Preparation method of high-purity gentamycin
A gentamicin, high-purity technology, applied in the field of separation and purification of gentamicin in fermentation broth, can solve the problems of unsatisfactory purification effect, difficulty in preparing pharmaceutical grade gentamicin, etc., and achieve product quality Good, low cost, and the effect of improving desorption efficiency
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Embodiment 1
[0023] The preparation of embodiment 1 high-purity gentamicin
[0024] a. Add 50 g of perlite to 10 L of gentamicin fermentation broth with a fermentation unit of 1960 μg / mL, stir for 30 min, and filter to obtain a filtrate containing gentamicin.
[0025] b. The above-mentioned filtrate containing gentamicin was introduced into a macroporous resin column LX-50 with a diameter-to-height ratio of 1:3 for adsorption, with a loading capacity of 2400 mL and a sample loading flow rate of 2400 mL / h. Then gradient elution was carried out with ammonia water. The concentration of ammonia water in gradient 1 was 0.1mol / L, the flow rate was 1200mL / h, and the dosage was 24L; the concentration of ammonia water in gradient 2 was 1mol / L, the flow rate was 1200mL / h, and the dosage was 7.2L. The eluate from gradient 2 was collected, concentrated under reduced pressure until the content of gentamicin was 10,000 units / ml, and the pH was adjusted to 4.0 with 3 mol / L sulfuric acid aqueous solution ...
Embodiment 2
[0028] The preparation of embodiment 2 high-purity gentamicin
[0029] a. Add 100 g of perlite to 10 L fermentation unit of 1800 μg / mL gentamicin fermentation liquid, stir for 30 min, and filter to obtain filtrate containing gentamicin.
[0030] b. The above-mentioned filtrate containing gentamicin was introduced into a macroporous resin column D113 with a diameter-to-height ratio of 1:3 for adsorption, with a loading capacity of 2400 mL and a sample loading flow rate of 2400 mL / h. Then carry out gradient elution with ammonia water, the concentration of ammonia water in gradient 1 is 0.2mol / L, the flow rate is 1200mL / h, and the dosage is 24L; the concentration of ammonia water in gradient 2 is 2.5mol / L, the flow rate is 1200mL / h, and the dosage is 7.2L . The eluate from gradient 2 was collected, concentrated under reduced pressure until the content of gentamicin was 13,000 units / ml, and the pH was adjusted to 5.0 with 3 mol / L sulfuric acid aqueous solution to obtain a crude e...
Embodiment 3
[0032] Embodiment 3 The preparation of high-purity gentamicin
[0033] a. Add 1 kg of diatomaceous earth to a 20 L fermentation unit of 2300 μg / mL gentamicin fermentation broth, stir for 30 min, and filter to obtain a filtrate containing gentamicin.
[0034] b. The above-mentioned filtrate containing gentamicin was introduced into a macroporous resin column HP20 with a diameter-to-height ratio of 1:3 for adsorption, with a loading capacity of 5000 mL and a sample loading flow rate of 5000 mL / h. Then carry out gradient elution with ammonia water, the concentration of ammonia water in gradient 1 is 0.3mol / L, the flow rate is 2500mL / h, and the dosage is 40L; the concentration of ammonia water in gradient 2 is 3mol / L, the flow rate is 2500mL / h, and the dosage is 15L. The eluate from gradient 2 was collected, concentrated under reduced pressure until the content of gentamicin was 15,000 units / ml, and the pH was adjusted to 6.0 with 3 mol / L sulfuric acid aqueous solution to obtain a...
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