1.0 ml, place it in a 10 ml volumetric flask, add ethanol to dilute to the scale, shake well, and obtain.

[0044] 2) Set the gas chromatography detection conditions: a capillary column with 6% hydropropylphenyl-94% dimethylpolysiloxane as the stationary liquid was used as the chromatographic column (DB-624 gas chromatography column, 30 m×0.530 mm, 3.00 μm); the carrier gas was high-purity nitrogen; the detector temperature was 250 ℃; the injection port temperature was 230 ℃; the detector was an FID detector; the split ratio was 4:1; the injection volume was 1 μL; the solvent was ethanol; the flow rate was 2.0 ml / min; the starting temperature of the column was 60 ℃, maintained for 5 minutes, then increased to 75 ℃ at a rate of 1 ℃ / min, maintained for 5 minutes, and then increased to 250 ℃ at a rate of 50 ℃ / min, and maintained for 6 minutes.

[0045] 3) Accurately pipette 1 μL of the test solution and reference solution respectively, inject them into the gas chromatograph, detect and record the chromatogram.

[0046] Example 2

[0047] The detection method disclosed in Example 1 was verified from the aspects of specificity, limit of quantification and limit of detection, linear range, accuracy, injection precision, repeatability, intermediate precision, solution stability and durability, as follows:

[0048] 1. Exclusivity

[0049] Blank solvent: ethanol;

[0050] Test sample stock solution: accurately weigh 200 mg of 2-ethoxyethyl chloride, place in a 10 mL volumetric flask, add solvent to dissolve and dilute to scale, shake well, and obtain;

[0051] Test solution: Accurately measure 1.0 mL of the test stock solution, place it in a 10 mL volumetric flask, add solvent to dilute to the mark, shake well, and obtain;

[0052] Reference stock solution: accurately weigh 200 mg of 2-ethoxyethyl chloride reference substance, place in a 10 mL volumetric flask, add solvent to dissolve and dilute to scale, shake well, and obtain;

[0053] Reference substance solution: accurately measure 1.0 mL of reference substance stock solution, place it in a 10 mL volumetric flask, add solvent to dilute to the mark, shake well, and obtain;

[0054] 2-Ethylene glycol ether stock solution: accurately weigh 100 mg of 2-Ethylene glycol ether reference substance, place it in a 100 mL volumetric flask, add solvent to dissolve and dilute to the scale, shake well, and obtain;

[0055] 2-Ethylene glycol ether positioning solution: accurately measure 1.0 mL of 2-Ethylene glycol ether stock solution, place it in a 10 mL volumetric flask, add solvent to dilute to the scale, shake well, and obtain;

[0056] Test sample + impurity mixed solution (100% limit concentration): Accurately measure 1.0 mL of 2-ethylene glycol ethyl ether fixation solution and 1.0 mL of test sample stock solution, place in a 10 mL volumetric flask, add solvent to dilute to the scale, shake well, and obtain;

[0057] Mixed reference substance solution (10 times the limit concentration): accurately measure 1.0 mL of 2-ethylene glycol ethyl ether stock solution and 200 mg of 2-ethoxyethyl chloride reference substance, place in a 10 mL volumetric flask, add solvent to dilute to the scale, shake well, and obtain;

[0058] Mixed reference solution: accurately measure 1.0 mL of mixed reference solution (10 times the limit concentration), place it in a 10 mL volumetric flask, add solvent to dilute to the mark, shake well, and obtain;

[0059] Accurately pipette 1 μL of each of the above solutions, inject into the gas chromatograph, and record the chromatogram. Accurately measure 1 μL of blank solvent, test solution, mixed reference solution, reference plus test solution, and positioning solution, and inject into the gas chromatograph, and record the chromatogram. The results are shown in Table 4. Figure 1-3 .

[0060] Table 4 Specificity test results

[0061]

[0062] As shown in Table 4, the blank solvent does not interfere with the determination, the separation degree between the analyte peak and the adjacent peak is greater than 1.5, and the peak positions of the reference and the test are consistent, which meets the verification requirements.

[0063] 2. Limit of quantification and detection limit

[0064] Accurately measure the reference solution and 2-ethylene glycol ethyl ether positioning solution, dilute the solution and inject according to the content detection chromatographic conditions, record the chromatogram, and use the signal-to-noise ratio S / N≈10 as the quantitative limit and the signal-to-noise ratio S / N≈3 as the detection limit. The results are shown in Tables 5 and 6. Figure 4 and Figure 5 .

[0065] Table 5 Results of limit of quantitation and limit of detection tests

[0066]

[0067] Table 6 Precision of quantitation limit

[0068]

[0069] It can be seen from Tables 5 and 6 that the quantitative limit solution was prepared in 6 portions by the same method, each was injected once, and the RSDs of the peak areas of the main peak and impurity peak were both <10.0%.

[0070] 3. Linear range

[0071] Take an appropriate amount of 2-ethoxyethyl chloride and 2-ethylene glycol ethyl ether reference substances, dilute with diluent to prepare a series of linear solutions (take 5 concentration points within the range of 120% of the quantitative limit of 2-ethoxyethyl chloride and 2-ethylene glycol ethyl ether), accurately measure 1 μL of each linear solution, inject it into the gas chromatograph, record the chromatogram, determine the peak area, and perform linear regression with the linear solution concentration as the horizontal axis and the peak area as the vertical axis. The results are shown in Table 7, Table 8, Figure 6 and Figure 7 shown.

[0072] Table 7 2-ethylene glycol ethyl ether linear determination results

[0073]

[0074] From Table 7 and Figure 6 It can be seen that 2-ethylene glycol ethyl ether is linear in the concentration range of 1.040 μg / ml~12.48 μg / ml, and the linear equation is y=1.8688x-0.1089, R 2 It is 0.9994, and the intercept is less than 10% of the y value when the concentration is 100%, which meets the requirements.

[0075] Table 8 Linear determination results of 2-ethoxyethyl chloride

[0076]

[0077] From Table 8 and Figure 7 It can be seen that 2-ethoxychloroethane is linear in the concentration range of 0.9943 μg / ml~2359 μg / ml, and the linear equation is y=1.7207x+7.4857, R 2 It is 0.9999, and the intercept is less than 10% when the y value is 10% of the concentration, which meets the requirements.

[0078] 4. Accuracy test

[0079] Quantitatively add 4 concentrations (10%, 80%, 100%, 120%) of 2-ethylene glycol ethyl ether to the test solution, and the content obtained after deducting the 2-ethylene glycol ethyl ether already in the test solution is the measured value, and the measured value divided by the added value is the recovery rate of each component. The results are shown in Table 9.

[0080] Table 9 Accuracy test results

[0081]

[0082] It can be seen from Table 9 that the recovery rate of 2-ethylene glycol ethyl ether is 104.9%, and the RSD is less than 10%, which meets the requirements and the accuracy of the method is high.

[0083] 5. Injection precision

[0084] The mixed reference solution was injected 6 times in succession, 1 μL each time, into the gas chromatograph, and the chromatogram was recorded. The results are shown in Table 10.

[0085] Table 10 Injection precision test results

[0086]

[0087] As shown in Table 10, the mixed reference solution was injected 6 times continuously, and the retention time and peak area RSD of the main peak and impurities met the requirements, and the system precision was good.

[0088] 6. Repeatability

[0089] Accurately measure 1 μL of the test solution (6 times in parallel), inject it into the gas chromatograph, and record the chromatogram. The results are shown in Table 11.

[0090] Table 11 Repeatability test results

[0091]

[0092] As shown in Table 11, among the 6 samples, the RSD of 2-ethylene glycol ethyl ether and the maximum unknown single impurity content met the requirements and had good repeatability.

[0093] 7. Intermediate precision

[0094] Different personnel used different instruments at different times, referring to the items under repeatability, to prepare blank solvent and test solution, accurately measured 1 μL of each solution, injected it into the gas chromatograph, and recorded the chromatogram. The results are shown in Table 12.

[0095] Table 12 Intermediate precision test results

[0096]

[0097] As shown in Table 12, the statistical repeatability, intermediate precision, RSD of impurity X, maximum unknown single impurity and total impurity content of 12 samples all meet the requirements, and the intermediate precision of this method is good.

[0098] 8. Solution stability

[0099] Take the test solution and the reference solution, place them at room temperature, examine their stability, and record the chromatograms. The results are shown in Table 13.

[0100] Table 13 Solution stability test results

[0101]

[0102] It can be seen from Table 13 that the test solution is stable within 58 h, and the mixed control solution is stable within 35 h.

[0103] 9. Durability

[0104] The effects of injection port temperature ±5 °C, detector temperature ±5 °C, column starting temperature ±5 °C, carrier gas flow rate ±0.1 ml / min, split ratio ±1, and different chromatographic columns of the same model and manufacturer on the system were investigated. The results are shown in Table 14.

[0105] Table 14 Durability test results

[0106]

[0107] It can be seen from Table 14 that the injection temperature is ±5 ℃; the detector temperature is ±5 ℃; the column starting temperature is ±5 ℃; the carrier gas flow rate is ±0.1 ml / min; the split ratio is ±1; for different chromatographic columns of the same model and manufacturer, except for slight changes in retention time, there is no difference in impurity content above the reporting limit, and the durability is good.

[0108] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A method for analyzing the content of 2-ethylene glycol ethyl ether and unknown impurities in 2-ethoxyethyl chloride, a starting material of bilastine, characterized in that: Using gas chromatography, the following steps are included: 1) Prepare the test solution and reference solution and set aside; 2) Set gas chromatography detection conditions; 3) Take the test solution and the reference solution respectively, inject them into the gas chromatograph, record the chromatogram, and calculate the content of 2-ethylene glycol ethyl ether and unknown impurities in 2-ethoxychloroethane.

2. The analytical method for the content of 2-ethylene glycol ethyl ether and unknown impurities in the bilastine starting material 2-ethoxyethyl chloride according to claim 1, characterized in that: The chromatographic column is a capillary column with 6% hydropropylphenyl-94% dimethylpolysiloxane as the stationary liquid, a length of 30 m, an inner diameter of 0.530 mm, and a filler particle size of 3.0 μm.

3. The analytical method of 2-ethylene glycol ethyl ether and unknown impurity content in the bilastine starting material 2-ethoxyethyl chloride according to claim 1, characterized in that: The carrier gas is high-purity nitrogen, the detector temperature is 250°C, the injection port temperature is 230°C, and the detector is a FID detector.

4. The analytical method for the content of 2-ethylene glycol ethyl ether and unknown impurities in the bilastine starting material 2-ethoxyethyl chloride according to claim 1, characterized in that: The split ratio is 4:

1.

5. The analytical method for the content of 2-ethylene glycol ethyl ether and unknown impurities in the bilastine starting material 2-ethoxyethyl chloride according to claim 1, characterized in that: The injection volume was 1 μL.

6. The method for analyzing the content of 2-ethylene glycol ethyl ether and unknown impurities in the bilastine starting material 2-ethoxyethyl chloride according to claim 1, characterized in that: The flow rate was 2.0 ml / min.

7. The method for analyzing the content of 2-ethylene glycol ethyl ether and unknown impurities in the bilastine starting material 2-ethoxyethyl chloride according to claim 1, characterized in that: The temperature rising program is as follows: the starting temperature is 60°C, maintained for 5 minutes, then raised to 75°C at a rate of 1°C per minute, maintained for 5 minutes, then raised to 250°C at a rate of 50°C per minute, maintained for 6 minutes.

8. The method for analyzing the content of 2-ethylene glycol ethyl ether and unknown impurities in the bilastine starting material 2-ethoxyethyl chloride according to claim 1, characterized in that: The test sample and reference sample were prepared using ethanol as solvent.

9. The method for analyzing the content of 2-ethylene glycol ethyl ether and unknown impurities in the bilastine starting material 2-ethoxyethyl chloride according to claim 1, characterized in that: Preparation of the test solution: Accurately weigh 200 mg of 2-ethoxyethyl chloride, place in a 100 mL volumetric flask, add ethanol to dissolve and dilute to the scale, shake well, and obtain.

10. The method for analyzing the content of 2-ethylene glycol ethyl ether and unknown impurities in 2-ethoxyethyl chloride, the starting material of bilastine according to claim 1, characterized in that: Preparation of the reference solution: accurately weigh 100 mg of 2-ethylene glycol ethyl ether reference substance, place it in a 100 mL volumetric flask, add ethanol to dissolve and dilute to the scale, shake well, then accurately measure 1.0 ml, place it in a 10 ml volumetric flask, add ethanol to dilute to the scale, shake well, and obtain.

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