A related substance of telamycin, its enrichment preparation method and application

Abstract

The invention discloses a tulathromycin related substance, an enriching and preparation method and an application thereof. The invention provides a tulathromycin related substance (3) or a salt thereof and also provides the enriching and preparation method of the tulathromycin related substance (3). The method includes the steps of: through high performance liquid chromatography, eluting a to-be-tested substance in a chromatographic column, and collecting the tulathromycin related substance (3). The tulathromycin related substance is a necessary substance of quality control of tulathromycin. The method can effectively separate the tulathromycin related substance (3) so as to control the quality of the tulathromycin, thus laying an excellent fundament for researching on unknown impurities of the tulathromycin.

Description

technical field

[0001] The invention relates to a telamycin related substance, its enrichment preparation method and application. Background technique

[0002] Tulathromycin is a semi-synthetic macrolide antibiotic that was launched in the US and EU in 2004. The drug is mainly used for the prevention and treatment of respiratory infectious diseases caused by sensitive bacteria in cattle and pigs and bovine infectious keratoconjunctivitis caused by Moraxella bovis, and its efficacy is stronger than that of macrolide antibiotics widely used in the market Tylosin and tilmicosin have broad prospects for use in livestock and poultry production.

[0003] Tyramycin is composed of isomers A and B (molecular formula C 41 h 79 N 3 o 12 , molecular weight 806.09) is a fifteen-membered ring macrolide antibiotic composed of 9:1, and the two isomers can be converted through the formation and cleavage of the lactone bond between C11 and C13. Tyramycin contains three polar amino group...

Examples

Embodiment 1

[0094] 5mg of telamycin API was dissolved in 1mL of acetonitrile, and the injection volume was 20μL.

[0095] Chromatographic parameters: Xbridge TM C 18 (250*4.6mm, 5μm) column, mobile phase A: 0.35% formic acid aqueous solution (ammonia water to adjust the pH value to 7.66), mobile phase B: methanol:acetonitrile=45:25, gradient elution 0→15min, A:B=65 :35; 15→40min, A:B=65:35→30:70; 40→55min, A:B=30:70, UV absorption wavelength 205nm, column temperature 35℃, flow rate 1.0ml / min, HPLC fraction After passing through the UV detector, it enters MS detection with a 3:1 split. mass spectrometric total ion current as figure 1 Shown, each impurity is as shown in table 1. When the salt concentration of the mobile phase was 0.35%, the retention time of the main peak was appropriate and the peak shape was good. Related substances can be well separated from the main peak. It can be seen from the figure that the method of the present invention can be used to detect telamycin and i...

Embodiment 2

[0105] 5mg of telamycin API was dissolved in 1mL of acetonitrile, and the injection volume was 20μL.

[0106] Chromatographic parameters: Xbridge TM C 18 (250*4.6mm, 5μm) column, mobile phase A: 0.3% formic acid aqueous solution (ammonia water to adjust the pH value to 7.66), mobile phase B: methanol:acetonitrile=45:25, gradient elution 0→15min, A:B=65 :35; 15→40min, A:B=65:35→30:70; 40→55min, A:B=30:70, UV absorption wavelength 205nm, column temperature 35℃, flow rate 1.0ml / min, HPLC fraction After passing through the UV detector, it enters MS detection with a 3:1 split. mass spectrometric total ion current as Figure 6 shown. When the salt concentration of the mobile phase was 0.3%, the retention time of the main peak was appropriate and the peak shape was good. Related substances can be well separated from the main peak.

Embodiment 3

[0108] 5mg of telamycin API was dissolved in 1mL of acetonitrile, and the injection volume was 20μL.

[0109] Chromatographic parameters: Xbridge TM C 18 (250*4.6mm, 5μm) column, mobile phase A: 0.4% formic acid aqueous solution (ammonia water adjusts the pH value to 7.66), mobile phase B: methanol:acetonitrile=45:25, gradient elution 0→15min, A:B=65 :35; 15→40min, A:B=65:35→30:70; 40→55min, A:B=30:70, UV absorption wavelength 205nm, column temperature 35℃, flow rate 1.0ml / min, HPLC fraction After passing through the UV detector, it enters MS detection with a 3:1 split. mass spectrometric total ion current as Figure 7 shown. When the salt concentration of the mobile phase was 0.4%, the retention time of the main peak was appropriate and the peak shape was good. Related substances can be well separated from the main peak.

technical field

[0001] The invention relates to a telamycin related substance, its enrichment preparation method and application. Background technique

[0002] Tulathromycin is a semi-synthetic macrolide antibiotic that was launched in the US and EU in 2004. The drug is mainly used for the prevention and treatment of respiratory infectious diseases caused by sensitive bacteria in cattle and pigs and bovine infectious keratoconjunctivitis caused by Moraxella bovis, and its efficacy is stronger than that of macrolide antibiotics widely used in the market Tylosin and tilmicosin have broad prospects for use in livestock and poultry production.

[0003] Tyramycin is composed of isomers A and B (molecular formula C 41 h 79 N 3 o 12 , molecular weight 806.09) is a fifteen-membered ring macrolide antibiotic composed of 9:1, and the two isomers can be converted through the formation and cleavage of the lactone bond between C11 and C13. Tyramycin contains three polar amino group...

Examples

Embodiment 1

[0094] 5mg of telamycin API was dissolved in 1mL of acetonitrile, and the injection volume was 20μL.

[0095] Chromatographic parameters: Xbridge TM C 18 (250*4.6mm, 5μm) column, mobile phase A: 0.35% formic acid aqueous solution (ammonia water to adjust the pH value to 7.66), mobile phase B: methanol:acetonitrile=45:25, gradient elution 0→15min, A:B=65 :35; 15→40min, A:B=65:35→30:70; 40→55min, A:B=30:70, UV absorption wavelength 205nm, column temperature 35℃, flow rate 1.0ml / min, HPLC fraction After passing through the UV detector, it enters MS detection with a 3:1 split. mass spectrometric total ion current as figure 1 Shown, each impurity is as shown in table 1. When the salt concentration of the mobile phase was 0.35%, the retention time of the main peak was appropriate and the peak shape was good. Related substances can be well separated from the main peak. It can be seen from the figure that the method of the present invention can be used to detect telamycin and i...

Embodiment 2

[0105] 5mg of telamycin API was dissolved in 1mL of acetonitrile, and the injection volume was 20μL.

[0106] Chromatographic parameters: Xbridge TM C 18 (250*4.6mm, 5μm) column, mobile phase A: 0.3% formic acid aqueous solution (ammonia water to adjust the pH value to 7.66), mobile phase B: methanol:acetonitrile=45:25, gradient elution 0→15min, A:B=65 :35; 15→40min, A:B=65:35→30:70; 40→55min, A:B=30:70, UV absorption wavelength 205nm, column temperature 35℃, flow rate 1.0ml / min, HPLC fraction After passing through the UV detector, it enters MS detection with a 3:1 split. mass spectrometric total ion current as Figure 6 shown. When the salt concentration of the mobile phase was 0.3%, the retention time of the main peak was appropriate and the peak shape was good. Related substances can be well separated from the main peak.

Embodiment 3

[0108] 5mg of telamycin API was dissolved in 1mL of acetonitrile, and the injection volume was 20μL.

[0109] Chromatographic parameters: Xbridge TM C 18 (250*4.6mm, 5μm) column, mobile phase A: 0.4% formic acid aqueous solution (ammonia water adjusts the pH value to 7.66), mobile phase B: methanol:acetonitrile=45:25, gradient elution 0→15min, A:B=65 :35; 15→40min, A:B=65:35→30:70; 40→55min, A:B=30:70, UV absorption wavelength 205nm, column temperature 35℃, flow rate 1.0ml / min, HPLC fraction After passing through the UV detector, it enters MS detection with a 3:1 split. mass spectrometric total ion current as Figure 7 shown. When the salt concentration of the mobile phase was 0.4%, the retention time of the main peak was appropriate and the peak shape was good. Related substances can be well separated from the main peak.

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