CN106632551A - Method for preparing fidaxomicin by flash chromatography - Google Patents
Method for preparing fidaxomicin by flash chromatography Download PDFInfo
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- C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
- C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
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Abstract
The invention discloses a method for preparing fidaxomicin by flash chromatography. The method comprises the following steps: firstly, extracting fidaxomicin fermentation mycelia with 60 to 80 percent ethanol, roughly separating extract liquid through polystyrene resin, and collecting a fraction which contains fidaxomicin; then, performing reverse-phase bonding silica gel rapid column chromatography separation twice on the fraction, wherein elution solvents of different selectivity are adopted; finally obtaining a non-fidaxomicin refined product of which the purity is over 98.5 percent. By adopting the method, the purity and yield of the obtained fidaxomicin are high; moreover, the method is relatively easy to operate, is relatively low in cost, and is suitable for industrial production.
Description
【Technical field】
The invention belongs to field of microbial pharmacy, and in particular to a kind of method that flash chromatography prepares feldamycin.
【Background technology】
Clostridium difficile is a kind of Gram-positive anaerobic spore-bearing bacilli, is distributed widely in the excrement of natural environment and animal and people
Just in.Its undue growth and when discharging toxin, can cause C. difficile infection (CDI), cause colitis, severe diarrhea even
It is dead.
Feldamycin (fidaxomicin) is the macrolides of a kind of new narrow spectrum of Optimer drugmakers research and development
Antibiotic, FDA approvals are obtained in May, 2011 in U.S.'s listing, are become first over 30 years and are mainly used in treating clostridium difficile phase
The antibiotic of closing property diarrhoea (CDAD).Feldamycin by suppress clostridium difficile RNA polymerase, blocking ribonucleic acid synthesis and
It is sterilized.Compared with conventional medicament vancomycin, it can significantly reduce the recurrence rate of C. difficile infection and improve overall cure rate.
The structure of feldamycin is as follows：
In prior art, the method for extraction purification feldamycin has from zymotic fluid：
Chinese patent CN104846043A discloses a kind of technique for separating from zymotic fluid and purifying feldamycin；The work
Skill extracts mycelium using polar organic solvent；Leaching liquor is by large pore resin absorption column and desorbs；Stripping liquid organic solvent
Extraction, then through normal phase column chromatography, obtain feldamycin crude product；Crude product Jing high pressure liquid chromatography equipment, with organic solvent/acid
Water elution, collects high-purity fidaxomicin efflux；Eluent Jing polystyrene microspheres resin column chromatography again；Eluent extraction,
Concentrate, be dried, obtain white powder feldamycin sterling, purity 96%～97%, yield about 73%.The processing step is numerous
It is trivial, and high pressure Liquid apparatus have been used, it is appropriate only for laboratory operation.
Chinese patent CN104418925A discloses a kind of method for preparing high-purity fidaxomicin；The method will reach from non-
The mycelium isolated in mycin zymotic fluid is extracted with polar organic solvent；Leaching liquor imports macroporous absorbent resin after being diluted with water
SP825L adsorbs, and then resin column is purified and is desorbed, and obtains the crude product stripping liquid I of feldamycin, after concentration, crystallization
Feldamycin crude product；After the aqueous dissolution of feldamycin crude product acetonitrile, UniPS40 resin columns are imported, Jing affords non-
Up to mycin eluent II, dissolving is evaporated off and obtains solid, then recrystallized with polar organic solvent, obtain feldamycin product.The party
Method yield 50%～60%, purity more than 98.5%, but need one or many to crystallize, operate relatively complicated.
Chinese patent CN104098637A discloses a kind of purification process of feldamycin；The method ferments feldamycin
Liquid carries out separation of solid and liquid and obtains mycelium, then soaks mycelium with organic solvent and obtains the solution containing feldamycin；This is molten
Liquid carries out nanofiltration concentration；Concentrate prepares column chromatography by the DAC200 equipped with C8 fillers, and with the sour water containing organic solvent
Solution is eluted, and is collected, is merged the cut containing feldamycin, is added water and is forced out feldamycin, is filtered, is dried to obtain feldamycin
Dry powder, and yield reaches more than 50%, purity and reaches more than 99.5%.The method is simple to operate, obtain feldamycin purity and receipts
Rate is high, but as a result of dynamic axial compression column (DAC) technology, cost input higher to equipment and filler specification requirement
It is larger.
Therefore, the method for extraction purification feldamycin still needs to be optimized improvement from zymotic fluid, is desirably to obtain a kind of tool
The method having the following advantages：Purity and high income, while operate relatively easy, cost input relatively low, and suitable for industrial metaplasia
Produce.
【The content of the invention】
The technical problem to be solved is to provide a kind of method that flash chromatography prepares feldamycin, not only institute
The purity and high income of feldamycin are obtained, and operates relatively easy, cost relatively low, and suitable for industrialized production.
The present invention is to solve above-mentioned technical problem by the following technical programs：A kind of flash chromatography prepares feldamycin
Method, including following concrete operation step：
(1), feldamycin zymotic fluid is carried out into separation of solid and liquid and obtains mycelium, be subsequently adding the immersion of 60%～80% ethanol
Mycelium is simultaneously centrifuged, and obtains feldamycin leaching liquor, stand-by；
(2) it is 10%～40%, the feldamycin leaching liquor obtained by step (1) to be diluted with water to into concentration of alcohol, afterwards
Import and adsorbed in polystyrene resin post, then gradient desorption is carried out using ethanol, collect feldamycin content 45% with
On stripping liquid；
(3), the stripping liquid collected by step (2) is merged, and it is 45%～55% to be diluted with water to concentration of alcohol, then
In importing reversed-phase silica gel chromatography post, gradient elution is carried out using methanol-water-acetic acid, collect feldamycin content more than 85%
Eluent；
(4), the eluent collected by step (3) is merged, and is adopted and is diluted with water to methanol concentration for 55%～65%,
Import again in reversed-phase silica gel chromatography post, gradient elution is carried out using acetonitrile-water-acetic acid, collect feldamycin content 98.5%
More than eluent simultaneously merge, concentrated, be dried after obtain highly purified feldamycin finished product.
Further, in the step (1), the addition of 60%～80% ethanol is to add in per kilogram of wet mycelium
1～8 liter；Extraction temperature is 10～50 DEG C；Extraction time is 2～8h.
Further, in the step (2), polystyrene resin post is DIAION HP-20 polystyrene resin posts, receives
Micro- NM-200 polystyrene resins post, D101 polystyrene resin posts, HPD-400 polystyrene resin posts, AmberliteXAD-
Any one in 8 polystyrene resin posts or HZ816 polystyrene resin posts.
Further, in the step (3), the filler of reversed-phase silica gel chromatography post is alkyl silica gel bonded from C4, C8 or C18
Any one in phase, and particle diameter is 50 μm.
Further, in the step (4), the filler of reversed-phase silica gel chromatography post is alkyl silica gel bonded from C4, C8 or C18
Any one in phase, and particle diameter is 50 μm.
The beneficial effects of the present invention is：
A kind of method that flash chromatography prepares feldamycin is provided, feldamycin can not only be quickly and easily obtained, and
And the purity of the feldamycin for being obtained is high with yield, additionally, whole method operates relatively easy, cost relatively low, Neng Goushi
For industrialized production.
【Description of the drawings】
With reference to the accompanying drawings in conjunction with the embodiments the invention will be further described.
Fig. 1 is the HPLC chromatogram of leaching liquor in embodiment of the present invention 1-3.
Fig. 2 is the HPLC chromatogram of stripping liquid in the embodiment of the present invention 1.
Fig. 3 is the HPLC chromatogram of the eluent of collected feldamycin content more than 85% in the embodiment of the present invention 1.
Fig. 4 is the HPLC chromatogram of the eluent of collected feldamycin content more than 98.5% in the embodiment of the present invention 1.
Fig. 5 is the HPLC chromatogram of stripping liquid in the embodiment of the present invention 2.
Fig. 6 is the HPLC chromatogram of the eluent of collected feldamycin content more than 85% in the embodiment of the present invention 2.
Fig. 7 is the HPLC chromatogram of the eluent of collected feldamycin content more than 98.5% in the embodiment of the present invention 2.
Fig. 8 is the HPLC chromatogram of stripping liquid in the embodiment of the present invention 3.
Fig. 9 is the HPLC chromatogram of the eluent of collected feldamycin content more than 85% in the embodiment of the present invention 3.
Figure 10 is the HPLC chromatogram of the eluent of collected feldamycin content more than 98.5% in the embodiment of the present invention 3
Figure.
【Specific embodiment】
The method that a kind of flash chromatography of the present invention prepares feldamycin, including following concrete operation step：
(1), feldamycin zymotic fluid is carried out into separation of solid and liquid and obtains mycelium, be subsequently adding the immersion of 60%～80% ethanol
Mycelium is simultaneously centrifuged, and obtains feldamycin leaching liquor, stand-by；
(2) it is 10%～40%, the feldamycin leaching liquor obtained by step (1) to be diluted with water to into concentration of alcohol, afterwards
Import and adsorbed in polystyrene resin post, then gradient desorption is carried out using ethanol, collect feldamycin content 45% with
On stripping liquid；
(3), the stripping liquid collected by step (2) is merged, and it is 45%～55% to be diluted with water to concentration of alcohol, then
In importing reversed-phase silica gel chromatography post, gradient elution is carried out using methanol-water-acetic acid, collect feldamycin content more than 85%
Eluent；
(4), the eluent collected by step (3) is merged, and is adopted and is diluted with water to methanol concentration for 55%～65%,
Import again in reversed-phase silica gel chromatography post, gradient elution is carried out using acetonitrile-water-acetic acid, collect feldamycin content 98.5%
More than eluent simultaneously merge, concentrated, be dried after obtain highly purified feldamycin finished product.
Wherein, in step (1), the addition of 60%～80% ethanol is that 1～8 liter is added in per kilogram of wet mycelium；
Extraction temperature is 10～50 DEG C；Extraction time is 2～8h.In step (2), polystyrene resin post is DIAION HP-20 polyphenyl
Vinyl post, receive micro- NM-200 polystyrene resins post, D101 polystyrene resin posts, HPD-400 polystyrene resin posts,
Any one in AmberliteXAD-8 polystyrene resins post or HZ816 polystyrene resin posts；Can preferably receive micro- NM-
200 polystyrene resin posts or HZ816 polystyrene resin posts.In step (3), the filler of reversed-phase silica gel chromatography post from C4,
Any one in the alkyl silica gel bonded phases of C8 or C18, and particle diameter is 50 μm；Can preferably the alkyl silica gel bonded phases of C18 (particle diameter is
50μm).In step (4), the filler of reversed-phase silica gel chromatography post from any one in the alkyl silica gel bonded phase of C4, C8 or C18, and
Particle diameter is 50 μm；Can preferred alkyl silica gel bonded phases of C18 (particle diameter be 50 μm).It should be noted that methyl alcohol is dense in the present invention
Degree, the percentage of concentration of alcohol are percentage by volume in the case of without specified otherwise.
In addition, actinoplanes used by the fermented and cultured of the feldamycin zymotic fluid mentioned in the present invention are
Actinoplanes deccanensis ATCC21983, purchased from American Type Culture collection warehousing (ATCC)；DIAION HP-
20 polystyrene resins are purchased from Mitsubishi chemical Co., Ltd；Receive micro- NM-200 polystyrene resins, D101 polystyrene resins,
HPD-400 polystyrene resins are purchased from Suzhou Nano-Micro Bio-technology Co., Ltd.；AmberliteXAD-8 polystyrene resins are purchased
From Beijing Hui De Easytech Inc.；HZ816 polystyrene resins are purchased from Shanghai Huazhen Science and Technology Co., Ltd.；Buy and come
Polystyrene resin using when insert corresponding polystyrene resin post formed in corresponding cylinder；Reversed-phase silica gel chromatography post
Filler is available from YMC companies；The reagents such as methyl alcohol, ethanol, acetonitrile are commercially available；Water is distilled water；High performance liquid chromatograph is island
Tianjin LC-20A.
Feldamycin detection adopts high performance liquid chromatography, chromatographic condition：Chromatographic column is moon rising sun Ultimate XB-C18
(ID4.6mm × 250mm, 5 μm), mobile phase is methyl alcohol：0.1% aqueous formic acid (73:27), flow velocity 1mL/min, Detection wavelength
266nm, the μ L of sample size 10,40 DEG C of column temperature；The retention time of feldamycin slightly fluctuates between different detection batches, and scope is 17
～18min.
The concrete preparation process of feldamycin zymotic fluid is as follows in the present invention：
A, the preparation of inclined-plane bacterium colony and culture：Actinoplanes thalline is connected on slant medium, and is trained at 30 DEG C
Support 10 days；Slant medium is that, using ISP culture mediums, its formula is：Yeast extract 4g/L, malt extract 10g/L, grape
Sugared 4g/L, agar 20g/L, pH7.2；
B, the preparation of seed culture medium and culture:The single bacterium colony obtained by culture on slant medium is moved into using platinum loop is planted
In sub- culture medium, and at 30 DEG C of cultivation temperature, 240rpm shaking table shaken cultivations 48h obtain the one-level kind of feldamycin fermentation
Sub- liquid；
Seed culture based formulas:Soluble starch 20g/L, glucose 5g/L, peptone 5g/L, dusty yeast 5g/L, soya bean
Powder 5g/L, ammonium sulfate 0.5g/L, dipotassium hydrogen phosphate 0.5g/L, bitter salt 0.5g/L, calcium carbonate 2g/L, pH7.2, shaking flask
Liquid amount is 50mL/500mL.
C, the preparation of fermentation medium and culture:Obtained step (b) by 5% inoculum concentration with percent by volume one
Level seed liquor moves into fermented and cultured in fermentation medium, and 28 DEG C of fermented and cultured temperature, 240rpm shaking table shaken cultivations 136h are then obtained
Obtain required feldamycin zymotic fluid of the invention；
Fermentative medium formula:Soluble starch 4g/L, glucose 10g/L, peptone 3g/L, yeast extract 1g/L,
Analysis for soybean powder 20g/L, sodium chloride 2.5g/L, ammonium sulfate 0.25g/L, dipotassium hydrogen phosphate 0.25g/L, bitter salt 0.25g/L,
Calcium carbonate 5g/L, micro- salt 1mL, pH7.5.
In order to clearly illustrate the detailed process of the inventive method, the applicant illustrates several as follows being embodied as
Example.
Embodiment 1
Feldamycin zymotic fluid 10L is taken, zymotic fluid is centrifuged 10 minutes with 4500rpm, obtain 2kg mycelium (weight in wet base)；
70% ethanol 4L is added in mycelium, under room temperature, electric mixer stirring (350rpm) is centrifuged after extracting 4 hours, and collection contains
The leaching liquor (4.5L, 1179mg containing feldamycin) of feldamycin, and the HPLC chromatogram of leaching liquor is as shown in Figure 1；Will extraction
Liquid is diluted with water to concentration of alcohol and adsorbed in micro- NM-200 polystyrene resins post absorption to import after 20% to receive, and resin is filled
Measure as 300mL, flow velocity is 2BV/h, and resin column carries out gradient desorption with the ethanol solution of concentration 40%, 50%, 60% successively, stream
Speed is 2BV/h, and the stripping liquid of Fractional Collections feldamycin content more than 45%, the HPLC chromatogram of stripping liquid is as shown in Figure 2.
The stripping liquid of the feldamycin content more than 45% collected is merged, concentration of alcohol is diluted with water to for after 50%
YMCGEL ODS-A-HG reversed-phase silica gel chromatography posts (column diameter 3cm, column length 38cm) are imported, methyl alcohol is then used successively：Water：Second
Acid=50：50：0.1 (V/V/V), methyl alcohol：Water：Acetic acid=70：30：0.1 (V/V/V) carries out gradient elution, elution volume difference
For 3BV, 10BV, flow velocity is 10mL/min, the eluent of Fractional Collections feldamycin content more than 85%, the HPLC of the eluent
Chromatogram is as shown in Figure 3.
The eluent of the feldamycin content more than 85% collected is merged, methanol concentration is diluted with water to for after 58%
YMCGEL ODS-A-HG reversed-phase silica gel chromatography posts (column diameter 3cm, column length 38cm) are imported again, and acetonitrile is then used successively：
Water：Acetic acid=30：70：0.1 (V/V/V), acetonitrile：Water：Acetic acid=45：55：0.1 (V/V/V) carries out gradient elution, elution volume
Respectively 3BV, 10BV, flow velocity is 10mL/min, the eluent of Fractional Collections feldamycin content more than 98.5%, the de- liquid
HPLC chromatogram is as shown in Figure 4；The eluent of the feldamycin content more than 98.5% that merging is collected simultaneously is concentrated, is dried
Obtain fidaxomicin 390mg.It is 33.1% that Jing determines the total recovery of this enforcement feldamycin.
The HPLC purity of feldamycin in each step of the present embodiment preparation process is calculated according to area normalization method：Extraction
Feldamycin purity is 33.7% in liquid；Feldamycin purity is the anti-phase silicon of 55.5%, YMCGEL ODS-A-HG in stripping liquid
Feldamycin purity is 92.9%, YMCGEL ODS-A-HG chromatographic columns in the methanol/water of glue chromatographic column/acetic acid eluent
Feldamycin purity is 98.8% in acetonitrile/water/acetic acid eluent.
Embodiment 2
Feldamycin zymotic fluid 20L is taken, zymotic fluid is centrifuged 10 minutes with 4500rpm, obtain 3.9kg mycelium (wet
Weight)；60% ethanol 8L is added in mycelium, under room temperature, electric mixer stirring (350rpm) is centrifuged after extracting 4 hours, receives
Leaching liquor (9L, containing feldamycin 2358mg) of the collection containing feldamycin, and the HPLC chromatogram of leaching liquor is as shown in Figure 1；Will leaching
Extract is diluted with water to concentration of alcohol and adsorbed in micro- NM-200 polystyrene resins post absorption to import after 40% to receive, resin
Loading amount is 500mL, and flow velocity is 2BV/h, and resin column carries out gradient desorption with the ethanol solution of concentration 40%, 50%, 60% successively,
Flow velocity is 2BV/h, and the stripping liquid of Fractional Collections feldamycin content more than 45%, the HPLC chromatogram of stripping liquid is as shown in Figure 5.
The stripping liquid of the feldamycin content more than 45% collected is merged, concentration of alcohol is diluted with water to for after 45%
YMCGEL ODS-A-HG reversed-phase silica gel chromatography posts (column diameter 3cm, column length 38cm) are imported, methyl alcohol is then used successively：Water：Second
Acid=50：50：0.1 (V/V/V), methyl alcohol：Water：Acetic acid=70：30：0.1 (V/V/V) carries out gradient elution, elution volume difference
For 4BV, 10BV, flow velocity is 10mL/min, the eluent of Fractional Collections feldamycin content more than 85%, the HPLC of the eluent
Chromatogram is as shown in Figure 6.
The eluent of the feldamycin content more than 85% collected is merged, methanol concentration is diluted with water to for after 55%
YMCGEL ODS-A-HG reversed-phase silica gel chromatography posts (column diameter 3cm, column length 38cm) are imported again, and acetonitrile is then used successively：
Water：Acetic acid=30：70：0.1 (V/V/V), acetonitrile：Water：Acetic acid=50：50：0.1 (V/V/V) carries out gradient elution, elution volume
Respectively 3BV, 10BV, flow velocity is 12mL/min, the eluent of Fractional Collections feldamycin content more than 98.5%, the de- liquid
HPLC chromatogram is as shown in Figure 7；The eluent of the feldamycin content more than 98.5% that merging is collected simultaneously is concentrated, is dried
Obtain fidaxomicin 806mg；It is 34.2% that Jing determines the total recovery of this enforcement feldamycin.
The HPLC purity of feldamycin in each step of the present embodiment preparation process is calculated according to area normalization method：Extraction
Feldamycin purity is 33.7% in liquid；Feldamycin purity is the anti-phase silicon of 58.8%, YMCGEL ODS-A-HG in stripping liquid
Feldamycin purity is 92.7%, YMCGEL ODS-A-HG chromatographic columns in the methanol/water of glue chromatographic column/acetic acid eluent
Feldamycin purity is 99.1% in acetonitrile/water/acetic acid eluent.
Embodiment 3
Feldamycin zymotic fluid 30L is taken, zymotic fluid is centrifuged 10 minutes with 4500rpm, obtain 5.9kg mycelium (wet
Weight)；80% ethanol 12L is added in above-mentioned mycelium, under room temperature, electric mixer stirring (350rpm) extract 4 hours after from
The heart, collects the leaching liquor (13.6L, 3531mg containing feldamycin) containing feldamycin, and the HPLC chromatogram of leaching liquor such as Fig. 1 institutes
Show；Leaching liquor is diluted with water to into concentration of alcohol and receives micro- NM-200 polystyrene resins post absorption to import after 10%, resin dress
Measure as 500mL, flow velocity is 2BV/h, and resin column carries out gradient desorption with the ethanol solution of concentration 40%, 50%, 60% successively, stream
Speed is 2BV/h, and Fractional Collections collects the stripping liquid of feldamycin content more than 45%, HPLC chromatogram such as Fig. 8 institutes of stripping liquid
Show.
The stripping liquid of the feldamycin content more than 45% collected is merged, concentration of alcohol is diluted with water to for after 55%
YMCGEL ODS-A-HG reversed-phase silica gel chromatography posts (column diameter 4.6cm, column length 40cm) are imported, methyl alcohol is then used successively：Water：
Acetic acid=50：50：0.1 (V/V/V), methyl alcohol：Water：Acetic acid=70：30：0.1 (V/V/V) carries out gradient elution, elution volume point
Not Wei 2BV, 10BV, flow velocity is 25mL/min, the eluent of Fractional Collections feldamycin content more than 85%, the eluent
HPLC chromatogram is as shown in Figure 9.
The eluent of the feldamycin content more than 85% collected is merged, methanol concentration is diluted with water to for after 65%
YMCGEL ODS-A-HG reversed-phase silica gel chromatography posts (column diameter 4.6cm, column length 40cm) are imported again, and acetonitrile is then used successively：
Water：Acetic acid=30：70：0.1 (V/V/V), acetonitrile：Water：Acetic acid=45：55：0.1 (V/V/V) carries out gradient elution, elution volume
Respectively 2BV, 10BV, flow velocity is 25mL/min, the eluent of Fractional Collections feldamycin content more than 98.5%, the eluent
HPLC chromatogram it is as shown in Figure 10；The eluent of feldamycin content more than 98.5% that merging is collected and concentrated,
It is dried to obtain fidaxomicin 1144mg；It is 32.4% that Jing determines the total recovery of this enforcement feldamycin.
The HPLC purity of feldamycin in each step of the present embodiment preparation process is calculated according to area normalization method：Extraction
Feldamycin purity is 33.7% in liquid；Feldamycin purity is the anti-phase silicon of 55.1%, YMCGEL ODS-A-HG in stripping liquid
Feldamycin purity is 88.5%, YMCGEL ODS-A-HG chromatographic columns in the methanol/water of glue chromatographic column/acetic acid eluent
Feldamycin purity is 98.9% in acetonitrile/water/acetic acid eluent.
To sum up, the inventive method, can not only quickly and easily obtain feldamycin, and the feldamycin for being obtained
Purity is high with yield, additionally, whole method operates relatively easy, cost relatively low, can be applied to industrialized production.
Claims (5)
1. a kind of method that flash chromatography prepares feldamycin, it is characterised in that：Including following concrete operation step：
(1), feldamycin zymotic fluid is carried out into separation of solid and liquid and obtains mycelium, be subsequently adding 60%～80% ethanol immersion mycelia
Body is simultaneously centrifuged, and obtains feldamycin leaching liquor, stand-by；
(2) it is 10%～40%, the feldamycin leaching liquor obtained by step (1) to be diluted with water to into concentration of alcohol, is imported afterwards
Adsorbed in polystyrene resin post, then gradient desorption is carried out using ethanol, collected feldamycin content more than 45%
Stripping liquid；
(3), the stripping liquid collected by step (2) is merged, and it is 45%～55% to be diluted with water to concentration of alcohol, is then directed into
In reversed-phase silica gel chromatography post, gradient elution is carried out using methanol-water-acetic acid, collect the wash-out of feldamycin content more than 85%
Liquid；
(4), the eluent collected by step (3) is merged, and is adopted and is diluted with water to methanol concentration for 55%～65%, again
In importing reversed-phase silica gel chromatography post, gradient elution is carried out using acetonitrile-water-acetic acid, collect feldamycin content more than 98.5%
Eluent and merge, concentrated, be dried after obtain highly purified feldamycin finished product.
2. a kind of method that according to claim 1 flash chromatography prepares feldamycin, it is characterised in that：The step (1)
In, the addition of 60%～80% ethanol is that 1～8 liter is added in per kilogram of wet mycelium；Extraction temperature is 10～50 DEG C；Leaching
The time is carried for 2～8h.
3. a kind of method that according to claim 1 flash chromatography prepares feldamycin, it is characterised in that：The step (2)
In, polystyrene resin post be DIAION HP-20 polystyrene resin posts, receive micro- NM-200 polystyrene resins post, D101 gather
Styrene resin post, HPD-400 polystyrene resin posts, AmberliteXAD-8 polystyrene resins post or HZ816 polyphenyl second
Any one in olefine resin post.
4. a kind of method that according to claim 1 flash chromatography prepares feldamycin, it is characterised in that：The step (3)
In, the filler of reversed-phase silica gel chromatography post is from any one in the alkyl silica gel bonded phase of C4, C8 or C18, and particle diameter is 50 μm.
5. a kind of method that according to claim 1 flash chromatography prepares feldamycin, it is characterised in that：The step (4)
In, the filler of reversed-phase silica gel chromatography post is from any one in the alkyl silica gel bonded phase of C4, C8 or C18, and particle diameter is 50 μm.
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| CN109251229A (en) * | 2017-07-14 | 2019-01-22 | 成都大学 | The method for isolating and purifying feldamycin |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109251229A (en) * | 2017-07-14 | 2019-01-22 | 成都大学 | The method for isolating and purifying feldamycin |
| CN109251229B (en) * | 2017-07-14 | 2021-08-31 | 成都大学 | Method for separating and purifying fidaxomicin |
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