Method for determining aureomycin premix content and related substances through high-performance liquid chromatography method

Abstract

The present invention relates to a method for determining the aureomycin premix content and related substances through a high-performance liquid chromatography method, wherein the method adopts the following chromatography conditions that the chromatographic column is (C18, 5 [mu]m) chromatographic column, a mixed solution of water, phosphoric acid and N,N-dimethylformamide according to a volume ratio of 63-77:1.8-2.2:25-30 is adopted as a mobile phase, the phosphoric acid concentration is 2 mol / L, the flow rate is 0.8-1.2 ml / min, the detection wavelength is 270-400 nm, the column temperature is 30-40 DEG C, the sample injection volume is 10-25 [mu]L, and the detector is an ultraviolet detector. According to the method, an aureomycin standard substance solution, a tetracycline standard substance solution, a chlorquatrimycin standard substance solution and an aureomycin premix sample solution are prepared, high-performance liquid chromatography analysis is performed according to the chromatography conditions, and the content of the related substances in the premix is calculated through the peak area. The detection method of the present invention has characteristics of relatively mild chromatography conditions (the mobile phase does not contain the easily-crystallizing salt and does not use the strong acid), easy equipment maintenance, and high test accuracy.

Description

technical field

[0001] The invention belongs to the field of analysis and testing, and relates to a method for detecting the content of a premix and related substances, in particular to a method for measuring the content of aureomycin premix and related substances by high performance liquid chromatography. Background technique

[0002] Aureomycin premix is ​​a chlortetracycline calcium salt made from the whole fermentation broth of the chlortetracycline-producing bacteria and an appropriate amount of calcium carbonate. The main drug ingredient is aureomycin, but it also contains some related fermentation by-products. Such as tetracycline, aureomycin and so on. The determination of the aureomycin content generally adopts the microbiological detection method (recorded in the national standard of veterinary medicine). Practice has shown that the use of microbiological methods to determine the content has several disadvantages: 1) the method is cumbersome and takes a long time,...

Examples

Embodiment Construction

[0038] The preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings.

[0039] Testing instruments and conditions:

[0040] Shimadzu LC-20 series high performance liquid chromatography, SPD-M20A detector;

[0041] Chromatographic column: C18, 5μ, 4.6×150mm;

[0042] Column temperature control: 35°C;

[0043] Detection wavelength: UV 365nm;

[0044] Mobile phase: water: 2mol / L phosphoric acid solution: N,N-dimethylformamide = 70:2:28 (volume ratio);

[0045] Pump flow: 1.00ml / min;

[0046] Quantitative volume: 20μL;

[0047] Purified water (HPLC grade), hydrochloric acid (excellent grade), phosphoric acid (excellent grade), N,N-dimethylformamide (chromatographic grade)

[0048] Preparation of mobile phase solution: Take 700ml of purified water, add 20ml of 2mol / L phosphoric acid solution and mix well, then add 280ml of N,N-dimethylformamide, filter through a 0.45μm membrane with a vacuum filter, and set as...

technical field

[0001] The invention belongs to the field of analysis and testing, and relates to a method for detecting the content of a premix and related substances, in particular to a method for measuring the content of aureomycin premix and related substances by high performance liquid chromatography. Background technique

[0002] Aureomycin premix is ​​a chlortetracycline calcium salt made from the whole fermentation broth of the chlortetracycline-producing bacteria and an appropriate amount of calcium carbonate. The main drug ingredient is aureomycin, but it also contains some related fermentation by-products. Such as tetracycline, aureomycin and so on. The determination of the aureomycin content generally adopts the microbiological detection method (recorded in the national standard of veterinary medicine). Practice has shown that the use of microbiological methods to determine the content has several disadvantages: 1) the method is cumbersome and takes a long time,...

Examples

Embodiment Construction

[0038] The preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings.

[0039] Testing instruments and conditions:

[0040] Shimadzu LC-20 series high performance liquid chromatography, SPD-M20A detector;

[0041] Chromatographic column: C18, 5μ, 4.6×150mm;

[0042] Column temperature control: 35°C;

[0043] Detection wavelength: UV 365nm;

[0044] Mobile phase: water: 2mol / L phosphoric acid solution: N,N-dimethylformamide = 70:2:28 (volume ratio);

[0045] Pump flow: 1.00ml / min;

[0046] Quantitative volume: 20μL;

[0047] Purified water (HPLC grade), hydrochloric acid (excellent grade), phosphoric acid (excellent grade), N,N-dimethylformamide (chromatographic grade)

[0048] Preparation of mobile phase solution: Take 700ml of purified water, add 20ml of 2mol / L phosphoric acid solution and mix well, then add 280ml of N,N-dimethylformamide, filter through a 0.45μm membrane with a vacuum filter, and set as...

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