Method for separating and purifying geldanamycin
A separation and purification technology of geldanamycin, which is applied in the field of drug separation and purification, can solve the problems of low purity of geldanamycin, and achieve the effects of low cost, easy scale-up, and simple operation
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Embodiment 1
[0042] Take 2L of the geldanamycin fermentation broth, adjust the pH to 4.5, centrifuge at 4000r / min for 30 minutes to get the supernatant, and obtain the pretreated geldanamycin fermentation broth.
[0043] Take 50ml of pretreated microsphere No. 1 resin and put it in a Φ30×350mm chromatography column, take 2L of the treated fermentation broth, the unit is 747μg / ml, put it directly on the column, control its flow rate to 3ml / min, after the adsorption is complete , after loading the column, first elute the resin with 250ml of deionized water to remove the salt in the resin, use 150ml of 20% methanol water, 40% methanol water, 80% methanol water, and methanol solution to elute, and collect and wash in steps dehydration. The detection found that its purity was 68%, and the recovery rate was about 40%.
[0044] Pack a 30 ml acidic alumina chromatography column (Φ20×250 mm), pass geldanamycin methanol eluate through the chromatography column, and collect the effluent.
[0045] S...
Embodiment 2
[0047] Take 2L of geldanamycin fermentation broth, adjust the pH to 4.5, centrifuge at 4000r / min for 30min to get the supernatant, and obtain the pretreated geldanamycin fermentation broth.
[0048]Take 50ml of the pretreated microsphere No. 1 resin and put it in a Φ30×350mm chromatography column, take 2L of the treated fermentation broth, the unit is 847μg / ml, put it directly on the column, control its flow rate to 3ml / min, after the adsorption is complete , after loading the column, use 250ml of deionized water to elute the resin to remove the salt in the resin, and use 150ml of 20% ethanol water, 40% ethanol water, 80% ethanol water, ethanol to elute, and the flow rate is 6ml / min , and collect the eluate step by step. The detection found that its purity was 72%, and the recovery rate was about 51%.
[0049] Pack a 30 ml acidic alumina chromatography column (Φ20×250 mm), pass geldanamycin ethanol eluate through the chromatography column, and collect the effluent.
[0050] ...
Embodiment 3
[0052] Take 2L of geldanamycin fermentation broth, adjust the pH to 4.5, centrifuge at 4000r / min for 30min to get the supernatant, and obtain the pretreated geldanamycin fermentation broth.
[0053] Take 50ml of pretreated HZ 803 resin and put it in a Φ30×350mm chromatographic column, take 2L of the treated fermentation broth, the unit is 822μg / ml, and put it directly on the column, and control its flow rate to 3ml / min. After the adsorption is complete, put it on the column. After the column, there is no need to wash with deionized water to remove salt, and directly use 150ml of 20% acetone water and acetone solution for elution, collect the eluent step by step, and control the flow rate at 6ml / min. The detection found that its purity was 67%, and the recovery rate was about 79%.
[0054] Pack a 30 ml acidic alumina chromatography column (Φ20×250 mm), pass the geldanamycin acetone eluate through the chromatography column, and collect the effluent.
[0055] The acetone collect...
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Abstract
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