CN115078555A - Method for determining content of main components in cefepime abamectin for injection - Google Patents

Method for determining content of main components in cefepime abamectin for injection Download PDF

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CN115078555A
CN115078555A CN202110263040.5A CN202110263040A CN115078555A CN 115078555 A CN115078555 A CN 115078555A CN 202110263040 A CN202110263040 A CN 202110263040A CN 115078555 A CN115078555 A CN 115078555A
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cefepime
abamectin
solution
phosphate buffer
mobile phase
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周玉梅
姚洛芫
李磊
张峰
朱素华
肖哲媛
马昀
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Nanjing Libowei Pharmaceutical Co ltd
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    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract

The invention belongs to the field of pharmaceutical analysis, and particularly relates to a method for determining the content of main components in cefepime abamectin for injection. According to the method, octadecylsilane chemically bonded silica is used as a filler of a chromatographic column, a certain proportion of phosphate buffer solution-acetonitrile is used as a mobile phase, and the contents of two main components in cefepime and abamectin for injection of the compound preparation can be simultaneously detected, so that the quality of cefepime and abamectin for injection can be effectively controlled. The determination method disclosed by the invention is high in accuracy, good in stability, simple and convenient to operate, strong in pertinence and good in application value.

Description

Method for determining content of main component in cefepime abamectin for injection
Technical Field
The invention belongs to the field of pharmaceutical analysis, and particularly relates to a method for determining the content of main components in cefepime abamectin for injection.
Background
Cefepime avibactam for injection is cefepimeA compound preparation of pyrixime (Cefepime) and Avibactam (Avibactam). Cefepime (molecular formula C) 19 H 24 N 6 O 5 S 2 Molecular weight 480.56) belongs to the fourth generation cephalosporin, and has strong antibacterial activity on various gram positive and negative bacteria including enterobacter, pseudomonas aeruginosa and other non-zymogenic bacillus, haemophilus, staphylococcus, etc. Abamebactam (molecular formula is C) 7 H 11 N 3 O 6 S, molecular weight 265.25) belongs to diazabicyclooctanone compounds, is a novel beta-lactamase inhibitor, can be bonded with enzyme reversibly and covalently for a long time, and does not induce the generation of beta-lactamase. The cefepime and abamectin are combined to enhance the antibacterial effect and broaden the antibacterial spectrum.
At present, the pharmacopoeia of each country only contains a single main component inspection method, and although the inspection method can effectively inspect a single component, the inspection method cannot simultaneously measure the respective contents of cefepime and abamectin.
Patent 2020109593309 discloses the use of cefepime abamectin composition in preparing a medicament for inhibiting a drug-resistant Klebsiella pneumoniae of A-class or D-class carbapenems, and cefepime abamectin has different mass ratios and different antibacterial effects. Therefore, a method capable of simultaneously determining the content of the main component in cefepime abamectin for injection is needed, so as to effectively control the quality of cefepime abamectin.
At present, no literature reports a method for simultaneously determining the content of main components in cefepime abamectin for injection.
Disclosure of Invention
Aiming at the fact that a simple, convenient and efficient detection method for cefepime, abamectin and related impurities does not exist at present, the invention provides a method for determining the content of main components (cefepime and abamectin) in cefepime abamectin for injection, and the blank in the prior art is made up.
In order to solve the technical problems, the invention adopts the following technical scheme:
a method for determining the content of main components in cefepime abamectin for injection comprises the following steps:
(1) chromatographic conditions are as follows:
octadecylsilane chemically bonded silica is used as a filler for the chromatographic column;
phosphate buffer solution-organic solvent is used as a mobile phase;
the flow rate of the mobile phase is 0.9-1.1 ml/min;
the column temperature is 25-35 ℃;
the sample injection volume is 10 mul;
the detection wavelength is 192-198 nm;
(2) and respectively injecting the test solution and the reference solution into a liquid chromatograph, and recording the chromatogram.
In a preferred embodiment, the ratio of cefepime to abamectin in the sample solution of the method is 1-8: 8-1.
In a preferred embodiment, the ratio of cefepime to abamectin is 2: 1.
In a preferred embodiment, the column in step (1) is an Inertsil ODS-3 column.
In a preferred embodiment, the phosphate buffer-organic solvent in step (1) is phosphate buffer-acetonitrile.
In a preferred embodiment, the volume ratio of the phosphate buffer solution to the acetonitrile is 85-95: 5-15.
In a preferred embodiment, the phosphate buffer to acetonitrile ratio is 90:10 by volume.
In a preferred embodiment, the molar concentration of the potassium dihydrogen phosphate in the phosphate buffer solution in the step (1) is 0.001 to 0.01 mol/L.
In a preferred embodiment, the molar concentration of potassium dihydrogen phosphate in the phosphate buffer solution in step (1) is 0.005 mol/L.
In a preferred embodiment, the pH value of the mobile phase phosphate buffer solution-acetonitrile in the step (1) is 4.8-5.2.
In a preferred embodiment, the mobile phase phosphate buffer-acetonitrile has a pH of 5.0.
In a preferred embodiment, the method is performed in an isocratic elution mode.
In a preferred embodiment, the flow rate of the mobile phase in step (1) is 1.0 ml/min.
In a preferred embodiment, the column temperature in step (1) is 30 ℃.
In a preferred embodiment, the detection wavelength in step (1) is 195 nm.
The technical scheme claimed by the invention is as follows:
1. a method for determining the content of main components in cefepime abamectin for injection is characterized by comprising the following steps:
(1) chromatographic conditions are as follows:
octadecylsilane chemically bonded silica is used as a filler for the chromatographic column;
phosphate buffer solution-organic solvent is used as a mobile phase;
the flow rate of the mobile phase is 0.9-1.1 ml/min;
the column temperature is 25-35 ℃;
the sample injection volume is 10 mul;
the detection wavelength is 192-198 nm;
(2) and respectively injecting the test solution and the reference solution into a liquid chromatograph, and recording the chromatogram. 2. The method according to claim 1, characterized in that it further comprises the steps of: preparing a test solution:
weighing a proper amount of cefepime abamectin, adding a mobile phase for dilution, and shaking up to obtain a test solution;
preparing a reference substance solution:
weighing appropriate amount of cefepime reference substance and abamectin reference substance, adding mobile phase to obtain cefepime abamectin mixed solution as reference solution.
3. The method according to scheme 2, wherein the ratio of cefepime to abamectin in the cefepime-abamectin mixed solution is 1-8: 8-1.
4. The method according to scheme 3, wherein the ratio of cefepime to abamectin is 2: 1.
5. The method according to scheme 1, wherein the column in step (1) is an Inertsil ODS-3 column.
6. The method according to scheme 1, wherein the phosphate buffer-organic solvent in step (1) is phosphate buffer-acetonitrile.
7. The method according to claim 6, wherein the volume ratio of the phosphate buffer solution to the acetonitrile is 85-95: 5-15.
8. The method of scheme 7, wherein the phosphate buffer-acetonitrile ratio is 90:10 by volume.
9. The method according to scheme 1, wherein the molar concentration of the potassium dihydrogen phosphate in the phosphate buffer solution in the step (1) is 0.001-0.01 mol/L.
10. The method according to claim 9, wherein the molar concentration of potassium dihydrogen phosphate in the phosphate buffer solution is 0.005 mol/L.
11. The method according to scheme 1, wherein the mobile phase phosphate buffer solution-acetonitrile in the step (1) has a pH value of 4.8-5.2.
12. The method of claim 11, wherein the mobile phase phosphate buffer-acetonitrile has a pH of 5.0.
13. The method according to scheme 1, wherein the method is characterized in that the elution mode is isocratic elution.
14. The process according to scheme 1, wherein the flow rate of the mobile phase in step (1) is 1.0 ml/min.
15. The process according to claim 1, wherein the column temperature in the step (1) is 30 ℃.
16. The method according to claim 1, wherein the detection wavelength in the step (1) is 195 nm.
The invention takes octadecylsilane chemically bonded silica as a filler of a chromatographic column, takes phosphate buffer solution-acetonitrile with a certain proportion as a mobile phase, and can simultaneously detect the contents of two main components in the compound preparation cefepime abamectin, thereby effectively controlling the quality of the cefepime abamectin. The determination method disclosed by the invention is high in accuracy, good in stability, simple and convenient to operate, strong in pertinence and good in application value.
Drawings
FIG. 1 is a liquid chromatogram of a control solution at a wavelength of 254nm in the first example.
FIG. 2 is a liquid chromatogram of the sample solution in the chromatographic condition with a wavelength of 254nm in the first example.
FIG. 3 is a liquid chromatogram of the control solution of example II under the chromatographic condition of 195nm wavelength.
FIG. 4 is a liquid chromatogram of the sample solution of example II under the chromatographic condition of 195nm wavelength.
Detailed Description
The technical solution of the present invention will be further explained with reference to the accompanying drawings and specific embodiments. Those who do not specify particular techniques or conditions in the following examples are conducted according to techniques or conditions described in the literature of the art. Reagents, drugs and instruments used in the following examples are all available by conventional commercial means unless otherwise indicated.
The first embodiment is as follows:
(1) chromatographic conditions
Referring to the chromatographic condition test of cefepime hydrochloride content in 2020 edition Chinese pharmacopoeia, wherein:
a chromatographic column: inertsil ODS-3(4.6 mm. times.250 mm, 5 μm) column;
mobile phase: phosphate buffer (0.68 g potassium dihydrogen phosphate, dissolved in water and diluted to 1000ml) -acetonitrile (90:10) (pH adjusted to 5.0 with 2% phosphoric acid solution or 2% potassium hydroxide solution);
and (3) an elution mode: isocratic elution;
flow rate of mobile phase: 1.0 ml/min;
sample introduction amount: 10 mu l of the mixture;
column temperature: 30 ℃;
detection wavelength: 254 nm.
(2) Solution preparation
Control solution: taking about 20mg of cefepime reference substance and about 10mg of abamectin reference substance, precisely weighing, placing in a 100ml measuring flask, adding a mobile phase for dissolving, diluting to a scale, and shaking up to obtain a reference substance solution;
test solution: weighing cefepime abamectin, adding water to dissolve the cefepime abamectin, transferring the cefepime abamectin to a 250ml measuring flask, adding water to dilute the cefepime abamectin to a scale, uniformly mixing, precisely measuring 1ml, placing the cefepime abamectin in a 100ml measuring flask, adding a mobile phase to dilute the cefepime abamectin to the scale, and shaking the cefepime abamectin to obtain a sample solution.
(3) Measurement results
And (3) respectively sucking the reference substance solution and the sample solution to be tested, and recording chromatograms as shown in figures 1-2 and tables 1-2.
TABLE 1 liquid chromatography results of control solutions at 254nm wavelength
Figure BDA0002970866450000081
TABLE 2 liquid chromatography results of the test solutions under the chromatography condition of 254nm wavelength
Figure BDA0002970866450000082
The results show that: under the chromatographic condition of 254nm wavelength, in the chromatogram of the test solution, the retention time of the abamectin and cefepime is 4.333 minutes and 6.460 minutes respectively, and the theoretical plate number is 14819 and 11610 respectively. However, under this chromatographic condition, there is a problem that avibactam responds less.
Example two:
(1) chromatographic conditions
A chromatographic column: inertsil ODS-3(4.6 mm. times.250 mm, 5 μm) column;
mobile phase: phosphate buffer (0.68 g potassium dihydrogen phosphate, dissolved in water and diluted to 1000ml) -acetonitrile (90:10) (pH adjusted to 5.0 with 2% phosphoric acid solution or 2% potassium hydroxide solution);
an elution mode: isocratic elution;
flow rate of mobile phase: 1.0 ml/min;
sample injection amount: 10 mu l of the mixture;
column temperature: 30 ℃;
detection wavelength: 195 nm.
(2) Solution preparation
Control solution: taking about 20mg of cefepime reference substance and about 10mg of abamectin reference substance, precisely weighing, placing in a 100ml measuring flask, adding a mobile phase for dissolving, diluting to a scale, and shaking up to obtain a reference substance solution;
test solution: weighing cefepime abamectin, adding water to dissolve the cefepime abamectin, transferring the cefepime abamectin to a 250ml measuring flask, adding water to dilute the cefepime abamectin to a scale, uniformly mixing, precisely measuring 1ml, placing the cefepime abamectin in a 100ml measuring flask, adding a mobile phase to dilute the cefepime abamectin to the scale, and shaking the cefepime abamectin to obtain a sample solution.
(3) Measurement results
And (3) respectively sucking the reference substance solution and the sample solution for sample injection, and recording chromatograms as shown in figures 3-4 and tables 3-5.
TABLE 3 liquid chromatography results of control solutions at 195nm wavelength
Figure BDA0002970866450000091
TABLE 4 liquid chromatography results of the test solutions at 195nm wavelength
Figure BDA0002970866450000092
TABLE 5 comparison of the peak areas of the two main components at different detection wavelengths
Figure BDA0002970866450000093
Figure BDA0002970866450000101
The results show that: under the chromatographic condition of wavelength of 195nm, in the chromatogram of the test solution, the retention time of abamectin and cefepime is 4.333 minutes and 6.460 minutes respectively, the theoretical plate number is 14904 and 11609 respectively, the separation degree between chromatographic peaks is better, and the detection time is shorter. Under the detection wavelength of 195nm, the response value of abamectin is obviously increased, and the peak area is increased by about 41 times.
Example three: system applicability
(1) Solution preparation
Blank solvent: phosphate buffer (0.68 g potassium dihydrogen phosphate dissolved in 1000ml water) -acetonitrile (90:10), pH 5.0 with phosphoric acid, shaking up, as a blank solvent.
Mixing impurity solution: weighing an appropriate amount of an impurity A (cefepime E isomer), an impurity D ((Z) -2- (2-aminothiazole-4-yl) -2- (methoxyimino) acetic acid), an impurity E ((6R,7R) -7-amino-3- ((1-methylpyrrolidinium) methyl) -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylate) and a control substance of the decarbonylation avibactam, adding a blank solvent to dissolve and dilute the mixture into a mixed impurity solution containing A, D, E of the impurity and 0.1 mu g of the decarbonylation avibactam in each 1 ml.
Positioning solution: taking appropriate amount of cefepime and abamectin reference substances, respectively adding blank solvent to dissolve and dilute to prepare positioning solution containing 0.1mg of cefepime and abamectin per 1 ml.
Test solution: dissolving 5 samples in water, transferring to a 250ml volumetric flask, taking a proper amount, diluting with a blank solvent to obtain 1ml of a sample solution containing 0.2mg of cefepime and 0.1mg of abamectin.
(2) Measurement results
Respectively and precisely measuring 10 mu l of each solution, carrying out HPLC detection, recording a chromatogram, and inspecting the system applicability of the method. Chromatograms were recorded and the results are shown in table 6.
TABLE 6 System suitability and impurity localization
Figure BDA0002970866450000111
The results show that: impurities are known to have no interference to the detection of the main component, and the separation degree between cefepime and abamectin meets the requirement.
Example four: methodological validation of assay methods
1. Specificity property
(1) Acid destruction
Blank solvent: phosphate buffer (0.68 g of potassium dihydrogen phosphate was weighed, dissolved in water and diluted to 1000ml) -acetonitrile (90:10), and the pH was adjusted to 5.0 with phosphoric acid (hereinafter referred to as "diluent").
Sample acid disruption solution: taking about 22.5mg of cefepime abamectin, precisely weighing, placing in a 10ml measuring flask, adding 20 mu L of 0.1mol/L hydrochloric acid, placing for 30min, adding 20 mu L of 0.1mol/L sodium hydroxide for neutralization, diluting to a scale with a diluent, precisely transferring 1ml into the 10ml measuring flask, diluting to the scale with the diluent, and shaking uniformly.
Cefepime acid disruption solution: taking about 20mg of cefepime hydrochloride/L-arginine raw material, precisely weighing, placing in a 100ml measuring flask, adding 20 mu L of 0.1mol/L hydrochloric acid, standing for 30min, adding 20 mu L of 0.1mol/L sodium hydroxide for neutralization, diluting with diluent to scale, and shaking up.
Abamebactam acid destroying solution: taking about 12.5mg of abamectin sodium as a raw material, placing the abamectin sodium in a 50ml measuring flask, adding 20 mu L of 0.1mol/L hydrochloric acid, standing for 30min, adding 20 mu L of 0.1mol/L sodium hydroxide for neutralization, diluting the abamectin sodium to a scale with a diluent, and shaking up. Precisely transferring 1ml, placing in a 10ml measuring flask, diluting with diluent to scale, and shaking.
Acid-breaking the blank solution: 20 mu L of 0.1mol/L hydrochloric acid is taken and put into a 10ml measuring flask, and is neutralized to be neutral by 20 mu L of 0.1mol/L sodium hydroxide, diluted to scale by diluent and shaken up.
(2) Alkali destruction
Sample alkali destruction solution: taking about 22.5mg of cefepime abamectin, accurately weighing, placing in a 10ml measuring flask, adding 20 mu L of 0.1mol/L sodium hydroxide, standing for 30min, adding 20 mu L of 0.1mol/L hydrochloric acid for neutralization, diluting to a scale with a diluent, accurately transferring 1ml, diluting to the scale with the diluent in the 10ml measuring flask, and shaking uniformly.
Cefepime base disruption solution: taking about 20mg of cefepime hydrochloride/L-arginine as a raw material, precisely weighing, putting into a 100ml measuring flask, adding 20 mu L of 0.1mol/L sodium hydroxide, standing for 30min, adding 20 mu L of 0.1mol/L hydrochloric acid for neutralization, diluting to a scale with a diluent, and shaking up.
Abamebactam alkali destroying solution: taking about 12.5mg of abamectin sodium as a raw material, putting the abamectin sodium into a 50ml measuring flask, adding 20 mu L of 0.1mol/L sodium hydroxide, standing for 30min, adding 20 mu L of 0.1mol/L hydrochloric acid for neutralization, diluting the abamectin sodium to a scale with a diluent, and shaking up. Precisely transferring 1ml, placing in a 10ml measuring flask, diluting with diluent to scale, and shaking.
Base destruction blank solution: 20 mu L of 0.1mol/L hydrochloric acid is taken and put into a 10ml measuring flask, and is neutralized to be neutral by 20 mu L of 0.1mol/L sodium hydroxide, diluted to scale by diluent and shaken up.
(3) Oxidative destruction
Sample oxidative destruction solution: taking about 22.5mg of cefepime abamectin, precisely weighing, placing in a 10ml measuring flask, adding 20 mu l of 0.03% hydrogen peroxide, placing for 90min, diluting to scale with diluent, precisely transferring 1ml into the 10ml measuring flask, diluting to scale with diluent, and shaking uniformly.
Oxidative destruction solution of cefepime: taking about 20mg of cefepime hydrochloride/L-arginine as a raw material, precisely weighing, putting into a 100ml measuring flask, adding 20 mu L of 0.03% hydrogen peroxide, standing for 90min, diluting to a scale with a diluent, and shaking uniformly.
Abamebactam oxidation destruction solution: taking about 12.5mg of the avibactam sodium raw material, putting the raw material into a 50ml measuring flask, adding 20 mu l of 0.03% hydrogen peroxide, standing for 90min, diluting to a scale with a diluent, and shaking up. Precisely transferring 1ml, placing in a 10ml measuring flask, diluting with diluent to scale, and shaking.
Oxidative destruction of the blank solution: and (3) taking 20 mu l of 0.03% hydrogen peroxide, placing the hydrogen peroxide into a 10ml measuring flask, diluting the hydrogen peroxide to the scale with the diluent, and shaking up the hydrogen peroxide.
(4) Destruction by light
Sample light destruction solution: taking about 22.5mg of cefepime abamectin, precisely weighing, placing in a 10ml measuring flask, placing in an ultraviolet illumination box for 24h after opening, diluting to a scale with a diluent, precisely transferring 1ml into the 10ml measuring flask, diluting to the scale with the diluent, and shaking uniformly.
Illumination disruption solution of cefepime: taking about 20mg of cefepime hydrochloride/L-arginine as a raw material, precisely weighing, placing the cefepime hydrochloride/L-arginine in a 100ml measuring flask, placing the cefepime/L-arginine in an ultraviolet illumination box for 24 hours after opening, diluting the cefepime/L-arginine to a scale with a diluent, and shaking up.
Avermebactam light damage solution: taking about 12.5mg of the avibactam sodium raw material, putting the raw material into a 50ml measuring flask, placing the measuring flask in an ultraviolet illumination box for 24 hours after opening, diluting the raw material to a scale with a diluent, and shaking up the raw material. Precisely transferring 1ml, placing in a 10ml measuring flask, diluting with diluent to scale, and shaking.
(5) High temperature destruction
Sample high temperature disruption solution: taking about 22.5mg of cefepime abamectin, precisely weighing, placing in a 10ml measuring flask, placing for 24h at 60 ℃, cooling, diluting to a scale with a diluent, precisely transferring 1ml to the 10ml measuring flask, diluting to the scale with the diluent, and shaking uniformly.
Cefepime high temperature disruption solution: taking about 20mg of cefepime hydrochloride/L-arginine as a raw material, precisely weighing, placing in a 100ml measuring flask, standing at 60 ℃ for 24h, cooling, diluting with diluent to scale, and shaking uniformly.
Abamebactam high-temperature destruction solution: taking about 12.5mg of the abamectin sodium raw material, placing the abamectin sodium raw material in a 50ml measuring flask, standing at 60 ℃ for 24h, cooling, diluting with a diluent to a scale, and shaking up. Precisely transferring 1ml, placing in a 10ml measuring flask, diluting with diluent to scale, and shaking.
(6) Analysis of results
And respectively and precisely measuring 10 mu l of each solution, detecting, recording a chromatogram, and inspecting the change condition of each main peak according to an area normalization method, wherein the results are shown in tables 7-8.
TABLE 7 Change in Abamebactam content in samples after disruption
Figure BDA0002970866450000141
TABLE 8 variation of cefepime content in samples after disruption
Figure BDA0002970866450000142
After the cefepime abamectin is damaged by acid, alkali, oxidation, illumination and high temperature, the separation degree between the main peaks meets the requirement, and the purity of the main peaks is 1000. The cefepime hydrochloride/L-arginine destruction does not interfere with the determination of the abamectin. The damage of the abamectin sodium does not interfere with the determination of cefepime.
Under the damage conditions of acid, high temperature and illumination, the content of the abamectin and the cefepime has no obvious change. Under base-destroying conditions, cefepime was reduced by about 3%; cefepime decreased by about 5% under oxidative damage conditions.
Under the destructive conditions of acid, alkali, oxidation, high temperature, illumination and the like, the separation degree between the abamectin and the cefepime in the sample is good, and the materials are basically conserved, which shows that the method can effectively detect the content of each component and has good specificity.
2. Verification of content analysis methods (detection limits and quantitation limits)
The impurity control solutions of known concentrations were diluted to low concentrations, respectively, and the measured signals were compared with those of the blank solvent (baseline noise) to calculate amounts that could be reliably detected. The limit of quantitation was determined with a signal-to-noise ratio of about 10:1 and the limit of detection was determined with a signal-to-noise ratio of about 3: 1.
Taking appropriate amount of reference substances of cefepime hydrochloride and abamectin respectively, precisely weighing, diluting with diluent step by step, precisely weighing 10 μ l, injecting sample, and determining quantitative limit according to signal-to-noise ratio S/N which is approximately equal to 10/1. Six parts are prepared in parallel to meet the requirements of accuracy and precision.
Taking appropriate amount of reference substances of cefepime hydrochloride and abamectin respectively, precisely weighing, diluting with diluent step by step, precisely weighing 10 μ l, injecting sample, and determining detection limit according to signal-to-noise ratio S/N which is approximately equal to 3/1. The results are shown in tables 9 to 10.
TABLE 9 quantitation Limit and detection Limit results
Figure BDA0002970866450000151
Figure BDA0002970866450000161
TABLE 10 repeatability results (peak area) for limits of quantitation
Figure BDA0002970866450000162
The results show that: the detection limit of the abamectin is 0.12 mu g/ml, and the quantification limit is 0.24 mu g/ml; the detection limit of cefepime is 0.22 mu g/ml, and the quantification limit is 0.45 mu g/ml.
Example five: determination of the content of the principal Components
(1) Chromatographic conditions
And (3) chromatographic column: inertsil ODS-3(4.6 mm. times.250 mm, 5 μm) column;
mobile phase: phosphate buffer (0.68 g potassium dihydrogen phosphate, dissolved in water and diluted to 1000ml) -acetonitrile (90:10) (pH adjusted to 5.0 with 2% phosphoric acid solution or 2% potassium hydroxide solution);
and (3) an elution mode: isocratic elution;
flow rate of mobile phase: 1.0 ml/min;
sample introduction amount: 10 mu l of the mixture;
detection wavelength: 195 nm.
(2) Solution preparation
Test solution: dissolving cefepime abamectin 5 in water, transferring to a 500ml measuring flask, adding water to dilute to a scale, mixing uniformly, precisely measuring 1ml, placing in a 100ml measuring flask, adding a mobile phase to dilute to the scale, and shaking uniformly.
Control solution: taking an avibactam reference substance of about 10mg, precisely weighing, placing in a 20ml measuring flask, dissolving with a mobile phase, diluting to a scale, and shaking up to obtain an avibactam reference substance stock solution; weighing cefepime reference substance about 10mg, precisely weighing, placing in a 100ml measuring flask, adding abamectin reference substance stock solution 5ml, precisely weighing, diluting with mobile phase to scale, and shaking.
(3) Measurement results
Precisely measuring the test solution and the reference solution, respectively injecting into a liquid chromatograph, and checking the results shown in Table 11.
TABLE 11 results of content measurement
Name (R) Content (%)
Abamebactam 95.4
Cefepime 106.1
As can be seen from the first to fifth embodiments, the method uses octadecylsilane chemically bonded silica as a filler of a chromatographic column, uses a phosphate buffer solution-acetonitrile with a certain proportion as a mobile phase, and can simultaneously detect the contents of two main components in cefepime and abamectin for injection of a compound preparation, thereby effectively controlling the quality of cefepime and abamectin for injection.
The protection content of the present invention is not limited to the above embodiments. Variations and advantages that may occur to those skilled in the art may be incorporated into the invention without departing from the spirit and scope of the inventive concept and the scope of the appended claims is intended to be protected.

Claims (10)

1. A method for determining the content of main components in cefepime abamectin for injection is characterized by comprising the following steps:
(1) chromatographic conditions are as follows:
octadecylsilane chemically bonded silica is used as a filler for the chromatographic column;
phosphate buffer solution-organic solvent is used as a mobile phase;
the flow velocity of the mobile phase is 0.9-1.1 ml/min;
the column temperature is 25-35 ℃;
the sample injection volume is 10 mul;
the detection wavelength is 192-198 nm;
(2) and respectively injecting the test solution and the reference solution into a liquid chromatograph, and recording the chromatogram.
2. The method according to claim 1, wherein the ratio of cefepime to abamectin in the sample solution is 1-8: 8-1;
preferably, the ratio of cefepime to abamectin is 2: 1.
3. The method as claimed in claim 1, wherein the chromatographic column in step (1) is an InertsilODS-3 chromatographic column.
4. The method according to claim 1, wherein the phosphate buffer-organic solvent in step (1) is phosphate buffer-acetonitrile;
preferably, the volume ratio of the phosphate buffer solution to the acetonitrile is 85-95: 5-15;
more preferably, the phosphate buffer-acetonitrile volume ratio is 90: 10.
5. The method according to claim 1, wherein the molar concentration of the potassium dihydrogen phosphate in the phosphate buffer solution in the step (1) is 0.001-0.01 mol/L;
preferably, the molar concentration of the potassium dihydrogen phosphate in the phosphate buffer is 0.005 mol/L.
6. The method according to claim 1, wherein the mobile phase phosphate buffer-acetonitrile in the step (1) has a pH value of 4.8-5.2;
preferably, the mobile phase phosphate buffer-acetonitrile has a pH of 5.0.
7. The method of claim 1, wherein the method is performed in an isocratic elution mode.
8. The method according to claim 1, wherein the flow rate of the mobile phase in step (1) is 1.0 ml/min.
9. The method according to claim 1, wherein the column temperature in step (1) is 30 ℃.
10. The method according to claim 1, wherein the detection wavelength in step (1) is 195 nm.
CN202110263040.5A 2021-03-11 2021-03-11 Method for determining content of main components in cefepime abamectin for injection Pending CN115078555A (en)

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