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Purification method of high-purity teicoplanin refined powder

A technology of coplanin refined powder and purification method, which is applied to the purification field of high-purity teicoplanin refined powder, can solve problems such as insufficient separation effect, and achieve the effects of reliable quality, controllable content and simple process

Inactive Publication Date: 2015-02-25
FUJIAN INST OF MICROBIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method uses nano-polymer microspheres for column chromatography. The skeleton structure of nano-polymer microspheres has higher mechanical strength and chemical stability, but the separation effect is not as good as that of reversed-phase silica gel.

Method used

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  • Purification method of high-purity teicoplanin refined powder
  • Purification method of high-purity teicoplanin refined powder
  • Purification method of high-purity teicoplanin refined powder

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] A. Adjust the pH of 60L of teicoplanin fermentation broth to 11.0, keep it at room temperature for about 15min, return to pH7.5, centrifuge at 4500rpm for 10min, take the supernatant and dilute it to 120L with 15L HP20 macroporous resin, first use 2BV (2 times Column volume) Pure water was eluted at a flow rate of 10L / h to remove impurities and pigments, and then eluted with 10%, 15%, 20%, and 25% acetone aqueous solution at a flow rate of 12L / h to remove impurities, and then 70% acetone The aqueous solution was eluted at a flow rate of 12 L / h, and the eluate with 70% acetone aqueous solution as the eluent was collected to obtain 216 L of the eluate, the content of teicoplanin was 822.03 g, the yield was 88.7%, and the potency was 13.70 g / L.

[0039] B. Concentrate the 216L eluate to a small volume under reduced pressure, put it on a 5L Sephadex gel chromatography column, first use 15% methanol water to elute at a flow rate of 8L / h, and then use a volume ratio of 35:65 m...

Embodiment 2

[0046] A. Adjust the pH of 60L of teicoplanin fermentation broth to 11.0, keep it at room temperature for about 15min, return to pH7.5, centrifuge at 4500rpm for 10min, take the supernatant and dilute it to 120L, take 20L of the diluted solution, and install it with D3520 macroporous resin for adsorption , first eluted with 2BV (column volume) pure water at a flow rate of 10L / h to remove impurities and pigments, and then eluted with 10%, 15%, 20%, and 25% acetone aqueous solution at a flow rate of 12L / h to remove impurities , and then eluted with 70% acetone aqueous solution at a flow rate of 12L / h, collecting the eluent with 70% acetone aqueous solution as the eluent, obtaining 81L eluent with a content of 270.8g and a titer of 13.54g / L, and recovering Teicoara Ning 245.34g, the yield is 90.6%, and the purity of the product by liquid chromatography is 83.01%.

[0047] B. Adsorb 81L of the eluate on a 5L agarose gel chromatography column with a sample loading flow rate of 6L / h...

Embodiment 3

[0054] The difference between this part and embodiment 1 is:

[0055] A. Take 20L of diluent and put it on HZ816 macroporous adsorption resin;

[0056] Obtain 75L eluent, teicoplanin content 210.4g, potency 14.04g / L, reclaim teicoplanin 192.30g, yield is 91.4%, product liquid chromatography purity 81.76% (purity is represented by teicoplanin A2 The sum of the components is the basis).

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Abstract

The invention provides a purification method of high-purity teicoplanin refined powder, which comprises the following steps: centrifuging a teicoplanin fermentation liquid under alkaline conditions to remove solids in the fermentation liquid, regulating the filtrate to a neutral state, adsorbing and eluting with a macroporous adsorbent resin to obtain a teicoplanin eluate with the concentration of 65% above, and filtering by gel chromatography to obtain a 80% teicoplanin eluate; concentrating the eluate, adding acetone to precipitate, and centrifuging to obtain a milk white teicoplanin crude product; and dissolving the crude product with a right amount of water, adding the sample to a chromatographic column filled with reversed phase silica gel, carrying out gradient elution with an acetonitrile-water mixed solution to obtain a high-purity teicoplanin eluate, adding activated carbon for decolorization, filtering, cooling to precipitate, removing the residual impurities, adding acetone to the clear solution to completely precipitate, and carrying out infrared drying on the wet product to obtain the teicoplanin pure product. The method has the advantages of simple technique, reliable quality, high purity of the teicoplanin refined powder and controllable single-component content, and is suitable for industrial large-scale production of high-purity teicoplanin.

Description

technical field [0001] The invention specifically relates to a method for purifying high-purity teicoplanin fine powder. Background technique [0002] Teicoplanin (teicoplanin) is a multi-component glycopeptide antibiotic produced by the fermentation of Actinoplanes teichomyceticus, mainly containing five compounds with similar chemical structures (A2-1, A2-2, A2-3, A2-4 and A2-5), such as figure 1 As shown, the difference between the five components is only the difference in the acyl side chain, of which A2-2 is the main component, accounting for 35%-55% of the relative content of each component, and contains a small amount of side chain degradation product A3-1 . [0003] The structural formula of teicoplanin: [0004] [0005] As a new glycopeptide antibiotic similar to vancomycin, teicoplanin has similar antibacterial spectrum and antibacterial activity to vancomycin, and has fewer adverse reactions in clinical research. With the continuous deepening of the resear...

Claims

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Application Information

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IPC IPC(8): C07K9/00C07K1/20
Inventor 陈丽连云阳罗婷江宁宇彭飞王传喜
Owner FUJIAN INST OF MICROBIOLOGY
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