A kind of method for separation and purification of veterinary antibiotic avelamycin
A technology for separation and purification of avelamycin, applied in the fields of biomedicine and industrial biotechnology, can solve the problems of deviation of analysis results, less research, immature technology, etc., and achieve the effect of easy operation and simple purification steps
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Embodiment 1
[0029] Accurately weigh 1.00 g of avelamycin-containing mycelia (20%) into a 100 mL volumetric flask, and add 30 mL of a mixture of methanol and chloroform with a volume ratio of 1:1. Sonicate for 90 min under the condition that the temperature does not exceed 40°C, dilute to 100 mL, centrifuge at 3000 r / min for 10 min to obtain the supernatant, rehydrate and crystallize according to the ratio of extract: water = 1:1, 4000 r / min Centrifuge for 15 min, and dry at 40°C to obtain the crude extract of avelamycin.
[0030] Weigh the crude avelamycin extract according to the ratio of avelamycin crude extract: silica gel = 1:70 (g:g), dissolve it in 3% 2-propanol chloroform and put it on the column. Elute with a volume of 3% 2-propanol in chloroform at a flow rate of 1 mL / min. Use silica gel HF254 plates (silica gel thickness 0.2 mm) for thin-layer chromatography (TLC) to track the spots, and collect them in segments according to the spot positions. The eluate of the target fractio...
Embodiment 2
[0034] Accurately weigh 1.00 g of avelamycin-containing mycelia (20%) into a 100 mL volumetric flask, and add 40 mL of a mixture of methanol and chloroform with a volume ratio of 1:1. Sonicate for 100 min under the condition that the temperature does not exceed 40°C, dilute to 100 mL, centrifuge at 3000 r / min for 10 min to obtain the supernatant, rehydrate and crystallize according to the ratio of extract: water = 1:1.5, 4000 r / min Centrifuge for 15 min, and dry at 40°C to obtain the crude extract of avelamycin.
[0035] Weigh the crude avelamycin extract according to the ratio of avelamycin crude extract: silica gel = 1:60 (g:g), dissolve it in 3% 2-propanol chloroform and put it on the column. Elute with a volume of 3% 2-propanol in chloroform at a flow rate of 1 mL / min. Use silica gel HF254 plates (silica gel thickness 0.2 mm) for thin-layer chromatography (TLC) to track the spots, and collect them in segments according to the spot positions. The eluate of the target frac...
Embodiment 3
[0039]Accurately weigh 1.00 g of avelamycin-containing mycelium (20%) into a 100 mL volumetric flask, and add 50 mL of a mixture of methanol and chloroform with a volume ratio of 1:1. Sonicate for 120 min under the condition that the temperature does not exceed 40°C, dilute to 100 mL, centrifuge at 3000 r / min for 10 min to obtain the supernatant, rehydrate and crystallize according to the ratio of extract: water = 1:2, 4000 r / min Centrifuge for 15 min, and dry at 40°C to obtain the crude extract of avelamycin.
[0040] Weigh the crude avelamycin extract according to the ratio of avelamycin crude extract: silica gel = 1:50 (g:g), dissolve it in 3% 2-propanol chloroform and put it on the column. Elute with a volume of 3% 2-propanol in chloroform at a flow rate of 1 mL / min. Use silica gel HF254 plates (silica gel thickness 0.2 mm) for thin-layer chromatography (TLC) to track the spots, and collect them in segments according to the spot positions. The eluate of the target fracti...
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