Method for detecting content of erythromycin lactobionate freeze-dried powder for injection
The content of erythromycin lactonic acid for injection was detected by high-performance liquid chromatography, and the problem of difficulty in efficiently detecting the content of erythromycin lactonic acid for injection in the prior art was solved, and efficient detection of lyophilized samples was achieved, ensuring the quality of the drug.
Patent Information
- Application Number
- CN202411876709.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-19
- Publication Date
- 2025-03-11
AI Technical Summary
The prior art is difficult to efficiently detect the content of erythromycin lactonate for injection, especially in lyophilized samples.
The detection was performed by high performance liquid chromatography, and a reverse phase chromatography column with bonded silica gel was used. Mobile phase A was 0.4% ammonia water and mobile phase B was acetonitrile. The detection was achieved by gradient elution.
It has achieved efficient detection of the lyophilized content of erythromycin lactonic acid for injection, and has the advantages of high specialization and sensitivity, effectively ensuring the quality of the drug.
Smart Images
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Abstract
Description
Technical Field
[0001] The invention belongs to the field of drug analysis, and particularly relates to a method for detecting the freeze-dried content of erythromycin lactobionate for injection. Background Art
[0002] Erythromycin lactobionate (Isoniazid) is an alternative drug for the treatment of the following infections in patients allergic to penicillin: acute tonsillitis, acute pharyngitis, and sinusitis caused by hemolytic streptococci, Streptococcus pneumoniae, etc.; scarlet fever and cellulitis caused by hemolytic streptococci; diphtheria and diphtheria carriers; gas gangrene, anthrax, and tetanus; actinomycosis; syphilis; monocytosis and listeriosis; Legionnaires' disease; Mycoplasma pneumoniae pneumonia; Chlamydia pneumoniae pneumonia; urogenital infections caused by other Chlamydia and Mycoplasma species; Chlamydia trachomatis conjunctivitis; gonococcal infections; oral infections caused by anaerobic bacteria; Campylobacter jejuni enteritis; and whooping cough.
[0003] The dosage form of erythromycin lactobionate is mainly erythromycin lactobionate for injection.
[0004] Erythromycin lactobionate for injection has flexible dosage and irreplaceable advantages in clinical practice.
[0005] Among the mainstream pharmacopoeias in various countries, only the British Pharmacopoeia and the Chinese Pharmacopoeia include erythromycin lactobionate for injection. The Chinese Pharmacopoeia uses a potency determination method, while the British Pharmacopoeia uses a related substance detection method to detect the content. Develop a high-performance liquid chromatography method for the content of erythromycin lactobionate for injection. Summary of the invention
[0006] Purpose of the invention: The purpose of the present invention is to provide a high performance liquid phase detection method for detecting the content of erythromycin lactobionate for injection.
[0007] The object of the present invention is achieved through the following technical scheme: a method for detecting the content of erythromycin lactobionate, characterized in that high performance liquid chromatography is used for detection, the chromatographic column is a reverse phase chromatographic column of bonded silica gel, the mobile phase A is 0.4% ammonia water, the mobile phase B is acetonitrile, and the mobile phase elution gradient is as follows: Time (min) Mobile phase A (%) Mobile phase B (%) 0 63 37 2 63 37 5 58 42 10 25 75 12 25 75 12.1 63 37 15 63 37 .
[0008] The chromatographic column is filled with an octadecylsilane bonded silica gel column.
[0009] The chromatographic column is preferably Waters Xbridge C18 (150 mm×4.6 mm, 3.5 μm).
[0010] The mobile phase A is prepared by the following method: 4 ml of aqueous ammonia is taken, dissolved in water and diluted to 1000 ml.
[0011] Other chromatographic conditions of the high performance liquid chromatography method are as follows: column temperature is 35°C to 45°C, injection volume is 50 to 100 μl, mobile phase flow rate is 0.9 to 1.5 ml / min, and detection wavelength is 200 to 210 nm.
[0012] Other chromatographic conditions of the HPLC method are as follows: the column temperature is preferably 40° C., the injection volume is preferably 100 μl, the mobile phase flow rate is preferably 1.0 ml / min, and the detection wavelength is preferably 210 nm.
[0013] Other chromatographic conditions of the high performance liquid chromatography method are as follows: the preparation concentration of the sample to be tested is 1.0-2.0 mg / ml.
[0014] Other chromatographic conditions of the high performance liquid chromatography method: the prepared concentration of the sample to be tested is preferably 2.0 mg / ml.
[0015] The present invention has the positive effects that the detection method of the present invention can effectively detect the content of lyophilized erythromycin lactobionate for injection, has the advantages of strong specificity and high sensitivity, and effectively guarantees the quality of erythromycin lactobionate for injection. BRIEF DESCRIPTION OF THE DRAWINGS
[0016] Figure 1 This is the blank solution chromatogram of Example 1.
[0017] Figure 2 This is the chromatogram of the sample solution in Example 1.
[0018] Figure 3 This is the blank solution chromatogram of Example 2.
[0019] Figure 4 This is the chromatogram of the sample solution of Example 2.
[0020] Figure 5 This is the blank solution chromatogram of Example 3.
[0021] Figure 6 This is the spectrum of the reference solution of Example 3.
[0022] Figure 7 This is the chromatogram of the sample solution of Example 3. DETAILED DESCRIPTION
[0023] The present invention is further described in detail below in conjunction with examples, but the embodiments of the present invention are not limited thereto.
[0024] Example 1 A method for detecting the content of erythromycin lactobionate for injection, the chromatographic conditions of which are shown in Table 1.
[0025] Table 1 Chromatographic conditions of Example 1 Chromatographic columns Waters Xterra RP C18 (250mm×4.6mm, 3.5μm) Mobile phase A Acetonitrile-0.2mol / L potassium dihydrogen phosphate solution (pH 7.0)-water = 35:5:60 Mobile phase B Acetonitrile-0.2mol / L potassium dihydrogen phosphate solution (pH 7.0)-water = 50:5:45 Elution gradient Time (min) Mobile phase A (%) Mobile phase B (%) 0 1000 44 1000 46 85 100 59 85 100 60 1000 65 1000 Flow rate 1.0ml / min Column temperature 65℃ Injection volume 100μl Detection wavelength 215nm Preparation of reference solution Take an appropriate amount of erythromycin reference substance and add solvent to quantitatively dilute it to about 4 mg / ml per ml Sample solution preparation Take an appropriate amount of this product and add solvent to quantitatively dilute it to about 4 mg / ml per ml
[0026] The experimental results of Example 1 are shown in Table 2.
[0027] Table 2 Experimental results of Example 1 Experimental solution Experimental Results Attached photos blank solution Blank without interference Figure 1 Test solution No impurity peaks interfere with erythromycin peak Figure 2 .
[0028] Example 2 A method for detecting the content of erythromycin lactobionate for injection, the chromatographic conditions of which are shown in Table 3.
[0029] Table 3 Chromatographic conditions of Example 2 Chromatographic columns Waters XBridge C18 (150mm×4.6mm, 3.5μm) Mobile phase A 0.4% ammonia water Mobile phase B Acetonitrile Elution gradient Time (min) Mobile phase A (%) Mobile phase B (%) 06337263375584210257512257512.16337156337 Flow rate 1.0ml / min Column temperature 40℃ Injection volume 35μl Detection wavelength 215nm Preparation of reference solution Take an appropriate amount of erythromycin reference substance and add solvent to quantitatively dilute it to about 4 mg / ml per ml Preparation of test solution Take an appropriate amount of this product and add solvent to quantitatively dilute it to about 4 mg / ml per ml .
[0030] The experimental results of Example 2 are shown in Table 4.
[0031] Table 4 Experimental results of Example 2 Experimental solution Experimental Results Attached photos blank solution Blank without interference Figure 3 Test solution No impurity peaks interfere with erythromycin peak Figure 4 .
[0032] Example 3 A method for detecting the content of erythromycin lactobionate for injection, the chromatographic conditions of which are shown in Table 5.
[0033] Table 5 Chromatographic conditions of Example 3 Chromatographic columns Waters XBridge C18 (150mm×4.6mm, 3.5μm) Mobile phase A 0.4% ammonia water Mobile phase B Acetonitrile Elution gradient Time (min) Mobile phase A (%) Mobile phase B (%) 06337263375584210257512257512.16337156337 Flow rate 1.0ml / min Column temperature 40℃ Injection volume 100μl Detection wavelength 210nm Preparation of reference solution Take an appropriate amount of erythromycin reference substance and add solvent to quantitatively dilute it to about 2 mg / ml per ml Preparation of test solution Take an appropriate amount of this product and add solvent to quantitatively dilute it to about 2mg / ml per ml .
[0034] The experimental results of Example 3 are shown in Table 6.
[0035] Table 6 Experimental results of Example 3 Experimental solution Experimental Results Attached photos blank solution Blank without interference Figure 5 Preparation of reference solution Erythromycin A theoretical plate number>3000 Figure 6 Preparation of test solution No impurity peaks interfere with erythromycin peak Figure 7 .
[0036] Comparative Examples 1-3: The chromatographic conditions of each comparative example are quite different between Example 1 and Example 2 and Example 3; Example 2 is basically the same as Example 3, except for the injection volume and sample concentration, as detailed below.
[0037] The preparation method of mobile phase A in Comparative Example 1 is as follows: acetonitrile-0.2 mol / L potassium dihydrogen phosphate solution (pH 7.0)-water = 35:5:60.
[0038] The preparation method of mobile phase A in Comparative Example 2 is as follows: take 4 ml of ammonia water, dissolve it in water and dilute it to 1000 ml.
[0039] In summary, the mobile phase preparation in Example 1 was complex and the running time was too long, the response in Example 2 was low, and the response in Example 3 was good and the running time was good.
Claims
1. A method for detecting the content of erythromycin lactobionate for injection, characterized in that: The detection was carried out by high performance liquid chromatography. The chromatographic column was a reverse phase chromatographic column bonded with silica gel. The mobile phase A was 0.4% ammonia water, the mobile phase B was acetonitrile, and the mobile phase elution gradient was as follows:
2. The method according to claim 1, characterized in that The chromatographic column is filled with octadecylsilane bonded silica gel.
3. The method according to claim 2, characterized in that The chromatographic column described is preferably Waters Xbridge C18 (150 mm×4.6 mm, 3.5 μm).
4. The method according to claim 1, characterized in that The mobile phase A is prepared by the following method: 4 ml of ammonia water is taken, dissolved in water and diluted to 1000 ml.
5. The method according to claim 1, characterized in that Other chromatographic conditions of the high performance liquid chromatography method are: column temperature of 35°C to 45°C, injection volume of 50 to 100 μl, mobile phase flow rate of 0.8 to 1.5 ml / min, and detection wavelength of 200 to 210 nm.
6. The method according to claim 5, characterized in that Other chromatographic conditions of the HPLC method are as follows: column temperature is 40° C., injection volume is 100 μl, mobile phase flow rate is 1.0 ml / min, and detection wavelength is 210 nm.
7. The method according to claim 1, characterized in that Other chromatographic conditions of the high performance liquid chromatography method are as follows: the preparation concentration of the sample to be tested is 1.0 to 4.0 mg / ml.
8. The method according to claim 7, characterized in that Other chromatographic conditions of the high performance liquid chromatography method: the preparation concentration of the sample to be tested is 2.0 mg / ml.