CN103421025B - Prepare the method for 7 α-methoxy cephalosporin C - Google Patents
Prepare the method for 7 α-methoxy cephalosporin C Download PDFInfo
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Abstract
The present invention discloses a kind of method of preparing 7 α-methoxy cephalosporin C, it is characterized in that described method comprises by conversion fluid pre-treatment, polymeric adsorbent Image processing and reversed-phase silica gel column chromatography processing, from by cephalosporin substrate conversion being the step of preparing 7 α-methoxy cephalosporin C the conversion fluid of 7 α-methoxy cephalosporin C. Described method not only technique is simple, can be good at removing the impurity such as the very similar conversion of substrate of structure, intermediate product, and yield and the purity of acquisition methoxy cephalosporin C are all very high.
Description
Technical field
The invention belongs to pharmaceutical engineering field, be specifically related to one and prepare high-purity 7 α-methoxyl group cephaloThe method of rhzomorph C (7 α-MethoxycephalosporinC).
Background technology
Cephalosporins is that in anti-infectives, development early, is applied a wider large class.7 α-methoxy cephalosporin (its chemical structural formula wherein growing up in early 1970sAs shown in the formula I), because there is a trans methoxyl group C7 position on beta-lactam nucleus, acyl in stopingApproaching of amine ring and enzyme molecule, and then improved the stability of medicine to beta-lactamase. For thisThe research of class medicine is latest developments Disciplinary Frontiers more rapidly.
The existing multiple product listing of methoxy cephalosporins, comprises Cefminox, cephalo U.S.Azoles etc. The common semi-synthetic intermediate of this class medicine is methoxy cephalosporin. Methoxyl group cephalo bacteriumThe synthetic of element generally can be by chemical synthesis and microbe transformation method. Chemical method is at present in the industryGenerally adopt method, main direction of studying is all to synthesize taking 7-ACA as raw material, for example inState patent CN200910075806.6 has reported taking 7-ACA as raw material, and the reaction of first mercapto tetrazoleObtain 7-TMCA, 7-TMCA and methyl sulphur bromine carry out imidization, then with hexichol diazoniumMethane reaction obtains intermediate, intermediate and methylating reagent methylation reaction, and obtaining can be forThe intermediate 7-MAC of synthetic methoxy cephalosporins medicine. In addition, Chinese patent CN101210019AReported with C4, the cepham compound of 7 protections, C3 position halomethyl is raw material, synthesizing methoxyCephalosporin intermediate. But the method ubiquity organic solvent use amount of these chemical syntheses is large,The problems such as energy consumption is serious. And can effectively address these problems by the method for bio-transformation, utilizeThe cephamycin C that bio-transformation obtains is as synthetic methoxy cephalosporins, forCleaner production, reducing the aspects such as energy consumption has significant advantage.
Document (TheConversionofCephalosporinsto7 α-MethoxycephalosporinsbyCell-FreeExtractofStreptomycesclavuligerus.Biochem.J，1980,186:613-616) report utilize clavuligerus (Streptomycesclavuligerus,ATCC27064) crude enzyme liquid after clasmatosis, adds co-factor and cephalosporin substrate, transformsObtain 7 α-methoxy cephalosporin C, but there is no at present report from transformation system by product 7 α-The method of methoxy cephalosporin C separation and purification. Separately there is document (Non-radioactiveassayandHPLCanalysisofcephalosporinC7α-methocxylationbycell-freeextractsofStreptomycesclavuligerus.ApplMicrobiolBiotechnol, 1991,35:793-797) reportDow Chemical is synthesized the method for 7 α-methoxy cephalosporin C, and α-benzhydryl-N-t-fourth is first coupledThe own diester of ketone-D-alpha-amido and benzhydryl-7-aminocephalosporinic acid ester, obtain N-t-butanone-bis-bis-The CPC of benzyl esters, then using methyl alcohol lithium salts as methyl donor, adds the anti-of methoxyl groupShould, finally slough hydroxyl and amino blocking group, obtain 7 α-methoxy cephalosporin C. ChangingLearn in building-up process, used more chemical reagent, reaction is complicated. Although in chemical synthesisCheng Zhong, by organic solvent extraction, revolve the steps such as steaming and can obtain the more than 90% 7 α-methoxy of purityBase cephalosporin, but because system after bio-transformation and chemical synthesis system have very big-difference,Thereby report separation methoxy cephalosporin C method and be not suitable for purifying from converted productMethoxy cephalosporin C.
Summary of the invention
For solving the deficiencies in the prior art, the present invention relates to one and prepare high-purity 7 α-methoxyThe method of base cephalosporin, the method not only technique is simple, can be good at removing structure veryThe similarly impurity such as conversion of substrate, intermediate product, and obtain methoxy cephalosporin C yield andPurity is all very high.
Therefore, the invention provides a kind of method of preparing 7 α-methoxy cephalosporin C, its feature existsComprise by conversion fluid pre-treatment, polymeric adsorbent Image processing and reverse phase silica gel post layer in described methodAnalyse processing, from being the conversion fluid of 7 α-methoxy cephalosporin C by cephalosporin substrate conversionPrepare the step of 7 α-methoxy cephalosporin C.
According to of the present invention one preferred embodiment, described method comprises the following steps:
A) in every milliliter of conversion fluid, add the glacial acetic acid of 30-70 μ l to mix, make the large portion in conversion fluidPoint albuminous degeneration precipitation, suction filtration is got the filtrate of clarification, with sour adjust pH be 3.0-5.0, bePost mother liquor;
B) the upper prop mother liquor obtaining is slowly passed through to macroporous adsorption resin chromatography post; With pH be 2.5-4.5Acid rinsing pillar 2-6CV (column volume); Then with containing 10%-40% (v: v), preferably15-30% (v: the v) aqueous solution of low fat alcohol, 2-5CV wash-out pillar, substep is collected and HPLCDetect efflux; The receiving flask merging that 7 α-methoxy cephalosporin CHPLC purity is greater than to 15%,Adjust pH3.0-5.0, reduced vacuum concentrated by rotary evaporation obtains concentrate.
C) sour water pretreatment octadecyl (C18) the bonded silica gel chromatographic column that is 3.0-5.0 by pH value,And by the concentrate obtaining in b) slowly by above-mentioned C18 reverse phase silica gel post; Then by pH value beThe acid rinsing pillar 3-4CV of 3.0-5.0; Again with adding (the v: v) methyl alcohol or ethanol containing 1%-3%The aqueous solution, 2-4CV wash-out pillar, substep collect and HPLC detect efflux, then with containingThe aqueous solution of 4%-8% methyl alcohol or ethanol, 3-8CV wash-out pillar, substep is collected and HPLC detects;The receiving flask merging that purity is greater than to 85%, freeze-drying after reduced vacuum concentrated by rotary evaporation, makes 7 α-methoxyThe unformed powder of base cephalosporin.
The inventive method a) in step acid used be hydrochloric acid or sulfuric acid, be preferably hydrochloric acid.
The inventive method a) in step conversion fluid suction filtration get after filtrate, preferably pH value is adjusted to 3.0-4.5.
The inventive method a) in step low fat alcohol be methyl alcohol, ethanol, acetone or isopropyl alcohol, preferablyFor methyl alcohol or ethanol.
The inventive method b) in step macroporous absorbent resin be commercially available unmodified polystyrene-divinylbenzeneType macrolattice middle polarity resin, its specific area >=300m2/ g, characteristic aperture isMediation particle diameter 250-840 μ m. The preferably HZ-816 of Shanghai Huazhen Science and Technology Co., Ltd., HZ-803,HZ-818 type resin, the XAD-16N of Rhom and Hass, XAD-16HP, XAD1600N,XAD1180, XAD-4 or XAD-18 type resin, most preferably be XAD-18 resin.
In the present invention, term " mediation particle diameter " is defined as in hybrid particles, all particles flatAll diameters.
The inventive method b), c) sour water in step refers to contain a small amount of aqueous acid, and this acid is sulphurAcid or hydrochloric acid, be preferably hydrochloric acid.
The inventive method b), c) sour water described in step is that pH is 2.5-4.5, preferred 3.0-4.0The aqueous solution.
The inventive method b) lower aliphatic of 10%-40% described in step alcohol solution be at distilled water orIt is prepared by the alcohol of 10%-40% in deionized water, adding volume; Described 15%-30% lower aliphatic alcoholsThe aqueous solution is that in distilled water or deionized water, to add volume be prepared by the alcohol of 15%-30%.
The inventive method c) in step octadecyl (C18) bonded silica gel filler be spherical hydrophilic, spyLevying aperture isThe particle diameter that is in harmonious proportion is 5-70 μ m. Preferably Suzhou Sepax Technologies, Inc. fills outMaterial Bio-C8, HP-C18; The filler DYB7130 of Shanghai great You chromatographic technique Services Co., Ltd,DYB7219。
The inventive method c) described in step containing 1%-3% (v: v) in the aqueous solution of methyl alcohol or ethanolPreferred alcohols concentration is the ethanol of 1%-2% or the methyl alcohol that determining alcohol is 2%-3%; Described 4%-8% (slightlyHigher than concentration before) preferred alcohols concentration is 4%-5% in the aqueous solution of methyl alcohol or ethanol ethanol orDetermining alcohol is the methyl alcohol of 4%-7%.
The chromatographic column ratio of height to diameter that the inventive method b), c) is used in step is more than or equal to 3: 1.
In the inventive method step b) Main Function be to remove the protein in conversion fluid, salt,Pigment etc. some not with the impurity of resin adsorption, this has side for improving step separating effect c) very muchHelp.
Step of the present invention is carried out wash-out object product with the low fat alcohol of low concentration in c), can be wellRemove other strong polar impurities, and can be good at by the substrate of methoxy cephalosporin C and conversion,Intermediate product separates, and the object product purity obtaining is higher.
Method of the present invention is simple, and resin and reverse phase silica gel filler can Reusabilities, regeneration sideJust, the few and recovery capable of circulation of the organic solvent amount of use, and the purity of the object product obtaining is very high.
Brief description of the drawings
Fig. 1 is the process that substrate cephalosporin is converted into 7 α-methoxy cephalosporin C;
Fig. 2 is 7 α-methoxy cephalosporin C (HPLC purity 97.6%) that embodiment 1 obtainsHigh performance liquid chromatography (HPLC) collection of illustrative plates (7 α in figure-methoxy cephalosporin C appearance time is17.4min)；
Fig. 3 is 7 α-methoxy cephalosporin C (HPLC purity 97.6%) that embodiment 1 obtainsMass-spectrogram;
Fig. 4 is 7 α-methoxy cephalosporin C (HPLC purity 97.6%) that embodiment 1 obtainsNuclear-magnetism H1 spectrum.
Detailed description of the invention
The present invention 7 α-methoxy cephalosporin CHPLC testing conditions is as follows:
Chromatographic column: post UltimateAQ-C18,5 μ m, 250 × 4.6mm;
Detect wavelength: 272nm;
Column temperature: 30 DEG C;
Flow velocity: 0.8ml/min;
Mobile phase: A phase: add 2.4ml formic acid in 1L water, 0.77g ammonium acetate
B phase: add 2ml formic acid in 1L acetonitrile
Thalli growth condition used in the present invention is as follows:
Bacterial classification: StreptomycesclavuligerusATCC27064 (NRRL3585)
Inclined-plane is cultivated
Culture medium (g/L): malt extract 10, yeast extract 4, glucose 4, agar 25,PH7.0-7.4,121 DEG C of sterilizing 30min;
Condition of culture: cultivate 7-10 days for 28 DEG C.
Seed culture
Culture medium (g/L): glucose 10, import peptone 10, import yeast extract 5,121 DEG C of sterilizing 20min;
Condition of culture: 28 DEG C, 250rpm, cultivates 2 days.
Conversion condition used in the present invention is as follows:
Clavuligerus StreptomycesclavuligerusATCC27064 trains in seed culture mediumSupport suction filtration after 2 days and collect thalline, and with after deionized water washing, put-20 DEG C frozen.
Every gram of frozen thalline is added to 10ml phosphate buffer (pH7.4), melt rear ultrasonic brokenWall, as first enzyme liquid, adds co-factor 2-oxoglutaric acid (final concentration 2mM), L-AA(final concentration 1mM), FeSO4(final concentration 1mM), S-adenosylmethionine (SAM,Final concentration 1mM) and substrate cephalosporin (final concentration 500 μ g/ml) mix constant temperature oscillationTransform (28 DEG C, 240r/min) 4 to 6h.
Embodiment 1
Get conversion fluid 500ml, the content of wherein 7 α-methoxy cephalosporin C is about 0.2mg/ml,HPLC purity is about 7%, adds 25ml glacial acetic acid in conversion fluid, mixes rear filtration, gets filterLiquid, recording pH value is 3.5, without regulating pH value again.
Fill a high about 20cm, the XAD-18 macroporous adsorption resin chromatography post of diameter 5cm, by upperState filtrate and cross post with the flow velocity of 7ml/min, the sour water 3CV column scrubber of the pH3.5 preparing with hydrochloric acidSon; With 20% ethanolic solution 3CV washing pillar, each 70ml collects one bottle again. Use HPLCDetect, receiving flask higher methoxy cephalosporin C content is merged, purity is 24%.
Fill a high about 10cm, the Bio-C18 chromatographic column of diameter 2cm, uses hydrochloric acid secure phBe that 3.5 sour water is processed pillar. The part reduced vacuum concentrated by rotary evaporation that previous step is merged, adjusts pHAfter value 3.5, upper prop, flow velocity is 1ml/min. The sour water 3CV that is 3.5 by the pH value of hydrochloric acid preparationWashing pillar, removes impurity; Then wash with 2% methanol solution 3CV, collect every 15mlOne pipe, HPLC detects; Wash with 4% methanol solution 4CV, every 10ml collects a pipe again,HPLC detects.
Part higher purity is merged, and the purity of last amalgamation liquid is 90%, and yield is 65%.
Embodiment 2
Get conversion fluid 500ml, the content of wherein 7 α-methoxy cephalosporin C is about 0.2mg/ml,HPLC purity is about 6%, adds 25ml glacial acetic acid in conversion fluid, mixes rear filtration, gets filterLiquid, recording pH value is 3.6, without regulating pH value again.
Fill a high about 20cm, the XAD-4 macroporous adsorption resin chromatography post of diameter 5cm, by upperState filtrate and cross post with the flow velocity of 7ml/min, the sour water 3CV column scrubber of the pH3.5 preparing with hydrochloric acidSon; With 30% ethanolic solution 3CV washing pillar, each 70ml collects one bottle again. Use HPLCDetect, the part that has more methoxy cephalosporin C is merged, purity is 23%.
Fill a high about 10cm, the HP-C18 chromatographic column of diameter 2cm, uses hydrochloric acid secure phBe that 3.5 sour water is processed pillar. The part reduced vacuum concentrated by rotary evaporation that previous step is merged, adjusts pHAfter value 3.5, upper prop, flow velocity is 1ml/min. The sour water 3CV that is 3.5 by the pH value of hydrochloric acid preparationWashing pillar, removes impurity; Then wash with 1% ethanolic solution 3CV, collect every 15mlOne pipe, HPLC detects; Wash with 4% ethanolic solution 4CV, every 10ml collects a pipe again,HPLC detects.
Part higher purity is merged, and the purity of last amalgamation liquid is 92%, and yield is 62%.
Embodiment 3
Get conversion fluid 500ml, the content of wherein 7 α-methoxy cephalosporin C is about 0.15mg/ml,HPLC purity is about 5%, adds 25ml glacial acetic acid in conversion fluid, mixes rear filtration, gets filterLiquid, recording pH value is 3.6, without regulating pH value again.
Fill a high about 20cm, the HZ-816 macroporous adsorption resin chromatography post of diameter 5cm, by upperState filtrate and cross post with the flow velocity of 6ml/min, the sour water 3CV column scrubber of the pH3.5 preparing with hydrochloric acidSon; With 15% ethanolic solution 3CV washing pillar, each 70ml collects one bottle again. Use HPLCDetect, the part that has more methoxy cephalosporin C is merged, purity is 20%.
Fill a high about 10cm, the DYB7130 of diameter 2cm (be in harmonious proportion m) chromatographic column of particle diameter 5 μ,The sour water that is 3.5 with hydrochloric acid secure ph is processed pillar. The part reduced vacuum that previous step is mergedConcentrated by rotary evaporation, after adjust pH 3.5, upper prop, flow velocity is 0.8ml/min. With the pH of hydrochloric acid preparationValue is 3.5 sour water 3CV washing pillar, removes impurity; Then use 3% methanol solution 3CVWashing, collects a pipe every 15ml, and HPLC detects; Wash with 6% methanol solution 4CV again,Every 10ml collects a pipe, and HPLC detects.
Part higher purity is merged, and the purity of last amalgamation liquid is 92%, and yield is 64%.
Embodiment 4
Get conversion fluid 500ml, the content of wherein 7 α-methoxy cephalosporin C is about 0.2mg/ml,HPLC purity is about 6%, adds 25ml glacial acetic acid in conversion fluid, mixes rear filtration, gets filterLiquid, recording pH value is 3.5, without regulating pH value again.
Fill a high about 20cm, the HZ-803 macroporous adsorption resin chromatography post of diameter 5cm, by upperState filtrate and cross post with the flow velocity of 7ml/min, the sour water 3CV column scrubber of the pH3.5 preparing with hydrochloric acidSon; With 20% ethanolic solution 3CV washing pillar, each 70ml collects one bottle again. Use HPLCDetect, the part that has more methoxy cephalosporin C is merged, purity is 22%.
Fill a high about 10cm, the DYB7130 of diameter 2cm (be in harmonious proportion m) chromatographic column of particle diameter 40 μ,The sour water that is 3.5 with hydrochloric acid secure ph is processed pillar. The part reduced vacuum that previous step is mergedConcentrated by rotary evaporation, after adjust pH 3.5, upper prop, flow velocity is 0.8ml/min. With the pH of hydrochloric acid preparationValue is 3.5 sour water 3CV washing pillar, removes impurity; Then use 3% methanol solution 3CVWashing, collects a pipe every 15ml, and HPLC detects; Wash with 7% methanol solution 4CV again,Each 10ml collects a pipe, and HPLC detects.
Part higher purity is merged, and the purity of last amalgamation liquid is 92%, and yield is 62%.
Embodiment 5
Get conversion fluid 500ml, the content of wherein 7 α-methoxy cephalosporin C is about 0.2mg/ml,HPLC purity is about 5%, adds 25ml glacial acetic acid in conversion fluid, mixes rear filtration, gets filterLiquid, recording pH value is 3.6, without regulating pH value again.
Fill a high about 20cm, the HZ-818 macroporous adsorption resin chromatography post of diameter 5cm, by upperState filtrate and cross post with the flow velocity of 7ml/min, the sour water 3CV column scrubber of the pH3.5 preparing with hydrochloric acidSon; With 35% ethanolic solution 3CV washing pillar, each 70ml collects one bottle again. Use HPLCDetect, the part that has more methoxy cephalosporin C is merged, purity is 20%.
Fill a high about 10cm, the DYB7130 of diameter 2cm (be in harmonious proportion m) chromatographic column of particle diameter 20 μ,The sour water that is 3.5 with hydrochloric acid secure ph is processed pillar. The part reduced vacuum that previous step is mergedConcentrated by rotary evaporation, after adjust pH 3.5, upper prop, flow velocity is 1ml/min. With the pH of hydrochloric acid preparationValue is 3.5 sour water 3CV washing pillar, removes impurity; Then use 2% ethanolic solution 3CVWashing, collects a pipe every 15ml, and HPLC detects; Wash with 4% ethanolic solution 4CV again,Every 10ml collects a pipe, and HPLC detects.
Part higher purity is merged, and the purity of last amalgamation liquid is 90%, and yield is 65%.
Embodiment 6
Get conversion fluid 500ml, the content of wherein 7 α-methoxy cephalosporin C is about 0.2mg/ml,HPLC purity is about 5%, adds 25ml glacial acetic acid in conversion fluid, mixes rear filtration, gets filterLiquid, recording pH value is 3.4, without regulating pH value again.
Fill a high about 20cm, the XAD-16N macroporous adsorption resin chromatography post of diameter 5cm, willAbove-mentioned filtrate is crossed post with the flow velocity of 7ml/min, the sour water 3CV washing of the pH3.5 preparing with hydrochloric acidPillar; With 30% ethanolic solution 3CV washing pillar, each 70ml collects one bottle again. Use HPLCDetect, the part that has more methoxy cephalosporin C is merged, purity is 19%.
Fill a high about 10cm, the DYB7219 of diameter 2cm (be in harmonious proportion m) chromatographic column of particle diameter 5 μ,The sour water that is 3.5 with hydrochloric acid secure ph is processed pillar. The part reduced vacuum that previous step is mergedConcentrated by rotary evaporation, after adjust pH 3.5, upper prop, flow velocity is 1ml/min. With the pH of hydrochloric acid preparationValue is 3.5 sour water 3CV washing pillar, removes impurity; Then use 2% methanol solution 3CVWashing, collects a pipe every 15ml, and HPLC detects; Wash with 5% methanol solution 4CV again,Every 10ml collects a pipe, and HPLC detects.
Part higher purity is merged, and the purity of last amalgamation liquid is 93%, and yield is 62%.
Embodiment 7
Get conversion fluid 500ml, the content of wherein 7 α-methoxy cephalosporin C is about 0.2mg/ml,HPLC purity is about 4%, adds 25ml glacial acetic acid in conversion fluid, mixes rear filtration, gets filterLiquid, recording pH value is 3.5, without regulating pH value again.
Fill a high about 20cm, the XAD-1600N macroporous adsorption resin chromatography post of diameter 5cm,Above-mentioned filtrate is crossed to post with the flow velocity of 7ml/min, and the sour water 3CV of the pH3.5 preparing with hydrochloric acid washesWash pillar; With 20% ethanolic solution 3CV washing pillar, each 70ml collects one bottle again. Use HPLCDetect, the part that has more methoxy cephalosporin C is merged, purity is 23%.
Fill a high about 10cm, the DYB7219 of diameter 2cm (be in harmonious proportion m) chromatographic column of particle diameter 50 μ,The sour water that is 3.5 with hydrochloric acid secure ph is processed pillar. The part reduced vacuum that previous step is mergedConcentrated by rotary evaporation, after adjust pH 3.5, upper prop, flow velocity is 1ml/min. With the pH of hydrochloric acid preparationValue is 3.5 sour water 3CV washing pillar, removes impurity; Then use 2% ethanolic solution 3CVWashing, collects a pipe every 15ml, and HPLC detects; Wash with 4% ethanolic solution 4CV again,Every 10ml collects a pipe, and HPLC detects.
Part higher purity is merged, and the purity of last amalgamation liquid is 91%, and yield is 64%.
Embodiment 8
Get conversion fluid 500ml, the content of wherein 7 α-methoxy cephalosporin C is about 0.2mg/ml,HPLC purity is about 7%, adds 25ml glacial acetic acid in conversion fluid, mixes rear filtration, gets filterLiquid, recording pH value is 3.5, without regulating pH value again.
Fill a high about 20cm, the XAD-1180N macroporous adsorption resin chromatography post of diameter 5cm,Above-mentioned filtrate is crossed to post with the flow velocity of 7ml/min, and the sour water 3CV of the pH3.5 preparing with hydrochloric acid washesWash pillar; With 25% ethanolic solution 3CV washing pillar, each 70ml collects one bottle again. Use HPLCDetect, the part that has more methoxy cephalosporin C is merged, purity is 20%.
Fill a high about 10cm, the DYB7219 of diameter 2cm (be in harmonious proportion m) chromatographic column of particle diameter 20 μ,The sour water that is 3.5 with hydrochloric acid secure ph is processed pillar. The part reduced vacuum that previous step is mergedConcentrated by rotary evaporation, after adjust pH 3.5, upper prop, flow velocity is 1ml/min. With the pH of hydrochloric acid preparationValue is 3.5 sour water 3CV washing pillar, removes impurity; Then use 3% methanol solution 3CVWashing, collects a pipe every 15ml, and HPLC detects; Wash with 6% methanol solution 4CV again,Every 10ml collects a pipe, and HPLC detects.
Part higher purity is merged, and the purity of last amalgamation liquid is 91%, and yield is 64%.
Claims (8)
1. the method for preparation 7 α-methoxy cephalosporin C, is characterized in that described method comprises logicalCross conversion fluid pre-treatment, polymeric adsorbent Image processing and reversed-phase silica gel column chromatography processing, from by cephaloRhzomorph C substrate conversion is to prepare 7 α-methoxyl group cephalo in the conversion fluid of 7 α-methoxy cephalosporin CThe step of rhzomorph C;
Described conversion fluid pre-treatment comprises the steps:
In every milliliter of conversion fluid, add the glacial acetic acid of 30-70 μ l to mix, suction filtration is got the filtrate of clarification,With sour adjust pH be 3.0-5.0, obtain upper prop mother liquor;
Described polymeric adsorbent Image processing comprises the steps:
Described upper prop mother liquor, by macroporous adsorption resin chromatography post, is obtained to concentrate; With pH beThe acid rinsing pillar 2-6CV of 2.5-4.5; Then with methyl alcohol, ethanol, acetone containing 10%-40%Or isopropanol water solution, 2-5CV wash-out pillar, substep is collected and HPLC detects efflux; By 7 α-The receiving flask merging that methoxy cephalosporin CHPLC purity is greater than 15%, adjusts pH3.0-5.0, subtractsPress vacuum concentrated by rotary evaporation to obtain concentrate;
Described reversed-phase silica gel column chromatography processing comprises the steps:
The sour water pretreatment octadecyl silane chromatographic column that is 3.0-5.0 by pH value, and by instituteState concentrate slowly by above-mentioned C18 reverse phase silica gel post; Then the sour water that is 3.0-5.0 by pH valueWashing pillar 3-4CV; Again with the aqueous solution adding containing 1%-3% methyl alcohol or ethanol, 2-4CV wash-outPillar, substep is collected and HPLC detects efflux, then with the water containing 4%-8% methyl alcohol or ethanolSolution, 3-8CV wash-out pillar, substep is collected and HPLC detects; The receipts that purity is greater than to 85%Collection bottle merges, and freeze-drying after reduced vacuum concentrated by rotary evaporation makes 7 α-methoxy cephalosporin C without fixedType powder.
2. the method for claim 1, is characterized in that described acid is hydrochloric acid or sulfuric acid.
3. the method for claim 1, is characterized in that described in described conversion fluid pre-treatmentPH value is adjusted to 3.0-4.5.
4. the method for claim 1, is characterized in that described macroporous absorbent resin is polyphenylEthene-divinylbenzene type macrolattice middle polarity macroporous absorbent resin, its specific area >=300m2/g，Characteristic aperture isMediation particle diameter 250-840 μ m.
5. method as claimed in claim 4, is characterized in that described reversed-phase silica gel column chromatography processingDescribed in the pH value of sour water be 3.0-4.0.
6. the method for claim 1, is characterized in that described octadecyl silaneFiller is spherical hydrophilic, and characteristic aperture isThe particle diameter that is in harmonious proportion is 5-70 μ m.
7. the method for claim 1, is characterized in that described containing 1%-3% methyl alcohol or ethanolThe aqueous solution in, determining alcohol is 1%-2% ethanol or 2%-3% methyl alcohol; Described containing 4%-8% methyl alcohol orIn the aqueous solution of ethanol, determining alcohol is 4%-5% ethanol or 4%-7% methyl alcohol.
8. the method for claim 1, is characterized in that described octadecyl silane layerAnalyse post ratio of height to diameter and be more than or equal to 3:1.
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