CN117990817A - Method for measuring impurity content in pharmaceutical intermediate acyl chloride iminodibenzyl by utilizing HPLC - Google Patents
Method for measuring impurity content in pharmaceutical intermediate acyl chloride iminodibenzyl by utilizing HPLC Download PDFInfo
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- 239000012535 impurity Substances 0.000 title claims abstract description 56
- 238000000034 method Methods 0.000 title claims abstract description 36
- 238000004128 high performance liquid chromatography Methods 0.000 title claims abstract description 15
- 239000012450 pharmaceutical intermediate Substances 0.000 title claims abstract description 7
- 239000012488 sample solution Substances 0.000 claims abstract description 31
- 238000001514 detection method Methods 0.000 claims abstract description 16
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 claims abstract description 12
- 239000007788 liquid Substances 0.000 claims abstract description 12
- 239000000543 intermediate Substances 0.000 claims abstract description 10
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 8
- 238000010828 elution Methods 0.000 claims abstract description 7
- 239000003814 drug Substances 0.000 claims abstract description 6
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- 238000004811 liquid chromatography Methods 0.000 claims abstract description 6
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims abstract description 4
- 239000000377 silicon dioxide Substances 0.000 claims abstract description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 87
- 239000000243 solution Substances 0.000 claims description 68
- ZSMRRZONCYIFNB-UHFFFAOYSA-N 6,11-dihydro-5h-benzo[b][1]benzazepine Chemical group C1CC2=CC=CC=C2NC2=CC=CC=C12 ZSMRRZONCYIFNB-UHFFFAOYSA-N 0.000 claims description 42
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 26
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 21
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 13
- 235000019253 formic acid Nutrition 0.000 claims description 13
- -1 iminodibenzyl acid chloride Chemical compound 0.000 claims description 11
- 239000000523 sample Substances 0.000 claims description 11
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- 239000003085 diluting agent Substances 0.000 claims description 8
- 238000002360 preparation method Methods 0.000 claims description 7
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- 238000012417 linear regression Methods 0.000 claims description 4
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- FFGPTBGBLSHEPO-UHFFFAOYSA-N carbamazepine Chemical compound C1=CC2=CC=CC=C2N(C(=O)N)C2=CC=CC=C21 FFGPTBGBLSHEPO-UHFFFAOYSA-N 0.000 description 5
- 229960000623 carbamazepine Drugs 0.000 description 5
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Abstract
The invention discloses a method for determining the impurity content in an intermediate of a pharmaceutical acid chloride iminodibenzyl by utilizing HPLC, and belongs to the technical field of medicine analysis. The invention comprises the following steps: 1) Preparing a sample solution, a control solution and a system applicability solution; 2) Setting liquid chromatography detection conditions: gradient elution is carried out by adopting a chromatographic column taking octadecylsilane chemically bonded silica as a filler; 3) Precisely sucking the sample solution, the control solution and the system applicability solution respectively, injecting into a liquid chromatograph, and starting to detect and record a chromatogram; 4) And drawing a standard curve to obtain a standard curve equation, and calculating the impurity content in the pharmaceutical intermediate, namely the imide dibenzyl. The method has good specificity, has no influence on the impurity separation and inspection results of the related substances of the imidodibenzyl chloride when the chromatographic condition parameters change slightly, and has good durability.
Description
Technical Field
The invention belongs to the technical field of medicine analysis, and particularly relates to a method for measuring impurity content in a pharmaceutical intermediate acyl chloride iminodibenzyl by utilizing HPLC.
Background
Iminodibenzyl formyl chloride is a key intermediate for synthesizing carbamazepine, and the carbamazepine is mainly used for treating epilepsy, is also commonly used for treating diseases such as peripheral neuralgia, manic depression, arrhythmia and the like, and is generally used as a first-choice medicament for treating epilepsy in clinic.
According to the preparation process of the iminodibenzyl of the acyl chloride, iminodibenzyl is taken as a key raw material, the iminodibenzyl is taken as a carbamazepine specific impurity, and residues of iminodibenzyl participate in subsequent reactions to form process impurities, so that the quality of the carbamazepine is influenced, and the quality of the carbamazepine is strictly controlled (less than 0.15%). Therefore, the method for determining the impurity content in the intermediate acyl chloride iminodibenzyl has important significance for the generation and storage of raw materials.
Disclosure of Invention
The invention aims at solving the problems in the prior art, and provides a method for measuring the impurity content in the pharmaceutical intermediate iminodibenzyl by utilizing HPLC, which can rapidly, effectively, accurately and reliably separate and detect the impurity iminodibenzyl in the iminodibenzyl by using the liquid chromatography, thereby being beneficial to improving the product quality of the iminodibenzyl.
In order to solve the technical problems, the technical scheme adopted by the invention is as follows:
A method for determining the impurity content in an intermediate of medicine, namely, imide dibenzyl by utilizing HPLC, which comprises the following steps:
1) Preparing a sample solution, a control solution and a system applicability solution;
2) Setting liquid chromatography detection conditions: adopting a chromatographic column taking octadecylsilane chemically bonded silica as a filler, wherein a mobile phase consists of a phase A and a phase B, wherein the phase A is a mixed solution of triethylamine, formic acid and water, and the phase B is a mixed solution of methanol and formic acid, and performing gradient elution;
3) Precisely sucking the sample solution, the control solution and the system applicability solution respectively, injecting into a liquid chromatograph, and starting to detect and record a chromatogram;
4) And drawing a standard curve to obtain a standard curve equation, and calculating the impurity content in the pharmaceutical intermediate, namely the imide dibenzyl.
Preferably, in the step 1), the control solution is prepared by the following steps: precisely measuring a sample solution, adding a diluent to dilute the sample solution into a solution containing 0.2ug of imidodibenzyl chloride per mL of the sample solution, and taking the diluted solution as a control solution; the preparation method of the system applicability solution comprises the following steps: the iminodibenzyl acid chloride sample and the iminodibenzyl reference substance were weighed, and dissolved and diluted by a diluent with ultrasound to a solution containing 0.3 μg iminodibenzyl and 0.2mg iminodibenzyl acid chloride per mL, as a system applicability solution.
Preferably, the diluent is methanol.
Preferably, in the step 2), the specification of the chromatographic column is 250mm×4.6mm, the particle size of the packing is 5 μm, and the column temperature is 35 ℃.
Preferably, in the step 2), the volume ratio of triethylamine, formic acid and water in the A phase is 0.5:0.5:1000, and the volume ratio of methanol and formic acid in the B phase is 1000:0.25.
Preferably, in the step 2), the volume ratio of the A phase to the B phase is 30:70, and the gradient elution flow rate is 1mL/min.
Preferably, in the step 3), the detection wavelength of the liquid chromatograph is 206nm.
Preferably, in the step 4), a linear regression equation is performed based on the chromatogram with the peak area a as the ordinate and the concentration C as the abscissa, the linear equation of iminodibenzyl is a=101.83c+0.2744, and the linear equation of acid chloride iminodibenzyl is a= 83.014C-0.4609.
Preferably, the linear concentration range of iminodibenzyl is 0.0601-0.6013 mug/mL, the quantitative limit is 0.0601 mug/mL, and the detection limit is 0.0150 mug/mL; the linear concentration range of the imidodibenzyl chloride is 0.0603-0.6033 mug/mL, the quantitative limit is 0.0608 mug/mL, and the detection limit is 0.0152 mug/mL.
Compared with the prior art, the invention has the beneficial effects that:
1) The method has good specificity, does not influence the impurity separation and inspection results of the related substances of the imidodibenzyl chloride when the chromatographic condition parameters change slightly, and has good durability;
2) The invention can rapidly, effectively, accurately and reliably separate and detect related substances in the intermediate acyl chloride iminodibenzyl by using the liquid chromatography, and is beneficial to improving the product quality of the intermediate acyl chloride iminodibenzyl.
Drawings
FIG. 1 is a diagram of the intermediate acid chloride iminodibenzyl-specific profile; wherein 2a is a blank solvent spectrogram; 2b is iminodibenzyl localization spectrogram; 2c is a system applicability spectrum; 2d is a spectrogram of the solution of the test sample;
FIG. 2 is a graph of iminodibenzyl standard;
FIG. 3 is a graph of the standard curve of the iminodibenzyl acid chloride.
Detailed Description
The present invention will be further described with reference to specific embodiments for the purpose of making the objects, technical solutions and advantages of the present invention more apparent. Unless otherwise indicated, all technical means used in the following examples are conventional means well known to those skilled in the art.
In the following examples, the instruments, reagents, controls and sample information used are as follows:
The model of the high performance liquid chromatograph (instrument 1) is Agilent1260 series (VWD);
the model of the high performance liquid chromatograph (instrument 2) is Agilent1260 series (DAD);
Methanol is a commercial product of chromatographic grade; triethylamine and formic acid are commercial products of analytical grade;
iminodibenzyl in the control is a commercial product with 98.49 percent and is purchased from QCC-USA;
The control and sample were commercially available products having an imidodibenzyl content of 99.82% and were purchased from the pharmaceutical company, inc. of lake Nanhua.
Example 1
A method for determining the impurity content in an intermediate of medicine, namely, imide dibenzyl by utilizing HPLC, which comprises the following steps:
1) Preparing a test solution, a control solution and a system applicability solution:
The preparation method of the sample solution comprises the following steps: precisely weighing 20mg of iminodibenzyl chloride, placing into a 100mL measuring flask, adding methanol for dissolution, and diluting to scale to obtain a sample solution;
The preparation method of the control solution comprises the following steps: precisely measuring a sample solution, adding a methanol diluent to dilute the sample solution into a solution containing 0.2ug of imidodibenzyl chloride per mL of the solution, and taking the solution as a control solution;
The preparation method of the system applicability solution comprises the following steps: weighing an imidodibenzyl sample and an imidodibenzyl reference substance, and ultrasonically dissolving and diluting the imidodibenzyl sample and the imidodibenzyl reference substance by using a methanol diluent to obtain a solution containing 0.3 mug of imidodibenzyl and 0.2mg of imidodibenzyl per ml, wherein the solution is used as a system applicability solution;
2) Setting liquid chromatography detection conditions: adopting a chromatographic column with octadecylsilane chemically bonded silica as filler, wherein the specification of the chromatographic column is 250mm multiplied by 4.6mm, the particle size of the filler is 5 mu m, and the column temperature is 35 ℃; the mobile phase consists of a phase A and a phase B, wherein the phase A is a mixed solution of triethylamine, formic acid and water, and 0.5mL of triethylamine and 0.5mL of formic acid are added into 1000mL of water to prepare the phase A; the phase B is a mixed solution of methanol and formic acid, and 0.25mL of formic acid is added into 1000mL of methanol to prepare the phase B; the solvent is methanol, gradient elution is carried out, the volume ratio of the phase A to the phase B is 30:70, and the gradient elution flow rate is 1mL/min;
3) Precisely sucking the sample solution, the control solution and the system applicability solution respectively, injecting into a liquid chromatograph, detecting and recording the chromatograms, wherein the detection wavelength of the liquid chromatograph is 206 nm;
4) According to a chromatogram, a linear regression equation is carried out by taking a peak area A as an ordinate and a concentration C as an abscissa, wherein the linear equation of iminodibenzyl is A=101.83C+0.2744, the linear equation of iminodibenzyl is A= 83.014C-0.4609, the linear concentration range of iminodibenzyl is 0.0601-0.6012 mug/mL, the quantitative limit is 0.0601 mug/mL, and the detection limit is 0.0150 mug/mL; the linear concentration range of the imidodibenzyl chloride is 0.0603-0.6033 mug/mL, the quantitative limit is 0.0603 mug/mL, and the detection limit is 0.0152 mug/mL.
Example 2
1. Specialization of
Precisely weighing 15mg of iminodibenzyl reference substance, placing into a 100mL measuring flask, adding methanol for dissolution and dilution to a scale, precisely weighing a 4 mL-20 mL measuring flask, adding methanol for dilution to a scale, and taking the methanol as an impurity positioning solution;
Precisely weighing 20mg of iminodibenzyl chloride, placing into a 100mL measuring flask, precisely adding 1mL of iminodibenzyl positioning solution, adding methanol for dissolution, and diluting to scale to obtain a system adaptive solution;
The iminodibenzyl chloride 20mg is precisely weighed, placed in a 100mL measuring flask, dissolved by adding methanol and diluted to a scale, and used as a test solution.
Precisely measuring 10 mu L of each solution, respectively injecting into a liquid chromatograph, and recording a chromatogram. The peak time and separation of each component were examined.
TABLE 5 impurity localization and separation degree
From the test results of table 5 and fig. 2, it is shown that the blank solvent does not interfere with the sample related substance determination; the separation degree between the main component and the adjacent impurities and between the iminodibenzyl and the adjacent impurities in the system applicability solution chromatogram is more than 1.5; the separation degree of the main component and the adjacent impurities in the sample solution is more than 1.5; the method has good specificity.
2. Quantitative limit and detection limit
The iminodibenzyl reference substance is taken as a proper amount, dissolved and diluted by adding methanol, and a solution containing 0.06 mug of each component per 1mL is prepared as a quantitative limit solution (about 0.03 percent of the concentration of the solution of the test sample), and a solution which is diluted by 3 times on the basis of the quantitative limit is prepared as a detection limit solution. The concentration and signal to noise ratio of each compound are shown in tables 6 and 7.
TABLE 6 detection limit test results
TABLE 7 quantitative limit test results
As can be seen from tables 6 and 7, iminodibenzyl and acid chloride iminodibenzyl correspond to 0.030% of the sample solution; the RSD (n=6) of the peak area is less than 10.0%, and the method can accurately quantify.
3. Linearity and range
Precisely weighing 15mg of each of iminodibenzyl and iminodibenzyl, dissolving with methanol, and diluting to obtain a series of linear solutions (7 concentration points are taken from the range of 0.03% to 0.3% of the corresponding sample concentration); the linear regression equation and correlation coefficient of each compound are calculated by drawing each component standard curve with the peak area A as ordinate and the corresponding concentration C as abscissa, and the results are shown in tables 8 and 9.
TABLE 8 results of the imide dibenzyl Linear test (instrument 1)
TABLE 9 iminodibenzyl Linear test results (instrument 1)
As can be seen from tables 8 and 9, the concentration and peak area of the imidodibenzyl chloride were in the range of 0.0603ug/mL to 0.6033ug/mL, and the concentration and peak area were excellent in linearity, and r=0.9997; iminodibenzyl is in the range of 0.0601ug/mL to 0.6012ug/mL, the concentration and the peak area are in good linearity, and r=0.9994.
4. Correction factor
Different experimenters test on different instruments to examine the response difference between the impurity and the main component, calculate the correction factor of the impurity by using a standard curve (the ratio of the slope of the main component line to the slope of the impurity line is the correction factor of the corresponding impurity), and the results are shown in tables 10-11.
TABLE 10 Acryliminodibenzyl Linear test results (instrument 2)
TABLE 11 iminodibenzyl Linear test results (instrument 2)
Table 12 correction factor results
5. Accuracy of
The iminodibenzyl reference substance is taken to be proper, and is added into the iminodibenzyl chloride according to the proportion of 50-150 percent of the impurity limit (0.15 percent), and the recovery rate of the iminodibenzyl is calculated according to the ratio of the measured quantity to the added quantity by an external standard method and a main component comparison method added with a correction factor respectively. The preparation process of the solution is as follows:
Precisely weighing 15mg of iminodibenzyl, placing into a 100mL measuring flask, adding methanol for dissolution and dilution to a scale, precisely weighing 5mL, placing into a 50mL measuring flask, adding methanol for dilution to a scale, and preparing a solution containing 15 mug per 1mL as a reference stock solution; precisely measuring 2mL of reference stock solution, placing in a 100mL measuring flask, and adding methanol to dilute to scale to obtain reference solution.
About 20mg of imidodibenzyl chloride is taken, precisely weighed, placed in a 100mL measuring flask, dissolved with methanol and diluted to a scale, used as an unlabeled test solution, and a corresponding control solution is prepared.
About 20mg of imidodibenzyl chloride is taken, precisely weighed, placed in a 100mL measuring flask, respectively and precisely added with 1mL, 2mL and 3mL of reference stock solution, dissolved and diluted to the scale by methanol to respectively serve as 50%, 100% and 150% horizontal recovery rate solutions, and the reference solutions are prepared, and 3 parts of reference solutions are prepared in parallel for each level;
precisely measuring 10 mu L of each solution, respectively injecting into a liquid chromatograph, and recording a chromatogram. The recovery results are shown in tables 13 and 14.
TABLE 13 recovery test of iminodibenzyl (correction factor method)
TABLE 14 recovery test of iminodibenzyl (external standard method)
As is clear from tables 13 and 14, the recovery rate was calculated by the correction factor method and by the external standard method, and the recovery rate of iminodibenzyl was in the range of 90.0% to 110.0% at three concentration levels, namely, high (limit of 150%), medium (limit of 100%) and low (limit of 50%), and RSD (n=9) was less than 5.0%, and the method was satisfactory in accuracy.
6. Precision of
Taking a proper amount of iminodibenzyl reference substance, adding the iminodibenzyl reference substance into an iminodibenzyl raw material of acyl chloride according to the proportion of 100 percent of the limit (0.15 percent) of impurities, checking the content change of the impurities, and checking the repeatability of the method; 6 parts of sample solution and control solution with the same concentration are prepared by the same method at different times and different testers and different instruments, the change condition of the impurity content is inspected, the intermediate precision of the method is inspected, the change of the impurity content is measured by two testers, and the precision of the method is inspected, wherein the result is shown in table 15.
TABLE 15 precision test results
As can be seen from table 15, the measurement was repeated 6 times, the RSD (n=6) of each impurity and the total impurity content was less than 20%, and the number of impurities (not less than 0.03%) was the same; the RSD (n=12) of the total impurity content of the 12 groups of single impurity contents measured by two laboratory workers is also less than 20%, which indicates that the method has better precision.
7. Solution stability
About 20mg of imidodibenzyl chloride is taken, precisely weighed, placed in a 100mL measuring flask, dissolved by adding methanol and diluted to a scale, and taken as a sample solution.
Precisely measuring 1mL of the sample solution, placing a 10mL measuring flask, adding methanol for dilution to a scale, precisely measuring 1mL, placing a 100mL measuring flask, adding methanol for dilution to a scale, and taking as a control solution
Taking a sample solution and a control solution at different time points, respectively precisely measuring 10 mu L, injecting the sample solution and the control solution into a liquid chromatograph, and examining the solution stability in different time by the control solution according to the change of peak area, wherein the sample solution examines the solution stability in different time according to the impurity content and the impurity number. The results are shown in tables 16 and 17.
TABLE 16 stability results of test solutions
| Name of the name | 0h | 160min | 320min | 8h |
| Unknown impurity (rrt=0.74) | Not detected | 0.02％ | 0.03％ | 0.05％ |
| Unknown impurity (rrt=1.6) | 0.03％ | 0.03％ | 0.03％ | 0.03％ |
| Total impurities | 0.09％ | 0.13％ | 0.13％ | 0.14％ |
| More than 0.03% of impurity number | 1 | 1 | 2 | 2 |
TABLE 17 stability test peak surface results for control solutions
| Name of the name | 0h | 2.5min | 5.3min | 8h | RSD |
| Peak area of control solution | 14.47 | 13.97 | 14.57 | 14.97 | 2.8％ |
As can be seen from tables 16 and 17, the sample solution was left at room temperature for 2.5min to generate new impurities, and the impurities were gradually increased with the prolonged leaving time, so that the sample solution was ready to be prepared; the control solution was relatively stable for 8 hours at room temperature.
8. Durability of
The method comprises the steps of preparing a sample solution, a control solution and a system applicability solution according to the method, slightly changing chromatographic conditions (flow rate, wavelength, column temperature and mobile phase proportion of chromatographic columns of different brands), precisely measuring 10 mu L, injecting the sample solution into a chromatograph, recording the chromatograph, detecting the number and content of impurities in the sample solution, changing the separation degree of main peaks and impurities in the system applicability solution, and examining the durability of the method. The results are shown in tables 18 and 19.
Column 2: welch Ultimate XS-C18 4.6X250 mm,5 μm
Column 3: YMC-Pack ODS-A4.6.250 mm,5 μm
TABLE 18 results of test of substances related to the test sample under different conditions
Table 19 results of System suitability test under different conditions
As can be seen from tables 18 and 19, there is no obvious difference in the impurity content and the number of detected impurities in the sample solutions when the chromatographic conditions are slightly changed; the separation degree between main peaks and impurities in the system applicability solution chromatogram meets the requirements, and the method has good durability.
The above examples are preferred embodiments of the present application, but the embodiments of the present application are not limited to the examples, and the examples and features of the examples may be arbitrarily combined with each other without collision. Any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principles of the application are intended to be equivalent substitutes for those that do not depart from the spirit and principles of the application. In addition, technical details not described in detail in the present specification belong to the known technology of those skilled in the art, so the present specification is not repeated.
Claims (9)
1. A method for determining the impurity content in an intermediate of medicine, namely, imide dibenzyl by utilizing HPLC, which is characterized by comprising the following steps:
1) Preparing a sample solution, a control solution and a system applicability solution;
2) Setting liquid chromatography detection conditions: adopting a chromatographic column taking octadecylsilane chemically bonded silica as a filler, wherein a mobile phase consists of a phase A and a phase B, wherein the phase A is a mixed solution of triethylamine, formic acid and water, the phase B is a mixed solution of methanol and formic acid, and a solvent is methanol for gradient elution;
3) Precisely sucking the sample solution, the control solution and the system applicability solution respectively, injecting into a liquid chromatograph, and starting to detect and record a chromatogram;
4) And drawing a standard curve to obtain a standard curve equation, and calculating the impurity content in the pharmaceutical intermediate, namely the imide dibenzyl.
2. The method for determining the impurity content in pharmaceutical acid chloride dibenzyl by using HPLC according to claim 1, wherein in the step 1), the control solution is prepared by the following method: precisely measuring a sample solution, adding a diluent to dilute the sample solution into a solution containing 0.2ug of imidodibenzyl chloride per mL of the sample solution, and taking the diluted solution as a control solution; the preparation method of the system applicability solution comprises the following steps: the iminodibenzyl acid chloride sample and the iminodibenzyl reference substance were weighed, and dissolved and diluted by a diluent with ultrasound to a solution containing 0.3 μg iminodibenzyl and 0.2mg iminodibenzyl acid chloride per mL, as a system applicability solution.
3. The method for determining the impurity content of pharmaceutical acid chloride dibenzyl by HPLC according to claim 2, wherein said diluent is methanol.
4. The method for determining the impurity content of pharmaceutical acid chloride dibenzyl by HPLC according to claim 1, wherein in said step 2), the specification of the chromatographic column is 250mm×4.6mm, the filler particle size is 5 μm, and the column temperature is 35 ℃.
5. The method for determining the impurity content of pharmaceutical acid chloride dibenzyl by HPLC according to claim 1, wherein in the step 2), the volume ratio of triethylamine, formic acid and water in the a phase is 0.5:0.5:1000, and the volume ratio of methanol and formic acid in the b phase is 1000:0.25.
6. The method for determining the impurity content in pharmaceutical acid chloride dibenzyl by utilizing HPLC according to claim 1, wherein in the step 2), the volume ratio of the A phase to the B phase is 30:70, and the gradient elution flow rate is 1mL/min.
7. The method for determining the impurity content of pharmaceutical acid chloride dibenzyl by HPLC according to claim 1, wherein in the step 3), the detection wavelength of the liquid chromatograph is 206nm.
8. The method according to claim 1, wherein in the step 4), a linear regression equation is performed based on the chromatogram with a peak area a as an ordinate and a concentration C as an abscissa, the linear equation of iminodibenzyl is a=101.83c+0.2744, and the linear equation of iminodibenzyl is a= 83.014C-0.4609.
9. The method for determining the impurity content in pharmaceutical acid chloride dibenzyl by using HPLC according to claim 9, wherein the linear concentration range of iminodibenzyl is 0.0601-0.6013 μg/mL, the quantitative limit is 0.0601 μg/mL, and the detection limit is 0.0150 μg/mL; the linear concentration range of the imidodibenzyl chloride is 0.0603-0.6033 mug/mL, the quantitative limit is 0.0608 mug/mL, and the detection limit is 0.0152 mug/mL.
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| CN202311795770.5A CN117990817A (en) | 2023-12-25 | 2023-12-25 | Method for measuring impurity content in pharmaceutical intermediate acyl chloride iminodibenzyl by utilizing HPLC |
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