CN116023356A - Preparation method of hydroxysafflor yellow A - Google Patents
Preparation method of hydroxysafflor yellow A Download PDFInfo
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- CN116023356A CN116023356A CN202111247575.XA CN202111247575A CN116023356A CN 116023356 A CN116023356 A CN 116023356A CN 202111247575 A CN202111247575 A CN 202111247575A CN 116023356 A CN116023356 A CN 116023356A
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- ZZMASNSDVDSYKO-UHFFFAOYSA-N hydroxysafflor yellow A Natural products OCC1OC(C(O)C(O)C1O)C2=C(O)C(O)(C3OC(CO)C(O)C(O)C3O)C(=O)C(=C2O)C(=O)C=Cc4ccc(O)cc4 ZZMASNSDVDSYKO-UHFFFAOYSA-N 0.000 title claims abstract description 49
- IAVUBSCVWHLRGE-UXEKTNMQSA-N (6e)-2,5-dihydroxy-6-[(e)-1-hydroxy-3-(4-hydroxyphenyl)prop-2-enylidene]-2,4-bis[(2s,3r,4r,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]cyclohex-4-ene-1,3-dione Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1C(C(C(O)([C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)C1=O)=O)=C(O)\C1=C(/O)\C=C\C1=CC=C(O)C=C1 IAVUBSCVWHLRGE-UXEKTNMQSA-N 0.000 title claims abstract description 48
- 238000002360 preparation method Methods 0.000 title claims abstract description 18
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- 229920005989 resin Polymers 0.000 claims abstract description 25
- 239000003814 drug Substances 0.000 claims abstract description 20
- 238000001035 drying Methods 0.000 claims abstract description 17
- 238000005189 flocculation Methods 0.000 claims abstract description 15
- 230000016615 flocculation Effects 0.000 claims abstract description 15
- 238000000605 extraction Methods 0.000 claims abstract description 11
- 238000004587 chromatography analysis Methods 0.000 claims abstract description 9
- 238000004519 manufacturing process Methods 0.000 claims abstract description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 51
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 43
- 239000003480 eluent Substances 0.000 claims description 34
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- KINGXFAMZNIVNL-SXQDSXCISA-N safflor yellow A Natural products OC[C@@H]1O[C@H]2[C@H](OC3=C2C(=O)C(=C(O)C=Cc4ccc(O)cc4)C(=O)[C@]3(O)[C@@H]5O[C@H](CO)[C@@H](O)[C@H](O)[C@H]5O)[C@@H](O)[C@H]1O KINGXFAMZNIVNL-SXQDSXCISA-N 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 14
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- 238000001914 filtration Methods 0.000 claims description 12
- 244000020518 Carthamus tinctorius Species 0.000 claims description 11
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- 239000000843 powder Substances 0.000 claims description 10
- 238000001291 vacuum drying Methods 0.000 claims description 10
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 9
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- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 7
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- 230000008569 process Effects 0.000 claims description 7
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- 238000010298 pulverizing process Methods 0.000 claims description 5
- 238000002791 soaking Methods 0.000 claims description 5
- DLYUQMMRRRQYAE-UHFFFAOYSA-N tetraphosphorus decaoxide Chemical compound O1P(O2)(=O)OP3(=O)OP1(=O)OP2(=O)O3 DLYUQMMRRRQYAE-UHFFFAOYSA-N 0.000 claims description 5
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 4
- 241000628997 Flos Species 0.000 claims description 4
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 4
- 239000003153 chemical reaction reagent Substances 0.000 claims description 3
- 238000010828 elution Methods 0.000 claims description 3
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- 239000005909 Kieselgur Substances 0.000 claims 1
- 238000004321 preservation Methods 0.000 claims 1
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- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 abstract description 2
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- DYQVDISPPLTLLR-HJQYTNQXSA-N Carthamin Natural products CC[C@H]1O[C@H]([C@H](O)[C@@H](O)[C@@H]1O)[C@]2(O)C(=C(C=C/3C(=O)C(=C(O)[C@](O)([C@@H]4O[C@H](CO)[C@@H](O)[C@H](O)[C@H]4O)C3=O)C(=O)C=Cc5ccc(O)cc5)C(=O)C(=C2O)C(=O)C=Cc6ccc(O)cc6)O DYQVDISPPLTLLR-HJQYTNQXSA-N 0.000 description 5
- WLYGSPLCNKYESI-RSUQVHIMSA-N Carthamin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1[C@@]1(O)C(O)=C(C(=O)\C=C\C=2C=CC(O)=CC=2)C(=O)C(\C=C\2C([C@](O)([C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)C(O)=C(C(=O)\C=C\C=3C=CC(O)=CC=3)C/2=O)=O)=C1O WLYGSPLCNKYESI-RSUQVHIMSA-N 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
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- 230000000694 effects Effects 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- HVAUUPRFYPCOCA-AREMUKBSSA-N 2-O-acetyl-1-O-hexadecyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCCOC[C@@H](OC(C)=O)COP([O-])(=O)OCC[N+](C)(C)C HVAUUPRFYPCOCA-AREMUKBSSA-N 0.000 description 2
- 208000002193 Pain Diseases 0.000 description 2
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Images
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention belongs to the field of natural medicine extraction, and particularly relates to a preparation method of hydroxysafflor yellow A. The preparation method of the hydroxysafflor yellow A comprises the following steps: extracting and concentrating; flocculation; HZ chromatography 3 # Separating by a resin column; HZ001 cation exchange resin; separating by a gel column; concentrating and drying. The preparation method solves the problems of long flow, low yield and high production cost of the existing hydroxysafflor yellow A extraction process. The invention provides the preparation method of the hydroxysafflor yellow A, which has the advantages of short preparation process flow, low energy consumption and high product purity, and effectively improves the yield of the hydroxysafflor yellow A.
Description
Technical Field
The invention belongs to the field of natural medicine extraction, and particularly relates to a preparation method of hydroxysafflor yellow A.
Background
Safflower is a dried flower of Compositae plant safflower Carthamus tinctorius L. The flowers are picked when the flowers turn from yellow to red, and dried in the shade or sun-dried. Has effects of promoting blood circulation, dredging channels, removing blood stasis, and relieving pain. Safflower is a common traditional Chinese medicine for activating blood and dissolving stasis, and can be used for treating a plurality of blood circulation disorders such as coronary heart disease, angina pectoris and the like. Also can be used for treating amenorrhea, dysmenorrhea, lochia, abdominal mass, chest pain, abdominal pain due to stagnation, thorny chest and hypochondrium, traumatic injury, and swelling and pain due to sore.
The safflower is rich in carthamin yellow, and the proportion of carthamin yellow in the safflower is 20% -30%. Hydroxysafflor yellow A is the main pharmacodynamic substance of safflower yellow. The hydroxyl safflower yellow A (Hydroxysafflor yellow A) is a compound with a monochalcone glycoside structure, is the most effective water-soluble part with the pharmacological efficacy of safflower, can inhibit platelet aggregation and release induced by platelet activating factor, can competitively inhibit the combination of platelet activating factor and platelet receptor, and is an active ingredient of safflower yellow for promoting blood circulation and removing blood stasis. Has antithrombin-induced platelet aggregation activity, antiinflammatory activity, cytoprotective activity, and antitumor activity, and has effects of resisting platelet and myocardial ischemia. Meanwhile, the hydroxy safflower yellow A can also be used for preparing health care and cosmetics, and is also a good food dye. The safflower yellow is a natural food coloring agent. The color is bright yellow, has extremely low toxicity, strong tinting strength and stable color tone, and is widely applied to food coloring of fruit juice type beverages, candies, cake color packages and the like.
Although the prior art has disclosed hydroxysafflor yellow A and a separation and purification method thereof, for example, chinese patent application CN201010606888.5 discloses a preparation method of hydroxysafflor yellow A, which rapidly prepares a large amount of high-purity hydroxysafflor yellow A from safflower by high-speed countercurrent chromatography, and provides a material basis for pharmacological and pharmacodynamics research of safflower preparations and hydroxysafflor yellow A. Chinese patent application CN02132781.5 discloses a method for preparing hydroxysafflor yellow a, which is prepared by extracting, separating, precipitating with ethanol, and purifying.
However, the purity of the existing hydroxysafflor yellow A product is not high. From the viewpoint of purity of the conventional safflower yellow, it is necessary to further purify the safflower yellow after separation because 10 to 20% or more of the impurity substances are substantially present. The structural properties of the impurities are not qualitative, and certain quality uncontrollability exists. In addition, the existing safflower yellow extraction process has the problems of long flow, high energy consumption, low yield, high production cost and the like.
The invention aims to provide a preparation method of hydroxysafflor yellow A, which adopts a more environment-friendly leaching method for extraction, and compared with an ultrasonic extraction method, the energy consumption level in the preparation process of hydroxysafflor yellow A is obviously reduced, and further provides a preparation method with low energy consumption and environment friendliness. The preparation method of the hydroxysafflor yellow A provided by the invention has the advantages of short process flow, low energy consumption, environmental protection, high product purity and effective reduction of the production cost of the safflower yellow.
Disclosure of Invention
In order to overcome the technical problems, the invention provides a preparation method of hydroxysafflor yellow A, which is obtained through optimized process extraction, separation and purification steps, and provides a method for preparing high-purity hydroxysafflor yellow A rapidly and at low cost.
In order to achieve the above object, the technical scheme provided by the invention is as follows:
a preparation method of hydroxysafflor yellow A comprises the following steps:
(1) Extracting and concentrating: extracting Carthami flos with 60-90% (volume concentration) ethanol, concentrating the extractive solution to obtain concentrated solution;
(2) Flocculation: flocculating the concentrated solution by using a deacetylated chitosan solution to prepare a flocculation treatment solution;
(3) HZ chromatography 3 # Resin column separation: subjecting the flocculation treatment liquid to HZ chromatography 3 # Separating with resin column, eluting with purified water at a speed of 0.5-1BV/h, and collecting eluate;
(4) HZ001 cation resin exchange: exchanging the eluent in the step (3) by using HZ001 cationic resin, eluting with purified water at a speed of 0.5-1BV/h after loading, and collecting the eluent;
(5) Gel column separation: separating the eluent in step (4) by LH-20 gel column, eluting with purified water after loading, wherein the loading elution speed is 0.2-0.4BV/h, and collecting eluent;
(6) Concentrating and drying: concentrating under reduced pressure to dry, pulverizing into fine powder, stirring at 50deg.C for 2 hr, vacuum filtering, soaking and washing the filter cake with ethanol for 2 times, vacuum filtering, and vacuum drying to obtain hydroxysafflor yellow A.
Preferably, in (1), the extraction is performed using 80-90% ethanol (by volume).
Preferably, in (1), the temperature of the extraction is 65-70 ℃.
Preferably, in (2), the reagent used in flocculation is a solution of 1% glacial acetic acid containing 2% chitosan.
Preferably, in (2), the flocculation treatment is: firstly, reducing the stirring speed to 15-60rpm, stirring, then adding diatomite with the weight of 4-6% of that of the safflower medicinal material, treating for 5-10min, and standing for 1-3h.
Preferably, in (2), the flocculation treatment is: firstly, reducing the stirring speed to 15-60rpm, stirring, then adding diatomite accounting for 5% of the safflower medicinal material, treating for 5min, and standing for 2h.
Preferably, in the step (2), the flocculation treatment process is to add 3.0-3.5% of 1% glacial acetic acid with the medicinal material amount to prepare 2% of deacetylated chitosan solution under the stirring speed of 80rpm, continuously stir for 1-5 minutes, reduce the stirring speed to 15-60rpm, then add 5% of diatomite with the medicinal material amount to stir for 5 minutes, and stand for 2 hours.
Preferably, in (3), the HZ chromatography 3 # Separating with resin column, eluting with purified water at a speed of 0.5-1BV/h, collecting 5-10 parts of eluent in a section of 0.02-0.04BV when hydroxysafflor yellow A yellow color band reaches the bottom of the column (when the eluent is detected to be strongly absorbed by an ultraviolet detector at 403nm absorption wavelength or the whiteness of the eluent is reduced from 10 DEG Bx to 8 DEG Bx or the eluent is obviously changed from pale yellow to pale red), respectively taking 1 mu sample, spreading in 35-40% acetic acid on the same polyamide plate, combining the eluent with obvious spots, and continuously collecting the eluent to 1-2BV.
Preferably, in (3), the HZ chromatography 3 # Separating with resin column, eluting with purified water at a rate of 0.5-1BV/h, collecting hydroxyl groupsWhen the yellow color band of carthamin A reached the bottom of the column, 8 parts of the eluent were collected at 0.04BV intervals, 1. Mu. Of each sample was spotted on the same polyamide plate and developed in 36% acetic acid, and the eluates with distinct spots were combined and the eluent was collected continuously to 1.5BV.
Preferably, in (4), the HZ001 cation resin is exchanged, purified water is used for eluting at the speed of 0.5-1BV/h after the sample is loaded, collection is started when effluent liquid starts to turn yellow, the collection amount is the sum of the loaded liquid medicine amount and the volume of a column bed, the pH value of the collected liquid is regulated to 4.2-4.7 by ammonia water, and the collected liquid is concentrated to the relative density of 1.07-1.10 at the temperature of 20-30 ℃ under reduced pressure below 45 ℃.
Preferably, in the step (5), the gel column is separated, the gel column is LH-20 column, purified water is used for eluting after sample loading, the sample loading eluting speed is 0.2-0.4BV/h, when the hydroxysafflor yellow A yellow color band reaches the bottom of the column, 6-8 parts of the hydroxysafflor yellow A yellow is collected by 0.02-0.04BV each part, 0.02-0.04BV each part is collected, the total collection amount is 0.3BV, 1 μl of the eluent is respectively sampled and spotted on the same polyamide plate, the eluent with obvious bright yellow spots is developed in 30-40% acetic acid.
Preferably, in (5), the acetic acid is 36% acetic acid.
Preferably, in (6), the concentration and drying are performed at 40-60 ℃, preferably 45 ℃, under reduced pressure.
Preferably, in the step (6), the product concentrated under reduced pressure is crushed into fine powder, then 8-10 times (V/W) of ethanol is added into the fine powder, the mixture is stirred for 1-3 hours at the temperature of 40-60 ℃, the suction filtration is carried out, and the filter cake is soaked and washed for 1-3 times by the ethanol; drying at 30-50deg.C and-0.090-0.050 MPa for 4-8 hr, and adding 1/3-2/3 of wet phosphorus pentoxide into the drying oven, and vacuum drying at 30-50deg.C and-0.090-0.050 MPa for 4-8 hr.
Preferably, in (6), the concentration and drying process is: concentrating the gel column separated sample under reduced pressure below 45deg.C to dry, pulverizing into fine powder, adding 9 times of (V/W) ethanol, stirring at 50deg.C for 2 hr, suction filtering, soaking the filter cake with ethanol for 2 times, each time with 2 times of (V/W) ethanol, suction filtering, and placing wet product in tray with thickness of about 0.3g/cm 2 ，30℃、-0.090Vacuum drying under-0.090 MPa for 5 hr, and vacuum drying under-50 deg.C with 1/3 of wet phosphorus pentoxide with equal thickness in a drying oven to obtain hydroxy safflower yellow A.
Compared with the prior art, the invention has the technical advantages that:
(1) The invention provides a novel method for preparing a hydroxysafflor yellow A compound, which is prepared by the steps of extraction, separation and purification by an optimized process, and provides a method for preparing the hydroxysafflor yellow A with high speed and low cost.
(2) The preparation method provided by the invention has the advantages of short process flow, low energy consumption and high product purity, effectively reduces the production cost of the carthamin yellow, and can rapidly prepare the high-purity hydroxy carthamin yellow A with low cost.
Drawings
Fig. 1: liquid chromatogram of the product of example 1.
Detailed Description
The present invention will be described by way of specific examples, to facilitate understanding and grasping of the technical solution of the present invention, but the present invention is not limited thereto. The experimental methods described in the following examples are all conventional methods unless otherwise specified; the reagents and materials, unless otherwise specified, are commercially available.
Example 1
1. Extracting and concentrating
Extracting Carthami flos with 80% ethanol at 68deg.C twice for 1.5 hr and 10 times (V/W) for 1 hr, filtering, centrifuging the residue, mixing the medicinal liquids, and concentrating under reduced pressure below 45deg.C to relative density of 1.03 (45deg.C).
2. Flocculation
Adding 3.3% of 1% glacial acetic acid with the medicinal material amount to prepare 2% of deacetylated chitosan solution under the stirring speed of 80rpm, continuously stirring for 5 minutes, reducing the stirring speed to 30rpm, adding 5% of diatomite with the medicinal material amount, stirring for 5 minutes, standing for 2 hours, and centrifuging.
HZ chromatography 3 # Resin column separation
Loading the flocculated sample onto HZ chromatograph 3 at a rate of 0.5BV/h # Resin column (purchased from Shanghai Huazhu technology Co., ltd.) (100-150 mesh, column bed height 800mm, 1ml resin sample loading liquid medicine amount is not more than [14.3 (mg/g)/medicinal material hydroxysafflor yellow A content (mg/g)/90% (transfer rate) x 80%]g or not more than 11mg calculated by hydroxy safflower yellow A, the temperature of the liquid medicine is normal temperature, 1BV/h of 1% sodium hydroxide, purified water and 70% ethanol are sequentially used for eluting 1BV respectively at the speed of 1BV/h, then purified water is used for eluting about 3BV till no alcohol is used, after the sample is applied, purified water is used for eluting at the speed of 0.75BV/h, when the yellow color band of the hydroxy safflower yellow A reaches the bottom of the column (when the eluent is detected to be strongly absorbed by an ultraviolet detector at the absorption wavelength of 403nm or the whiteness of the eluent is reduced from 10 DEG Bx to 8 DEG Bx or the color of the eluent is obviously changed from pale yellow to pale red), 8 parts of the eluent is firstly collected in a segmented mode at about 0.04BV each part, 1 mu point is respectively applied to the same polyamide plate and is developed in 36% acetic acid, and the eluent with obvious spots are combined and the eluent is continuously collected till 1.5BV.
HZ001 cation exchange resin
Loading the mixture on an HZ001 cation resin exchange column (50-80 meshes, the height of a column bed is not less than 800mm, the liquid medicine amount of each 1ml resin loaded is not more than 2.1g based on medicinal materials, the temperature of the liquid medicine is normal temperature, 4BV is eluted by 4% hydrochloric acid solution at the speed of 1BV/h and then purified water of about 4BV is used for eluting to pH of about 4) at the speed of about 4BV/h, the purified water is used for eluting at the speed of 0.5BV/h after loading, collection is started when effluent begins to turn yellow, the collection amount is the sum of the liquid medicine loaded and the volume of the column bed, the pH of the collection liquid is regulated to about 4.5 by ammonia water, and the relative density is reduced to about 1.08 (25 ℃), wherein the pH of the collection liquid is reduced to below 45 ℃.
5. Gel column separation
Loading onto LH-20 column (Fine, column bed height 800mm, the liquid medicine amount of each 1ml gel is not more than 0.7g calculated by medicinal materials, the liquid medicine volume is not more than 10% of the gel column volume, the temperature of the loaded liquid medicine is normal temperature, 0.2BV is eluted by 0.3% sodium carbonate solution at the speed of 0.3BV/h, then purified water is used for eluting to neutrality, the elution speed of loading is 0.3BV/h after loading, when hydroxysafflor yellow A yellow color band reaches the bottom of the column, 7 parts are collected by about 0.02BV each part, then about 0.03BV each part is collected, the total amount is about 0.3BV, 1 μl sample is respectively taken and spread in 36% acetic acid of the same polyamide plate, and the eluent with obvious bright yellow spots is combined.
6. Concentrating and drying
Concentrating the gel column separated sample under reduced pressure below 45deg.C to dry, pulverizing into fine powder, adding 9 times of (V/W) ethanol, stirring at 50deg.C for 2 hr, suction filtering, soaking the filter cake with ethanol for 2 times, each time with 2 times of (V/W) ethanol, suction filtering, and placing wet product in tray with thickness of about 0.3g/cm 2 Vacuum drying at 30 deg.C and-0.090 MPa for about 5 hr, and vacuum drying at 50 deg.C and-0.090 MPa for about 5 hr to obtain hydroxy safflower yellow A. The purity of hydroxysafflor yellow A was 98.7%.
Example 2
1. Extracting and concentrating
Extracting Carthami flos with 80% ethanol at 68deg.C twice for 1.5 hr and 10 times (V/W) for 1 hr, filtering, centrifuging the residue, mixing the medicinal liquids, and concentrating under reduced pressure below 45deg.C to relative density of 1.03 (45deg.C).
2. Flocculation
Adding 3.3% of 1% glacial acetic acid with the medicinal material amount to prepare 2% of deacetylated chitosan solution under the stirring speed of 80rpm, continuously stirring for 5 minutes, reducing the stirring speed to 30rpm, adding 5% of diatomite with the medicinal material amount, stirring for 5 minutes, standing for 2 hours, and centrifuging.
HZ-816 resin column separation
Loading the flocculated sample onto HZ-816 resin column (100-150 mesh, column height 800mm, 1ml resin loading liquid medicine amount is not more than [14.3 (mg/g)/(90% (transfer rate) ×80% ] g based on medicinal material) or not more than 11mg based on hydroxysafflor yellow A, loading at room temperature, eluting with 30% ethanol at 0.5BV/h, when hydroxysafflor yellow A color band reaches the bottom of the column (when strong absorption of eluent is detected by ultraviolet detector at 403nm absorption wavelength or whiteness of eluent is reduced from 10 DEG Bx to 8 DEG Bx, or eluent color is obviously changed from pale yellow to pale red), starting to collect 8 parts of eluent in sections of about 0.04BV each part, respectively taking 1 mu sample and spreading in 36% acetic acid, combining obvious eluent and continuing to collect eluent until 1.5BV.
4.724 resin column separation
Loading onto 724 resin column (50-80 mesh, column bed height not less than 800mm, liquid medicine amount per 1ml resin is not more than 2.1g calculated by medicinal materials, liquid medicine temperature is normal temperature, 4BV is eluted by 4% hydrochloric acid solution at 1BV/h speed, and then purified water of about 4BV is eluted to pH about 4), after loading, purified water is eluted at 0.75BV/h speed, collection is started when effluent begins to turn yellow, collection amount is the sum of liquid medicine amount loaded and column bed volume, collected liquid is adjusted to pH about 4.5 by ammonia water, and reduced pressure concentration is carried out to relative density about 1.07 (45 ℃).
5. Gel column separation
Loading onto LH-20 column (Fine, column bed height 800mm, the liquid medicine amount of each 1ml gel is not more than 0.7g calculated by medicinal materials, the liquid medicine volume is not more than 10% of the gel column volume, the temperature of the loaded liquid medicine is normal temperature, about 0.2BV is eluted by 0.3% sodium carbonate solution at the speed of 0.3BV/h, then about 1.5BV purified water is used for eluting to neutrality), the purified water is used for eluting after loading, the eluting speed of loading is 0.3BV/h, 7 parts are collected by about 0.02BV each part when hydroxysafflor yellow A color band reaches the bottom of the column, then about 0.03BV each part is collected, the total amount is about 0.3BV, 1 μl sample is respectively taken and applied on the same polyamide plate to be developed in 36% acetic acid, and the eluent with obvious bright yellow spots is combined.
6. Concentrating and drying
Concentrating the gel column separated sample under reduced pressure below 45deg.C to dry, pulverizing into fine powder, adding 9 times of (V/W) ethanol, stirring at 50deg.C for 2 hr, suction filtering, soaking the filter cake with ethanol for 2 times, each time with 2 times of (V/W) ethanol, suction filtering, and placing wet product in tray with thickness of about 0.3g/cm 2 Vacuum drying at 30 ℃ and-0.090 MPa of about 5After the hour, putting phosphorus pentoxide with the thickness of 1/3 of the wet product quantity and the like into a drying box, and continuing to perform vacuum drying at 50 ℃ and minus 0.090MPa for about 5 hours to obtain the hydroxy safflower yellow A. The purity of hydroxysafflor yellow A was 90.1%.
The foregoing detailed description is directed to one of the possible embodiments of the present invention, which is not intended to limit the scope of the invention, but is to be accorded the full scope of all such equivalents and modifications so as not to depart from the scope of the invention.
Claims (10)
1. A preparation method of hydroxysafflor yellow A comprises the following steps:
(1) Extracting and concentrating: extracting Carthami flos with 60-90% ethanol, concentrating the extractive solution to obtain concentrated solution;
(2) Flocculation: flocculating the concentrated solution by using chitosan and/or diatomite to prepare flocculation treatment liquid;
(3) HZ chromatography 3 # Resin column separation: subjecting the flocculation treatment liquid to HZ chromatography 3 # Separating with resin column, eluting with purified water at a speed of 0.5-1BV/h, and collecting eluate;
(4) HZ001 cation resin exchange: exchanging the eluent in the step (3) by using HZ001 cationic resin, eluting with purified water at a speed of 0.5-1BV/h after loading, and collecting the eluent;
(5) Gel column separation: separating the eluent in step (4) by LH-20 gel column, eluting with purified water after loading, wherein the loading elution speed is 0.2-0.4BV/h, and collecting eluent;
(6) Concentrating and drying: concentrating and drying the eluent in the step (5) to obtain hydroxysafflor yellow A.
2. The process for producing hydroxysafflor yellow a according to claim 1, wherein in (1), 80-90% ethanol is used for the extraction; the extraction temperature is 65-70 ℃.
3. The process for producing hydroxysafflor yellow A according to claim 1, wherein in (2), the flocculation reagent is a solution of 1% glacial acetic acid and 2% chitosan deacetylate.
4. The method for preparing hydroxysafflor yellow A as claimed in claim 3, wherein (2) the flocculation treatment process further comprises reducing the stirring speed to 15-60rpm after the chitosan solution treatment, adding diatomaceous earth with an amount of 4-6% of safflower medicinal material for treatment for 5-10min, and standing for 1-3h.
5. The method for producing hydroxysafflor yellow A according to claim 1, wherein in (3), the HZ chromatography is 3 # Separating with resin column, eluting with purified water at a speed of 0.5-1BV/h, collecting eluate at a ratio of 0.02-0.04BV when hydroxysafflor yellow A yellow color band reaches the bottom of column, spreading 1 μ sample on the same polyamide plate in 35-40% acetic acid, mixing the eluates with obvious spots, and continuously collecting eluate to 1-2BV.
6. The process for preparing hydroxysafflor yellow A as claimed in claim 1, wherein in (4), HZ001 cation resin is exchanged, purified water is used for eluting at a speed of 0.5-1BV/h after loading, collection is started when effluent liquid starts to turn yellow, the collection amount is the sum of the loading liquid medicine and the column bed volume, pH of the collection liquid is adjusted to 4.2-4.7 by ammonia water, and the collection liquid is concentrated to a relative density of 1.07-1.10 at a temperature of 20-30 ℃ under reduced pressure below 45 ℃.
7. The process for preparing hydroxysafflor yellow A as claimed in claim 1, wherein in (5), the gel column is separated, the gel column is LH-20 column, purified water is used for eluting after loading, the loading eluting speed is 0.2-0.4BV/h, 6-8 parts of hydroxysafflor yellow A yellow is collected by 0.02-0.04BV each part when the yellow color band of hydroxysafflor yellow A reaches the bottom of the column, 0.02-0.04BV each part is collected, the total amount of collection is 0.3BV, 1 μl of the gel column is respectively spotted on the same polyamide plate and developed in 30-40% acetic acid, and the eluents with obvious bright yellow spots are combined.
8. The process for producing hydroxysafflor yellow A as claimed in claim 1, wherein in (6), the concentration and drying are carried out at 40-60℃under reduced pressure.
9. The process for preparing hydroxysafflor yellow A as claimed in any one of claims 1 to 8, wherein (6) the concentrated product is crushed into fine powder, then 8 to 10 times of ethanol is added to the fine powder, the mixture is stirred for 1 to 3 hours at 40 to 60 ℃ with heat preservation, suction filtration is carried out, and the filter cake is soaked and washed with ethanol for 1 to 3 times; drying at 30-50deg.C and-0.090-0.050 MPa for 4-8 hr, and adding 1/3-2/3 of wet phosphorus pentoxide with thickness into the drying oven, and continuing at 30-50deg.C and-0.090-
Vacuum drying at 0.050MPa for 4-8 hours.
10. The method for producing hydroxysafflor yellow a of claim 1, wherein (6) the concentration and drying process is: concentrating the gel column separated sample under reduced pressure below 45deg.C to dry, pulverizing into fine powder, adding 9 times of ethanol, stirring at 50deg.C for 2 hr, vacuum filtering, soaking and washing the filter cake with ethanol for 2 times, each time 2 times of the amount of the fine powder, vacuum filtering, and placing wet product in tray with thickness of about 0.3g/cm 2 Vacuum drying at 30 deg.C and 0.090MPa for 5 hr, and vacuum drying at 50 deg.C and 0.090MPa for 5 hr to obtain hydroxy safflower yellow A.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101195647A (en) * | 2006-12-06 | 2008-06-11 | 杨炜 | Hydroxyl carthamus tinctorius yellow colour A, preparation method and application thereof |
| CN102127124A (en) * | 2010-12-27 | 2011-07-20 | 中国科学院新疆理化技术研究所 | Method for preparing hydroxysafflor yellow A |
| CN102702150A (en) * | 2012-06-19 | 2012-10-03 | 浙江永宁药业股份有限公司 | Preparation method and application of hydroxysafflor yellow A |
-
2021
- 2021-10-26 CN CN202111247575.XA patent/CN116023356A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101195647A (en) * | 2006-12-06 | 2008-06-11 | 杨炜 | Hydroxyl carthamus tinctorius yellow colour A, preparation method and application thereof |
| CN102127124A (en) * | 2010-12-27 | 2011-07-20 | 中国科学院新疆理化技术研究所 | Method for preparing hydroxysafflor yellow A |
| CN102702150A (en) * | 2012-06-19 | 2012-10-03 | 浙江永宁药业股份有限公司 | Preparation method and application of hydroxysafflor yellow A |
Non-Patent Citations (4)
| Title |
|---|
| 徐森玲;万海同;金湛;何昱;: "PHPLC法同时分离纯化红花中2种成分", 中成药, vol. 39, no. 02, 20 February 2017 (2017-02-20), pages 436 - 439 * |
| 李栋;马秉智;梁莹莹;张淑君;张相林;赫军;: "红花中羟基红花黄色素A的提取方法与含量测定研究进展", 中日友好医院学报, vol. 32, no. 01, 15 February 2018 (2018-02-15), pages 39 - 41 * |
| 杨辉;阿依吐伦・斯马义;吴桂荣;杨晓君;: "羟基红花黄色素A的提取及含量测定", 新疆医科大学学报, vol. 31, no. 10, 30 October 2008 (2008-10-30), pages 1358 - 1360 * |
| 肖艳华等: ""红花中羟基红花黄色素A的提取工艺"", 武汉工程大学学报, vol. 37, no. 3, 31 December 2015 (2015-12-31), pages 15 - 19 * |
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