CN110146621B - Method for determining content of polymer in cephalosporin antibiotic medicine - Google Patents
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Abstract
The invention relates to a method for measuring the content of a polymer in a cephalosporin antibiotic drug. A method for measuring the content of polymers in cephalosporin antibiotic drugs comprises the following steps: respectively detecting a test solution and a control solution according to a high performance liquid chromatography to obtain a chromatogram of the test solution and a chromatogram of the control solution, wherein the test solution is a solution containing cephalosporin antibiotic drugs, the control solution is a diluent of the test solution, and the chromatographic conditions are as follows: the chromatographic column is an octadecylsilane bonded silica gel column, the mobile phase comprises a mobile phase A and a mobile phase B, the mobile phase A comprises water, and the mobile phase B comprises methanol; and calculating the content of the polymer in the cephalosporin antibiotic medicament by adopting a main component self-comparison method according to the chromatogram of the test solution and the chromatogram of the control solution. Experiments prove that the content of the polymer in the cephalosporin antibiotic medicine is more accurately determined.
Description
Technical Field
The invention relates to the field of pharmaceutical analysis, in particular to a method for determining the content of a polymer in a cephalosporin antibiotic drug.
Background
Cephalosporin antibiotics have become one of the most widely used antibacterial drugs in clinical application by virtue of the advantages of wide antibacterial spectrum, strong antibacterial activity, small side effect and the like. The most common adverse reaction in clinical application is anaphylactic reaction. The 2016 national adverse drug reaction monitoring annual report indicates that in suspected drugs related to the annual adverse drug reaction event report, the number of anti-infective drugs accounts for 43.9 percent of the total times of chemical drugs. The antibiotic allergen is not the drug itself, but a high molecular polymer formed by degrading the antibiotic allergen is irreversibly combined with a macromolecular carrier in vivo to cause antigen-antibody reaction, thereby a series of anaphylactic reaction symptoms occur. The sources of high molecular polymers are generally classified into exogenous sources (conjugates of proteins, polypeptides, etc. derived from fermentation processes) and endogenous sources (polymerization of drugs themselves and polymerization of drug-degrading impurities). The content of high molecular polymer in the medicine directly affects the incidence rate of anaphylactic reaction, so the content of high molecular polymer in the cephalosporin antibiotic medicine is strictly controlled.
The method for detecting the high molecular polymer of more than 10 cephalosporin antibiotic medicine raw materials and preparations thereof is recorded in the 'Chinese pharmacopoeia' 2015 edition, and the traditional molecular exclusion chromatography is basically adopted, but the molecular exclusion chromatography is inaccurate in measuring the content of the polymer in the cephalosporin antibiotic medicine.
Disclosure of Invention
Based on the above, there is a need for a method for determining the content of polymer in cephalosporin antibiotic drugs with high accuracy.
A method for measuring the content of polymers in cephalosporin antibiotic drugs comprises the following steps:
respectively detecting a test solution and a control solution according to a high performance liquid chromatography to obtain a chromatogram of the test solution and a chromatogram of the control solution, wherein the test solution is a solution containing cephalosporin antibiotic drugs, the control solution is a diluent of the test solution, and the chromatographic conditions are as follows: the chromatographic column is an octadecylsilane chemically bonded silica column, the mobile phase comprises a mobile phase A and a mobile phase B, the mobile phase A comprises water, the mobile phase B comprises methanol, and the elution gradient is as follows: 0min-20 min: 33% to 43% of said mobile phase B; 20min-35 min: 33% to 43% of said mobile phase B is increased to 48% to 55% of said mobile phase B; 35min-36 min: from 48% to 55% of said mobile phase B decreasing to from 33% to 43% of said mobile phase B; 36min-45 min: 33% to 43% of said mobile phase B;
and calculating the content of the polymer in the cephalosporin antibiotic drug by adopting a main component self-comparison method according to the chromatogram of the test solution and the chromatogram of the control solution.
Experiments prove that the problems of poor separation degree and low column efficiency exist in the determination of the polymer in the cephalosporin antibiotic drugs by adopting the molecular exclusion method, so that the determination result is inaccurate. The method for measuring the content of the polymer in the cephalosporin antibiotic drug has the advantages of good separation degree and high column efficiency, can ensure that the accuracy of measuring the content of the polymer in the cephalosporin antibiotic drug is higher, and is used for controlling the quality of the cephalosporin antibiotic drug.
In one embodiment, the mobile phase a further comprises a pH adjuster selected from at least one of ammonium dihydrogen phosphate and formic acid.
In one embodiment, the pH regulator is ammonium dihydrogen phosphate, and the molar concentration of the pH regulator is less than or equal to 0.2 mol/L.
In one embodiment, the pH adjusting agent is formic acid, and the volume percentage concentration of the pH adjusting agent is 0.2% or less.
In one embodiment, the step of detecting the test solution and the control solution according to high performance liquid chromatography respectively has a flow rate of 0.8mL/min to 1.2 mL/min.
In one embodiment, in the step of detecting the test solution and the control solution respectively according to high performance liquid chromatography, the detection wavelength is 278 nm.
In one embodiment, the step of separately detecting the sample solution and the control solution according to high performance liquid chromatography comprises a step of column temperature of 30-40 ℃.
In one embodiment, the step of detecting the test solution and the control solution respectively according to the high performance liquid chromatography comprises the step of sampling 10 to 20 μ L.
In one embodiment, the cephalosporin antibiotic drug is selected from one of cefuroxime axetil raw material and cefuroxime axetil preparation.
In one embodiment, the cefuroxime axetil preparation is one selected from the group consisting of cefuroxime axetil tablets, cefuroxime axetil capsules and cefuroxime axetil granules.
Drawings
FIG. 1 is a high performance liquid chromatogram of a test solution in example 1;
FIG. 2 is a high performance liquid chromatogram of the test solution of example 2;
FIG. 3 is a high performance liquid chromatogram of the test solution of example 3;
FIG. 4 is a high performance liquid chromatogram of the test solution of example 4;
FIG. 5 is a high performance liquid chromatogram of the test solution of example 5;
FIG. 6 is a high performance liquid chromatogram of the test solution in comparative example 1.
Detailed Description
To facilitate an understanding of the invention, the invention will now be described more fully with reference to the accompanying drawings. Preferred embodiments of the present invention are shown in the drawings. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.
The method for determining the content of the polymer in the cephalosporin antibiotic drug comprises the following steps:
step S110: and respectively detecting the test solution and the control solution according to the high performance liquid chromatography so as to obtain a chromatogram of the test solution and a chromatogram of the control solution.
Wherein the test solution is a solution containing cephalosporin antibiotic drugs. Further, the cephalosporin antibiotic drug is selected from one of cefuroxime axetil raw materials and cefuroxime axetil preparations. Further, the cefuroxime axetil preparation is one selected from the group consisting of cefuroxime axetil tablets, cefuroxime axetil capsules and cefuroxime axetil granules.
Cefuroxime axetil is a second-generation cephalosporin antibiotic, is a prodrug of cefuroxime, is rapidly hydrolyzed by nonspecific esterase in gastrointestinal mucosal cells after oral administration to release cefuroxime to exert drug effect, is suitable for infection caused by various sensitive bacteria, and has become an anti-infective drug which is popular in the world. The structure of cefuroxime axetil is shown in the following diagram:
cefuroxime axetil was developed by GlaxoSmithKline (glatiramer), and cefuroxime axetil tablets were first marketed in the united kingdom on 3/9 in 1987 and subsequently sold in many countries worldwide. The product was approved by FDA in the United states in 1987, 12 and 28 days, and was marketed in the United states in 1988, Japanese in the same year, and first imported in China in 2000. Cefuroxime axetil and related preparations thereof are recorded in the second part of the 2015 edition of the Chinese pharmacopoeia, but a polymer control method of cefuroxime axetil and preparations thereof is not recorded, and the polymer cannot be detected by the existing related substance detection method in the pharmacopoeia.
Specifically, the preparation method of the test solution comprises the following steps: dissolving cephalosporin antibiotic medicine in methanol to obtain test solution. More specifically, the preparation method of the test solution comprises the following steps: precisely weighing a proper amount of cephalosporin antibiotic drugs, placing the cephalosporin antibiotic drugs in a measuring flask, adding 10% methanol for dissolving, diluting the cephalosporin antibiotic drugs to a scale by using a mobile phase, shaking up, preparing a solution containing 0.5mg of cephalosporin antibiotic drugs per mL, and filtering the solution to obtain a test solution.
Wherein the control solution is a diluent of the test solution. Further, the preparation method of the control solution comprises the following steps: diluting the test solution by 50-1000 times with water to obtain a control solution. Specifically, the test solution was diluted 100-fold with water to obtain a control solution.
Wherein, the chromatographic conditions are as follows: the chromatographic column is an octadecylsilane chemically bonded silica column, so that the separation column efficiency of the cephalosporin antibiotic drugs is higher. Specifically, the length of the column was 150mm, the inner diameter of the column was 4.6mm, and the particle size of the packing of the column was 5 μm. More specifically, the chromatographic column is Waters X-bridge C18。
Wherein the mobile phase comprises a mobile phase A and a mobile phase B. Specifically, mobile phase a includes water and mobile phase B includes methanol.
Further, the mobile phase a also includes a pH adjuster. Further, the pH adjuster is at least one selected from ammonium dihydrogen phosphate and formic acid.
In one embodiment, the pH regulator is ammonium dihydrogen phosphate, and the molar concentration of the pH regulator is less than or equal to 0.2 mol/L.
In another embodiment, the pH adjusting agent is formic acid and the volume percent concentration of the pH adjusting agent is less than 0.2%.
Wherein the elution gradient is:
0min-20 min: 33 to 43 percent of mobile phase B;
20min-35 min: increasing the content of the mobile phase B from 33 percent to 43 percent to the content of the mobile phase B from 48 percent to 55 percent;
35min-36 min: reducing the flow rate of the mobile phase B from 48 percent to 55 percent to the flow rate of the mobile phase B from 33 percent to 43 percent;
36min-45 min: 33 to 43 percent of mobile phase B.
Further, the elution gradient was:
0min-20 min: 38% of mobile phase B;
20min-35 min: increasing from 38% mobile phase B to 50% mobile phase B;
35min-36 min: 50% of mobile phase B was decreased to 38% of mobile phase B;
36min-45 min: 38% of mobile phase B.
Wherein, in the step of respectively detecting the sample solution and the control solution according to the high performance liquid chromatography, the flow rate is 0.8mL/min to 1.2 mL/min. Further, the flow rate was 1.0 mL/min.
Wherein, in the step of detecting the test solution and the control solution respectively according to the high performance liquid chromatography, the detection wavelength is 278 nm.
Wherein, in the step of respectively detecting the sample solution and the reference solution according to the high performance liquid chromatography, the column temperature is 30-40 ℃. Further, the column temperature was 35 ℃.
Wherein, in the step of detecting the sample solution and the control solution respectively according to the high performance liquid chromatography, the sample amount is 10-20 muL. Further, the amount of the sample was 20. mu.L.
Step S120: and calculating the content of the polymer in the cephalosporin antibiotic medicament by adopting a main component self-comparison method according to the chromatogram of the test solution and the chromatogram of the control solution.
Specifically, the formula C% ═ r is adoptedu/rsX 100/n, calculating the content of the polymer in the cephalosporin antibiotic medicine, wherein C% is the mass percentage content of the polymer in the cephalosporin antibiotic, ruIs the peak area value, r, of the polymer in the chromatogram of the test solutionsThe peak area value of the main peak of the cephalosporin antibiotic drug in the chromatogram of the control solution is shown, and n is the dilution multiple of the control solution.
The main component self-comparison method has the advantages of no need of comparison products, low cost, simple operation and the like.
Wherein, the polymer in the cephalosporin antibiotic drug is generally a high molecular polymer.
The method for measuring the content of the polymer in the cephalosporin antibiotic medicament at least has the following advantages:
1) experiments prove that the problems of poor separation degree and low column efficiency exist in the determination of the polymer in the cephalosporin antibiotic drugs by adopting the molecular exclusion method, so that the determination result is inaccurate. The method for measuring the content of the polymer in the cephalosporin antibiotic drug has the advantages of good separation degree and high column efficiency, can ensure that the accuracy of measuring the content of the polymer in the cephalosporin antibiotic drug is higher, and is used for controlling the quality of the cephalosporin antibiotic drug.
2) Compared with the traditional molecular exclusion chromatography, the method for measuring the content of the polymer in the cephalosporin antibiotic medicine has the advantages of simplicity, rapidness, good durability, good reproducibility and the like.
3) The content determination method of the polymer in the cephalosporin antibiotic drugs realizes the determination of the content of the cefuroxime axetil polymer in the cefuroxime axetil and the preparation thereof by adopting an RP-HPLC method, and simultaneously can control the content of other impurities, such as cefuroxime and delta3Isomer, E isomer and the like, and is suitable for the quality control of cefuroxime axetil and the preparation thereof. The traditional size exclusion chromatographic column can only measure the polymer and cannot measure the content of known impurities and unknown impurities in a sample.
The following are specific examples:
instrument and reagent
The instrument comprises the following steps: agilent 1260 high performance liquid chromatograph; a McLer ME204 electronic balance; a mettler XP6 parts per million electronic balance;
reagent testing: cefuroxime axetil, cefuroxime axetil granules, cefuroxime axetil capsules, cefuroxime axetil tablets; ammonium dihydrogen phosphate (national drug group chemical agents limited); methanol (j.t.baker).
Example 1
The method for determining the content of the polymer in the cefuroxime axetil raw material in this example is as follows:
(1) precisely weighing a proper amount of cefuroxime axetil raw material (about 50mg equivalent to cefuroxime), placing the cefuroxime axetil raw material into a 100ml measuring flask, adding 10ml of methanol to dissolve the cefuroxime axetil raw material, diluting the cefuroxime axetil raw material to a scale by using a mobile phase, shaking up, and filtering to obtain a test solution;
(2) precisely transferring 1ml of the test solution, and diluting to 100ml with a mobile phase to obtain a control solution;
(3) respectively detecting the test solution and the control solution according to a high performance liquid chromatography to obtain a chromatogram of the test solution and a chromatogram of the control solution, wherein the chromatographic conditions are as follows:
the chromatographic column is a chromatographic column (Waters X-bridge C) with octadecylsilane chemically bonded silica as filler18150X 4.6mm, 5 μm); the mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is 0.1mol/L ammonium dihydrogen phosphate waterThe solution, mobile phase B is methanol; the detection wavelength is 278 nm; the flow rate is 1.0 ml/min; the sample injection amount is 20 mu L; the column temperature was 35 ℃. The elution gradient was: 0min-20 min: 38% methanol; 20min-35 min: 38% methanol was increased to 50% methanol; 35min-36 min: the 50% methanol was decreased to 38% methanol; 36min-45 min: 38% methanol;
(4) calculating the polymer content, cefuroxime content and delta content of the cephalosporin antibiotic drug by a main component self-comparison method according to the chromatogram of the test solution and the chromatogram of the control solution3The results are shown in Table 1, wherein the chromatogram of the test solution is shown in FIG. 1, the peak 1 is cefuroxime, the peaks 2 and 3 are cefuroxime axetil peaks, and the peak 4 is Δ3-isomer, peaks 5 and 6 are all E-isomer peaks, and peaks 7, 8 and 9 are all polymer peaks.
Example 2
The method for determining the content of the polymer in the cefuroxime axetil raw material in this example is as follows:
(1) precisely weighing a proper amount of cefuroxime axetil raw material (about 50mg equivalent to cefuroxime), placing the cefuroxime axetil raw material into a 100ml measuring flask, adding 10ml of methanol to dissolve the cefuroxime axetil raw material, diluting the cefuroxime axetil raw material to a scale by using a mobile phase, shaking up, and filtering to obtain a test solution;
(2) precisely transferring 1ml of the test solution, and diluting to 100ml with a mobile phase to obtain a control solution;
(3) respectively detecting the test solution and the control solution according to a high performance liquid chromatography to obtain a chromatogram of the test solution and a chromatogram of the control solution, wherein the chromatographic conditions are as follows:
the chromatographic column is a chromatographic column (Waters X-bridge C) with octadecylsilane chemically bonded silica as filler18150X 4.6mm, 5 μm); the mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is a 0.1% formic acid aqueous solution, and the mobile phase B is methanol; the detection wavelength is 278 nm; the flow rate is 1.0 ml/min; the sample injection amount is 20 mu L; the column temperature was 35 ℃; the elution gradient was: 0min-20 min: 38% methanol; 20min-35 min: 38 percent ofMethanol was ramped up to 50% methanol; 35min-36 min: the 50% methanol was decreased to 38% methanol; 36min-45 min: 38% methanol;
(4) calculating the polymer content, cefuroxime content and delta content of the cephalosporin antibiotic drug by a main component self-comparison method according to the chromatogram of the test solution and the chromatogram of the control solution3The results are shown in Table 1, wherein the chromatogram of the test solution is shown in FIG. 2, the peak 1 is cefuroxime, the peaks 2 and 3 are cefuroxime axetil peaks, and the peak 4 is Δ3-isomer, peaks 5 and 6 are all E-isomer peaks, and peaks 7, 8 and 9 are all polymer peaks.
Example 3
The method for determining the content of the polymer in the cefuroxime axetil particles of this example is as follows:
(1) taking cefuroxime axetil particles, grinding, mixing uniformly, precisely weighing an appropriate amount (about equivalent to 50mg of cefuroxime), placing in a 100ml measuring flask, adding 10ml of methanol for dissolving, diluting to a scale with a mobile phase, shaking uniformly, and filtering to obtain a test solution.
(2) Precisely transferring 1ml of the test solution, and diluting to 100ml with a mobile phase to obtain a control solution;
(3) respectively detecting the test solution and the control solution according to a high performance liquid chromatography to obtain a chromatogram of the test solution and a chromatogram of the control solution, wherein the chromatographic conditions are as follows:
the chromatographic column is a chromatographic column (Waters X-bridge C) with octadecylsilane chemically bonded silica as filler18150X 4.6mm, 5 μm); the mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is 0.2mol/L ammonium dihydrogen phosphate aqueous solution, and the mobile phase B is methanol; the detection wavelength is 278 nm; the flow rate is 1.2 ml/min; the sample injection amount is 10 mu L; the column temperature was 40 ℃; the elution gradient was: 0min-20 min: 33% methanol; 20min-35 min: 33% methanol was increased to 55% methanol; 35min-36 min: 55% methanol was decreased to 33% methanol; 36min-45 min: 33% methanol;
(4) according to supply ofThe chromatogram of the test solution and the chromatogram of the control solution are obtained by calculating the polymer content, cefuroxime content and delta content of the cephalosporin antibiotic drug by a main component self-control method3The results are shown in Table 1, wherein the chromatogram of the test solution is shown in FIG. 3, the peak 1 is cefuroxime, the peaks 2 and 3 are cefuroxime axetil peaks, and the peak 4 is Δ3-isomer, peaks 5 and 6 are all E-isomer peaks, and peaks 7, 8 and 9 are all polymer peaks.
Example 4
The method for determining the content of the polymer in the cefuroxime axetil capsule in this example is as follows:
(1) taking cefuroxime axetil capsules, grinding, uniformly mixing, precisely weighing an appropriate amount (about equivalent to 50mg of cefuroxime), putting into a 100ml measuring flask, adding 10ml of methanol for dissolving, diluting to a scale by using a mobile phase, shaking uniformly, and filtering to obtain a test solution;
(2) precisely transferring 1ml of the test solution, and diluting to 100ml with a mobile phase to obtain a control solution;
(3) respectively detecting the test solution and the control solution according to a high performance liquid chromatography to obtain a chromatogram of the test solution and a chromatogram of the control solution, wherein the chromatographic conditions are as follows:
chromatographic column using octadecylsilane bonded silica gel as filler (Waters X-bridge C)18150X 4.6mm, 5 μm); the mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is water, and the mobile phase B is methanol; the detection wavelength is 278 nm; the flow rate is 0.8 ml/min; the sample injection amount is 20 mu L; the column temperature is 30 ℃; the elution gradient was: 0min-20 min: 43% methanol; 20min-35 min: 43% methanol was increased to 48% methanol; 35min-36 min: the 48% methanol was decreased to 43% methanol; 36min-45 min: 43% methanol;
(4) calculating the polymer content, cefuroxime content and delta content of the cephalosporin antibiotic drug by a main component self-comparison method according to the chromatogram of the test solution and the chromatogram of the control solution3Of the isomersThe results are shown in Table 1, wherein the chromatogram of the test solution is shown in FIG. 4, the peak 1 is cefuroxime, the peaks 2 and 3 are cefuroxime axetil peaks, and the peak 4 is Delta3-isomer, peaks 5 and 6 are all E-isomer peaks, and peaks 7, 8 and 9 are all polymer peaks.
Example 5
The method for determining the content of the polymer in the cefuroxime axetil tablet of this example is as follows:
(1) taking cefuroxime axetil tablets, grinding, mixing uniformly, precisely weighing a proper amount (about equivalent to 50mg of cefuroxime), putting into a 100ml measuring flask, adding 10ml of methanol for dissolving, diluting to a scale by using a mobile phase, shaking uniformly, and filtering to obtain a test solution;
(2) precisely transferring 1ml of the test solution, and diluting to 100ml with a mobile phase to obtain a control solution;
(3) respectively detecting the test solution and the control solution according to a high performance liquid chromatography to obtain a chromatogram of the test solution and a chromatogram of the control solution, wherein the chromatographic conditions are as follows:
the chromatographic column is a chromatographic column (Waters X-bridge C) with octadecylsilane chemically bonded silica as filler18150X 4.6mm, 5 μm); the mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is a 0.2% formic acid aqueous solution in volume concentration, and the mobile phase B is methanol; the detection wavelength is 278 nm; the flow rate is 1.0 ml/min; the sample injection amount is 20 mu L; the column temperature was 35 ℃; the elution gradient was: 0min-20 min: 38% methanol; 20min-35 min: 38% methanol was increased to 50% methanol; 35min-36 min: the 50% methanol was decreased to 38% methanol; 36min-45 min: 38% methanol;
(4) calculating the polymer content, cefuroxime content and delta content of the cephalosporin antibiotic drug by a main component self-comparison method according to the chromatogram of the test solution and the chromatogram of the control solution3The contents of isomers, E-isomers, and the degrees of separation of the polymer peak from the adjacent peak and the theoretical plate number of the cefuroxime axetil peak were calculated, and the results are shown in Table 1, in whichThe chromatogram of the sample solution is shown in FIG. 5, in which peak 1 is cefuroxime, peaks 2 and 3 are cefuroxime axetil peaks, and peak 4 is Δ3-isomer, peaks 5 and 6 are all E-isomer peaks, and peaks 7, 8 and 9 are all polymer peaks.
Comparative example 1
The method for determining the content of the polymer in cefuroxime axetil of the present comparative example is as follows:
(1) precisely weighing a proper amount of cefuroxime axetil (about equivalent to 25mg of cefuroxime), putting the cefuroxime axetil into a 25ml measuring flask, adding 5ml of methanol to dissolve, adding methanol to dilute to a scale, shaking up, and filtering to obtain a test solution;
(2) precisely transferring 1ml of the test solution, and diluting to 100ml with a mobile phase to obtain a control solution;
(3) respectively detecting the test solution and the control solution according to a high performance liquid chromatography to obtain a chromatogram of the test solution and a chromatogram of the control solution, wherein the chromatographic conditions are as follows:
the chromatographic column is a chromatographic column (300 multiplied by 7.8mm, 7 μm) with polystyrene copolymer as a filler; the mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is methanol, the mobile phase B is water, and the volume ratio of the mobile phase A to the mobile phase B is 95: 5; the flow rate is 1.0ml/min, the detection wavelength is 278nm, the column temperature is 35 ℃, and the sample injection amount is 10 mu L;
(4) according to the chromatogram of the test solution and the chromatogram of the control solution, the content of the polymer in the cephalosporin antibiotic drug is calculated by adopting a main component self-control method, and the separation degree of a polymer peak and adjacent peaks and the theoretical plate number of a cefuroxime axetil peak are calculated, and the result is shown in table 1, wherein the chromatogram of the test solution is shown in fig. 6, the peak 1 and the peak 2 are both polymer peaks, and the peak 3 is a cefuroxime axetil peak.
TABLE 1
As can be seen from Table 1, compared with comparative example 1, the degree of separation between the polymer peak and the adjacent peak is higher in the determination method of the polymers of examples 1 to 5, and the theoretical plate number of the cefuroxime axetil peak is higher, which shows that the degree of separation is better and the column efficiency is higher in the determination method of the polymers of examples 1 to 5, so that the content determination of the polymer is more accurate.
The polymer can only be measured using the size exclusion method of comparative example 1 and the content of known and unknown impurities in the sample cannot be determined.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. The method for measuring the content of the polymer in the cephalosporin antibiotic medicine is characterized by comprising the following steps:
respectively detecting a test solution and a control solution according to a high performance liquid chromatography to obtain a chromatogram of the test solution and a chromatogram of the control solution, wherein the test solution is a solution containing cephalosporin antibiotic drugs, the control solution is a diluent of the test solution, and the chromatographic conditions are as follows: the chromatographic column is an octadecylsilane chemically bonded silica column, the mobile phase consists of a mobile phase A and a mobile phase B, the mobile phase A is an aqueous solution of a pH regulator, the pH regulator is ammonium dihydrogen phosphate or formic acid, the mobile phase B is methanol, and the elution procedure is as follows: 0min-20 min: the mobile phase B keeps 33% -43% of isocratic elution; 20min-35 min: the mobile phase B is increased from 33% -43% to 48% -55%; 35min-36 min: the mobile phase B is decreased from 48% -55% to 33% -43%; 36min-45 min: the mobile phase B keeps 33% -43% of isocratic elution;
calculating the content of the polymer in the cephalosporin antibiotic drug by adopting a main component self-comparison method according to the chromatogram of the test solution and the chromatogram of the control solution;
wherein the cephalosporin antibiotic drug is selected from one of cefuroxime axetil raw materials and cefuroxime axetil preparations.
2. The method for determining the content of the polymer in the cephalosporin antibiotic drug according to claim 1, wherein the step of separately detecting the test solution and the control solution according to high performance liquid chromatography comprises a column temperature of 30 ℃ to 40 ℃.
3. The method for determining the content of the polymer in the cephalosporin antibiotic drug according to claim 1, wherein the pH regulator is ammonium dihydrogen phosphate, and the molar concentration of the pH regulator is less than or equal to 0.2 mol/L.
4. The method for determining the content of the polymer in the cephalosporin antibiotic drug according to claim 1, wherein the pH regulator is formic acid, and the volume percentage concentration of the pH regulator is less than 0.2%.
5. The method for determining the content of the polymer in the cephalosporin antibiotic drug according to claim 1, wherein the step of separately detecting the test solution and the control solution according to high performance liquid chromatography has a flow rate of 0.8mL/min to 1.2 mL/min.
6. The method for determining the content of polymers in cephalosporin antibiotic drugs according to claim 1, wherein the step of detecting the test solution and the control solution separately according to high performance liquid chromatography has a detection wavelength of 278 nm.
7. The method for determining the content of the polymer in the cephalosporin antibiotic drug according to claim 1, wherein in the step of separately detecting the test solution and the control solution according to high performance liquid chromatography, the column temperature is 30 ℃ to 40 ℃; the elution procedure was: 0min-20 min: the mobile phase B keeps 33% -38% of isocratic elution; 20min-35 min: the mobile phase B is increased from 33% -38% to 50% -55%; 35min-36 min: the mobile phase B is decreased from 50% -55% to 33% -38%; 36min-45 min: the mobile phase B keeps 33% -38% of isocratic elution.
8. The method for determining the content of the polymer in the cephalosporin antibiotic drug according to claim 1, wherein the sample amount in the step of separately detecting the test solution and the control solution according to high performance liquid chromatography is 10 μ L to 20 μ L.
9. The method for determining the content of the polymer in the cephalosporin antibiotic drug according to claim 1, wherein the step of separately detecting the test solution and the control solution according to high performance liquid chromatography has a column temperature of 35 ℃ to 40 ℃.
10. The method for determining the content of polymer in cephalosporin antibiotic drug according to claim 1, wherein the cefuroxime axetil preparation is selected from one of cefuroxime axetil tablet, cefuroxime axetil capsule and cefuroxime axetil granule.
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