CN101935323A - Purification method of deuterohemin - Google Patents
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Abstract
The invention provides a purification method of deuterohemin, comprising the steps of: (1) dissolving crude deuterohemin in one type or multiple types of first solvents, adding one type or multiple types of second solvents into the solution to generate precipitates, and collecting the precipitates, and (2) dissolving the obtained precipitates in one type or multiple types of the first solvents, adding one type or multiple types of the second solvents to generate precipitates, and collecting the precipitates. The invention further discloses the deuterohimin prepared with the purification method of the invention.
Description
Technical field
The present invention relates to a kind of method of purifying secondary heme, also relate to the secondary heme that obtains by this method purifying.
Background technology
Secondary heme (Deuterohemin) is protoheme (Heme, Hame, homologue Protoheme).It has the biological function similar with protoheme, can be used as the prothetic group of some enzymes with biocatalysis such as catalase, peroxidase and cytopigment enzyme etc.When synthetic mimics of peroxidase for example,, make protoheme self to the superoxide instability because the vinyl on the protoheme is easy to by peroxide oxidation.Therefore, when the stand-in of design peroxidase, must eliminate this unstable of protoheme prothetic group.Existing report, thereby utilize Resorcinol to form secondary heme (Liu Yali etc., the improvement of the synthetic and purification process of deuterohemin, Jilin University's natural science journal by the vinyl that the C-alkylated reaction can make H replace on the protoheme, January calendar year 2001, the 91-92 page or leaf).Because secondary heme homeostasis, the enzyme that contains the protoheme prothetic group unstable to superoxide has been eliminated in the application that contains the enzyme simulation thing of this secondary heme prothetic group.
Secondary heme is a kind of ferric iron porphine derivative, and its chemical name is 3,7,12,17-tetramethyl--21H, and 23H-porphines-2,18-dipropionic acid iron(ic) chloride, molecular formula is [C
30H
28FeN
4O
4]
+Cl
-, molecular weight is 599.88, structural formula is as follows
It is reported that existing secondary heme separation purifying technique adopts column chromatography technology usually, for example the column chromatography of polymeric amide carrier, silica gel column chromatography technology (Liu Yali etc. see above).But often come with some shortcomings when chromatographic technique is applied to the separation and purification of secondary heme, for example, the usage quantity of chromatography carrier is big, needs to use deleterious eluting solvent usually, for example benzene etc.Purification by silica gel column chromatography method with people such as Liu Yali (seeing above) is an example, and per minute just needs carrier silica gel 2400 grams from purifying 1 gram secondary heme crude product, and the pure product secondary heme that can be purified into is 0.4 gram, and yield is about 40%.Calculate in this ratio, the pure product of preparation 1000 gram secondary hemes just need 6000 kilograms of column chromatography silica gels (6 tons).If the usage ratio of chromatography carrier and eluting solvent was calculated by 1: 10, prepare 60 tons of eluting solvents of secondary heme needs of above-mentioned output.Therefore, the existing method of carrying out the secondary heme purifying with chromatographic technique is unsuitable for large-scale low-cost suitability for industrialized production.
Therefore, still need to seek a kind of easy, fast and be easy to the secondary heme purification process of large-scale production.
Summary of the invention
The invention provides a kind of method of purifying secondary heme, described method comprises:
(1) with the secondary heme dissolving crude product in one or more first solvents, in this solution, add one or more second solvents producing precipitation, and the collecting precipitation thing and
(2) throw out that obtains is dissolved in one or more first solvents, in this solution, adds one or more second solvents producing precipitation, and collecting precipitation thing once more.
One aspect of the present invention, in described method, step (2) is carried out 2 times at least; Preferably, described step (2) is carried out 2-4 time.
The present invention also provides a kind of secondary heme that is obtained by method purifying of the present invention.The purity of secondary heme of the present invention can reach more than 90%, preferably can reach more than 95%.
Purifying process of the present invention is easy, need not special equipment and chromatography carrier (for example polymeric amide, silica gel etc.), also avoids using a large amount of noxious solvents, can not pollute environment.Technology of the present invention makes the purifying cycle significantly shorten, and product purity can reach more than 90%, and yield can reach more than 85%, even also can realize the preparation of the secondary heme of feather weight in common lab.This shows that the more existing chromatography purification method of the purification efficiency of technology of the present invention is significantly increased.
Therefore, the present invention provides the good technology that a kind of cost is low, be easy to suitability for industrialized production for the production purifying of secondary heme.
Embodiment
The disclosed compound of the application, composition, method, reagent are not limited in the literary composition specifically listed, and they also can have multiple version.For the description of the application's specific embodiments, especially embodiment, be intended to help for the understanding of the present invention, but not limitation of the scope of the invention.
Secondary heme crude product of the present invention is generally purity at the secondary heme below 25%.Described secondary heme crude product can be the mix products of chemical method preparation back without separation and purification.Preferably, described secondary heme crude product is made by chemosynthesis reaction; More preferably, described secondary heme crude product is by the chemical reaction preparation of protoheme and Resorcinol.
First solvent of the present invention is meant the solvent that secondary heme is had the excellent dissolution characteristic, and its available example includes but not limited to: diluted alkaline, pyridine, dimethyl formamide, dimethyl sulfoxide (DMSO), N-Methyl pyrrolidone, formic acid, acetate, butyric acid and their suitable mixture.
Second solvent of the present invention is meant can be with the sedimentary solvent of secondary heme that is dissolved in described first solvent.For example, factors such as polarity that described second solvent can be by changing the residing solvent environment of secondary heme or pH value precipitate the secondary heme that is dissolved in first solvent.Its available example includes but not limited to diluted acid, water, ethers (for example ether, methyl tertiary butyl ether), acetone, following alcohols (for example methyl alcohol, ethanol), ester class (for example ethyl acetate), halohydrocarbon and their suitable mixture of six carbon.
The concentration of diluted alkaline of the present invention is generally 0.01%-35%; Preferably, the concentration of described diluted alkaline is 0.05%-20%; Again preferably, the concentration range of described diluted alkaline is 0.1%-10%; More preferably, described diluted alkaline concentration is 0.1%-3%; Again more preferably, described diluted alkaline concentration is 1%-3%.Operable diluted alkaline for example is the aqueous solution, the metal hydroxides (for example sodium hydroxide, potassium hydroxide etc.) of ammoniacal liquor, the following aliphatic amide of six carbon, the aqueous solution (for example yellow soda ash, salt of wormwood, sodium bicarbonate, saleratus etc.) and their suitable mixture of carbonate among the present invention.
The concentration of diluted acid of the present invention is generally 3%-20%; Preferably, the concentration of described diluted acid is 5%-10%.Operable diluted acid for example has dilute hydrochloric acid, dilute sulphuric acid, rare nitric acid, the following fatty aqueous acid (for example dilute formic acid, acetic acid,diluted) of six carbon and their suitable mixture among the present invention.
In one embodiment of the invention, employed first solvent of each step that described method comprises each other can be identical or different, and each step uses second solvent each other can also be identical or different.For example, step (1) and employed first solvent phase of step (2) are same, and employed second solvent is also identical; Perhaps first solvent phase of step (1) and step (2) use is same, but the employed second solvent difference; Perhaps step (1) is different with first solvent of step (2) use, but employed second solvent phase together; Or first solvent and second solvent of step (1) and step (2) use are all inequality.The above-mentioned solvent use situation that exemplifies also is applicable to and comprises in the embodiment of repeatedly carrying out step (2), that is to say, employed first solvent each other can be identical in the step that each time carried out (2), also can be different, and employed second solvent each other also can be identical or different.For example, use pyridine as first solvent in step (1), water is as second solvent; Still can use pyridine as first solvent in step (2), water perhaps also can use for example dimethyl formamide etc. of other first solvents as second solvent, and other second solvents are ethanol etc. for example; In the step of further carrying out (2), also can use pyridine as first solvent, water is made second solvent, and first solvent that perhaps uses other is N-Methyl pyrrolidone etc. for example, and other second solvent is ethanol etc. for example.
In an embodiment of the inventive method, first solvent described in described step (1) and/or (2) is that diluted alkaline and described second solvent are diluted acid.In another embodiment of the inventive method, first solvent described in step (1) and/or (2) is not a diluted alkaline, and described second solvent is not a diluted acid.In another embodiment of the inventive method, first solvent described in the described step (1) is that diluted alkaline and described second solvent are diluted acid, and first solvent described in the described step (2) is not that diluted acid and described second solvent are not diluted alkaline.In another embodiment of the inventive method, first solvent of described step (1) is not that the diluted alkaline and second solvent are not diluted acid, and first solvent of described step (2) is that the diluted alkaline and second solvent are diluted acid.
" suitable mixture " mentioned among the application can easily be determined according to the routine techniques knowledge of its grasp by those skilled in the art.For example, can use the mixture of dimethyl formamide and dimethyl sulfoxide (DMSO) to be used to dissolve secondary heme as first solvent.
In addition, the percentage of addressing among the application is weight/volume percent concentration.
Usually after settling step, also can comprise the sedimentary step of centrifugal gained in the method for the present invention.Preferably, after each settling step, all carry out the sedimentary step of centrifugal gained.After described centrifugation step, also can comprise the sedimentary step of the centrifugal gained of washing, usually washings such as available suitable ether (for example ether), alcohol, ester, acetone or water.
The product that method of the present invention also can comprise usually with last settling step gained carries out the exsiccant step.Described drying can be carried out under 60 ℃-110 ℃ temperature.
The secondary heme that the method according to this invention purifying obtains has higher degree, for example reaches more than 90%, even 95% above purity.The analytical procedure of described purity is known for those skilled in the art, for example can adopt the RP-HPLC method that the purity of the secondary heme sample of purifying is analyzed.This method for example can be used following chromatographic separation condition and carry out:
Detect wavelength: 390nm; Chromatographic column: C1825 μ m post (150mm * 4.4mmI.D); Moving phase: acetonitrile: water: trifluoroacetic acid (35: 65: 0.1); Flow velocity: 1mL/min.Calculate the purity of secondary heme with the peak area normalization method.
Below in conjunction with specific embodiment, the present invention is explained in further detail in the mode of example.Compound that uses in following examples or reagent can be buied by commercial sources, perhaps prepare by ordinary method well known by persons skilled in the art; Employed laboratory apparatus can be buied by commercial sources.
Embodiment
The preparation of secondary heme crude product:
100g protoheme (available from Tianjin life science Applied Research Laboratory) and 300g Resorcinol are placed the 1000ml there-necked flask that has air set pipe and whipping appts, behind the thorough mixing, in oil bath, be heated to 140～150 ℃, and under this temperature, keep about 30min, be warming up to 180～190 ℃ and under this temperature, keep 15min with the velocity slope of about 2 ℃/min again.Remove oil bath, naturally cool to about 100 ℃ and make secondary heme crude mixture (mixture that promptly contains all reactants).
The purifying of secondary heme:
The 400 gram secondary heme crude mixture that make are added in the dilute alkaline soln of 10～30L 0.1%～10%, fully stirred warm its dissolving that makes through 10～20 minutes.In dilute alkaline soln, add 3%～10% diluted acid again, the pH value of solution is transferred to neutrality.Precipitation appears in the aqueous solution.Leave standstill make the precipitation sedimentation, incline most of supernatant liquor after, lower floor is centrifugal, remove mother liquor.Throw out is colourless to the ether washings with ether washing (1000ml * 4).To precipitate again with 500～1000ml pyridine and dissolve.In pyridine solution, add 1000ml ethanol, adding 20～30L 70-90 ℃ distilled water again under fully stirring in solution slowly separates out secondary heme from solution, leave standstill and make the precipitation natural subsidence, incline most of supernatant liquor after, centrifugal lower sediment and collecting precipitation thing, drying precipitated acquisition purity reaches the secondary heme more than 90%.Can the throw out of above-mentioned collection further be dissolved as required, the operation steps of precipitation, centrifugal and collecting precipitation, to obtain more highly purified secondary heme.
The 400 gram secondary heme crude mixture that make are added in the pyridine of 1～3L, fully stirred warm its dissolving that makes through 10～20 minutes.In pyridine solution, add 20～30L 70-90 ℃ distilled water again.Precipitation appears in solution.Leave standstill make the precipitation sedimentation, incline most of supernatant liquor after, lower floor is centrifugal, remove mother liquor.Throw out is colourless to the ether washings with ether washing (1000ml * 4).Use the dilute alkaline soln dissolution precipitation thing of 10～20L 0.1%～10% again.In solution, add 1000ml ethanol, fully stir and in solution, add 3%～10% dilute hydrochloric acid down again, the pH value of solution is transferred to neutrality.Precipitation appears in the aqueous solution.Leave standstill and make the precipitation natural subsidence, incline most of supernatant liquor after, centrifugal lower sediment and collecting precipitation thing, drying precipitated acquisition purity reaches the secondary heme more than 90%.Can the throw out of above-mentioned collection further be dissolved as required, the operation steps of precipitation, centrifugal and collecting precipitation, to obtain more highly purified secondary heme.
The 400 gram secondary heme crude mixture that make are added in the pyridine of 1～3L, fully stirred warm its dissolving that makes through 10～20 minutes.In pyridine solution, add 20～30L 70-90 ℃ distilled water again.Precipitation appears in solution.Leave standstill make the precipitation sedimentation, incline most of supernatant liquor after, with the centrifugal mother liquor of removing of lower floor.Throw out is colourless to washings with distilled water wash (10L * 4).Use the pyridine dissolution precipitation thing of 1L again.Fully stir and in solution, add 20～30L 70-90 ℃ distilled water down again, solution appearance precipitation.Leave standstill and make the precipitation natural subsidence, incline most of supernatant liquor after, centrifugal lower sediment and collecting precipitation thing, drying precipitated acquisition purity reaches the secondary heme more than 90%.Can further dissolve above-mentioned throw out as required, the operation steps of precipitation, centrifugal and collecting precipitation, to obtain more highly purified secondary heme.
Embodiment 1
Get secondary heme crude mixture 400 grams of above-mentioned chemical reaction preparation, it is added in the sodium hydroxide solution of 20L0.5%, fully stir and warmly made its dissolving in 10 minutes.In this sodium hydroxide solution, add 3% dilute hydrochloric acid again, the pH value of solution is transferred to neutrality, and the placement naturally cooling.Precipitation appears in solution.Leave standstill and make the precipitation sedimentation, incline most of supernatant liquor after, centrifugal lower floor solution is removed mother liquor.Throw out is colourless to the ether washings with ether washing (1000ml * 4).To precipitate again with the dissolving of 500ml pyridine.In pyridine solution, add 1000ml ethanol, fully stir and in solution, add 60-80 ℃ of distilled water of 30L down again secondary heme is slowly separated out from solution, leave standstill and make the precipitation natural subsidence, incline most of supernatant liquor after, centrifugal lower sediment and collecting precipitation thing.As mentioned above, the gained throw out is further fully dissolved with the 500ml pyridine solution, and adding 1L ethanol fully precipitates secondary heme in pyridine solution, and the collecting precipitation thing.At last gained is deposited in 90 ℃ of dryings and obtains pure product 77.98 grams of secondary heme.Product yield (promptly with the chemical reaction conversion rate of the secondary heme chemical preparation and the total recovery in two steps of purifying by 100% secondary heme that calculates) is 85%, and purity is 95%.
Embodiment 2
Get secondary heme crude mixture 400 grams of above-mentioned chemical reaction preparation, it is added in the ammoniacal liquor of 15L2%, fully stir and warmly made its dissolving in 10 minutes.In this ammonia soln, add 5% dilute hydrochloric acid again, the pH value of solution is transferred to neutrality.Precipitation appears in the aqueous solution.Leave standstill and make the precipitation sedimentation, incline most of supernatant liquor after, centrifugal lower floor solution is removed mother liquor.Precipitation washes with water, will precipitate after centrifugal with the dissolving of 500ml pyridine again.Fully stir and in solution, add 50-70 ℃ of distilled water of 30L down secondary heme is slowly separated out from solution, leave standstill and make the precipitation natural subsidence, incline most of supernatant liquor after, centrifugal lower sediment collecting precipitation thing.As mentioned above, gained precipitation is further fully dissolved, added 50-70 ℃ of distilled water of 20L in solution secondary heme is fully precipitated with the 500ml pyridine, at last gained is deposited in 90 ℃ of dryings and obtains the pure product 79.82 of secondary heme and restrain.Product yield is 87%, and purity is 94.5%.
Embodiment 3
Get secondary heme crude mixture 400 grams of above-mentioned chemical reaction preparation.It is fully being dissolved under the stirring thermal condition with the 1L dimethyl formamide.In this solution, add ethanol 1000ml, under well-beaten condition, add 40-60 ℃ of distilled water 20L.Placement makes its naturally cooling, and precipitation appears in solution.Leave standstill and make the precipitation sedimentation, incline most of supernatant liquor after, centrifugal lower floor solution is removed mother liquor.Throw out is colourless to the ether washings with ether washing (1000ml * 4).To precipitate the warm dissolving of sodium carbonate solution with 15L 3% again, in solution, add 5% dilute hydrochloric acid, the pH value of solution will be transferred to neutrality.Place solution and precipitation occurs.Leave standstill and make the precipitation sedimentation, incline most of supernatant liquor after, centrifugal lower floor and collecting precipitation thing.The gained throw out is fully dissolved with the 500ml pyridine.In this solution, add 1000ml ethanol, fully stir and in solution, add 20L70-90 ℃ of distilled water down again secondary heme is slowly separated out from solution, leave standstill and make the precipitation natural subsidence, incline most of supernatant liquor after, centrifugal lower sediment and collecting precipitation thing.At last gained is deposited in 105 ℃ of dryings and obtains pure product 82.57 grams of secondary heme.Product yield is 90%, and purity is 95%.
Embodiment 4
Get secondary heme crude mixture 400 grams of above-mentioned chemical reaction preparation.Fully stirring under the thermal condition with the 1L pyridine its dissolving.Under well-beaten condition, add 50-70 ℃ of distilled water 20L.Place naturally cooling, precipitation appears in solution.Leave standstill and make the precipitation sedimentation, incline most of supernatant liquor after, centrifugal lower floor solution is removed mother liquor.Throw out is colourless to the ether washings with ether washing (1000ml * 4).To precipitate the warm dissolving of potassium hydroxide solution with 15L 1% again, in solution, add 2% dilute sulphuric acid, the pH value of solution will be transferred to neutrality.Place solution, precipitation occurs.Leave standstill and make the precipitation sedimentation, incline most of supernatant liquor after, centrifugal lower floor and collecting precipitation thing.As mentioned above, precipitation further use 1L pyridine dissolving, fully stirred and add 50-70 ℃ of distilled water 20L down and secondary heme is fully precipitated and the operation steps of centrifugal collecting precipitation 2 times, at last gained is deposited in 95 ℃ of dryings and obtains the pure product 83.49 of secondary heme and restrain.Product yield is 91%, and purity is 95.2%.
Embodiment 5
Get secondary heme crude mixture 400 grams of above-mentioned chemical reaction preparation, be poured in the sodium hydroxide solution of 15L7%, fully stir and warmly made its dissolving in 10 minutes.In this sodium hydroxide solution, add 10% dilute hydrochloric acid again, the pH value of solution is transferred to neutrality.Place solution and make its naturally cooling.Precipitation appears in the aqueous solution.Leave standstill and make the precipitation sedimentation, incline most of supernatant liquor after, centrifugal lower floor solution is removed mother liquor.Precipitation adds 30-50 ℃ of distilled water 20L with the warm dissolving of dimethyl formamide of 1L in solution under the violent stirring, place naturally cooling, and precipitation appears in solution.Leave standstill and make the precipitation sedimentation, incline most of supernatant liquor after, mother liquor is removed by centrifugal lower floor.Precipitation is washed with distillation.Repeat above-mentioned with dimethyl formamide dissolving, add the operation steps 2 times that distilled water makes secondary heme precipitation, centrifugal and collecting precipitation, at last gained is deposited in 105 ℃ of dryings and obtains pure product 80.73 grams.Product yield is 88%, and purity is 94.2%.
Embodiment 6
Get secondary heme crude mixture 400 grams of above-mentioned chemical reaction preparation, wherein add the dimethyl formamide of 1L and the mixed solvent of dimethyl sulfoxide (DMSO) (1: 1), fully dissolving under the stirring thermal condition, under well-beaten condition, adding 50-70 ℃ of distilled water 20L.Place naturally cooling, precipitation appears in solution.Leave standstill and make the precipitation sedimentation, incline most of supernatant liquor after, centrifugal lower floor solution is removed mother liquor.Throw out is colourless to washings with distilled water wash (10L * 4).To precipitate again with the dimethyl formamide of 1L and mixed solvent (1: the 1) dissolving of dimethyl sulfoxide (DMSO).Add 1000ml ethanol in this solution, fully stirring adds 60-80 ℃ of distilled water of 30L down again in solution, places to make it naturally cool to room temperature, secondary heme is slowly separated out from solution, leave standstill and make the precipitation natural subsidence, incline most of supernatant liquor after, centrifugal lower floor and collecting precipitation thing.As mentioned above, the gained precipitation is further dissolved with the dimethyl formamide of 1L and the mixed solvent (1: 1) of dimethyl sulfoxide (DMSO), and, in solution, add 20L50-70 ℃ of distilled water under fully stirring again and make secondary heme precipitation, centrifugal and collecting precipitation to wherein adding 1000ml ethanol.At last gained is deposited in 100 ℃ of dryings and obtains pure product 87.16 grams of secondary heme.Product yield is 95%, and purity is 96.2%.
Embodiment 7
Get secondary heme crude mixture 400 grams of above-mentioned chemical reaction preparation, it is added in the sodium hydroxide solution of 20L1%, fully stir and warmly made its dissolving in 10 minutes.In this sodium hydroxide solution, add 5% dilute hydrochloric acid again, the pH value of solution is transferred to neutrality, and the placement naturally cooling.Precipitation appears in solution.Leave standstill and make the precipitation sedimentation, incline most of supernatant liquor after, centrifugal lower floor solution is removed mother liquor.Throw out is colourless to washings with distilled water wash (10L * 4).To precipitate the warm dissolving of sodium carbonate solution with 15L 3% again, in solution, add 5% dilute hydrochloric acid, the pH value of solution will be transferred to neutrality.Place solution and precipitation occurs.Leave standstill and make the precipitation sedimentation, incline most of supernatant liquor after, centrifugal lower floor and collecting precipitation thing.Repeat the dissolving of above-mentioned usefulness 3% yellow soda ash, precipitate and the sedimentary operation steps of centrifugal collection gained 2 times, at last gained is deposited in 105 ℃ of dryings and obtains pure product 81.65 grams of secondary heme by 5% dilute hydrochloric acid.Product yield is 89%, and purity is 95%.
Embodiment 8
Get secondary heme crude mixture 400 grams of above-mentioned chemical reaction preparation.Fully stirring under the thermal condition with the 1L pyridine its dissolving.Under well-beaten condition, add 50-70 ℃ of distilled water 20L.Place naturally cooling, precipitation appears in solution.Leave standstill and make the precipitation sedimentation, incline most of supernatant liquor after, centrifugal lower floor solution is removed mother liquor.Throw out is colourless to washings with distilled water wash (10L * 4).To precipitate again with the 1L pyridine and fully stir under the thermal condition its dissolving.Under well-beaten condition, add 50-70 ℃ of distilled water 20L.Place naturally cooling, precipitation appears in solution.Leave standstill and make the precipitation sedimentation, incline most of supernatant liquor after, centrifugal lower floor and collecting precipitation thing.As mentioned above, use the 1L pyridine further dissolve, by making the operation steps 2 times of secondary heme precipitation and centrifugal collecting precipitation to wherein adding 50-70 ℃ of distilled water 20L, at last gained is deposited in 100 ℃ of dryings and obtains pure product 83.94 grams of secondary heme.Product yield is 91.5%, and purity is 95.2%.
Claims (10)
1. the method for a purifying secondary heme comprises:
(1) with the secondary heme dissolving crude product in one or more first solvents, in this solution, add one or more second solvents producing precipitation, and the collecting precipitation thing and
(2) throw out that obtains is dissolved in one or more first solvents, in this solution, adds one or more second solvents producing precipitation, and collecting precipitation thing once more.
2. the method for claim 1 is characterized in that, described step (2) is carried out 2 times at least, preferably carries out 2-4 time.
3. claim 1 or 2 method is characterized in that first solvent that uses in each step is identical or different each other, and second solvent that uses in each step is identical or different each other.
4. claim 1 or 2 method is characterized in that described first solvent is selected from diluted alkaline, pyridine, dimethyl formamide, dimethyl sulfoxide (DMSO), N-Methyl pyrrolidone and their suitable mixture.
5. claim 1 or 2 method is characterized in that, described second solvent is selected from diluted acid, water, ethers, acetone, following alcohols, ester class, halohydrocarbon and their suitable mixture of six carbon.
6. claim 1 or 2 method is characterized in that described first solvent is that diluted alkaline and described second solvent are diluted acid.
7. the method for claim 4 is characterized in that, the concentration range of described diluted alkaline is 0.01%-35%; Preferably, the concentration range of described diluted alkaline is 0.05%-20%; Again preferably, the concentration range of described diluted alkaline is 0.1%-10%; More preferably, the concentration range of described diluted alkaline is 0.1%-3%; Again more preferably, the concentration range of described diluted alkaline is 1%-3%.
8. the method for claim 5 is characterized in that, the concentration range of described diluted acid is 3%-20%; Preferably, the concentration range of described diluted acid is 5%-10%.
9. secondary heme that obtains by each method of aforementioned claim.
10. the secondary heme of claim 9 is characterized in that, the purity of described secondary heme is at least 90%.
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| CN102584839A (en) * | 2012-01-17 | 2012-07-18 | 长春大政药业科技有限公司 | Preparation process of deuterohemin |
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| CN1424322A (en) * | 2002-12-11 | 2003-06-18 | 吉林大学 | Trivalent iron porphrin and its derivative-short peptide compound and its synthesis |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102584839A (en) * | 2012-01-17 | 2012-07-18 | 长春大政药业科技有限公司 | Preparation process of deuterohemin |
| CN102584839B (en) * | 2012-01-17 | 2014-08-06 | 长春大政药业科技有限公司 | Preparation process of deuterohemin |
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