CN105367586A - Method of extracting tacrolimus from fermentation liquor - Google Patents
Method of extracting tacrolimus from fermentation liquor Download PDFInfo
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- CN105367586A CN105367586A CN201410435044.7A CN201410435044A CN105367586A CN 105367586 A CN105367586 A CN 105367586A CN 201410435044 A CN201410435044 A CN 201410435044A CN 105367586 A CN105367586 A CN 105367586A
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Abstract
The invention relates to a method of extracting tacrolimus from fermentation liquor. According to the method, a ceramic membrane, a nanofiltration membrane and other devices are used in a combined mode, and an organic solvent is used for extracting the tacrolimus from the fermentation liquor at an abnormally high yield.
Description
Technical field
The present invention relates to medicinal chemistry art.Specifically, the present invention relates to a kind of method extracting tacrolimus from fermented liquid, more specifically, the present invention relates to and be a kind ofly combined ceramic membrane, nanofiltration membrane be combined the method that the tacrolimus in fermented liquid extracts with abnormal high yield by organic solvent.
Background technology
Tacrolimus (Tacrolimus) has another name called FK506, and be the tunning separated from streptomyces (streptomycestsukubaensis), its chemical structure belongs to 23 membered macrolide microbiotic.For a kind of neotype immunosuppressant of brute force, become a line medication of liver, renal transplantation in recent years, also in autoimmune disorder, played active effect.
Substantially to be (1) realize being separated of mycelium and aqueous phase with plate-and-frame filter press or whizzer to the method extracting at present tacrolimus from fermented liquid, soaks mycelium extract fat-soluble target components with organic solvent.The shortcoming of aforesaid method is that the part fat-soluble target components be free in aqueous phase enters in waste water when mycelium and aqueous phase separation with aqueous phase; Operate unfriendly to environment, organic solvent consumption is large; Because of complicated component in mycelium, in organic solvent extraction process, easily cause a large amount of emulsification, and then affect yield and quality; The cleaning of the mounting or dismounting of sheet frame, filter cake discharging and filter cloth is all manual operations, and non-cutting time is long, and greatly, production efficiency is low, thus affects the progress of whole production, lower with the degree of conformity of environmental requirement for labour intensity.(2) with organic solvent directly lixiviate tacrolimus in fermented liquid.The shortcoming of aforesaid method is that solvent consumption is large, easy emulsification, quality and yield instability.
Modern Membrane Technology originates from the sixties in 20th century, is a kind of novel fluid separation element operative technique.It take polymeric membrane for separation as separating medium, with pressure difference, concentration difference, potential difference etc. for mass transfer driving force, the difference of the different substances in fluid to be separated according to molecular weight, particle or electric charge is separated.Because membrane separation technique has the features such as energy-efficient, since first set industrial equipment is introduced to the market, obtain fast development, at present throughout nearly all industrial circle.According to the different principle of membrane sepn, film can be divided into the different sortses such as microfiltration membrane, ultra-filtration membrane, nanofiltration membrane, reverse osmosis membrane, dialysis membrane, electrodialytic membranes, infiltrating and vaporizing membrane and gas separation membrane.
Ceramic membrane is the one in microfiltration membrane, primarily of Al
2o
3, ZrO
2, TiO
2and SiO
2etc. porous-film prepared by inorganic materials, its aperture is 1-200nm.There is chemical stability good, acidproof, alkaline-resisting, the organic solvent-resistant of energy; Physical strength is large, can counterflush; Anti-microbe ability is strong, and high temperature resistant, pore size distribution is narrow, separation efficiency high, in foodstuffs industry, biotechnology, environmental engineering, chemical industry, petrochemical complex, controls the fields such as metal working industry and is widely used.
Ceramic membrane filter be dissolved with effective constituent vat liquor ceaselessly to the process appearing side and appear, from traditional by different for solid-liquid separation filtered model, thus whole filtration procedure does not need to add flocculating aids.
Nanofiltration membrane is the functional semi-permeable membranes of one allowing solvent molecule or some low molecular weight solutes or low price ion permeable, it retains organic molecular weight and is approximately about 150-500, be used to organism and the colourity of removing surface water, remove underground water hardness, part removes solvability salt, fruit juice concentrate and the useful matter etc. be separated in medicine.
Chinese patent CN1297475 discloses the method for a kind of ceramic membrane separating water-soluble product from fermented liquid; Chinese patent CN1699358 discloses a kind of method of ceramic membrane separation theoflavin; Chinese patent CN1775729 discloses a kind of method that ceramic membrane extracts high-purity citric acid; Chinese patent CN101565395 discloses a kind of method that ceramic membrane extracts L-Trp, above-mentioned patent is all carry out the water miscible target product of filtering separation with ceramic membrane, also discovery ceramic membrane does not carry out patent and the report of collecting by filtration fat-soluble component at present, does not find to combine with nanofiltration membrane the patent and the report that are separated tacrolimus with ceramic membrane.
Summary of the invention
The present invention be devoted to provide a kind of simpler effectively, environmental protection, the method for extracting tacrolimus from fermented liquid with high yield that cost is low, even if the poor quality of fermented liquid.
Method of the present invention for extracting tacrolimus from fermented liquid comprises:
A) ceramic membrane concentrated broth is used, then
B) continue to add in fermented liquid concentrated solution one or several be selected from alcohol, glycol, ketone, ester class, carboxylic-acid organic solvent or above-mentioned solvent and water mixture and after using ceramic membrane lixiviate tacrolimus vat liquor, then
C) concentrate tacrolimus vat liquor by nanofiltration membrane and obtain tacrolimus nanofiltration concentrated solution.
The present invention the present invention relates to a kind of method extracting tacrolimus from fermented liquid, more specifically, the present invention relates to one and is combined the equipment such as ceramic membrane, nanofiltration membrane, the method extracted with abnormal high yield by the tacrolimus in fermented liquid with organic solvent.
Generally speaking, the method extracting tacrolimus from fermented liquid comprises uses ceramic membrane concentrated broth, continue to add in fermented liquid concentrated solution one or several be selected from alcohol, glycol, ketone, ester class, carboxylic-acid organic solvent or above-mentioned solvent and water mixture and after using ceramic membrane lixiviate tacrolimus vat liquor, concentrate vat liquor by nanofiltration membrane.
Can add in fermented liquid in ceramic membrane concentration process water, tensio-active agent, acid, alkali, organic solvent and water mixture or change broth temperature to change fermented liquid character.
Extraction solvent can be selected from alcohol, such as: methyl alcohol, ethanol, Virahol; Glycol, such as: 1,3-PD; Ketone, such as: ethanol; Ester class, such as: ethyl acetate, butylacetate; Carboxylic-acid, such as: acetic acid.
The solvent of lixiviate directly can apply that one or several are used in combination, and also can be that one or several and water are used in combination, wherein the ratio of water be 1% to 70%.
In a preferred embodiment, the organic solvent that lixiviate uses is methyl alcohol, ethanol, ethyl acetate, hydrochloric acid.
In a preferred embodiment, lixiviate use organic solvent be ethanol, lixiviate with an organic solvent feature be containing have an appointment 30% ~ 60% water.
According to preferred embodiment, the effluent liquid used in step c) can overlap and be back in step b), carries out ceramic membrane lixiviate as extraction solvent.Thus reduce costs, and there is the effect of environmental protection.
The present invention measures tacrolimus content method and adopts high performance liquid chromatography, and the following examples will illustrate the preferred embodiment of the invention, but the meaning do not limited the scope of the invention.
The present invention is further illustrated below by embodiment.It should be understood that embodiments of the invention are for illustration of the present invention instead of limitation of the present invention.Essence according to the present invention all belongs to the scope of protection of present invention to the simple modifications that the present invention carries out.
systems axiol-ogy condition
1) chromatographic system
Chromatographic column: ZORBAXSBC8150 × 3mm, 3.5 μm or quite
Moving phase: acetonitrile-methyl tertiary butyl ether-water-trifluoroacetic acid (35:4:61:0.01)
Sample temperature: 2 ~ 10 DEG C
Flow velocity: 1.2ml/min
Determined wavelength: 210nm
Sample size: 10 μ l
Column temperature: 60 DEG C
2) solution preparation
Thinner: acetonitrile (Calcium Chloride Powder Anhydrous drying and dehydrating must be used before use)
Standardized solution: it is appropriate that precision takes tacrolimus standard substance, is mixed with the solution that concentration is 1.0mg/ml.(such as: 50mg → 50ml)
Sample solution: it is appropriate that precision takes tacrolimus sample, is mixed with the solution that concentration is 1.0mg/ml.(such as: 50mg → 50ml)
Injection procedure:
Enter blank (acetonitrile), answer noiseless peak; Enter sample solution containing isomer I and isomer II (as in sample solution not containing isomer I or isomer II, then degrade by adding water), record collection of illustrative plates, in color atlas, the peak sequence of tacrolimus and isomer is followed successively by isomer I, II and tacrolimus peak, relative retention time is respectively 0.5,0.7 and 1.0, isomer II and the peak-to-peak resolution of tacrolimus should be greater than 3.0, and number of theoretical plate is pressed tacrolimus peak and calculated, and should be not less than 1000.Continuous sample introduction standardized solution 6 times, RSD should more than 2.0%.
Calculate:
According to the peak area of standardized solution and sample solution, calculate as follows and tire:
R
f=standardized solution main peak average area/concentration of standard solution (μ g/ml)
Sample is tired=(sample solution peak area/R
f) × extension rate
embodiment 1
Tacrolimus fermented liquid 38L, tire 227 μ g/ml, chromatographic purity 87.1%.Containing tacrolimus 8.6g.Fermented liquid filtered on buchner funnel is to flowing out without obvious drop, wet cake is put into 40L70% ethanol soaking and stirring more than 6 hours, filter, collect filtrate, obtain filtrate 36.6L, tiring is 172 μ g/ml, and chromatographic purity 60.4%, containing tacrolimus 6.26g, residual tacrolimus 0.9g in waste residue, part tacrolimus doubly destroys.This process tacrolimus extract yield is 73%.
embodiment 2
Tacrolimus fermented liquid 153L, tire 234 μ g/ml, chromatographic purity 86.6%, containing tacrolimus 35.8g, with Plate Filtration tacrolimus fermented liquid to flowing out without obvious drop, takes sheet frame apart and collect filter cake.Filter cake is immersed in the ethanol of 150L70%, stir more than 6 hours.Plate Filtration, collect filtrate, obtain vat liquor 143L, tiring is 176.5 μ g/ml, chromatographic purity 60.7%, and containing tacrolimus 25.2g, containing tacrolimus 4.3g in waste residue, this process tacrolimus extract yield is 70.4%.
embodiment 3
Tacrolimus fermented liquid 70L, tire 246 μ g/ml, chromatographic purity 86.9%.Containing tacrolimus 17.2g.(ceramic membrane specification is as follows: PALL1960 type film core, 19 passages, passage aperture d=6mm, membrane pore size D=50nm, the long L=1m of film core, filtration area S=0.358m fermented liquid to be put into ceramic membrane treatment tank
2, trier impeller pump: H=36m, Q=10T/h, P=3Kw), start is filtered, and ceramic membrane appears after end appears 35L water and obtains fermented liquid concentrated solution 35L, starts the washing fermented liquid that adds water, maintains fermentating liquid volume 35
+2L reaches 35L until washing appears volume.In fermented liquid concentrated solution, add 33L ethanol, after stirring, open ceramic membrane filter, appear side at ceramic membrane and obtain tacrolimus alcohol extract.When the material liquid volume prepared in tank is less than 30L, continues in ceramic membrane treatment tank, add 70% ethanol (or nanofiltration peritoneal effluent) prepared and carry out lixiviate.Vat liquor is transferred to nanofiltration membrane treatment tank, (nanofiltration membrane specification is as follows: GE2540 type film core, molecular weight cut-off 500, membrane filtration area S=2.5m to open nanofiltration membrane
2, trier ram pump: △ P=70bar, Q=11.9L/min, P=2.2Kw) and concentrated.It is 70% ethanol that nanofiltration membrane appears end, and cover is back to ceramic membrane lixiviate.Follow the tracks of leaching process with high performance liquid phase, stop adding 70% ethanol (or nanofiltration peritoneal effluent), feed liquid is concentrated into 25L, stops ceramic membrane filter when appearing vat liquor volume and being 5 times that maintain volume, residue feed liquid is that waste residue processes further.All vat liquors are transferred to nanofiltration membrane treatment tank concentrate, last concentrated solution volume is 15L.Detect through HPLC, it is 1043.5 μ g/ml that concentrated solution is tired, chromatographic purity 67%, and containing tacrolimus 15.65g, whole process tacrolimus extract yield is 91%.
embodiment 4
Tacrolimus fermented liquid 70L, tire 250 μ g/ml, chromatographic purity 87.6%.Containing tacrolimus 17.5g.(ceramic membrane specification is as follows: PALL1960 type film core, 19 passages, passage aperture d=6mm, membrane pore size D=50nm, the long L=1m of film core, filtration area S=0.358m fermented liquid to be put into ceramic membrane treatment tank
2, trier impeller pump: H=36m, Q=10T/h, P=3Kw), start is filtered, and ceramic membrane appears after end appears 35L water and obtains fermented liquid concentrated solution 35L, starts the washing fermented liquid that adds water, maintains fermentating liquid volume 35
+2L reaches 35L until washing appears volume.In fermented liquid concentrated solution, add 14L ethanol, after stirring, open ceramic membrane filter, appear side at ceramic membrane and obtain tacrolimus alcohol extract.When the material liquid volume prepared in tank is less than 30L, continues in ceramic membrane treatment tank, add 50% ethanol (or nanofiltration peritoneal effluent) prepared and carry out lixiviate.Vat liquor is transferred to nanofiltration membrane treatment tank, (nanofiltration membrane specification is as follows: GE2540 type film core, molecular weight cut-off 500, membrane filtration area S=2.5m to open nanofiltration membrane
2, trier ram pump: △ P=70bar, Q=11.9L/min, P=2.2Kw) and concentrated.It is 50% ethanol that nanofiltration membrane appears end, and cover is back to ceramic membrane lixiviate.Follow the tracks of leaching process with high performance liquid phase, stop adding 50% ethanol (or nanofiltration peritoneal effluent), feed liquid is concentrated into 27L, stops ceramic membrane filter when appearing vat liquor volume and being 5 times that maintain volume, residue feed liquid is that waste residue processes further.All vat liquors are transferred to nanofiltration membrane treatment tank concentrate, last concentrated solution volume is 15L.Detect through HPLC, it is 1069.8 μ g/ml that concentrated solution is tired, chromatographic purity 66%, and containing tacrolimus 16g, whole process tacrolimus extract yield is 91.7%.
embodiment 5
Tacrolimus fermented liquid 70L, tire 250 μ g/ml, chromatographic purity 87.6%.Containing tacrolimus 17.5g.(ceramic membrane specification is as follows: PALL1960 type film core, 19 passages, passage aperture d=6mm, membrane pore size D=50nm, the long L=1m of film core, filtration area S=0.358m fermented liquid to be put into ceramic membrane treatment tank
2, trier impeller pump: H=36m, Q=10T/h, P=3Kw), start is filtered, and ceramic membrane appears after end appears 35L water and obtains fermented liquid concentrated solution 35L, starts the washing fermented liquid that adds water, maintains fermentating liquid volume 35
+2L reaches 35L until washing appears volume.In fermented liquid concentrated solution, add 10L ethanol, after stirring, open ceramic membrane filter, appear side at ceramic membrane and obtain tacrolimus alcohol extract.When the material liquid volume prepared in tank is less than 30L, continues in ceramic membrane treatment tank, add 40% ethanol (or nanofiltration peritoneal effluent) prepared and carry out lixiviate.Vat liquor is transferred to nanofiltration membrane treatment tank, (nanofiltration membrane specification is as follows: GE2540 type film core, molecular weight cut-off 500, membrane filtration area S=2.5m to open nanofiltration membrane
2, trier ram pump: △ P=70bar, Q=11.9L/min, P=2.2Kw) and concentrated.It is 40% ethanol that nanofiltration membrane appears end, and cover is back to ceramic membrane lixiviate.Follow the tracks of leaching process with high performance liquid phase, stop adding 40% ethanol (or nanofiltration peritoneal effluent), feed liquid is concentrated into 27L, stops ceramic membrane filter when appearing vat liquor volume and being 5 times that maintain volume, residue feed liquid is that waste residue processes further.All vat liquors are transferred to nanofiltration membrane treatment tank concentrate, last concentrated solution volume is 15L.Detect through HPLC, it is 1065 μ g/ml that concentrated solution is tired, chromatographic purity 67.8%, and containing tacrolimus 15.98g, whole process tacrolimus extract yield is 91.3%.
embodiment 6
Tacrolimus fermented liquid 70L, tire 250 μ g/ml, chromatographic purity 87.6%.Containing tacrolimus 17.5g.(ceramic membrane specification is as follows: PALL1960 type film core, 19 passages, passage aperture d=6mm, membrane pore size D=50nm, the long L=1m of film core, filtration area S=0.358m fermented liquid to be put into ceramic membrane treatment tank
2, trier impeller pump: H=36m, Q=10T/h, P=3Kw), start is filtered, and ceramic membrane appears after end appears 35L water and obtains fermented liquid concentrated solution 35L, starts the washing fermented liquid that adds water, maintains fermentating liquid volume 35
+2L reaches 35L until washing appears volume.In fermented liquid concentrated solution, add 6L ethanol, after stirring, open ceramic membrane filter, appear side at ceramic membrane and obtain tacrolimus alcohol extract.When the material liquid volume prepared in tank is less than 30L, continues in ceramic membrane treatment tank, add 30% ethanol (or nanofiltration peritoneal effluent) prepared and carry out lixiviate.Vat liquor is transferred to nanofiltration membrane treatment tank, (nanofiltration membrane specification is as follows: GE2540 type film core, molecular weight cut-off 500, membrane filtration area S=2.5m to open nanofiltration membrane
2, trier ram pump: △ P=70bar, Q=11.9L/min, P=2.2Kw) and concentrated.It is 30% ethanol that nanofiltration membrane appears end, and cover is back to ceramic membrane lixiviate.Follow the tracks of leaching process with high performance liquid phase, stop adding 30% ethanol (or nanofiltration peritoneal effluent), feed liquid is concentrated into 27L, stops ceramic membrane filter when appearing vat liquor volume and being 5 times that maintain volume, residue feed liquid is that waste residue processes further.All vat liquors are transferred to nanofiltration membrane treatment tank concentrate, last concentrated solution volume is 15L.Detect through HPLC, it is 624 μ g/ml that concentrated solution is tired, chromatographic purity 67%, and containing tacrolimus 9.3g, whole process tacrolimus extract yield is 53.1%.
embodiment 7
Tacrolimus fermented liquid 72L, tire 250 μ g/ml, chromatographic purity 87.6%.Containing tacrolimus 18g.(ceramic membrane specification is as follows: PALL1960 type film core, 19 passages, passage aperture d=6mm, membrane pore size D=50nm, the long L=1m of film core, filtration area S=0.358m fermented liquid to be put into ceramic membrane treatment tank
2, trier impeller pump: H=36m, Q=10T/h, P=3Kw), start is filtered, and ceramic membrane appears after end appears 35L water and obtains fermented liquid concentrated solution 37L, starts the washing fermented liquid that adds water, maintains fermentating liquid volume 37
+2L reaches 37L until washing appears volume.In fermented liquid concentrated solution, add 28L ethanol, after stirring, open ceramic membrane filter, appear side at ceramic membrane and obtain tacrolimus alcohol extract.When the material liquid volume prepared in tank is less than 30L, continues in ceramic membrane treatment tank, add 65% ethanol (or nanofiltration peritoneal effluent) prepared and carry out lixiviate.Vat liquor is transferred to nanofiltration membrane treatment tank, (nanofiltration membrane specification is as follows: GE2540 type film core, molecular weight cut-off 500, membrane filtration area S=2.5m to open nanofiltration membrane
2, trier ram pump: △ P=70bar, Q=11.9L/min, P=2.2Kw) and concentrated.It is 65% ethanol that nanofiltration membrane appears end, and cover is back to ceramic membrane lixiviate.Follow the tracks of leaching process with high performance liquid phase, stop adding 65% ethanol (or nanofiltration peritoneal effluent), feed liquid is concentrated into 27L, stops ceramic membrane filter when appearing vat liquor volume and being 5 times that maintain volume, residue feed liquid is that waste residue processes further.All vat liquors are transferred to nanofiltration membrane treatment tank concentrate, last concentrated solution volume is 15L.Detect through HPLC, it is 1116 μ g/ml that concentrated solution is tired, chromatographic purity 67%, and containing tacrolimus 16.7g, whole process tacrolimus extract yield is 92.8%.
embodiment 8
Tacrolimus fermented liquid 70L, tire 250 μ g/ml, chromatographic purity 87.6%.Containing tacrolimus 17.5g.(ceramic membrane specification is as follows: PALL1960 type film core, 19 passages, passage aperture d=6mm, membrane pore size D=50nm, the long L=1m of film core, filtration area S=0.358m fermented liquid to be put into ceramic membrane treatment tank
2, trier impeller pump: H=36m, Q=10T/h, P=3Kw), start is filtered, and ceramic membrane appears after end appears 35L water and obtains fermented liquid concentrated solution 35L, starts the washing fermented liquid that adds water, maintains fermentating liquid volume 35
+2L reaches 35L until washing appears volume.In fermented liquid concentrated solution, add 56L ethanol, after stirring, open ceramic membrane filter, appear side at ceramic membrane and obtain tacrolimus alcohol extract.When the material liquid volume prepared in tank is less than 30L, continues in ceramic membrane treatment tank, add 80% ethanol (or nanofiltration peritoneal effluent) prepared and carry out lixiviate.Vat liquor is transferred to nanofiltration membrane treatment tank, (nanofiltration membrane specification is as follows: GE2540 type film core, molecular weight cut-off 500, membrane filtration area S=2.5m to open nanofiltration membrane
2, trier ram pump: △ P=70bar, Q=11.9L/min, P=2.2Kw) and concentrated.It is 80% ethanol that nanofiltration membrane appears end, and cover is back to ceramic membrane lixiviate.Follow the tracks of leaching process with high performance liquid phase, stop adding 80% ethanol (or nanofiltration peritoneal effluent), feed liquid is concentrated into 27L, stops ceramic membrane filter when appearing vat liquor volume and being 5 times that maintain volume, residue feed liquid is that waste residue processes further.All vat liquors are transferred to nanofiltration membrane treatment tank concentrate, last concentrated solution volume is 15L.Detect through HPLC, it is 1067.5 μ g/ml that concentrated solution is tired, chromatographic purity 64.8%, and containing tacrolimus 16g, whole process tacrolimus extract yield is 91.5%.
embodiment 9
Tacrolimus fermented liquid 70L, tire 250 μ g/ml, chromatographic purity 87.6%.Containing tacrolimus 17.5g.(ceramic membrane specification is as follows: tami type film core, 19 passages, passage aperture d=6mm, membrane pore size D=50nm, the long L=1.178m of film core, filtration area S=0.25m fermented liquid to be put into ceramic membrane treatment tank
2, trier impeller pump: H=36m, Q=10T/h, P=3Kw), start is filtered, when fermentating liquid volume still remaining 50L time, TMP rapidly increases to 0.25Mpa, test failure.
embodiment 10
Tacrolimus fermented liquid 193L, tire 210.8 μ g/ml, and chromatographic purity 87%, containing tacrolimus 40.7g.(ceramic membrane specification is as follows: PALL1960 type film core, 19 passages, passage aperture d=6mm, membrane pore size D=50nm, the long L=1m of film core, filtration area S=0.358m fermented liquid to be put into ceramic membrane treatment tank
2, trier impeller pump: H=36m, Q=10T/h, P=3Kw), start is filtered, and ceramic membrane appears after end appears 103L water and obtains fermented liquid concentrated solution 90L, starts the washing fermented liquid that adds water, maintains fermentating liquid volume, 90
+5L reaches 90L until washing appears volume.Add after 32L ethanol stirs in fermented liquid concentrated solution, open ceramic membrane filter, appear side at ceramic membrane and obtain tacrolimus alcohol extract.When the material liquid volume prepared in tank is less than 80L, continues in ceramic membrane treatment tank, add 50% ethanol (or nanofiltration peritoneal effluent) prepared and carry out lixiviate.Vat liquor is transferred to nanofiltration membrane treatment tank, (nanofiltration membrane specification is as follows: GE2540 type film core, molecular weight cut-off 500, membrane filtration area S=2.5m to start nanofiltration
2, trier ram pump: △ P=70bar, Q=11.9L/min, P=2.2Kw) and concentrated.It is 50% ethanol that nanofiltration membrane appears end, and cover is back to ceramic membrane lixiviate.Follow the tracks of leaching process with high performance liquid phase, stop adding 50% ethanol (or nanofiltration peritoneal effluent), feed liquid is concentrated into 70L, stops ceramic membrane filter when appearing vat liquor volume and being 5 times that maintain volume, residue feed liquid is that waste residue processes further.All vat liquors are transferred to nanofiltration membrane treatment tank concentrate, last concentrated solution volume is 30L.Detect through HPLC, it is 1250 μ g/ml that concentrated solution is tired, chromatographic purity 65%, and containing tacrolimus 37.5g, whole process tacrolimus extract yield is 92.1%.
embodiment 11
Tacrolimus fermented liquid 12m
3, tire 238 μ g/ml, and chromatographic purity 88%, containing tacrolimus 2.856kg.(ceramic membrane specification is as follows: PALL1960 type film core, 19 passages, passage aperture d=6mm, membrane pore size D=50nm, the long L=1.0m of film core, single filtration area S=0.37m fermented liquid to be put into ceramic membrane treatment tank
2, have 6 film cylinders, have 37 film cores in each film cylinder, total area S=82.14, charging pump: H=30m, Q=88T/h, P=18.5Kw, force (forcing) pump: H=20-25m, Q=900m3/h, P=90Kw), start is filtered, and ceramic membrane appears end and appears 6m
3fermented liquid concentrated solution 6m is obtained after water
3,start the washing fermented liquid that adds water, maintain fermentating liquid volume, 6
+0.5m
3until washing appears volume reach 6m
3.2.4m is added in fermented liquid concentrated solution
3ethanol, after stirring, opens ceramic membrane filter, appears side obtain tacrolimus alcohol extract at ceramic membrane.Continue in ceramic membrane treatment tank, add 50% ethanol (or nanofiltration peritoneal effluent) lixiviate, maintain the material liquid volume 6 prepared in tank
+0.5m
3.Vat liquor is transferred to nanofiltration membrane treatment tank, (nanofiltration membrane specification is as follows: GE8540 type film core, molecular weight cut-off 500, membrane filtration area S=820m to start nanofiltration
2, charging pump: H=53m, Q=65m3/h, P=15Kw) and concentrated.It is 50% ethanol that nanofiltration membrane appears end, and cover is back to ceramic membrane lixiviate.Follow the tracks of leaching process with high performance liquid phase, stop adding 50% ethanol (or nanofiltration peritoneal effluent) when appearing vat liquor volume and being 5 times that maintain volume, feed liquid is concentrated into 5m
3, stop ceramic membrane filter, residue feed liquid is that waste residue processes further.All vat liquors are transferred to nanofiltration membrane treatment tank concentrate, last concentrated solution volume is 1.8m
3.Detect through HPLC, it is 1504 μ g/ml that concentrated solution is tired, chromatographic purity 64%, and containing tacrolimus 2.7kg, whole process tacrolimus extract yield is 95.0%.
Claims (9)
1. from fermented liquid, extract a method for tacrolimus, comprising:
A) ceramic membrane concentrated broth is used.
2. method according to claim 1, also comprises:
B) continue to add in fermented liquid concentrated solution one or several be selected from alcohol, glycol, ketone, ester class, carboxylic-acid organic solvent or above-mentioned solvent and water mixture and after using ceramic membrane lixiviate tacrolimus vat liquor.
3. method according to claim 2, also comprises:
C) concentrate tacrolimus vat liquor by nanofiltration membrane and obtain tacrolimus nanofiltration concentrated solution.
4. method according to claim 1, the ceramic membrane wherein described in step a) refers to the porous-film prepared by aluminum oxide, zirconium white, titanium dioxide, silicon-dioxide, has the forms such as multi-pore channel formula, flat and duct type.
5. method according to claim 2, the organic solvent wherein added in step b) is that one or several are selected from the mixture of alcohol, glycol, ketone, ester class, carboxylic-acid organic solvent or above-mentioned solvent and water, and wherein the ratio of water is 1% to 70%(V:V).
6. method according to claim 3, wherein the nanofiltration membrane of step c) is the organic membrane be made up of macromolecular material, control tunica fibrosa, rolled film, plate and frame film, tubular membrane etc. preferably.
7. the method according to any one of claim 1 ~ 3, wherein in step a), can add water, tensio-active agent, acid, alkali, organic solvent or change broth temperature in fermented liquid in ceramic membrane concentration process.
8. the method according to any one of claim 1 ~ 3, wherein in step b), tacrolimus is dissolved into one or several and is selected from alcohol, glycol, ketone, ester class, carboxylic-acid organic solvent.
9. the method according to any one of claim 1 ~ 3, wherein in step c), nanofiltration peritoneal effluent can directly be applied mechanically back in step b).
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106046023A (en) * | 2016-06-10 | 2016-10-26 | 山东新时代药业有限公司 | Extraction method of tacrolimus |
| CN107056814A (en) * | 2017-02-10 | 2017-08-18 | 广州市微生物研究所 | A kind of preparation method of the crude tacrolimus of low stain |
| CN112390817A (en) * | 2019-08-19 | 2021-02-23 | 鲁南制药集团股份有限公司 | Method for extracting tacrolimus fermentation liquor by salting out |
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| CN112390817A (en) * | 2019-08-19 | 2021-02-23 | 鲁南制药集团股份有限公司 | Method for extracting tacrolimus fermentation liquor by salting out |
| CN114315860A (en) * | 2021-12-27 | 2022-04-12 | 无锡福祈制药有限公司 | Preparation method of sirolimus crude product |
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