CS200022B1 - Method for the isolation of cephalosporin c from fermentation substrate - Google Patents
Method for the isolation of cephalosporin c from fermentation substrate Download PDFInfo
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- CS200022B1 CS200022B1 CS369078A CS369078A CS200022B1 CS 200022 B1 CS200022 B1 CS 200022B1 CS 369078 A CS369078 A CS 369078A CS 369078 A CS369078 A CS 369078A CS 200022 B1 CS200022 B1 CS 200022B1
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- Prior art keywords
- cephalosporin
- isolation
- aqueous
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- HOKIDJSKDBPKTQ-GLXFQSAKSA-N cephalosporin C Chemical compound S1CC(COC(=O)C)=C(C(O)=O)N2C(=O)[C@@H](NC(=O)CCC[C@@H](N)C(O)=O)[C@@H]12 HOKIDJSKDBPKTQ-GLXFQSAKSA-N 0.000 title claims description 43
- 238000000034 method Methods 0.000 title claims description 18
- 238000000855 fermentation Methods 0.000 title claims description 15
- 230000004151 fermentation Effects 0.000 title claims description 15
- 238000002955 isolation Methods 0.000 title claims description 7
- 239000000758 substrate Substances 0.000 title 1
- HOKIDJSKDBPKTQ-GLXFQSAKSA-M cephalosporin C(1-) Chemical compound S1CC(COC(=O)C)=C(C([O-])=O)N2C(=O)[C@@H](NC(=O)CCC[C@@H]([NH3+])C([O-])=O)[C@@H]12 HOKIDJSKDBPKTQ-GLXFQSAKSA-M 0.000 claims description 41
- 239000002594 sorbent Substances 0.000 claims description 15
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 12
- 239000000243 solution Substances 0.000 claims description 10
- 239000007864 aqueous solution Substances 0.000 claims description 7
- 229920005989 resin Polymers 0.000 claims description 5
- 239000011347 resin Substances 0.000 claims description 5
- 150000003839 salts Chemical class 0.000 claims description 5
- 239000003480 eluent Substances 0.000 claims description 3
- 238000010828 elution Methods 0.000 claims description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 claims description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 2
- 239000008351 acetate buffer Substances 0.000 claims description 2
- 239000000908 ammonium hydroxide Substances 0.000 claims description 2
- 238000003795 desorption Methods 0.000 claims description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N acetone Substances CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 17
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 239000000706 filtrate Substances 0.000 description 7
- 239000011701 zinc Substances 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 230000003115 biocidal effect Effects 0.000 description 5
- 238000000746 purification Methods 0.000 description 5
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 description 4
- 239000005695 Ammonium acetate Substances 0.000 description 4
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 4
- ZOIORXHNWRGPMV-UHFFFAOYSA-N acetic acid;zinc Chemical compound [Zn].CC(O)=O.CC(O)=O ZOIORXHNWRGPMV-UHFFFAOYSA-N 0.000 description 4
- 229940043376 ammonium acetate Drugs 0.000 description 4
- 235000019257 ammonium acetate Nutrition 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 239000004246 zinc acetate Substances 0.000 description 4
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 239000004202 carbamide Substances 0.000 description 3
- 229920001429 chelating resin Polymers 0.000 description 3
- 150000002500 ions Chemical class 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 229930186147 Cephalosporin Natural products 0.000 description 2
- 150000007513 acids Chemical class 0.000 description 2
- 229940124587 cephalosporin Drugs 0.000 description 2
- 150000001780 cephalosporins Chemical class 0.000 description 2
- 229910001385 heavy metal Inorganic materials 0.000 description 2
- WQYVRQLZKVEZGA-UHFFFAOYSA-N hypochlorite Chemical compound Cl[O-] WQYVRQLZKVEZGA-UHFFFAOYSA-N 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 238000009413 insulation Methods 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- HSHGZXNAXBPPDL-HZGVNTEJSA-N 7beta-aminocephalosporanic acid Chemical compound S1CC(COC(=O)C)=C(C([O-])=O)N2C(=O)[C@@H]([NH3+])[C@@H]12 HSHGZXNAXBPPDL-HZGVNTEJSA-N 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- 102100021935 C-C motif chemokine 26 Human genes 0.000 description 1
- 101000897493 Homo sapiens C-C motif chemokine 26 Proteins 0.000 description 1
- 230000001594 aberrant effect Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 239000002156 adsorbate Substances 0.000 description 1
- 229910052783 alkali metal Inorganic materials 0.000 description 1
- -1 alkali metal salts Chemical class 0.000 description 1
- 229910052784 alkaline earth metal Inorganic materials 0.000 description 1
- 150000001342 alkaline earth metals Chemical class 0.000 description 1
- 239000012670 alkaline solution Substances 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- 238000005349 anion exchange Methods 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000002199 base oil Substances 0.000 description 1
- 229940088623 biologically active substance Drugs 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 229920001577 copolymer Polymers 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 238000004042 decolorization Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 150000002484 inorganic compounds Chemical class 0.000 description 1
- 229910001410 inorganic ion Inorganic materials 0.000 description 1
- 229910010272 inorganic material Inorganic materials 0.000 description 1
- 229910052748 manganese Inorganic materials 0.000 description 1
- 229910052753 mercury Inorganic materials 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 150000007530 organic bases Chemical class 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 229920003002 synthetic resin Polymers 0.000 description 1
- 239000000057 synthetic resin Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Landscapes
- Cephalosporin Compounds (AREA)
Description
Vynález sa týká sposobu izolácie cefalosporinu C z fermentačnej pody pri súčasnom snížení obsahu nežiadúcich nečistot v Izolovaném produkte.The invention relates to a method of isolating cephalosporin C from a fermentation pod while reducing the content of undesirable impurities in the isolated product.
Izoláola a Čistenia cefalosporinu C navazuje na jeho přípravu fsrmentačnou cestou. Je známe, že keá fsrmsntujeme živná mádlům určitými mlkroorganlzmarni například kmeňom Cephaloeporium acrsmonlum ATCC 14553 obsahuje mádlům biologicky aktívnu látku /Craeford K. a kol.: Zntlbiotio productlon by a species of Cephaloeporium. J. Cen. Mlcrobiol. 6 /1952/, str. 47 - 59} Newton Q. G, F., Abraham E. P.j Cephaloeporium C a new antibiotlc containlng sulphur and 0-aminoadlpic acld. Nátuře 175 /1955/, str. 548; Hewitt W, L.s The čepil alospor ins. The Journal of Infeetlons Dissares 128 /1973/, str. 312. Tieto ferraentačná postupy sú uvedené tiež vo Svajčiarakych patentech 565246, 570459, 570460, v NSR pat. č.Isolaol and purification of cephalosporin C follows its preparation by a fermentation route. It is known that we cultivate nutrients with certain microorganisms such as Cephaloeporium acrsmonlum ATCC 14553 containing the biologically active substance (Craeford K. et al., Zntlbiotio productlon by a species of Cephaloeporium). J. Cen. Mlcrobiol. 6 (1952), p. 47-59} Newton Q.G, F., Abraham E.P. Nature 175 (1955), p. 548; Hewitt W, L.s The capped alospore ins. The Journal of Infeetlons Dissares 128 (1973), p. 312. These ferraentation processes are also disclosed in U.S. Patent Nos. 5,624,456, 5,7459, 5,70460, in German Pat. no.
492 053, 2 239 321, USA pat. č. 3 082 155, v brit. pat. č. 820 422. Tieto patenty obsahujú tisž postupy získavanla a čistenia cefalosporinu C zo surověj fermentačnsj pody, zahrnu júce tieto následné operácie: acldifikácia, flltrácla, dekolorizácia, odstránenie anorganických lontov, adsorpclu antibiotika a elúclu. Bodla iných izolačnýeh postupov napr. US pat. č. 3 739 002, 3 931 161, 3 835 129, NSR pat. č. 2 157 693, 2 255 973, pat.No. 492,053, 2,239,321, U.S. Pat. no. 3,082,155, in British. pat. no. 820,422. These patents include one or more processes for recovering and purifying cephalosporin C from a crude fermentation pod, including the following operations: acdification, filtration, decolorization, removal of inorganic ions, adsorption of the antibiotic, and elution. Stabs of other isolation procedures e.g. US Pat. no. 3,739,002, 3,931,161, 3,835,129, German Pat. no. 2 157 693, 2 255 973, U.S. Pat.
ZSSR č. 282 174 aa cefalosporín C z fermentačnsj pody získává vo formo roznych N-halogenalkanoyl čl N-benzoyl dsrlvátov, ktoré sa áalsj prevedú na soli.USSR no. 282 174 aa cephalosporin C from the fermentation pod obtains in the form of various N-haloalkanoyl or N-benzoyl salts, which are further converted into salts.
Významnou skupinou izolačnýeh postupov sú postupy založená na principe delenia bázických a aoidických látok pomocou syntetických ionexov. Tieto Izolačné postupy a roznýmiAn important group of isolation processes are those based on the principle of separation of basic and azidic substances using synthetic ion exchangers. These Insulation Procedures and Miscellaneous
200 022200 022
200 022 úpravami fermentačnej pody, obměnami použitých ionexov a kombinácii ionexov rozneho stupňa bázicity, resp. acidity popisuje například franc. pat. o, 1 210 823, USe«a»t. č. 3 467 654, pat. NSR č. 1 125 564 a 1 617 456 a švéd. pat. č. 215 561.200 022 by adjusting the fermentation tray, changing the ion exchangers used and combining ion exchangers of various degrees of basicity, respectively. acidity is described, for example, in French. pat. 0, 1 210 823, U.S. Pat. no. No. 3,467,654, U.S. Pat. NSR no. 1 125 564 and 1 617 456 and Swedish. pat. no. 215 561.
Hydrofilné antibiotiká, medzi ktoré patří tiež cefalosporín C sa móžu zo zložitých zmesi napr. zo surověj fermentačnej pody obsahujúoej velké množstvo organických a anorganických zlúčenín, získal pomocou makroporéznych neionogénnych aorbentov typu Amberlite XAD-2, resp. XAD-4, Diaion HP-20, resp. Ostion SP-1, reep. SP-2.Hydrophilic antibiotics, including cephalosporin C, can be formulated from complex mixtures e.g. from a crude fermentation broth containing a large number of organic and inorganic compounds, obtained by means of macroporous non-ionic aberrants of the Amberlite XAD-2 type, respectively. XAD-4, Diaion HP-20, respectively. Ostion SP-1, reep. SP-2.
Izolácia týmito spoeobmi je uvedené napr. v US pat. č. 3 725 400, švéd. pat, č.The insulation by these spoilers is given e.g. in US Pat. no. 3,725,400, Swedish. Pat, no.
541 579 a v pat. ZSSR č, 350 265. Podía uvedených spósobov sa antibiotikum získané pomocou neionogénnych sorbentov áalej purifikuje na slabo béziokýeh anexoch /napr. typu Amberlite IRA-68, IR-4B, IR-45, XB-265 alebo IMAC A 131 a A 17 i?/.· Z takto přečištěného filtrátu sa može po jeho prípadnom zahuštění Izolovat cefalosporín’0 buá vo formě voínej kyseliny, alebo vo formě solí s alkalickými kovmi, reep. kovmi alkalických zemin, amonných solí, či eolí s organickými bázemi, alebo tiež vo formě lažkorozpuetných komplexov e tažkými kovmi ako sú Cu, Hg, Mn a hlavně Zn, Kvalita Zn komplexu a rýchlosl jeho zrážania eú vo veíkej miere závislé od obsahu znečistenín, hlavně rozkladných tiobuténových kyselin, vznikajúcich v priebehu bioeynťézy cefalosporínu C z metloníau /F, Benz aj.: Metionine Metaboliem and CFS-C Synthesis in Cephaloeporium acremonium. Eur. J„ Bleohem. 20 /1971/, etr. 81-88}541,579 and in U.S. Pat. USSR No. 350,265. According to the above methods, the antibiotic obtained with non-ionic sorbents is further purified on weakly bare anion exchangers (e.g. of the Amberlite type IRA-68, IR-4B, IR-45, XB-265 or IMAC A 131 and A 17 I. The cephalosporin can be isolated from the purified filtrate after concentration, if necessary, in the form of a free acid, or in the form of alkali metal salts, reep. alkaline earth metals, ammonium salts, or organic base oils, or also in the form of lysine-spread complexes with heavy metals such as Cu, Hg, Mn and especially Zn, the Zn quality of the complex and its precipitation of decomposition thiobutenic acids, formed during the bioeynesis of cephalosporin C from metlonium / F, Benz et al. Eur. J “Bleoh. 20 (1971), etr. 81-88}
J, Nueeoh H, J. Treichleť: Genet. Ind, Macroorg., Prague, 1973, etr. 324-325/.J, Nueeoh H, J. Treichlete: Genet. Ind. Macroorg., Prague, 1973, etr. 324-325 /.
Purifikácia cefalosporínu C si doteraz vyžadovala dvojstupňový postup, pozostávajúol z čistenia na neionogénnych živiciach a néslednom čistění na snexe.The purification of cephalosporin C has hitherto required a two-step procedure, consisting of purification on non-ionic resins and subsequent purification on a snex.
Túto zložitosl doterajších izolačných postupov odstraňuje předložený vynález, ktorého predmetom je spóeob Izolácie cefalosporínu C z fermentačnej pódy upravenej močovinou, spracováním filtrátu okyalenej fermentačnej pódy neionogénnymi sorbentami, deeorpoiou vodným roztokom n-butanolu alebo acetátovým tlmivým roztokom a vyzréžaním cefalosporínu C z eluátov do formy voínej kyseliny, nerozpustných solí, alebo zrážaním v podobě lažko rozpustných komplexov,This complexity of the prior art isolation processes is overcome by the present invention, which relates to a method for the isolation of cephalosporin C from a urea-treated fermentation broth, treating the acidified fermentation broth with non-ionic sorbents, deeorpoia with an aqueous solution of n-butanol or acetate cephalosporate , insoluble salts, or precipitation in the form of sparingly soluble complexes,
Týmto sa celý postup zjednodušuje na jednoetupňovú purifikáciu na živiciach bez dočieíovánia na anexe.This simplifies the procedure for a one-step purification on resins without finishing the anion exchange.
Cefaloeporin C ea eorbuje na živicu z roztoku - obvykle z filtrátu fermentačnej pódy - kde je přítomné aj veíké množstvo látok podobných tomuto antibiotiku z híadiska fyzikálně j a chemlckej povahy, Cefaloeporin C ea adsorbuje kvantitativné, neadeorbuje ea však vačšia časl nečistot, ktoré přešli ešte do filtrátu zo surověj fermentačnej pódy.Cefaloeporin C ea absorbs to the resin from the solution - usually from the fermentation broth filtrate - where a large number of substances similar to this antibiotic are present from a physical and chemical point of view, Cefaloeporin C ea adsorbs quantitatively, but does not adsorb and more of the impurities from the crude fermentation stage.
Antibiotikum naadeorbované na eorbente sa z něho desorbuje 3 až 6,5% vodným roztokem n-butanolu, alebo 0,05 až 1 M vodnými roztokmi aoetátového tlmivého roztoku o pH 4,0 až 7,0.The antibiotic adsorbed on eorbent is desorbed from it with a 3 to 6.5% aqueous solution of n-butanol, or with 0.05 to 1 M aqueous solutions of an acetate buffer of pH 4.0 to 7.0.
Vhodná pracovně oblasl pH neionogénnych eorbentov je 1 až 8 s výhodou 2 až 3. Na túto hodnotu sa surová fermentačná póda, predom upravená s 5 až 10 mg/ml močoviny, obeahujúca cefalosporín C može upravil buá katexami v H* formě, alebo róznymi kyselinami, najČaetej šie kyselinou sírovou, chlorovodíkovou, fosforečnou, šlavelovou, Ako neionogenné sorbenty pre izoláoiu cefalosporínu C dobré použiteíná eú syntetické živice o veíkoeti pórov 4 až 20 mm, mernom povrchu 100-1000 m2/g a veíkoeti častíc 0,3 až 1 mm. Sú to například eorben3A suitable working range for the pH of the non-ionic eorbents is 1 to 8, preferably 2 to 3. To this value, the crude fermentation stage, pretreated with 5 to 10 mg / ml urea, enriching cephalosporin C may be adjusted either by cation exchangers in H * form or by various acids As the nonionic sorbents for the isolation of cephalosporin C, eu synthetic resins having a pore size of 4 to 20 mm, a surface area of 100-1000 m 2 / g and a particle size of 0.3 to 1 mm are good. For example, eorben3
200 022 ty typu Oetion SP-1 a SP-2, Amberlité XAD-2 a XAD-4, Diaion HP-20, ktoré aú kopolymérmi styrénu s divinylbenzénom. Sorbenty sa regenerujú vodnými alebo vodno-acetonovými alkalickými roztokmi napr. 25% vodným roztokom acetonu, obsahujúcim 2 % NaOH. Výhodná je tiež regenerácia vodným roztokom NaOH s následným premývaním živice acetónom. Na regeneráciu sa móže tiež použil vodný roztok alkalického chlornanu a zbytok oxidovadla odstránit vhodným redukčným činidlom.200 022 those of the type Oetion SP-1 and SP-2, Amberlite XAD-2 and XAD-4, Diaion HP-20, which are styrene-divinylbenzene copolymers. Sorbents are regenerated with aqueous or aqueous-acetone alkaline solutions e.g. 25% aqueous acetone solution containing 2% NaOH. Preference is also given to regeneration with an aqueous solution of NaOH followed by washing the resin with acetone. An aqueous alkaline hypochlorite solution may also be used for recovery and the remainder of the oxidant removed with a suitable reducing agent.
Z takto získaných eluátov obsahujúcich rozpuštěný cefalosporín C sa vyzráža cefalosporín C vo formě nerozpustných solí alebo komplexov z lažkými kovmi,From the eluates thus obtained containing dissolved cephalosporin C, cephalosporin C precipitates in the form of insoluble salts or complexes of heavy metals,
Prednoslou postupu izolácie cefalosporínu C podía předloženého vynálezu je zjednodušené získanie antibiotika sposobom zaistujúcim súčasne získanie produktu z dostatoČnou čistotou potřebnou napr. pre chemickú konverziu na 7-aminocefalosporínovú kyselinu, Predmet vynálezu je áalej podložený príkladmi, ktoré však rozsah vynálezu neobmedzujú.A preferred method of isolating cephalosporin C according to the present invention is to simplify the recovery of the antibiotic in a manner ensuring simultaneously the product of sufficient purity required e.g. for the chemical conversion to 7-aminocephalosporinic acid. The present invention is further illustrated by the following non-limiting examples.
Příklad 1Example 1
K surověj fermentačnej pode sa přidá 10 mg/ml močoviny a hodnota pH sa upraví s kyselinou sírovou zriedenou s vodou v pomere 1 »4 na pH 2,6, Biomasa a áalšie zložky sa oddelia a získá sa číry roztok z obsahom 3000/x4g/ml cefalosporínu C. 5000 ml tohoto filtrátu sa podrobí perkolácii cez 4000 ml Ostionu SP-1 pri memom zaležení 1 hodina*1. Zbytok adsorbátu sa odstráni dvomi litrami vody. Zo sorbentu ea cefalosporín C eluuje 6,2% vodným roztokom n-butanolu. Získá sa 2540 ml hlavných eluétových frakcii obsahujúcich 4300,/tg cefalosporínu C v 1 ml eluátu. K takto získanému eluátu sa za chladenia a miešania přidá 5 mg octanu zinočnatého v pevnej formě na 1 mg cefalosporínu C přítomného v eluáte. Po 30 minutách miešania sa ešte přidá 2500 ml acetonu. Po 5 hodinách miešania pri teplote 3 až 5 °C sa v matečných lúhoch nameralo 200^/tg cefalosporínu C v 1 ml. Vylúčený, lažko rozpustný Zn-komplex cefalosporínu C sa oddělí, premyje studenou vodou, acetonem a vysuší. Týmto postupom sa získá 11,8 g Zn-komplexu CPS-C o čistotě 85,3 %.10 mg / ml urea is added to the crude fermentation broth and the pH is adjusted to pH 2.6 with 1: 4 dilute sulfuric acid with water. The biomass and other ingredients are separated to give a clear solution of 3000 ( x4g) content. ml of cephalosporin C. 5000 ml of this filtrate was percolated through 4000 ml of Ostion SP-1 at a meage of 1 hour * 1 . The adsorbate residue is removed with two liters of water. It eluted from sorbent e and cephalosporin C with a 6.2% aqueous solution of n-butanol. 2540 ml of major eluate fractions containing 4300 [mu] g of cephalosporin C in 1 ml of eluate were obtained. To the eluate thus obtained, 5 mg of solid zinc acetate per 1 mg of cephalosporin C present in the eluate are added with cooling and stirring. After stirring for 30 minutes, 2500 ml of acetone were added. After stirring at 3-5 ° C for 5 hours, 200 µg of cephalosporin C in 1 ml was measured in the mother liquors. The precipitated, slightly soluble Zn complex of cephalosporin C is separated, washed with cold water, acetone and dried. 11.8 g of Zn complex CPS-C with a purity of 85.3% are obtained.
Použitý sorbent Ostion SP-1 sa regeneruje 1212% vodného roztoku NaOH. Náplň sa potom premýva 2 1 vody, 411% kyseliny sírovej a potom znovu 8 1 vody. Za týmto sa náplň premyje 151 acetonu a nakoniec sa dokonale premyje vodou. Měrné zalaženie vo všetkýchThe used Ostion SP-1 sorbent is regenerated with 1212% aqueous NaOH solution. The cartridge is then washed with 2 L of water, 411% sulfuric acid and then again with 8 L of water. After this, the cartridge is washed with 151 acetone and finally washed thoroughly with water. Specific distribution in all
Μ — 1 stupnoch sa pohybovalo v rozmédzí 1 až 2 hodiny .Μ - 1 degree ranged from 1 to 2 hours.
Příklad 2Example 2
Postupuje sa ako v příklade 1, s tým rozdielom, že na elúciu cefalosporínu C zo sorbentu sa použije 3,2% vodný roztok n-butanolu a filtrát sa spracuje na sorbente Diaion HP-20, Na 250 ml náplně Diaion HP-20 sa adsorbuje 500 ml kyslého filtrátu obsahujúceho 3400/Ag/ml cefalosporínu C. Po desorpcii cefalosporínu C zo sorbentu za použitia vyššie uvedeného eluentu sa namsria v eluétoch 73 % z povodně purifikovaného množstva cefalosporínu C, K eluátu, ktorý obsahuje 7500/cg cefalosporínu C v 1 ml, Ba přidá 8 mg octanu zinočnatého na 1 mg cefalosporínu C a potom 1,25 ml acetonu na 1 ml eluátu. Získá sa 1,3 g Zn-komplexu cefalosporínu C o čistotě 80 %.The procedure is as in Example 1 except that a 3.2% aqueous n-butanol solution is used to elute cephalosporin C from the sorbent and the filtrate is treated with Diaion HP-20 sorbent. 500 ml of an acidic filtrate containing 3400 / Ag / ml of cephalosporin C. After desorption of cephalosporin C from the sorbent using the above eluent, 73% of the flood-purified amount of cephalosporin C is eluted in the eluates. Ba add 8 mg of zinc acetate per 1 mg of cephalosporin C and then 1.25 ml of acetone per ml of eluate. 1.3 g of Zn complex of cephalosporin C with a purity of 80% are obtained.
200 022200 022
Použitý sorbent Diaion HP-20 sa regeneruje postupom uvedeným v příklade 1,The Diaion HP-20 sorbent used is regenerated as described in Example 1,
Příklad 3Example 3
Postupuje sa ako v příklade 1, s tým rozdielom, že na desorpciu cefalosporínu C zo sorbentu SP-1 sa použije 0,2 M vodný roztok octanu amónneho o pH 5,5. 5000 ml filtrátu získaného postupom v příklade 1 a obsahujúcom 2850 z«g/ml cefalosporínu C sa purifikuje na sorbente Ostion SP-1. Po dssorpcii cefalosporínu C za použitia hoře uvedeného roztoku octanu amónneho sa získá 3100 ml hlavných eluátových frakcií, ktoré obsahovaly δΟδΟ^β/ιηΙ cefalosporínu C. V sekundárných eluátových frakoiách sa nameria 5,2 % z povodně purifikovaného. množstva cefalosporínu C. Hlavné eluátové frakcie sa rozdelia na dve časti a připraví saThe procedure is as in Example 1, except that a 0.2 M aqueous ammonium acetate solution of pH 5.5 is used to desorb the cephalosporin C from the SP-1 sorbent. 5000 ml of the filtrate obtained in Example 1 and containing 2850 of 'g / ml cephalosporin C was purified on SP-sorbent Ostion first After absorption of cephalosporin C using the above ammonium acetate solution, 3100 ml of the main eluate fractions containing cephalosporin C were obtained. In the secondary eluate fractions 5.2% of the flood purified was measured. The main eluate fractions were divided into two portions and prepared
AA
Zn-komplex cefalosporínu C nižáie uvedenými sposobmi.The cephalosporin C zn complex is as described below.
Jedna část eluátov sa zráža 8 mg octanu zinočnatého na 1 ml cefalosporínu C a 1 ml acetonu na 1 ml eluátu. Získá sa 8,6 g Zn-komplexu cefalosporínu C o čistotě 81,5One portion of the eluates is precipitated with 8 mg of zinc acetate per ml of cephalosporin C and 1 ml of acetone per ml of eluate. 8.6 g of cephalosporin C Zn complex having a purity of 81.5 are obtained
V druhej časti eluátov sa hodnota pH z povodněj 4,7 upraví s hydroxidom amonným na hodnotu 5,3 a potom sa zráža ako v prvej časti. Získá sa 8,1 g Zn-komplexu cefalosporínu C o čistotě 77 %.In the second part of the eluates, the pH from the flood 4.7 was adjusted to 5.3 with ammonium hydroxide and then precipitated as in the first part. 8.1 g of cephalosporin C zn complex of 77% purity are obtained.
Příklad 4Example 4
500 ml filtrátu sa purifikuje na 250 ml náplně sorbentu Diaion HP-20 sposobom uvedeným v příklade 3, s tým rozdielom, že na elúciu sa použije 0,05 M octan amónny o pH 4,2. Získá sa sluát obsahujúci VSOO^g/ml cefalosporínu 0, v ktorom js přítomné 75 % z purifikovaného množstva cefalosporínu C. Z takto získaného eluátu sa postupom uvedeným v příklade 1 připraví Zn-komplex cefalosporínu C o čistotě 87 %.500 ml of the filtrate are purified on a 250 ml diaion HP-20 sorbent cartridge as described in Example 3, except that 0.05 M ammonium acetate pH 4.2 is used for elution. A salt containing VS00 µg / ml of cephalosporin 0 is obtained, in which 75% of the purified amount of cephalosporin C is present.
Příklad 5Example 5
Postupuje sa ako v příklade 3, s tým rozdielom, že cefalosporín C sa zo sorbentu SP-1 desorbuje 1 M vodným roztokom octanu amónneho o pH 6,9 a na vyzrážanie Zn-komplexu cefalosporínu C z eluátu sa použije 10 mg octanu zinočnatého na 1 mg cefalosporínu C a 1,5 ml acetonu na 1 ml eluátu. Získá sa jemná zrazenina Zn-komplexu cefalosporínu C o čistoto 80 % a množstve 68 % z purifikovaného množstva cefalosporínu C.The procedure is as in Example 3, except that cephalosporin C is desorbed from the SP-1 sorbent with a 1 M aqueous ammonium acetate solution of pH 6.9 and 10 mg of zinc acetate per l is used to precipitate the cephalosporin C zn complex from the eluate. mg of cephalosporin C and 1.5 ml of acetone per ml of eluate. A fine precipitate of Zn complex of cephalosporin C of 80% purity and 68% of the purified amount of cephalosporin C is obtained.
PREDMET VYNÁLEZUOBJECT OF THE INVENTION
Claims (3)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS369078A CS200022B1 (en) | 1978-07-07 | 1978-07-07 | Method for the isolation of cephalosporin c from fermentation substrate |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS369078A CS200022B1 (en) | 1978-07-07 | 1978-07-07 | Method for the isolation of cephalosporin c from fermentation substrate |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CS200022B1 true CS200022B1 (en) | 1980-08-29 |
Family
ID=5377695
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CS369078A CS200022B1 (en) | 1978-07-07 | 1978-07-07 | Method for the isolation of cephalosporin c from fermentation substrate |
Country Status (1)
| Country | Link |
|---|---|
| CS (1) | CS200022B1 (en) |
-
1978
- 1978-07-07 CS CS369078A patent/CS200022B1/en unknown
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