A method for determining the purity of ceftaroline fosamil side chain acyl chloride

Abstract

The present invention provides a method for determining the purity of the side chain of ceftaroline fosamil, which is characterized by: the side chain of ceftaroline fosamil is amidated by an amine compound, and then the purity is determined by an instrumental analysis method. The method is simple and easy to implement, the derivatization is complete, and the detection is accurate.

Description

Technical Field

[0001] The present invention relates to the field of chemical analysis, and in particular to a method for determining the purity of ceftaroline fosamil side chain acyl chloride. Background Art

[0002] Cefuroxime is a fifth-generation cephalosporin drug with good antibacterial activity against Gram-positive bacteria including methicillin-resistant Staphylococcus aureus infection, multidrug-resistant Streptococcus pneumoniae infection and common Gram-negative bacteria infection. At present, Cefuroxime for Injection was approved by the US FDA on October 29, 2010 and is produced by AstraZeneca. There are no products that have been applied for or marketed in China. Today, when bacterial resistance is a global problem, Cefuroxime, as a new drug against multidrug-resistant bacteria, has broad application prospects.

[0003] Ceftaroline fosamil side chain acyl chloride is an important starting material in its synthesis process. Due to its active nature, it is easily hydrolyzed when it comes into contact with water. When its content is directly measured by reverse phase chromatography, the acyl chloride group is partially hydrolyzed, and the result is low, which cannot accurately reflect the quality of the starting material. Patent publication number CN107621512A uses methanol as a derivatization reagent, but it is unable to completely derivatize the acyl chloride side chain, and there is a small amount of hydrolysis product, which affects the detection accuracy.

[0004] The reaction mechanism of hydrolysis of the acyl chloride of the side chain of cefuroxime is as follows:

[0005] Summary of the invention

[0006] The invention aims to overcome the shortcomings of direct determination of the side chain acyl chloride of the starting material of cefuroxime and methanol as a derivatization reagent, and provides a method that can react quickly and completely and accurately quantitatively analyze the purity of the side chain of cefuroxime.

[0007] The present invention provides a method for determining the purity of the side chain of cefuroxime, characterized in that:

[0008] After the side chain of ceftaroline fosamil was amidated with an amine compound, its purity was determined by instrumental analysis.

[0009] The side chain acyl chloride of the ceftaroline fosamil starting material of the ceftaroline fosamil side chain, that is,

[0010] Furthermore, the present invention provides a method for determining the purity of the side chain of ceftaroline fosamil, which is also characterized in that the above-mentioned amine compound is selected from aromatic amines, such as aniline, naphthylamine, p-methylaniline, etc.

[0011] Furthermore, the present invention provides a method for determining the purity of the side chain of ceftaroline fosamil, which is also characterized in that: the above-mentioned instrumental analysis method is selected from high performance liquid chromatography.

[0012] Furthermore, the present invention provides a method for determining the purity of the side chain of ceftaroline fosamil, which is also characterized in that: the amidation process is as follows:

[0013] After quantitatively taking the side chain of ceftaroline fosamil, dilute it with an amine compound to the target volume, mix it evenly, keep it in a water bath at 20-40° C. for not less than 30 minutes, filter it, wash it with a solvent, and dry it to obtain an amide product.

[0014] The specific reaction equation is as follows:

[0015]

[0016] Furthermore, the present invention provides a method for determining the purity of a ceftaroline fosamil side chain, which is also characterized in that the ratio of the ceftaroline fosamil side chain to the amine compound is 0.5-2 g ceftaroline fosamil side chain / 10 ml amine compound.

[0017] Furthermore, the present invention provides a method for determining the purity of the side chain of ceftaroline fosamil, which is also characterized in that the temperature of the water bath is 30°C.

[0018] Furthermore, the present invention provides a method for determining the purity of the side chain of ceftaroline fosamil, which is also characterized in that: the specific parameters of the above-mentioned high performance liquid chromatography method are:

[0019] The mobile phase is selected from a mixture of dipotassium hydrogen phosphate and acetonitrile;

[0020] The mobile phase is 0.05-0.2 mol / L;

[0021] Column temperature 25-35°C;

[0022] Flow rate 0.5-2ml per minute;

[0023] Detection wavelength: 254nm.

[0024] Furthermore, the present invention provides a method for determining the purity of the side chain of ceftaroline fosamil, which is also characterized in that the amount of the analyte is 80-150 mg / 50 ml of mobile phase.

[0025] Furthermore, the present invention provides a method for determining the purity of the side chain of ceftaroline fosamil, which is also characterized in that the volume ratio of the above-mentioned dipotassium hydrogen phosphate to acetonitrile is 1-4:1.

[0026] Furthermore, the present invention provides a method for determining the purity of the side chain of ceftaroline fosamil, which is also characterized in that the concentration of the above-mentioned dipotassium hydrogen phosphate is 0.08-0.12 mol / L.

[0027] Functions and effects of the present invention:

[0028] The present invention adopts a pre-column derivatization method to measure the purity of the side chain acyl chloride of ceftaroline fosamil, thereby avoiding the hydrolysis of the side chain acyl chloride when directly measuring it. The technical solution of the present invention is completely derivatized, and the purity of the side chain acyl chloride of the ceftaroline fosamil starting material can be accurately measured, providing high-quality basic data for the preparation of ceftaroline fosamil. The derivatization reagent is a common solvent, the method is simple and easy, the derivatization is complete, and the detection is accurate. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 , high performance liquid chromatogram of Example 1;

[0030] Figure 2 , High performance liquid chromatogram of reference example 1;

[0031] Figure 3 High performance liquid chromatogram of comparative example 2. DETAILED DESCRIPTION

[0032] The present embodiment relates to a method for determining the purity of the side chain acyl chloride of a ceftaroline fosamil starting material, comprising the following steps:

[0033] S1. Take about 0.5-2 g of the side chain acyl chloride of the starting material of ceftaroline fosamil, add it to a 10 ml volumetric flask, dilute it to the scale with aniline, shake it well and keep it in a water bath at 25-40°C for 20-60 minutes, filter it, and wash the filter cake with isopropyl ether and dry it.

[0034] S2. Accurately measure 80-150 mg of the solid prepared in the first step and add it to a 50 ml volumetric flask, dilute it to the mark with the mobile phase, and use it as the test solution;

[0035] Mobile phase: a mixture of 0.08-0.12 mol / L potassium hydrogen phosphate dihydrate and acetonitrile in a volume ratio of 1:0.8-1.2.

[0036] The third step is to use a chromatographic column with a mobile phase of 0.08-0.12 mol / L potassium dihydrogen phosphate solution-acetonitrile (volume ratio 1-4:1, 3:1 is optimal), a column temperature of 25-35°C, a flow rate of 0.5-2 ml per minute, a detection wavelength of 254 nm, and high performance liquid chromatography to determine its purity.

[0037] Based on the above S1, in this example, the effects of water bath temperature and reaction time on the conversion rate of the derivatization reaction were investigated.

[0038] Investigation experiment 1

[0039] Under the derivatization condition of 30 minutes of water bath time, the effects of 25℃, 30℃, and 40℃ water bath temperatures on the reaction were investigated, indicating that the peak area had little change at 30℃ water bath temperature;

[0040] Investigation experiment 2

[0041] Under the derivatization condition of a water bath temperature of 30°C, the effects of reaction times of 20 minutes, 30 minutes, 40 minutes, 50 minutes and 60 minutes on the reaction conversion rate were investigated, indicating that the reaction was complete in 30 minutes.

[0042] Based on the above investigation experiments 1 and 2, the optimal conditions for S1 are: a water bath temperature of 30°C and a water bath time of 30 minutes.

[0043] Based on the test results under the above conditions, a specific comparative test was carried out using the following preferred embodiment 1.

[0044] Example 1.

[0045] Take about 1g of the side chain acyl chloride of ceftaroline fosamil starting material with batch number 20210205, add it to a 10ml volumetric flask, dilute it to the mark with aniline, keep it in a 30℃ water bath for 30 minutes, filter, wash the filter cake with isopropyl ether and dry it, take 100mg and add it to a 50ml volumetric flask, dilute it to the mark with the mobile phase as the test solution. Use Waters SymmetryShel dRP18 (250mm×4.6mm, 5μm) chromatographic column, the mobile phase is 0.1mol / L potassium dihydrogen phosphate solution-acetonitrile (volume ratio 3:1), the column temperature is 30℃, the flow rate is 1.0ml per minute, and the detection wavelength is 254nm;

[0046] Under this chromatographic condition, accurately measure 20 μL of the test solution, inject it into the liquid chromatograph, and record the chromatogram (such as Figure 1 The purity was calculated by peak area normalization method.

[0047] also, Figure 1 The peak at 2.7 minutes is the hydrolysis product of the acyl chloride, and the absence of a peak at 2.7 minutes indicates that the hydrolysis is complete.

[0048] Cumulative number of peaks: 11 Cumulative area sum: 285222685 Cumulative area percentage sum: 100.000%

[0049] Serial number Retention time Peak area Peak height Analysis results % 1 3.570 339067 2842 0.119 2 3.743 322999 3240 0.113 3 4.067 46196 354 0.016 4 4.397 1244015 7008 0.436 5 4.710 40930 383 0.014 6 5.090 66411 284 0.023 7 6.227 278712853 998325 97.718 8 10.803 37020 121 0.013 9 21.827 3396959 5970 1.191 10 25.520 353323 656 0.124 11 27.893 662912 752 0.232

[0050] The purity is 97.72%.

[0051] Comparative Example 1. Direct determination of the purity of the starting material side chain acyl chloride by liquid phase

[0052] Take an appropriate amount of the side chain acyl chloride of the starting material of ceftaroline fosamil with batch number 20210205, dissolve it in acetonitrile and dilute it to make a solution containing about 1 mg per 1 ml as the test solution, and the test solution needs to be prepared before use; use a Waters Symmetry SH ield RP18 (250mm×4.6mm, 5μm) chromatographic column, column temperature 30°C, detection wavelength 254nm, flow rate 1.0ml per minute, mobile phase 0.1mol / L potassium dihydrogen phosphate solution-acetonitrile (volume ratio 3:1), column;

[0053] Under this chromatographic condition, accurately measure 20 μL of the test solution, inject it into the liquid chromatograph, and record the chromatogram (such as Figure 2 The purity was calculated by peak area normalization method.

[0054] Cumulative number of peaks: 7 Cumulative area sum: 180303185 Cumulative area percentage sum: 100.000%

[0055] Serial number Retention time Peak area Peak height Analysis results % 1 2.700 30353164 148646 16.835 2 3.727 87909 696 0.049 3 4.690 2251408 10973 1.249 4 6.683 244582 976 0.136 5 7.850 1774523 5784 0.984 6 11.263 193448 1041 0.107 7 13.427 145398151 268714 80.641

[0056] The purity is 80.64%, of which the chromatographic peak with a retention time of 2.7min is the main hydrolysis product, accounting for 16.84%.

[0057] Comparative Example 2. The purity of the side chain acyl chloride of cefuroxime was detected by using the technical solution of the invention of patent CN 107621512.

[0058] Take about 100 mg of the side chain acyl chloride of ceftaroline fosamil starting material with batch number 20210205, add 5 ml of triethylamine solution, shake for 1 minute, transfer to a 50 ml volumetric flask, dilute to the scale with methanol, and shake well; accurately measure 5 ml of the solution prepared in the first step and place it in a suitable container, keep it in a 40 ° C water bath for 30 minutes, cool to room temperature, transfer it to a 50 ml volumetric flask, dilute to the scale with the mobile phase as the test solution; use Waters SymmetrySh ie ld RP18 (250 mm × 4.6 mm, 5 μm) chromatographic column, the mobile phase is 0.1 mol / L potassium dihydrogen phosphate solution-acetonitrile (volume ratio 3:1), the column temperature is 30 ° C, the flow rate is 1.0 ml per minute, and the detection wavelength is 254 nm;

[0059] Under this chromatographic condition, accurately measure 20 μL of the test solution, inject it into the liquid chromatograph, and record the chromatogram (such as Figure 3 The purity was calculated by peak area normalization method.

[0060] Cumulative number of peaks: 7 Cumulative area sum: 202923677 Cumulative area percentage sum: 100.000%

[0061] Serial number Retention time Peak area Peak height Analysis results % 1 2.687 3067261 21838 1.512 2 3.680 7803 128 0.004 3 4.693 2086906 10005 1.028 4 6.587 46167 256 0.023 5 8.003 2202495 7252 1.085 6 11.097 759526 2183 0.374 7 13.650 194753519 351576 95.974

[0062] The purity is 95.94%. The chromatographic peak with a retention time of 2.7min is the main hydrolysis product, accounting for 1.52%.

[0063] By comparing the results of the above examples with the control examples, it can be found that the purity of the direct determination method is significantly lower, and the methanol derivatization method shows that the derivatization is incomplete and the determination is inaccurate, indicating that the pre-column derivatization method has a strong accuracy and is suitable for the determination of the purity of the side chain acyl chloride of ceftaroline fosamil.

Claims

1. A method for determining the purity of the side chain of cefuroxime, characterized in that: After the side chain of ceftaroline fosamil is amidated by an amine compound, its purity is determined by instrumental analysis; The amine compound is selected from aniline; The instrumental analysis method is high performance liquid chromatography; The amidation process is as follows: After quantitatively taking the side chain of ceftaroline fosamil, dilute it with an amine compound to the target volume, mix it evenly, keep it in a water bath at 20-40° C. for not less than 30 minutes, filter it, wash it with a solvent, and dry it to obtain an amide product; The specific parameters of the high performance liquid chromatography are: The mobile phase was a mixture of dipotassium hydrogen phosphate and acetonitrile; Column temperature 25-35°C; Flow rate 0.5-2ml per minute; Detection wavelength: 254nm; The volume ratio of the dipotassium hydrogen phosphate to acetonitrile is 1-4:

1.

2. The method for determining the purity of the side chain of ceftaroline fosamil according to claim 1, characterized in that: The usage ratio of the ceftaroline fosamil side chain to the amine compound is 0.5-2 g of ceftaroline fosamil side chain / 10 ml of the amine compound.

3. The method for determining the purity of the side chain of ceftaroline fosamil according to claim 1, characterized in that: The temperature of the water bath was 30°C.

4. The method for determining the purity of the side chain of ceftaroline fosamil according to claim 1, characterized in that: The amount of the analyte used is 80-150 mg / 50 ml of mobile phase.

5. The method for determining the purity of the side chain of ceftaroline fosamil according to claim 1, characterized in that: The concentration of the dipotassium hydrogen phosphate is 0.08-0.12 mol / L.

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