CN101423547A - Purification method of teicoplanin - Google Patents
Purification method of teicoplanin Download PDFInfo
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- CN101423547A CN101423547A CNA2007101660155A CN200710166015A CN101423547A CN 101423547 A CN101423547 A CN 101423547A CN A2007101660155 A CNA2007101660155 A CN A2007101660155A CN 200710166015 A CN200710166015 A CN 200710166015A CN 101423547 A CN101423547 A CN 101423547A
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Abstract
The invention relates to a method for preparing glycopeptides antibiotic teicoplanin, which comprises the following steps: after teicoplanin fermented solution is subjected to filtration, adsorption by macroporous resin and decoloring by active carbon, the decoloring solution is concentrated and desolventizing by nanofilteration or hyperfiltration to obtain the teicoplanin water solution with high concentration, and then the teicoplanin is deposited from the solution by adjusting pH, adding salt or adjusting pH and adding salt simultaneously, and the mixture is filtered to obtain teicoplanin. The effective ingredients of the teicoplanin obtained through deposition are obviously improved, and other impurities are reduced. Meanwhile, the appearance color of the product is improved obviously.
Description
Technical field
The present invention relates to a kind of method of purification of separating the glycopeptide antibiotics teicoplanin, specifically, relate to a kind of method of purification of from the teicoplanin aqueous solution of 2～20W/V%, separating teicoplanin.
Background technology
In the technology patent about the teicoplanin preparation that exists now, preparation technology is mainly fermented liquid through the alkalization after-filtration, and filtrate is after resin absorption, to carry out desorb to resin with strippant, stripping liquid is through activated carbon decolorizing, and filtrate solubilizing agent precipitation is filtered and promptly got the wet product of teicoplanin.Carry out further making with extra care with resin or chromatography agent then, resin wherein is mainly macroporous adsorbent resin, and strippant is the aqueous solution of organic solvent, adds a large amount of solvents at last and precipitates.
Above method is used in concrete production fully, and proving effect is also good.But wherein Zui Da problem is exactly last this step of precipitation need use a large amount of organic solvents, can increase the weight of the pressure of environmental protection so widely, and more because remove the operation of impurity simultaneously, yield is low.If this intermediate processing can be changed, not not with an organic solvent or reduce the use of organic solvent, product yield and quality are improved simultaneously, that not only can reduce the raw materials cost of product, also can reduce environmental pollution, thereby economic benefit of enterprises and social benefit can be greatly improved all.
As a rule, material can crystallize out from solution, and basic reason is exactly that its solubleness in solution has diminished.And to change its solubleness, method generally change temperature, regulate pH value, with salt, to add organic solvent etc. several, wherein add this method of organic solvent owing to using a large amount of organic solvents, can strengthen the pressure to environmental protection, so now do not consider.Rangeability is not very big to the solubleness of teicoplanin along with variation of temperature in addition, so if that it is crystallized out is very difficult by changing temperature.And the present invention has selected adjusting pH and with this dual mode of salt.
Summary of the invention
Therefore, the invention provides a kind of method of purification of from the teicoplanin aqueous solution of 2～20W/V%, separating teicoplanin.
According to an aspect of the present invention, a kind of method of purification of separating teicoplanin comprises the teicoplanin aqueous solution of 2～20W/V% is regulated pH=2.5～6.0 with hydrochloric acid soln.
Wherein, the teicoplanin aqueous solution of above-mentioned 2～20W/V% makes as follows:
(1) the teicoplanin fermented liquid filters with strainer under alkaline condition, obtains filtrate;
(2) filtrate is carried out column chromatography by macroporous resin, washes with water respectively, the acidic aqueous solution of methyl alcohol or acetone organic solvent carries out wash-out, obtains elutriant;
(3) elutriant obtains the filtrate of decolouring by activated carbon decolorizing, filtration;
(4) be that the ultra-filtration membrane of 200Da～1000Da nanofiltration membrane or 5000Da～30000Da concentrates in pH=7.0～11.0 time, and constantly add deionized water and remove organic solvent, make the teicoplanin aqueous solution of 2～20W/V% by molecular weight.
Best, the teicoplanin aqueous solution of 2～20W/V% is by the filter element filtering of 1 μ m.
Best, be under 5～25 ℃ in temperature, mixed liquid 30～90 minutes, left standstill then 12～36 hours.
Best, employing pH=2.5～6.0, filtration back, temperature are that 0～10 ℃ water top is washed.
Best, described concentration of hydrochloric acid is 1M.
According to a further aspect in the invention, a kind of method of separating teicoplanin comprises add 5～20W/V% basic metal inorganic salt or alkaline-earth metal inorganic salt or ammonium salt in the teicoplanin aqueous solution of 2～20W/V%; After treating above-mentioned salt dissolving, transfer to pH=2.5～11 with hydrochloric acid or sodium hydroxide.
Wherein, the teicoplanin aqueous solution of 2～20W/V% makes as follows:
(1) the teicoplanin fermented liquid filters with strainer under alkaline condition, obtains filtrate;
(2) filtrate is carried out column chromatography by macroporous resin, washes with water respectively, the acidic aqueous solution of methyl alcohol or acetone organic solvent carries out wash-out, obtains elutriant;
(3) elutriant obtains the filtrate of decolouring by activated carbon decolorizing, filtration;
(4) be that the ultra-filtration membrane of 200Da～1000Da nanofiltration membrane or 5000Da～30000Da concentrates in pH=7.0～11.0 time, and constantly add deionized water and remove organic solvent, make the teicoplanin aqueous solution of 2～20W/V% by molecular weight.
Best, the teicoplanin aqueous solution of 2～20W/V% is by the filter element filtering of 1 μ m.
Best, be under 5～25 ℃ in temperature, mixed liquid 30～90 minutes, left standstill then 20～40 hours.
Best, employing pH=2.5～6.0, filtration back, temperature are that 0～10 ℃ water top is washed.
Best, described basic metal inorganic salt comprise the basic metal inorganic halides; Described alkaline-earth metal inorganic salt comprise the alkaline-earth metal inorganic halides.
Wherein, described basic metal inorganic halides comprises sodium-chlor, Repone K, lithium chloride, Sodium Bromide, Potassium Bromide, lithiumbromide, sodium iodide, potassiumiodide.
Wherein, described alkaline-earth metal inorganic halides comprises magnesium chloride, calcium chloride, bariumchloride.
Wherein, described ammonium salt is an ammonium chloride.
Best, described hydrochloric acid or naoh concentration are 1M.
Technical process provided by the present invention specifically describes as follows.It at first is the acquisition of teicoplanin concentrated aqueous solution.The teicoplanin fermented liquid is strainer such as the ceramic membrane filter of 0.02 μ m～0.5 μ m with the aperture under the condition of pH9.1～9.7, obtains clarifying filtrate, and filtrate readjustment pH adsorbs filtrate again by macroporous resin, and teicoplanin is adsorbed on the resin.Resin carries out wash-out through after washing with the acidic aqueous solution that contains organic solvent such as methyl alcohol or acetone, and teicoplanin is eluted from resin.In the elutriant that contains teicoplanin, add activated carbon decolorizing, filtering the back is that the nanofiltration membrane of 200Da～1000Da or the ultra-filtration membrane of 5000Da～30000Da concentrate with molecular weight, and constantly add deionized water and remove organic solvent, control process pH value is between 7.0～11.0, finally obtain containing the teicoplanin aqueous solution of minute quantity solvent, strength of solution is 2～20W/V%.
Next two kinds of methods of precipitation, a kind of is to regulate pH.Concentrated aqueous solution is regulated pH to 2.5～6.0 with the hydrochloric acid soln of 1M, have a large amount of light color precipitations in the solution and produce, stirred 30～90 minutes, left standstill then 12～36 hours, process temperature is controlled within 5～25 ℃.Pass through solid-liquid separation then, wash on 0～10 ℃ water top, get the wet product of teicoplanin with pH=2.5～6.0, temperature.
Another method is with salt.The inorganic salt that add 5～20W/V% in the concentrated solution stir and make its dissolving.Hydrochloric acid or sodium hydroxide solution with 1M transfer between pH=2.5～11.Continue stirring and stop after 30～90 minutes stirring, left standstill 5～25 ℃ of omnidistance controlled temperature 20～40 hours.Solid-liquid separation is washed on 0～10 ℃ water top with pH=2.5～6.0, temperature then, gets the wet product of teicoplanin.
The teicoplanin active princlple that obtains by the present invention is improved, and other impurity reduces, and the product appearance color obtains remarkable improvement simultaneously.
Embodiment
Come the present invention is further described in detail by following example, but be not limited in process parameters range among following examples and the embodiment.
Embodiment 1
Teicoplanin fermented liquid 100L, the 1268 μ g/ml that tire contain teicoplanin 126.8g, and the NaOH accent pH to 9.5 with 1M advances ceramic membrane filter, and the aperture of ceramic membrane is 0.02 μ m.After filtration, after water top washes, must mix filtrate 110L, tiring is 928 μ g/ml, contains teicoplanin 102.08g.Filtrate is gone up macroporous adsorbent resin adsorb, the resin volume is 4L, and flow velocity is 10L/hr.Wash resin after the end with water, to be washed behind the water of 6 times of column volumes, carry out wash-out with the aqueous solution that contains 60% acetone, pH3.5, flow velocity is 8L/hr, obtains elutriant 3.8L, and tiring is 14105 μ g/ml, and pH6.9 contains teicoplanin 53.599g.After transferring pH5.0 with the HCl of 1M, adding 50g gac in the elutriant stirred 30 minutes, filter destainer 3.7L, tiring is 12680 μ g/ml, pH4.8 contains teicoplanin 46.916g.Destainer is transferred to pH7.4 with the NaOH of 1M, puts in the nanofiltration membrane system and filter, nanofiltration membrane to see through molecular weight be 800Da.Earlier destainer is concentrated into 1/3 volume, is added to original volume with deionized water again, reconcentration to 1/3 volume, finish after so having repeated 4 times, the concentrated solution volume is 1.1L, and tiring is 42107 μ g/ml, contain teicoplanin 46.318g, measuring wherein according to gas-chromatography, acetone content is 0.9%.
Embodiment 2
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 52000 μ g/ml, and active princlple HPLC peak area ratio is 75.8%, and other impurity HPLC peak area ratio is 9.2%, volume 200ml, pH=8.2.HCl solution by adding 1M behind the filter element filtering of 1 μ m while stirring is stabilized in 2.5 with regulator solution pH up to pH, and teicoplanin is therefrom separated out, and forms suspension liquid.Continue stirring and stop after 30 minutes stirring, left standstill 12 hours, process temperature is controlled at about 5 ℃.Mixed solution is through filtering, and is that 50ml is washed on 0 ℃ water top with pH=2.5, temperature at last, and the top is washed and obtained the teicoplanin crude product 18.6g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 83.6%, and other impurity HPLC peak area ratio is 4.2%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 1.
Embodiment 3
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 52000 μ g/ml, active princlple HPLC peak area ratio be 75.8% (, other impurity HPLC peak area ratio is 9.2%, volume 200ml, pH=8.2.HCl solution by adding 1M behind the filter element filtering of 1 μ m while stirring is stabilized in 2.5 with regulator solution pH up to pH, and teicoplanin is therefrom separated out, and forms suspension liquid.Continue stirring and stop after 60 minutes stirring, left standstill 24 hours, process temperature is controlled at about 15 ℃.Mixed solution is through filtering, and is that 50ml is washed on 10 ℃ water top with pH=2.5, temperature at last, and the top is washed and obtained the teicoplanin crude product 18.2g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 84.7%, and other impurity HPLC peak area ratio is 4.0%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 1.
Embodiment 4
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 52000 μ g/ml, and active princlple HPLC peak area ratio is 75.8%, and other impurity HPLC peak area ratio is 9.2%, volume 200ml, pH=8.2.HCl solution by adding 1M behind the filter element filtering of 1 μ m while stirring is stabilized in 4.0 with regulator solution pH up to pH, and teicoplanin is therefrom separated out, and forms suspension liquid.Continue stirring and stop after 30 minutes stirring, left standstill 24 hours, process temperature is controlled at about 5 ℃.Mixed solution is through filtering, and is that 50ml is washed on 5 ℃ water top with pH=4.0, temperature at last, and the top is washed and obtained the teicoplanin crude product 18.8g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 84.1%, and other impurity HPLC peak area ratio is 4.3%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 1.
Embodiment 5
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 52000 μ g/ml, and active princlple HPLC peak area ratio is 75.8%, and other impurity HPLC peak area ratio is 9.2%, volume 200ml, pH=8.2.HCl solution by adding 1M behind the filter element filtering of 1 μ m while stirring is stabilized in 4.0 with regulator solution pH up to pH, and teicoplanin is therefrom separated out, and forms suspension liquid.Continue stirring and stop after 90 minutes stirring, left standstill 36 hours, process temperature is controlled at about 25 ℃.Mixed solution is through filtering, and is that 50ml is washed on 10 ℃ water top with pH=4.0, temperature at last, and the top is washed and obtained the teicoplanin crude product 18.5g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 83.8%, and other impurity HPLC peak area ratio is 4.5%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 1.
Embodiment 6
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 52000 μ g/ml, and active princlple HPLC peak area ratio is 75.8%, and other impurity HPLC peak area ratio is 9.2%, volume 200ml, pH=8.2.HCl solution by adding 1M behind the filter element filtering of 1 μ m while stirring is stabilized in 6.0 with regulator solution pH up to pH, and teicoplanin is therefrom separated out, and forms suspension liquid.Continue stirring and stop after 60 minutes stirring, left standstill 12 hours, process temperature is controlled at about 25 ℃.Mixed solution is through filtering, and is that 50ml is washed on 0 ℃ water top with pH=6.0, temperature at last, and the top is washed and obtained the teicoplanin crude product 18.0g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 84.6%, and other impurity HPLC peak area ratio is 4.3%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 1.
Embodiment 7
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 52000 μ g/ml, and active princlple HPLC peak area ratio is 75.8%, and other impurity HPLC peak area ratio is 9.2%, volume 200ml, pH=8.2.HCl solution by adding 1M behind the filter element filtering of 1 μ m while stirring is stabilized in 6.0 with regulator solution pH up to pH, and teicoplanin is therefrom separated out, and forms suspension liquid.Continue stirring and stop after 90 minutes stirring, left standstill 36 hours, process temperature is controlled at about 15 ℃.Mixed solution is through filtering, and is that 50ml is washed on 5 ℃ water top with pH=6.0, temperature at last, and the top is washed and obtained the teicoplanin crude product 18.7g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 84.1%, and other impurity HPLC peak area ratio is 4.1%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 1.
Embodiment 8
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 51800 μ g/ml, and active princlple HPLC peak area ratio is 74.5%, and other impurity HPLC peak area ratio is 10.5%, volume 200ml, pH=7.4.Behind the filter element filtering by 1 μ m, add 10g NaCl while stirring, treat NaCl all after the dissolving, transfer to pH=2.5, as seen have teicoplanin therefrom to separate out with the hydrochloric acid soln of 1M.Continue to stir 60 minutes, form soup compound, left standstill 30 hours, process temperature is controlled at about 15 ℃.Mixed solution is by filtering separation, is that 50ml is washed on 10 ℃ water top with pH=2.5, temperature at last, and the top is washed and obtained the teicoplanin crude product 17.8g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 82.4%, and other impurity HPLC peak area ratio is 6.9%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 2.
Embodiment 9
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 51800 μ g/ml, and active princlple HPLC peak area ratio is 74.5%, and other impurity HPLC peak area ratio is 10.5%, volume 200ml, pH=7.4.Behind the filter element filtering by 1 μ m, add 10g NaCl while stirring, treat NaCl all after the dissolving, transfer to pH=2.5, as seen have teicoplanin therefrom to separate out with the hydrochloric acid soln of 1M.Continue to stir 90 minutes, form soup compound, left standstill 40 hours, process temperature is controlled at about 15 ℃.Mixed solution is by filtering separation, is that 50ml is washed on 5 ℃ water top with pH=2.5, temperature at last, and the top is washed and obtained the teicoplanin crude product 17.4g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 81.6%, and other impurity HPLC peak area ratio is 6.5%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 2.
Embodiment 10
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 51800 μ g/ml, and active princlple HPLC peak area ratio is 74.5%, and other impurity HPLC peak area ratio is 10.5%, volume 200ml, pH=7.4.Behind the filter element filtering by 1 μ m, add 20g NaCl while stirring, treat NaCl all after the dissolving, transfer to pH=7.0, as seen have teicoplanin therefrom to separate out with the hydrochloric acid soln of 1M.Continue to stir 30 minutes, form soup compound, left standstill 40 hours, process temperature is controlled at about 5 ℃.Mixed solution is by filtering separation, is that 50ml is washed on 5 ℃ water top with pH=4.0, temperature at last, and the top is washed and obtained the teicoplanin crude product 17.5g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 82.1%, and other impurity HPLC peak area ratio is 7.1%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 2.
Embodiment 11
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 51800 μ g/ml, and active princlple HPLC peak area ratio is 74.5%, and other impurity HPLC peak area ratio is 10.5%, volume 200ml, pH=7.4.Behind the filter element filtering by 1 μ m, add 20g NaCl while stirring, treat NaCl all after the dissolving, transfer to pH=7.0, as seen have teicoplanin therefrom to separate out with the hydrochloric acid soln of 1M.Continue to stir 90 minutes, form soup compound, left standstill 20 hours, process temperature is controlled at about 25 ℃.Mixed solution is by filtering separation, is that 50ml is washed on 0 ℃ water top with pH=4.0, temperature at last, and the top is washed and obtained the teicoplanin crude product 17.6g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 82.2%, and other impurity HPLC peak area ratio is 7.0%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 2.
Embodiment 12
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 51800 μ g/ml, and active princlple HPLC peak area ratio is 74.5%, and other impurity HPLC peak area ratio is 10.5%, volume 200ml, pH=7.4.Behind the filter element filtering by 1 μ m, add 40g NaCl while stirring, treat NaCl all after the dissolving, transfer to pH=11.0, as seen have teicoplanin therefrom to separate out with the sodium hydroxide solution of 1M.Continue to stir 60 minutes, form soup compound, left standstill 30 hours, process temperature is controlled at about 5 ℃.Mixed solution is by filtering separation, is that 50ml is washed on 10 ℃ water top with pH=6.0, temperature at last, and the top is washed and obtained the teicoplanin crude product 17.9g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 82.1%, and other impurity HPLC peak area ratio is 6.3%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 2.
Embodiment 13
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 51800 μ g/ml, and active princlple HPLC peak area ratio is 74.5%, and other impurity HPLC peak area ratio is 10.5%, volume 200ml, pH=7.4.Behind the filter element filtering by 1 μ m, add 40g NaCl while stirring, treat NaCl all after the dissolving, transfer to pH=11.0, as seen have teicoplanin therefrom to separate out with the sodium hydroxide solution of 1M.Continue to stir 30 minutes, form soup compound, left standstill 20 hours, process temperature is controlled at about 25 ℃.Mixed solution is by filtering separation, is that 50ml is washed on 0 ℃ water top with pH=6.0, temperature at last, and the top is washed and obtained the teicoplanin crude product 18.2g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 81.7%, and other impurity HPLC peak area ratio is 6.5%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 2.
Embodiment 14
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 51800 μ g/ml, and active princlple HPLC peak area ratio is 74.5%, and other impurity HPLC peak area ratio is 10.5%, volume 200ml, pH=7.4.Behind the filter element filtering by 1 μ m, add 40g KI while stirring, treat KI all after the dissolving, transfer to pH=11.0, as seen have teicoplanin therefrom to separate out with the sodium hydroxide solution of 1M.Continue to stir 30 minutes, form soup compound, left standstill 20 hours, process temperature is controlled at about 25 ℃.Mixed solution is by filtering separation, is that 50ml is washed on 0 ℃ water top with pH=6.0, temperature at last, and the top is washed and obtained the teicoplanin crude product 17.6g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 81.3%, and other impurity HPLC peak area ratio is 7.2%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 3.
Embodiment 15
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 51800 μ g/ml, and active princlple HPLC peak area ratio is 74.5%, and other impurity HPLC peak area ratio is 10.5%, volume 200ml, pH=7.4.Behind the filter element filtering by 1 μ m, add 40g LiCl while stirring, treat LiCl all after the dissolving, transfer to pH=11.0, as seen have teicoplanin therefrom to separate out with the sodium hydroxide solution of 1M.Continue to stir 30 minutes, form soup compound, left standstill 20 hours, process temperature is controlled at about 25 ℃.Mixed solution is by filtering separation, is that 50ml is washed on 0 ℃ water top with pH=6.0, temperature at last, and the top is washed and obtained the teicoplanin crude product 17.4g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 81.8%, and other impurity HPLC peak area ratio is 6.9%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 3.
Embodiment 16
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 51800 μ g/ml, and active princlple HPLC peak area ratio is 74.5%, and other impurity HPLC peak area ratio is 10.5%, volume 200ml, pH=7.4.Behind the filter element filtering by 1 μ m, add 40g NaBr while stirring, treat NaBr all after the dissolving, transfer to pH=11.0, as seen have teicoplanin therefrom to separate out with the sodium hydroxide solution of 1M.Continue to stir 30 minutes, form soup compound, left standstill 20 hours, process temperature is controlled at about 25 ℃.Mixed solution is by filtering separation, is that 50ml is washed on 0 ℃ water top with pH=6.0, temperature at last, and the top is washed and obtained the teicoplanin crude product 18.0g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 81.5%, and other impurity HPLC peak area ratio is 7.0%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 3.
Embodiment 17
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 51800 μ g/ml, and active princlple HPLC peak area ratio is 74.5%, and other impurity HPLC peak area ratio is 10.5%, volume 200ml, pH=7.4.Behind the filter element filtering by 1 μ m, add 40g KBr while stirring, treat KBr all after the dissolving, transfer to pH=11.0, as seen have teicoplanin therefrom to separate out with the sodium hydroxide solution of 1M.Continue to stir 30 minutes, form soup compound, left standstill 20 hours, process temperature is controlled at about 25 ℃.Mixed solution is by filtering separation, is that 50ml is washed on 0 ℃ water top with pH=6.0, temperature at last, and the top is washed and obtained the teicoplanin crude product 18.2g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 82.0%, and other impurity HPLC peak area ratio is 6.8%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 3.
Embodiment 18
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 51800 μ g/ml, and active princlple HPLC peak area ratio is 74.5%, and other impurity HPLC peak area ratio is 10.5%, volume 200ml, pH=7.4.Behind the filter element filtering by 1 μ m, add 40g LiBr while stirring, treat LiBr all after the dissolving, transfer to pH=11.0, as seen have teicoplanin therefrom to separate out with the sodium hydroxide solution of 1M.Continue to stir 30 minutes, form soup compound, left standstill 20 hours, process temperature is controlled at about 25 ℃.Mixed solution is by filtering separation, is that 50ml is washed on 0 ℃ water top with pH=6.0, temperature at last, and the top is washed and obtained the teicoplanin crude product 17.9g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 82.3%, and other impurity HPLC peak area ratio is 6.5%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 3.
Embodiment 19
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 51800 μ g/ml, and active princlple HPLC peak area ratio is 74.5%, and other impurity HPLC peak area ratio is 10.5%, volume 200ml, pH=7.4.Behind the filter element filtering by 1 μ m, add 40g NaI while stirring, treat NaI all after the dissolving, transfer to pH=11.0, as seen have teicoplanin therefrom to separate out with the sodium hydroxide solution of 1M.Continue to stir 30 minutes, form soup compound, left standstill 20 hours, process temperature is controlled at about 25 ℃.Mixed solution is by filtering separation, is that 50ml is washed on 0 ℃ water top with pH=6.0, temperature at last, and the top is washed and obtained the teicoplanin crude product 17.5g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 81.4%, and other impurity HPLC peak area ratio is 7.1%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 3.
Embodiment 20
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 51800 μ g/ml, and active princlple HPLC peak area ratio is 74.5%, and other impurity HPLC peak area ratio is 10.5%, volume 200ml, pH=7.4.Behind the filter element filtering by 1 μ m, add 40g KI while stirring, treat KI all after the dissolving, transfer to pH=11.0, as seen have teicoplanin therefrom to separate out with the sodium hydroxide solution of 1M.Continue to stir 30 minutes, form soup compound, left standstill 20 hours, process temperature is controlled at about 25 ℃.Mixed solution is by filtering separation, is that 50ml is washed on 0 ℃ water top with pH=6.0, temperature at last, and the top is washed and obtained the teicoplanin crude product 17.6g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 81.9%, and other impurity HPLC peak area ratio is 6.7%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 3.
Embodiment 21
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 51800 μ g/ml, and active princlple HPLC peak area ratio is 74.5%, and other impurity HPLC peak area ratio is 10.5%, volume 200ml, pH=7.4.Behind the filter element filtering by 1 μ m, add 40g MgCl while stirring
2, treat MgCl
2All after the dissolving, transfer to pH=11.0, as seen have teicoplanin therefrom to separate out with the sodium hydroxide solution of 1M.Continue to stir 30 minutes, form soup compound, left standstill 20 hours, process temperature is controlled at about 25 ℃.Mixed solution is by filtering separation, is that 50ml is washed on 0 ℃ water top with pH=6.0, temperature at last, and the top is washed and obtained the teicoplanin crude product 17.2g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 81.1%, and other impurity HPLC peak area ratio is 6.9%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 3.
Embodiment 22
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 51800 μ g/ml, and active princlple HPLC peak area ratio is 74.5%, and other impurity HPLC peak area ratio is 10.5%, volume 200ml, pH=7.4.Behind the filter element filtering by 1 μ m, add 40g CaCl while stirring
2, treat CaCl
2All after the dissolving, transfer to pH=11.0, as seen have teicoplanin therefrom to separate out with the sodium hydroxide solution of 1M.Continue to stir 30 minutes, form soup compound, left standstill 20 hours, process temperature is controlled at about 25 ℃.Mixed solution is by filtering separation, is that 50ml is washed on 0 ℃ water top with pH=6.0, temperature at last, and the top is washed and obtained the teicoplanin crude product 18.3g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 82.1%, and other impurity HPLC peak area ratio is 6.4%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 3.
Embodiment 23
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 51800 μ g/ml, and active princlple HPLC peak area ratio is 74.5%, and other impurity HPLC peak area ratio is 10.5%, volume 200ml, pH=7.4.Behind the filter element filtering by 1 μ m, add 40g BaCl while stirring
2, treat BaCl
2All after the dissolving, transfer to pH=11.0, as seen have teicoplanin therefrom to separate out with the sodium hydroxide solution of 1M.Continue to stir 30 minutes, form soup compound, left standstill 20 hours, process temperature is controlled at about 25 ℃.Mixed solution is by filtering separation, is that 50ml is washed on 0 ℃ water top with pH=6.0, temperature at last, and the top is washed and obtained the teicoplanin crude product 17.5g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 81.7%, and other impurity HPLC peak area ratio is 6.6%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 3.
Embodiment 24
Teicoplanin concentrated aqueous solution concentration according to embodiment 1 method gained is 51800 μ g/ml, and active princlple HPLC peak area ratio is 74.5%, and other impurity HPLC peak area ratio is 10.5%, volume 200ml, pH=7.4.Behind the filter element filtering by 1 μ m, add 40g NH while stirring
4Cl treats NH
4Cl is all after the dissolving, transfers to pH=11.0 with the sodium hydroxide solution of 1M, as seen has teicoplanin therefrom to separate out.Continue to stir 30 minutes, form soup compound, left standstill 20 hours, process temperature is controlled at about 25 ℃.Mixed solution is by filtering separation, is that 50ml is washed on 0 ℃ water top with pH=6.0, temperature at last, and the top is washed and obtained the teicoplanin crude product 17.1g that wets after the end.The teicoplanin active princlple HPLC peak area ratio that obtains by precipitation is 81.6%, and other impurity HPLC peak area ratio is 7.4%, and the product appearance color obtains remarkable improvement simultaneously.Referring to table 3.
Above technology can be used for the preparation of teicoplanin intermediate and is used for the preparation of teicoplanin finished product, also can be used for teicoplanin through post precipitation and dissolves again obtain teicoplanin finished product DS again after freeze-drying, also can be by repeatedly precipitating the combined purifying teicoplanin.
Table 1
Table 2
Table 3
The present invention is illustrated by top embodiment, still, should be appreciated that the present invention is not limited to particular example as described herein and embodiment.The purpose that comprises these particular example and embodiment here is to help those of skill in the art to put into practice the present invention.Any those of skill in the art are easy to be further improved without departing from the spirit and scope of the present invention and perfect, therefore the present invention only is subjected to the restriction of the content and the scope of claim of the present invention, and its intention contains all and is included in alternatives and equivalent in the spirit and scope of the invention that is limited by appendix claim.
Claims (16)
1, a kind of method of purification of teicoplanin, described method comprise regulates pH=2.5～6.0 with the teicoplanin aqueous solution of 2～20W/V% with hydrochloric acid soln.
2, method of purification as claimed in claim 1, wherein, the teicoplanin aqueous solution of 2～20W/V% makes as follows:
(1) the teicoplanin fermented liquid filters with strainer under alkaline condition, obtains filtrate;
(2) filtrate is carried out column chromatography by macroporous resin, washes with water respectively, the acidic aqueous solution of methyl alcohol or acetone organic solvent carries out wash-out, obtains elutriant;
(3) elutriant obtains the filtrate of decolouring by activated carbon decolorizing, filtration;
(4) be that the ultra-filtration membrane of 200Da～1000Da nanofiltration membrane or 5000Da～30000Da concentrates in pH=7.0～11.0 time, and add deionized water and remove organic solvent, make the teicoplanin aqueous solution of 2～20W/V% by molecular weight.
3, method of purification as claimed in claim 1, wherein, the teicoplanin aqueous solution of 2～20W/V% is by the filter element filtering of 1 μ m.
4, as the arbitrary described method of purification of claim 1～3, wherein, be under 5～25 ℃ in temperature, mixed liquid 30～90 minutes, left standstill then 12～36 hours.
5, method of purification as claimed in claim 4, wherein, employing pH=2.5～6.0, filtration back, temperature are that 0～10 ℃ water top is washed.
6, method of purification as claimed in claim 1, wherein, described concentration of hydrochloric acid is 1M.
7, a kind of method of purification of teicoplanin, described method comprise add 5～20W/V% basic metal inorganic salt or alkaline-earth metal inorganic salt or ammonium salt in the teicoplanin aqueous solution of 2～20W/V%; After treating above-mentioned salt dissolving, transfer to pH=2.5～11 with hydrochloric acid or sodium hydroxide.
8, method of purification as claimed in claim 7, wherein, the teicoplanin aqueous solution of 2～20W/V% makes as follows:
(1) the teicoplanin fermented liquid filters with strainer under alkaline condition, obtains filtrate;
(2) filtrate is carried out column chromatography by macroporous resin, washes with water respectively, the acidic aqueous solution of methyl alcohol or acetone organic solvent carries out wash-out, obtains elutriant;
(3) elutriant obtains the filtrate of decolouring by activated carbon decolorizing, filtration;
(4) be that the ultra-filtration membrane of 200Da～1000Da nanofiltration membrane or 5000Da～30000Da concentrates in pH=7.0～11.0 time, and constantly add deionized water and remove organic solvent, make the teicoplanin aqueous solution of 2～20W/V% by molecular weight.
9, method of purification as claimed in claim 7, wherein, the teicoplanin aqueous solution of 2～20W/V% is by the filter element filtering of 1 μ m.
10, as the arbitrary described method of purification of claim 7～9, wherein, be under 5～25 ℃ in temperature, mixed liquid 30～90 minutes, left standstill then 20～40 hours.
11, method of purification as claimed in claim 10, wherein, employing pH=2.5～6.0, filtration back, temperature are that 0～10 ℃ water top is washed.
12, method of purification as claimed in claim 7, wherein, described basic metal inorganic salt comprise the basic metal inorganic halides; Described alkaline-earth metal inorganic salt comprise the alkaline-earth metal inorganic halides.
13, method of purification as claimed in claim 12, wherein, described basic metal inorganic halides comprises sodium-chlor, Repone K, lithium chloride, Sodium Bromide, Potassium Bromide, lithiumbromide, sodium iodide, potassiumiodide.
14, method of purification as claimed in claim 12, wherein, described alkaline-earth metal inorganic halides comprises magnesium chloride, calcium chloride, bariumchloride.
15, method of purification as claimed in claim 7, wherein, described ammonium salt is an ammonium chloride.
16, method of purification as claimed in claim 7, wherein, described hydrochloric acid or naoh concentration are 1M.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102010462A (en) * | 2009-09-04 | 2011-04-13 | 四川金稞生物科技有限公司 | Method for preparing ramoplanin from fermentation liquor by utilizing nanofiltration concentration purification technology |
| CN102690333A (en) * | 2012-06-30 | 2012-09-26 | 华北制药集团新药研究开发有限责任公司 | Preparation method of high-purity teicoplanin |
| CN102718843A (en) * | 2012-06-30 | 2012-10-10 | 华北制药集团新药研究开发有限责任公司 | Preparation method of single teicoplanin components |
| CN102964430A (en) * | 2011-09-01 | 2013-03-13 | 丽珠集团福州福兴医药有限公司 | Purification method of teicoplanin |
| EP2592089A1 (en) * | 2011-11-11 | 2013-05-15 | LEK Pharmaceuticals d.d. | Process of purification of teicoplanin |
| CN103776913A (en) * | 2012-10-25 | 2014-05-07 | 鲁南新时代生物技术有限公司 | HPLC method for quick teicoplanin fermentation liquor unit detection |
| CN104371011A (en) * | 2014-10-21 | 2015-02-25 | 福建省微生物研究所 | Purification method of high-purity teicoplanin refined powder |
| CN105985411A (en) * | 2015-02-03 | 2016-10-05 | 浙江医药股份有限公司新昌制药厂 | Method for improving clarity of teicoplanin for injection |
| CN112641926A (en) * | 2020-12-31 | 2021-04-13 | 华北制药股份有限公司 | Teicoplanin preparation for injection and its preparing method |
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| CN102010462A (en) * | 2009-09-04 | 2011-04-13 | 四川金稞生物科技有限公司 | Method for preparing ramoplanin from fermentation liquor by utilizing nanofiltration concentration purification technology |
| CN102964430B (en) * | 2011-09-01 | 2015-01-21 | 丽珠集团福州福兴医药有限公司 | Purification method of teicoplanin |
| CN102964430A (en) * | 2011-09-01 | 2013-03-13 | 丽珠集团福州福兴医药有限公司 | Purification method of teicoplanin |
| EP2592089A1 (en) * | 2011-11-11 | 2013-05-15 | LEK Pharmaceuticals d.d. | Process of purification of teicoplanin |
| WO2013068538A1 (en) | 2011-11-11 | 2013-05-16 | Lek Pharmaceuticals D.D. | Process of purification of teicoplanin |
| CN104066747A (en) * | 2011-11-11 | 2014-09-24 | 力奇制药公司 | Process of purification of teicoplanin |
| CN102718843A (en) * | 2012-06-30 | 2012-10-10 | 华北制药集团新药研究开发有限责任公司 | Preparation method of single teicoplanin components |
| CN102718843B (en) * | 2012-06-30 | 2014-05-14 | 华北制药集团新药研究开发有限责任公司 | Preparation method of single teicoplanin components |
| CN102690333A (en) * | 2012-06-30 | 2012-09-26 | 华北制药集团新药研究开发有限责任公司 | Preparation method of high-purity teicoplanin |
| CN103776913A (en) * | 2012-10-25 | 2014-05-07 | 鲁南新时代生物技术有限公司 | HPLC method for quick teicoplanin fermentation liquor unit detection |
| CN104371011A (en) * | 2014-10-21 | 2015-02-25 | 福建省微生物研究所 | Purification method of high-purity teicoplanin refined powder |
| CN105985411A (en) * | 2015-02-03 | 2016-10-05 | 浙江医药股份有限公司新昌制药厂 | Method for improving clarity of teicoplanin for injection |
| CN105985411B (en) * | 2015-02-03 | 2019-12-06 | 浙江医药股份有限公司新昌制药厂 | Method for improving clarity of teicoplanin for injection |
| CN112641926A (en) * | 2020-12-31 | 2021-04-13 | 华北制药股份有限公司 | Teicoplanin preparation for injection and its preparing method |
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