CN112946138A

CN112946138A - Method for determining ceftazidime related substances

Info

Publication number: CN112946138A
Application number: CN202110200980.XA
Authority: CN
Inventors: 张洁; 陈海刚; 王玉娟; 傅超婷; 李富志
Original assignee: Hainan General Sanyang Pharmaceutical Co ltd
Current assignee: Hainan General Sanyang Pharmaceutical Co ltd
Priority date: 2021-02-23
Filing date: 2021-02-23
Publication date: 2021-06-11

Abstract

The invention relates to a method for measuring ceftazidime related substances, which adopts a high performance liquid chromatography to detect a ceftazidime sample solution, wherein a mobile phase of the high performance liquid chromatography contains: mobile phase a, phosphate buffer, and mobile phase B, acetonitrile. The invention provides a detection method of ceftazidime related substances capable of eliminating ghost peaks.

Description

Method for determining ceftazidime related substances

Technical Field

The invention relates to the field of ceftazidime detection, in particular to a method for determining ceftazidime related substances.

Background

Ceftazidime is a semi-synthetic third generation cephalosporin. Similar to other third-generation cephalosporins, they have a broad response against both gram-positive and gram-negative bacteria. In contrast, it is effective against pseudomonas aeruginosa, but is less resistant to gram-positive bacteria and therefore not useful as such an infection. C₂₂H₂₂N₆O₇S₂Ceftazidime has good stability to beta-lactamase, low drug resistance probability under clinical reasonable use, less side reaction, third-generation broad-spectrum cephalosporin, is stable to a plurality of lactamase, has strong bactericidal action on gram positive bacteria, gram negative bacteria and anaerobic strains, has high efficiency on pseudomonas aeruginosa, is the only cephalosporin antibiotic capable of replacing aminoglycosides, and is called as the fourth-generation cephalosporin antibiotic. The traditional Chinese medicine composition is clinically used for treating severe infection (such as septicemia, meningitis, bacteremia and the like) caused by sensitive bacteria, respiratory tract infection (such as pneumonia, bronchitis and the like), ear-nose-throat infection, skin and soft tissue infection, urinary system infection, gastrointestinal, biliary and abdominal infection, bone and joint infection and the like.

During the detection process of related substances of ceftazidime, ghost peaks are generated, and the accuracy of the detection result is influenced.

Disclosure of Invention

The invention mainly aims to provide a detection method of ceftazidime related substances capable of eliminating ghost peaks.

In order to solve the technical problems, the invention adopts the technical scheme that: a method for determining ceftazidime related substances is characterized in that a ceftazidime sample solution is detected by adopting a high performance liquid chromatography, wherein a mobile phase of the high performance liquid chromatography contains: mobile phase a, phosphate buffer, and mobile phase B, acetonitrile.

Further, a ghost trap was used.

Further, the preparation method of the mobile phase A comprises the following steps: the mobile phase A was prepared by dissolving 3.6g of sodium hydrogen phosphate and 1.4g of potassium dihydrogen phosphate in water, diluting to 1000ml, and adjusting the pH with phosphoric acid solution.

Further, the pH was adjusted with a 10% phosphoric acid solution.

Further, the pH of the mobile phase A was 3.4. + -. 0.05.

Further, the high performance liquid chromatography was performed with gradient elution under the following conditions:

gradient of mobile phase

T(min)	A(％)	B(％)
			0	96	4
4	89	11
			5	89	11
8	84	16
			11	80	20
15	50	50
			18	20	80
25	20	80
			26	96	4
35	96	4

Furthermore, the specification of the chromatographic column is that octadecylsilane chemically bonded silica is used as a filler, the column temperature is 40 ℃, the flow rate is 1.3mL/min, the detection wavelength is 254nm, and the sample injection volume is 10 muL.

The invention has the beneficial effects that: the method of the invention can reduce the interference of ghost peaks to the main peak, and lead the detection result to be accurate and reliable. The mobile phase A phosphate buffer is mainly used for adjusting the pH value without influencing the experimental reaction so as to ensure that the chemical reaction of the experiment is carried out under the optimal condition. The viscosity of the acetonitrile in the mobile phase B is small, and the mass transfer impedance of the mobile phase is small from the perspective of a rate equation, so that the improvement of column efficiency is facilitated, the peak width is narrow, and the peak shape is good.

Drawings

The present invention will be described in further detail with reference to the accompanying drawings and specific embodiments.

FIG. 1 is a map of preparatory example 1;

FIG. 2 is a map of preparatory example 2.

Detailed Description

The technical solution in the embodiments of the present invention is clearly and completely described below with reference to the drawings in the embodiments of the present invention. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, but the present invention may be practiced in other ways than those specifically described and will be readily apparent to those of ordinary skill in the art without departing from the spirit of the present invention, and therefore the present invention is not limited to the specific embodiments disclosed below.

Preparative example 1:

a ghost trap was used.

Preparation of a test solution: taking about 0.15g of ceftazidime product for injection, putting the ceftazidime product for injection into a measuring flask, adding 5ml of acetonitrile, adding a water solution, diluting to 100ml, and shaking uniformly to obtain the ceftazidime injection.

Preparation of a blank solvent: measuring water to 100ml, measuring 5ml, and placing in another measuring bottle;

the detection method comprises the following steps: high performance liquid chromatography using the standard of the present invention with octadecylsilane bonded silica gel as a packing (Thermo Acclaim TM 120250 mm X4.6 mm, 5 μm).

Mobile phase A: phosphate buffer (taking 3.6g of dodecahydrate and disodium hydrogen phosphate and 1.4g of monopotassium phosphate, adding water to dissolve and dilute the mixture to 1000ml, and adjusting the pH value to 3.4 +/-0.05 by using 10% phosphoric acid solution); mobile phase B: and (3) acetonitrile.

Detection wavelength: 254 nm; column temperature: 40 ℃; flow rate: 1.3ml/min calculation method: 1% self-control injection volume: 10 μ l.

FIG. 1 is a graph showing the results of the experiment in preparative example 1. The position near the front of the test solution is the impurity in the solution, the highest peak is ceftazidime, and the position near the back of the atlas shows a peak. And the peak appears at the later position in the blank solvent, and is similar to the peak at the later position of the test solution, so that the later peak of the test solution can be judged as a ghost peak.

Preparative example 2:

the ceftazidime lot number used in preparative example 2 was the same as that used in preparative example 1, except that the detection method used was different.

The detection method comprises the following steps: conventional high performance liquid chromatography is used.

FIG. 2 is a graph showing the results of the experiment in preparative example 2. The blank solvent is relatively flat and has no obvious peak, while the sample solution has only impurity peak and main peak near the front position and no peak near the back position.

The results of the pre-preparation examples 1 and 2 show that the influence of the ghost peak on the experiment is eliminated in the process of detecting the ceftazidime related substance by using the method, so that the detection result is more accurate.

Example 1:

Preparation of a test solution: taking and injecting 0.15g of ceftazidime product, putting the ceftazidime product into a measuring flask, adding 5ml of acetonitrile, adding a water solution, diluting to 100ml, and shaking up to obtain the ceftazidime injection.

Preparation of control solution: precisely measuring 1ml of the test solution, placing in a measuring flask, adding 5ml of acetonitrile, diluting with water to 100ml, shaking, precisely measuring 5ml, placing in another measuring flask, diluting with water to 50ml, and shaking.

Preparation of system applicability solution: taking about 3mg of mixed impurity reference substance (containing impurities A, G and F), adding acetonitrile 0.5ml, adding water 1.5ml for dissolving to obtain the final product.

Detection wavelength: 254 nm; column temperature: 40 ℃; flow rate: 1.3ml/min injection volume: 10 mu l of the mixture; the linear elution gradient is shown in the following table:

gradient of mobile phase

Example 2:

example 2 is consistent with the assay method of example 1, except that the drug product is from a different batch.

gradient of mobile phase

The results of the study were compared with the data to show that: compared with the standard of Chinese pharmacopoeia 2020, the method can reduce the occurrence of ghost peaks and ensure the accuracy of the detection result.

The foregoing description is only of the preferred embodiments of the present invention, and it should be understood that the described embodiments are only a few, and not all, of the embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Claims

1. A method for determining ceftazidime related substances is characterized in that a ceftazidime sample solution is detected by adopting a high performance liquid chromatography, wherein a mobile phase of the high performance liquid chromatography contains a mobile phase A and a mobile phase B, the mobile phase A is phosphate buffer solution, and the mobile phase B is acetonitrile.

2. The method of claim 1, wherein the ghost peak trap is used for trapping ghost peaks.

3. The method for assaying ceftazidime-related substance according to claim 1, wherein the mobile phase a is prepared by: 3.6g of disodium hydrogen phosphate dodecahydrate and 1.4g of potassium dihydrogen phosphate are taken, dissolved in water and diluted to 1000ml, and the pH value is adjusted by using phosphoric acid solution to be used as a mobile phase A.

4. The method for assaying ceftazidime-related substance according to claim 3, wherein the pH is adjusted by using a 10% phosphoric acid solution.

5. The method for assaying ceftazidime-related substance according to claim 3, wherein the pH of the mobile phase A is 3.4 ± 0.05.

6. The method for assaying ceftazidime-related substance according to claim 1, wherein the high performance liquid chromatography is performed with gradient elution under the following conditions:

gradient of mobile phase

。

7. The method according to claim 1, wherein the column used in the HPLC method comprises octadecylsilane chemically bonded silica as a filler, a column temperature of 40 ℃, a flow rate of 1.3mL/min, a detection wavelength of 254nm, and a sample injection volume of 10 μ L.