CN115097051B - Method for separating and detecting fosfomycin sodium and related substances in preparation thereof by using high performance liquid chromatography - Google Patents
Method for separating and detecting fosfomycin sodium and related substances in preparation thereof by using high performance liquid chromatography Download PDFInfo
- Publication number
- CN115097051B CN115097051B CN202210891678.8A CN202210891678A CN115097051B CN 115097051 B CN115097051 B CN 115097051B CN 202210891678 A CN202210891678 A CN 202210891678A CN 115097051 B CN115097051 B CN 115097051B
- Authority
- CN
- China
- Prior art keywords
- substance
- solution
- fosfomycin sodium
- mass concentration
- fosfomycin
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N2030/042—Standards
- G01N2030/045—Standards internal
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The invention relates to a method for separating and detecting fosfomycin sodium and related substances in a preparation thereof by using high performance liquid chromatography. The high performance liquid chromatography uses an electrospray detector (CAD) for detection, and adopts a chromatographic column using octadecyl bonded silica gel as a filler and formic acid aqueous solution as a mobile phase. The method can effectively separate the fosfomycin sodium and the related substances in the preparation thereof, has good specificity, high sensitivity, high accuracy and low detection cost, can eliminate citric acid interference in the fosfomycin sodium preparation, and provides effective guarantee for the quality control of the fosfomycin sodium and the preparation thereof.
Description
Technical Field
The invention belongs to the technical field of medical analysis, and relates to a method for separating and detecting related substances of a medicament. In particular to a method for separating and detecting fosfomycin sodium and related substances in a preparation thereof by using high performance liquid chromatography.
Background
Fosfomycin sodium, chemical name is (-) - (1R, 2S) -1,2-epoxypropylphosphonic acid disodium salt, molecular formula is C 3 H 5 Na 2 O 4 P has the following specific structural formula:
phosphomycin sodium was originally isolated from Streptomyces fragilis and was effective against both gram-positive and gram-negative bacteria. The action mechanism is to inhibit the synthesis of bacterial cell walls, and the antibacterial agent is a promising antibacterial agent. Specifically, fosfomycin sodium can bind to bacterial cell wall synthase, preventing the first step reaction of bacteria to synthesize cell wall using related substances, thereby playing a bactericidal role. The fosfomycin sodium is mainly suitable for the infection of urinary tract, skin, soft tissue, intestinal tract and other parts caused by sensitive gram-negative bacteria.
A fosfomycin sodium formulation is prepared from fosfomycin sodium and citric acid in a ratio of about 97:3, and the weight ratio of the prepared fosfomycin sodium for injection.
The content determination of fosfomycin sodium and related substances in the preparation thereof is an important part of the quality control. Specifically, fosfomycin sodium may contain impurities A-E, and the preparation further contains citric acid, wherein the molecular formula and structural formula of substances A-E are shown in Table 1 below.
Table 1: fosfomycin sodium and molecular formula and structural formula of related substances in preparation thereof
Substance A, listed in Table 1 above, had no UV absorption, and substance B, C, D, E had a weaker UV absorption. In addition, the main component fosfomycin sodium has no ultraviolet absorption.
At present, the quality control of fosfomycin sodium and the preparation thereof in Chinese Pharmacopoeia (CP), british Pharmacopoeia (BP), european Pharmacopoeia (EP) and the like only controls a substance A, but does not control other related substances. In addition, the quality standard of pharmacopoeia of various countries about fosfomycin sodium adopts a titration method for detecting the substance A, which lacks accuracy for trace detection.
Chinese patent CN 110887925a describes a high performance liquid chromatography method for detecting fosfomycin sodium substance, wherein the detector is an ultraviolet detector, and can only be used for detecting related substances with ultraviolet absorption, but not for detecting related substances without ultraviolet absorption.
Chinese patent CN 102841155A describes a method for detecting related substances in fosfomycin sodium by using high performance liquid chromatography, wherein a detector is a differential refraction detector, but the differential refraction detector has the problems of low sensitivity, poor repeatability, incapability of walking gradient conditions and the like.
Chinese patent CN 113916927A describes a method for measuring related substances in fosfomycin sodium by using nuclear magnetic resonance hydrogen spectroscopy. Although the method has high sensitivity and high accuracy, the method has poor universality and high detection cost due to the accessibility problem of detection instruments and equipment.
None of the reported methods for detecting fosfomycin sodium related substances can meet the quality control requirements for fosfomycin sodium related substances. In addition, as the fosfomycin sodium preparation contains citric acid as a pH regulator, the interference of the citric acid is also required to be eliminated when relevant substances are detected. In view of the fact that the related substances are important quality control indexes influencing the safety of the medicine, the development of the fosfomycin sodium and the detection method of the related substances in the preparation thereof, which have the advantages of high sensitivity, high accuracy and low detection cost and can eliminate citric acid interference, has very important significance for the quality control.
Disclosure of Invention
Aiming at the defects in the prior art, the invention aims to provide a method for separating and detecting fosfomycin sodium and related substances in a preparation thereof by using a high performance liquid chromatography, which has the advantages of high sensitivity, high accuracy and low detection cost, and can eliminate citric acid interference.
The purpose of the invention is realized as follows: a method for separating and detecting fosfomycin sodium and related substances in its preparation by high performance liquid chromatography for simultaneous separation and detection
Substance A and the structural formula is->
Substance B of formula (II)>
Substance C and structural formula->
In (b) and a structural formula>
Substance E according to (1), characterized in that:
the chromatographic conditions are as follows:
detecting using an electrospray detector (CAD); adopting a chromatographic column with octadecyl bonded silica gel as a filler, wherein the temperature of the chromatographic column is 25-35 ℃; taking a formic acid aqueous solution with the volume concentration of 0.1-0.4% as a mobile phase, wherein the flow rate of the mobile phase is 0.4-0.6mL/min, and isocratically eluting;
the method comprises the following steps:
(1) Solution preparation
Preparing a test solution:
preparing fosfomycin sodium or a preparation thereof to be measured, and dissolving in water;
preparation of control solution:
transferring a certain amount of test solution, and adding water for quantitative dilution so that the mass concentration of fosfomycin sodium in the diluted solution is 1% of the mass concentration of fosfomycin sodium in the test solution;
(2) Assay method
Measuring a test solution and a control solution with the same volume, respectively injecting the test solution and the control solution into a liquid chromatograph, recording a chromatogram according to chromatographic conditions, and calculating the contents of a substance A, a substance B, a substance C, a substance D and a substance E in fosfomycin sodium or a preparation thereof by adopting a main component self-control method, wherein the specific calculation formula is as follows:
percent substance (%) = (Au/As) × (1/100) × 100%
Wherein:
au is the peak area of each substance peak in the test solution;
as is the peak area of fosfomycin in the control solution.
Compared with the prior art, the invention has the advantages and beneficial effects that:
1. the chromatographic condition of the invention is detected by an electrospray detector, can detect related substances without ultraviolet absorption, has high sensitivity, higher accuracy than a titration method and a differential refraction detection method, and lower cost than a nuclear magnetic resonance hydrogen spectrum detection method.
2. The high performance liquid chromatography adopts a chromatographic column with octadecyl bonded silica gel as a filling agent, and takes a formic acid aqueous solution as a mobile phase, so that the main peak of the fosfomycin can be effectively separated from the peaks A, B, C, D and E of related substances and the peak of citric acid, the separation degree is more than 1.5, and the method has good specificity.
Drawings
FIG. 1 is an HPLC chromatogram of a blank solution according to the chromatographic conditions of example 1;
FIG. 2 is an HPLC chromatogram of a solution of a sodium fosfomycin system applicability according to the chromatographic conditions of example 1;
FIG. 3 is an HPLC chromatogram of a test solution of fosfomycin sodium according to the chromatographic conditions of example 1;
FIG. 4 is an HPLC chromatogram of a test solution of a fosfomycin sodium preparation according to the chromatographic conditions of example 1;
FIG. 5 is an HPLC chromatogram of a sensitive solution of fosfomycin sodium according to the chromatographic conditions of example 1;
FIG. 6 is an HPLC chromatogram of a localized solution of fosfomycin sodium and each of related substances A-E according to the chromatographic conditions of example 1;
FIG. 7 is an HPLC chromatogram of a citric acid-localized solution according to the chromatographic conditions of example 1.
Detailed Description
In the present invention, the "test sample" refers to fosfomycin sodium or a preparation thereof to be tested for the relevant substance.
In the present invention, a "fosfomycin sodium preparation" refers to a mixture of fosfomycin sodium and citric acid in a ratio of about 97:3 weight ratio of prepared fosfomycin sodium for injection.
All numbers expressing quantities, percentages or proportions used in the specification and claims are to be understood as being modified in all instances by the term "about". In this regard, the term "about" as used herein may include ranges of the numerical values ± 10%, ± 8%, ± 6%, ± 5%, ± 4%, ± 3%, ± 2%, ± 1% or ± 0.5%.
The invention provides a method for separating and detecting fosfomycin sodium and related substances in a preparation thereof by using high performance liquid chromatography, which is used for simultaneously separating and detecting substances with structural formulas of
Substance A and the structural formula is->
Of (2)B. Structural formula is->
Is substance C and has a structural formula of>
In (b) and a structural formula>
Substance E according to (1), characterized in that:
the chromatographic conditions are as follows:
detecting using an electrospray detector (CAD); a chromatographic column using octadecyl bonded silica gel as a filler is adopted, and the temperature of the chromatographic column is 25-35 ℃; taking a formic acid aqueous solution with the volume concentration of 0.1-0.4% as a mobile phase, wherein the flow rate of the mobile phase is 0.4-0.6mL/min, and isocratically eluting;
the method comprises the following steps:
(1) Solution preparation
Preparing a test solution:
preparing fosfomycin sodium or a preparation thereof to be measured, and dissolving in water;
preparation of control solution:
transferring a certain amount of test solution, and adding water for quantitative dilution to ensure that the mass concentration of the fosfomycin sodium in the diluted solution is 1% of the mass concentration of the fosfomycin sodium in the test solution;
(2) Assay method
Measuring a test solution and a control solution with the same volume, respectively injecting the test solution and the control solution into a liquid chromatograph, recording a chromatogram according to chromatographic conditions, and calculating the contents of a substance A, a substance B, a substance C, a substance D and a substance E in fosfomycin sodium or a preparation thereof by adopting a main component self-control method, wherein the specific calculation formula is as follows:
percent substance (%) = (Au/As) × (1/100) × 100%
Wherein:
au is the peak area of each substance peak in the test solution;
as is the peak area of fosfomycin in the control solution.
In a specific embodiment of the invention, the chromatographic column using octadecyl bonded silica gel as a filler is selected from Waters
T3 (150X 4.6mm,3 μm). Preferably, the column temperature of the chromatography column is 25 ℃, 30 ℃ or 35 ℃.
Preferably, an aqueous solution of formic acid with a concentration of 0.2 to 0.4%, in particular 0.2%, by volume is used as the mobile phase. Further preferably, the mobile phase flow rate is 0.45-0.55mL/min, such as 0.45mL/min, 0.5mL/min or 0.55mL/min. Preferably, the injection volume of the mobile phase is 10-40. Mu.L, preferably 15-30. Mu.L, especially 20. Mu.L.
In a preferred embodiment of the invention, the concentration by mass of fosfomycin sodium in the fosfomycin sodium test solution is 0.8-2mg/mL, preferably 1-1.5mg/mL, especially 1mg/mL; the mass concentration of the fosfomycin sodium in the test solution of the fosfomycin sodium preparation is 2-10mg/mL, preferably 4-8mg/mL, and particularly 5mg/mL.
In a specific embodiment of the invention, the sample injection volume of the fosfomycin sodium and the preparation test solution and the corresponding control solution thereof is 5-40. Mu.L, preferably 10-30. Mu.L, more preferably 15-25. Mu.L, especially 20. Mu.L.
In the process of the invention, a mixed aqueous solution comprising fosfomycin sodium and substance a is also used as the system-adapting solution. Preferably, in the system-adapted solution, the ratio by mass of fosfomycin sodium to substance a is 80-120. In particular, in the systemically adaptive solution, the concentration by mass of fosfomycin sodium is from 0.8 to 2mg/mL, preferably from 1 to 1.5mg/mL, especially 1mg/mL, and the concentration by mass of substance A is from 5 to 20. Mu.g/mL, preferably from 10 to 15. Mu.g/mL, especially 10. Mu.g/mL. The system adaptive solution is used and the separation effect between the fosfomycin sodium peak and the substance A peak is observed, so that a chromatographic system can be tested and adjusted, and comprehensive characteristics in the aspects of electrical signals, analysis operation, samples and the like can be comprehensively detected.
In the method of the invention, a fosfomycin sodium sensitivity solution is also used to detect the sensitivity of the method of the invention. In the invention, the mass concentration of the fosfomycin sodium in the sensitivity solution is 0.05 percent of the mass concentration of the fosfomycin sodium in the test solution. Specifically, the sensitivity solution was prepared by quantitatively diluting a fosfomycin sodium control solution 20-fold with water. In a specific embodiment, the concentration by mass of fosfomycin sodium in the sensitivity solution is 0.5 μ g/mL.
In the method of the invention, a localization solution is also used to determine the peak position of each substance. The positioning solution is prepared by respectively adding water into stock solutions of a certain amount of fosfomycin sodium, a substance A, a substance B, a substance C, a substance D or a substance E for quantitative dilution. Preferably, the concentration by mass of fosfomycin sodium in the fosfomycin sodium locating solution is 0.1-0.6. Mu.g/mL, preferably 0.2-0.4. Mu.g/mL, especially 0.3. Mu.g/mL; in the substance A localization solution, the mass concentration of the substance A is 0.2-1. Mu.g/mL, preferably 0.4-0.8. Mu.g/mL, in particular 0.7. Mu.g/mL; the mass concentration of substance B in the substance B localization solution is 0.6-2. Mu.g/mL, preferably 0.8-1.5. Mu.g/mL, in particular 1. Mu.g/mL; in the substance C localization solution, the mass concentration of the substance C is 0.1-0.8. Mu.g/mL, preferably 0.2-0.6. Mu.g/mL, in particular 0.5. Mu.g/mL; the mass concentration of substance D in the substance D localization solution is 0.1-0.6. Mu.g/mL, preferably 0.2-0.5. Mu.g/mL, in particular 0.4. Mu.g/mL; in the substance E localization solution, the mass concentration of the substance E is 0.1 to 1. Mu.g/mL, preferably 0.2 to 0.8. Mu.g/mL, in particular 0.6. Mu.g/mL.
In the method of the present invention, a citric acid localization solution is also formulated to determine the peak position of citric acid. The citric acid positioning solution is prepared by adding water to a certain amount of citric acid for dissolving and quantitatively diluting. Preferably, the mass concentration of the citric acid positioning solution is 0.8-2mg/mL, preferably 1-1.5mg/mL, especially 1mg/mL.
In a particular embodiment of the invention, the system adaptation solution, the sensitivity solution and the corresponding localization solution are each introduced in a volume of 10-40. Mu.L, preferably 10-30. Mu.L, more preferably 15-25. Mu.L, in particular 20. Mu.L.
The process of the present invention will be described and illustrated in more detail with reference to specific examples. Those skilled in the art will appreciate that these examples are provided for illustrative purposes only and do not constitute any limitation on the scope of the present invention.
1. Chromatographic conditions
Example 1
The instrument comprises the following steps: high performance liquid chromatograph-electrospray detector (available from Saimer Feishale technologies Co., ltd., type Corona Veo CAD)
And (3) chromatographic column: waters
T3(150*4.6mm，3μm)/>
Column temperature: 30 ℃;
mobile phase: 0.2% by volume aqueous formic acid, isocratic elution
Flow rate: 0.5mL/min;
sample injection volume: 20 μ L.
Example 2
The chromatographic conditions were the same as in example 1 except that the column temperature was 25 ℃.
Example 3
The chromatographic conditions were the same as in example 1 except that the column temperature of the column was 35 ℃.
Example 4
The chromatographic conditions were the same as in example 1 except that the flow rate of the mobile phase was 0.45mL/min.
Example 5
The chromatographic conditions were the same as in example 1 except that the flow rate of the mobile phase was 0.55mL/min.
2. Sample solution preparation
2.1 Phosphomycin sodium
Test solution: taking a proper amount of fosfomycin sodium, precisely weighing, adding water to dissolve, quantitatively diluting to prepare a solution containing about 1mg of fosfomycin sodium in every 1ml, shaking up, filtering, and taking a subsequent filtrate as a test solution of fosfomycin sodium (sample batch No. FFE 10411).
Control solution: precisely transferring a proper amount of the test solution, adding water for quantitative dilution so that the mass concentration of the fosfomycin sodium in the diluted solution is 1% of the mass concentration of the fosfomycin sodium in the test solution, and shaking up to be used as a control solution of the fosfomycin sodium.
2.2 Fosfomycin sodium preparation
Test solution: taking a proper amount of fosfomycin sodium preparation, precisely weighing, dissolving in water, quantitatively diluting to obtain a solution containing 5mg fosfomycin sodium per 1ml, shaking, filtering, and taking the subsequent filtrate as the test solution (sample batch No. 02018) of fosfomycin sodium preparation.
Control solution: precisely transferring a proper amount of the test solution, adding water for quantitative dilution so that the mass concentration of the fosfomycin sodium in the diluted solution is 1% of the mass concentration of the fosfomycin sodium in the test solution, and shaking up to be used as a control solution of the fosfomycin sodium preparation.
3. Determination of related substances
20. Mu.l of the test solution and the corresponding control solution of fosfomycin sodium and the preparation thereof prepared as above were precisely measured, and injected into a liquid chromatograph, and chromatograms were recorded according to the chromatographic conditions of examples 1 to 5, respectively, wherein the chromatograms of example 1 are shown in FIGS. 3 and 4. Then, the content of each related substance in the test solution is calculated according to the main component self-contrast method, and the specific calculation formula is as follows:
percent content of substance (%) = (Au/As) × (1/100) × 100%
Wherein
Au: peak area of peak of each substance in the test solution;
as: peak area of fosfomycin in control solution.
The measurement results of the contents of the related substances in the test solution of fosfomycin sodium and the preparation thereof are shown in the following table 2:
TABLE 2 measurement results of contents of related substances in test solutions of fosfomycin sodium and preparation thereof
Remarking: "/" indicates that the relevant substance in the test solution of the lot is below the detection limit.
4. Verification of the inventive method
4.1 preparation of related solution:
(1) Blank solution: water (W)
(2) System applicability solution:
taking a proper amount of fosfomycin sodium and the substance A, adding water to dissolve, quantitatively diluting to prepare a solution containing about 1mg of fosfomycin sodium and 10 mu g of the substance A in each 1ml, and shaking up to obtain a system applicability solution.
(3) Sensitivity solution:
precisely transferring a proper amount of fosfomycin sodium control solution prepared in the step 2.1, adding water for quantitatively diluting by 20 times, and shaking up to obtain a sensitivity solution.
(4) Localization solution of fosfomycin sodium and related substances a-E:
precisely weighing appropriate amounts of fosfomycin sodium, a substance A, a substance B, a substance C, a substance D and a substance E stock solution, and respectively adding water to quantitatively dilute to prepare a solution containing about 0.3 mu g of fosfomycin sodium, 0.7 mu g of the substance A, 1 mu g of the substance B, 0.5 mu g of the substance C, 0.4 mu g of the substance D and 0.6 mu g of the substance E in each 1ml, wherein the solution is used as a positioning solution of the fosfomycin sodium and related substances A-E.
(5) Citric acid localization solution:
dissolving appropriate amount of citric acid in water, quantitatively diluting to obtain solution containing citric acid 1mg per 1ml, shaking, and filtering.
4.2 verification method
And (3) respectively injecting the blank solution, the system applicability solution, the sensitivity solution and the corresponding positioning solution into a liquid chromatograph, recording a chromatogram according to the chromatographic conditions of the embodiment 1, and specifically referring to figures 1-2 and figures 5-7, and calculating the separation degree of the fosfomycin and various related substances in the chromatogram. The results are shown in tables 3-4 below.
TABLE 3 System applicability and sensitivity results
TABLE 4 results on the localization of substances
From the above results, the separation degree of the substance A and the fosfomycin peak in the system applicability solution is more than 1.5, the separation degree between each peak and the adjacent chromatographic peak in each related substance location map is more than 1.5, no peak is interfered in the blank solution, the signal-to-noise ratio of the sensitivity solution is more than 10, and the system is in accordance with the regulations. The method for determining the fosfomycin sodium related substances by the high performance liquid chromatography has good sensitivity and specificity.
Claims (31)
1. A method for separating and detecting fosfomycin sodium and related substances in its preparation by high performance liquid chromatography for simultaneous separation and detection
The substance A has a structural formula
Substance B with the structural formula
Substance C, structural formula
Substance D and structural formula
Substance E according to (1), characterized in that:
the chromatographic conditions are as follows:
detecting using an electrospray detector (CAD); a chromatographic column using octadecyl bonded silica gel as a filler is adopted, and the temperature of the chromatographic column is 25-35 ℃; taking a formic acid aqueous solution with the volume concentration of 0.1-0.4% as a mobile phase, wherein the flow rate of the mobile phase is 0.4-0.6mL/min, and isocratic elution is carried out;
the method comprises the following steps:
(1) Solution preparation
Preparing a test solution:
preparing fosfomycin sodium or a preparation thereof to be measured, and dissolving in water;
preparation of control solution:
transferring a certain amount of test solution, and adding water for quantitative dilution so that the mass concentration of fosfomycin sodium in the diluted solution is 1% of the mass concentration of fosfomycin sodium in the test solution;
(2) Assay method
Measuring a test solution and a control solution with the same volume, respectively injecting the test solution and the control solution into a liquid chromatograph, recording a chromatogram according to chromatographic conditions, and calculating the contents of a substance A, a substance B, a substance C, a substance D and a substance E in fosfomycin sodium or a preparation thereof by adopting a main component self-control method, wherein the specific calculation formula is as follows:
percent substance (%) = (Au/As) × (1/100) × 100%
Wherein:
au is the peak area of each substance peak in the test solution;
as is the peak area of fosfomycin in the control solution.
2. The method according to claim 1, characterized in that a formic acid aqueous solution with a volume concentration of 0.2-0.4% is used as the mobile phase.
3. The method according to claim 2, characterized in that a 0.2% strength by volume aqueous formic acid solution is used as the mobile phase.
4. The method of claim 1, wherein the flow rate of the mobile phase is 0.45-0.55mL/min.
5. The method of claim 4, wherein the flow rate of the mobile phase is 0.45mL/min, 0.5mL/min, or 0.55mL/min.
6. The method of claim 1, wherein, in the case where the test solution is prepared from fosfomycin sodium, the mass concentration of fosfomycin sodium in the test solution is 0.8-2mg/mL; in the case where the test solution is prepared from a fosfomycin sodium preparation, the mass concentration of fosfomycin sodium in the test solution is 2-10mg/mL.
7. The method of claim 6, wherein where the test solution is prepared from fosfomycin sodium, the mass concentration of fosfomycin sodium in the test solution is 1-1.5mg/mL.
8. The method of claim 7, wherein, in the case where the test solution is prepared from fosfomycin sodium, the mass concentration of fosfomycin sodium in the test solution is 1mg/mL.
9. The method of claim 6, wherein, in the case where the test solution is prepared from a fosfomycin sodium formulation, the mass concentration of fosfomycin sodium in the test solution is 4-8mg/mL.
10. The method of claim 9, wherein, in the case where the test solution is prepared from a fosfomycin sodium formulation, the mass concentration of fosfomycin sodium in the test solution is 5mg/mL.
11. The method according to any one of claims 1 to 10, characterized in that a mixed aqueous solution comprising fosfomycin sodium and substance a is used as the system-adapting solution.
12. The method of claim 11, wherein in the system-adapted solution, the ratio of fosfomycin sodium to substance a by mass is 80-120.
13. The method of claim 12, wherein in the system-adapted solution, the ratio of fosfomycin sodium to substance a by mass is 100.
14. The method according to any one of claims 1 to 10, wherein a fosfomycin sodium sensitivity solution is also used, the mass concentration of fosfomycin sodium in the sensitivity solution being 0.05% of the mass concentration of fosfomycin sodium in the test solution.
15. The method of any one of claims 1 to 10, wherein a localization solution of fosfomycin sodium, substance a, substance B, substance C, substance D or substance E is also used to determine the peak position of each substance.
16. The method of claim 15, wherein the concentration by mass of fosfomycin sodium in the fosfomycin sodium locating solution is 0.1-0.6 μ g/mL; in the substance A positioning solution, the mass concentration of the substance A is 0.2-1 mug/mL; in the substance B positioning solution, the mass concentration of the substance B is 0.6-2 mug/mL; in the substance C positioning solution, the mass concentration of the substance C is 0.1-0.8 mug/mL; in the substance D positioning solution, the mass concentration of the substance D is 0.1-0.6 mug/mL; in the substance E positioning solution, the mass concentration of the substance E is 0.1-1 mu g/mL.
17. The method of claim 16 wherein the concentration of fosfomycin sodium by mass in the fosfomycin sodium locating solution is 0.2-0.4 μ g/mL.
18. The method of claim 17 wherein the concentration of fosfomycin sodium by mass in the fosfomycin sodium locating solution is 0.3 μ g/mL.
19. The method of claim 16, wherein the mass concentration of substance a in the substance a localization solution is 0.4-0.8 μ g/mL.
20. The method of claim 19, wherein the mass concentration of substance a in the substance a localization solution is 0.7 μ g/mL.
21. The method of claim 16, wherein the mass concentration of substance B in the substance B localization solution is 0.8-1.5 μ g/mL.
22. The method of claim 21, wherein the mass concentration of substance B in the substance B localization solution is 1 μ g/mL.
23. The method of claim 16, wherein the mass concentration of substance C in the substance C localization solution is 0.2-0.6 μ g/mL.
24. The method of claim 23, wherein the mass concentration of substance C in the substance C localization solution is 0.5 μ g/mL.
25. The method of claim 16, wherein the mass concentration of substance D in the substance D localization solution is 0.2-0.5 μ g/mL.
26. The method of claim 25, wherein the mass concentration of substance D in the substance D localization solution is 0.4 μ g/mL.
27. The method of claim 16, wherein the mass concentration of substance E in the substance E localization solution is 0.2-0.8 μ g/mL.
28. The method of claim 27, wherein the mass concentration of substance E in the substance E localization solution is 0.6 μ g/mL.
29. The method according to any one of claims 1 to 10, further formulating a citric acid localization solution at a mass concentration of 0.8-2mg/mL to determine the peak position of citric acid.
30. The method of claim 29, wherein the citric acid positioning solution has a mass concentration of 1-1.5mg/mL.
31. The method of claim 30, wherein the citric acid positioning solution has a mass concentration of 1mg/mL.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210891678.8A CN115097051B (en) | 2022-07-27 | 2022-07-27 | Method for separating and detecting fosfomycin sodium and related substances in preparation thereof by using high performance liquid chromatography |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210891678.8A CN115097051B (en) | 2022-07-27 | 2022-07-27 | Method for separating and detecting fosfomycin sodium and related substances in preparation thereof by using high performance liquid chromatography |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115097051A CN115097051A (en) | 2022-09-23 |
| CN115097051B true CN115097051B (en) | 2023-03-24 |
Family
ID=83298974
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210891678.8A Active CN115097051B (en) | 2022-07-27 | 2022-07-27 | Method for separating and detecting fosfomycin sodium and related substances in preparation thereof by using high performance liquid chromatography |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115097051B (en) |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102841155B (en) * | 2012-09-25 | 2014-11-26 | 东北制药集团股份有限公司 | Method for detecting related substances in fosfomycin sodium by high-performance liquid chromatography |
| ES2784076B2 (en) * | 2019-03-21 | 2021-04-12 | Laboratorios Ern S A | METHOD FOR THE DETECTION AND QUANTIFICATION OF PHOSPHOMYCIN AND ITS IMPURITIES AND / OR DEGRADATION PRODUCTS |
| CN110398548B (en) * | 2019-07-03 | 2022-04-19 | 山西仟源医药集团股份有限公司 | High performance liquid chromatography separation and determination method for related impurities in fosfomycin trometamol raw material medicine and preparation thereof and application |
-
2022
- 2022-07-27 CN CN202210891678.8A patent/CN115097051B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN115097051A (en) | 2022-09-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102798678B (en) | Detection method and content determining method of sodium calcium edetate in pantoprazole sodium for injecting | |
| CN109060973B (en) | Method for detecting ethylenediamine in lipoic acid injection | |
| CN114047271B (en) | Method for detecting related substances in ceftazidime preparation for injection | |
| CN113484430B (en) | Method for measuring related substances of L-alanine isopropyl ester hydrochloride by adopting high performance liquid chromatography | |
| CN115097051B (en) | Method for separating and detecting fosfomycin sodium and related substances in preparation thereof by using high performance liquid chromatography | |
| CN113588837A (en) | Detection method of moxifloxacin hydrochloride related substances | |
| CN111024831A (en) | Method for separating moxifloxacin hydrochloride and impurities thereof by high performance liquid chromatography | |
| CN113533569A (en) | Method for detecting related substances in antibacterial eye drops | |
| CN111965273A (en) | HPLC (high Performance liquid chromatography) method for detecting genotoxic impurities in candesartan cilexetil | |
| CN110988167A (en) | Method for detecting genotoxic impurities in gatifloxacin | |
| CN112924601B (en) | Method for detecting lincomycin impurity E in lincomycin hydrochloride injection by high performance liquid chromatography-evaporative light method | |
| CN115266998A (en) | Method for detecting content of sodium ions in sodium salt molecules | |
| CN114354810A (en) | Method for detecting impurity N in clindamycin phosphate and method for separating impurity | |
| CN111351886B (en) | Method for determining impurity and main medicine content in phenol sulfoethylamine medicine | |
| CN113607842A (en) | Method for identifying and measuring content of lactic acid in levofloxacin lactate | |
| CN112881538A (en) | Detection method for impurities and enantiomers in fudosteine and fudosteine tablets | |
| CN117890496B (en) | Method for detecting related substances of compound preparation of novel oral solution of guaifenesin | |
| CN114137102B (en) | Method for measuring clenbuterol hydrochloride content in ambroxol oral solution | |
| CN113777180B (en) | Method for simultaneously detecting sitafloxacin and various isomer impurities thereof | |
| CN111307962B (en) | Method for detecting 3-dimethylamino ethyl acrylate in moxifloxacin hydrochloride | |
| CN115166083B (en) | Method for detecting fosfomycin trometamol related substances and application thereof | |
| CN111157647B (en) | Method for detecting arbekacin sulfate and impurities thereof | |
| CN114577944B (en) | Detection method for related substances in 5-bromo-2-chloro-4' -ethoxydiphenylmethane | |
| CN114295750B (en) | Method for detecting related substances in ursodeoxycholic acid synthesis | |
| CN115993412A (en) | HPLC detection method for colchicine impurity F |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |