CN114644649A - Method for extracting high-purity lysophosphatidylcholine from egg yolk powder - Google Patents
Method for extracting high-purity lysophosphatidylcholine from egg yolk powder Download PDFInfo
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- CN114644649A CN114644649A CN202011516027.8A CN202011516027A CN114644649A CN 114644649 A CN114644649 A CN 114644649A CN 202011516027 A CN202011516027 A CN 202011516027A CN 114644649 A CN114644649 A CN 114644649A
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- lysophosphatidylcholine
- yolk powder
- deoiled
- egg yolk
- alcohol
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- RYCNUMLMNKHWPZ-SNVBAGLBSA-N 1-acetyl-sn-glycero-3-phosphocholine Chemical compound CC(=O)OC[C@@H](O)COP([O-])(=O)OCC[N+](C)(C)C RYCNUMLMNKHWPZ-SNVBAGLBSA-N 0.000 title claims abstract description 101
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- 238000000034 method Methods 0.000 title claims abstract description 39
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 122
- 238000000605 extraction Methods 0.000 claims abstract description 36
- 239000003480 eluent Substances 0.000 claims abstract description 34
- 238000001035 drying Methods 0.000 claims abstract description 27
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 27
- 238000004440 column chromatography Methods 0.000 claims abstract description 25
- 239000000284 extract Substances 0.000 claims abstract description 23
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- 238000000926 separation method Methods 0.000 claims abstract description 17
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- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 66
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- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 claims description 16
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- JQWAHKMIYCERGA-UHFFFAOYSA-N (2-nonanoyloxy-3-octadeca-9,12-dienoyloxypropoxy)-[2-(trimethylazaniumyl)ethyl]phosphinate Chemical compound CCCCCCCCC(=O)OC(COP([O-])(=O)CC[N+](C)(C)C)COC(=O)CCCCCCCC=CCC=CCCCCC JQWAHKMIYCERGA-UHFFFAOYSA-N 0.000 claims description 15
- 238000011049 filling Methods 0.000 claims description 14
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- 150000001335 aliphatic alkanes Chemical class 0.000 claims description 7
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- 229940079593 drug Drugs 0.000 description 3
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- -1 phospholipid compound Chemical class 0.000 description 3
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- 206010018910 Haemolysis Diseases 0.000 description 2
- NTYJJOPFIAHURM-UHFFFAOYSA-N Histamine Chemical compound NCCC1=CN=CN1 NTYJJOPFIAHURM-UHFFFAOYSA-N 0.000 description 2
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- 238000003860 storage Methods 0.000 description 2
- ASWBNKHCZGQVJV-UHFFFAOYSA-N (3-hexadecanoyloxy-2-hydroxypropyl) 2-(trimethylazaniumyl)ethyl phosphate Chemical compound CCCCCCCCCCCCCCCC(=O)OCC(O)COP([O-])(=O)OCC[N+](C)(C)C ASWBNKHCZGQVJV-UHFFFAOYSA-N 0.000 description 1
- UCTWMZQNUQWSLP-VIFPVBQESA-N (R)-adrenaline Chemical compound CNC[C@H](O)C1=CC=C(O)C(O)=C1 UCTWMZQNUQWSLP-VIFPVBQESA-N 0.000 description 1
- 229930182837 (R)-adrenaline Natural products 0.000 description 1
- JLPULHDHAOZNQI-ZTIMHPMXSA-N 1-hexadecanoyl-2-(9Z,12Z-octadecadienoyl)-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/C\C=C/CCCCC JLPULHDHAOZNQI-ZTIMHPMXSA-N 0.000 description 1
- ZIIUUSVHCHPIQD-UHFFFAOYSA-N 2,4,6-trimethyl-N-[3-(trifluoromethyl)phenyl]benzenesulfonamide Chemical compound CC1=CC(C)=CC(C)=C1S(=O)(=O)NC1=CC=CC(C(F)(F)F)=C1 ZIIUUSVHCHPIQD-UHFFFAOYSA-N 0.000 description 1
- 101800004538 Bradykinin Proteins 0.000 description 1
- JZNWSCPGTDBMEW-UHFFFAOYSA-N Glycerophosphorylethanolamin Natural products NCCOP(O)(=O)OCC(O)CO JZNWSCPGTDBMEW-UHFFFAOYSA-N 0.000 description 1
- QXZGBUJJYSLZLT-UHFFFAOYSA-N H-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg-OH Natural products NC(N)=NCCCC(N)C(=O)N1CCCC1C(=O)N1C(C(=O)NCC(=O)NC(CC=2C=CC=CC=2)C(=O)NC(CO)C(=O)N2C(CCC2)C(=O)NC(CC=2C=CC=CC=2)C(=O)NC(CCCN=C(N)N)C(O)=O)CCC1 QXZGBUJJYSLZLT-UHFFFAOYSA-N 0.000 description 1
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- QXZGBUJJYSLZLT-FDISYFBBSA-N bradykinin Chemical compound NC(=N)NCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(=O)NCC(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CO)C(=O)N2[C@@H](CCC2)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)CCC1 QXZGBUJJYSLZLT-FDISYFBBSA-N 0.000 description 1
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- 238000001514 detection method Methods 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 239000008344 egg yolk phospholipid Substances 0.000 description 1
- 239000012156 elution solvent Substances 0.000 description 1
- 229960005139 epinephrine Drugs 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 231100000171 higher toxicity Toxicity 0.000 description 1
- 229960001340 histamine Drugs 0.000 description 1
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- 239000007924 injection Substances 0.000 description 1
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- 230000001575 pathological effect Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 150000008104 phosphatidylethanolamines Chemical class 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
- C07F9/02—Phosphorus compounds
- C07F9/06—Phosphorus compounds without P—C bonds
- C07F9/08—Esters of oxyacids of phosphorus
- C07F9/09—Esters of phosphoric acids
- C07F9/10—Phosphatides, e.g. lecithin
- C07F9/103—Extraction or purification by physical or chemical treatment of natural phosphatides; Preparation of compositions containing phosphatides of unknown structure
-
- 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
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
Abstract
The invention relates to the field of natural medicine extraction and separation, and discloses a method for extracting high-purity lysophosphatidylcholine from egg yolk powder, which comprises the following steps: A. extracting deoiled yolk powder with alcohol, filtering to remove residue, mixing filtrates, concentrating, and drying to obtain deoiled yolk powder alcohol extract; B. completely dissolving the deoiled yolk powder alcohol extract, separating by alumina column chromatography, collecting the filtrate obtained by separation, concentrating and drying to obtain the deoiled yolk powder extract; C. completely dissolving the deoiled yolk powder extract, adopting a chromatographic column filled with reverse packing to elute by adopting a short-chain alcohol water solution a as an eluent, combining the eluents, concentrating and drying to obtain a crude product of the lysophosphatidylcholine; D. and completely dissolving the crude lysophosphatidylcholine product, performing secondary column chromatography, eluting by adopting an alkane-short chain alcohol system, collecting eluent, concentrating and freeze-drying to obtain a lysophosphatidylcholine product.
Description
Technical Field
The invention relates to the field of natural medicine extraction and separation, and particularly relates to a method for extracting high-purity lysophosphatidylcholine from egg yolk powder.
Background
Lecithin is the most commonly used phospholipid compound, so lysophospholipid refers to lysolecithin, also called Lysophosphatidylcholine (LPC), which is a degradation product generated by hydrolysis or enzymolysis of 1-position or 2-position ester bond of Phosphatidylcholine (PC) under the action of phospholipase a, and part of lecithin is decomposed to generate lysophosphatidylcholine in the processes of raw material preparation, purification, storage and the like. LPC has a strong surface activity and thus can rupture red blood cells and other cell membranes, causing hemolysis or cell necrosis. After the LPC causes hemolysis, the permeability of capillary vessels is increased to cause bleeding, and tissues release histamine, 5-hydroxytryptamine, epinephrine, bradykinin and the like, so that the functions of the cardiovascular system and the nervous system are indirectly influenced, and a series of complex pathological reactions are stimulated. Therefore, the LPC content is an important quality control index for drugs, especially for injections containing phospholipid excipients, and is directly related to the clinical medication safety of drugs, and the content of the LPC content needs to be strictly controlled.
Lysophosphatidylcholine is mainly produced by phospholipid hydrolysis in the processes of raw and auxiliary materials, preparation process, product storage and the like, and because the content of the lysophosphatidylcholine in nature is very low, most of the lysophosphatidylcholine for quality control at present is directly hydrolyzed and enzymolyzed by phospholipid or synthesized by glycerol and fatty acid, so that the natural property of the lysophosphatidylcholine is changed. Because lysophosphatidylcholine is a degradation product obtained by hydrolyzing ester bonds at 1-position or 2-position of phospholipid and has certain randomness, namely lysophospholipid is a mixture of 1-position and 2-position, the natural property of lysophosphatidylcholine cannot be completely ensured by artificial hydrolysis, enzymolysis or synthesis means, and great hidden danger exists in the quality control process. Most of lysophosphatidylcholine standard products purchased from middle-school hospitals or abroad are artificially hydrolyzed, enzymolyzed or synthesized, and the reference product of natural lysophosphatidylcholine is very expensive due to the low content of the natural lysophosphatidylcholine, so that the detection cost of enterprise products is greatly increased.
Therefore, the natural source lysophosphatidylcholine with low cost and stable quality is urgently needed by various preparation enterprises in China at present, and the preparation process of the natural source lysophosphatidylcholine with low cost and stable quality is not reported at present. Through search, the Guangzhou Baiyunshan prescription modern pharmaceutical industry Co., Ltd applies for 2014 a preparation method of natural source lysophosphatidylcholine, which takes egg yolk or soybean phospholipid as a raw material and halogenated alkane-low carbon alcohol-water as an eluent to carry out elution twice, and finally, the lysophosphatidylcholine with the content of 98-99.9% is obtained. The technical scheme can obtain the lysophosphatidylcholine with natural sources and reliable quality, but a large amount of eluent, including halogenated alkane with high toxicity, is used in the preparation process. Meanwhile, the technical scheme adopts soybean or yolk phospholipid as a starting material, the soybean or yolk phospholipid has higher cost, and the content of lysophosphatidylcholine in the soybean or yolk phospholipid is lower, so that the extraction cost of LPC is higher.
Disclosure of Invention
The invention aims to solve the technical problems of high extraction cost, low elution efficiency, high toxicity of extraction solvent and the like of high-purity lysophosphatidylcholine in the prior art, and provides the natural lysophosphatidylcholine with lower cost and more stable quality, wherein the lysophosphatidylcholine with the purity of more than 98% is finally obtained by taking yolk powder as a starting material through deoiling, extraction, column chromatography separation and the like, meanwhile, the yolk lecithin with the PC content of more than 70% can be obtained, and the production of two varieties of LPCs can be realized by one process, so that the production cost of LPCs is greatly reduced.
In order to solve the technical problems, the invention adopts the following technical scheme:
a method for extracting high-purity lysophosphatidylcholine from egg yolk powder comprises the following steps:
A. extracting deoiled yolk powder with alcohol, filtering to remove residue, mixing filtrates, concentrating, and drying to obtain deoiled yolk powder alcohol extract;
B. completely dissolving the deoiled yolk powder alcohol extract, separating by alumina column chromatography, collecting the filtrate obtained by separation, concentrating and drying to obtain the deoiled yolk powder extract;
C. completely dissolving the deoiled yolk powder extract, adopting a short-chain alcohol water solution a as an eluent, eluting through a chromatographic column filled with reverse packing, combining the eluents, concentrating and drying to obtain a crude product of lysophosphatidylcholine;
D. and completely dissolving the crude lysophosphatidylcholine product, performing secondary column chromatography, eluting by adopting an alkane-short chain alcohol system, collecting eluent, concentrating and freeze-drying to obtain a lysophosphatidylcholine product.
Preferably, in step D, the alkane-short chain alcohol system comprises: alkane and short-chain alcohol are mixed according to the volume ratio of 1: 1, the alkane comprises one or more of chloroform and dichloromethane, and the short-chain alcohol comprises one or more of methanol, ethanol, isopropanol and the like.
With alkanes and short-chain alcohols 1: the system 1 can reduce the dosage of elution solvent as much as possible and accelerate the elution efficiency when elution is carried out.
Preferably, in the step D, the crude lysophosphatidylcholine is completely dissolved by chloroform, the filler of the secondary column chromatography comprises one or more of silica gel and alumina, and the mass ratio of the crude lysophosphatidylcholine to the filler of the secondary column chromatography is 1: 1-5 filling columns, wherein the diameter-height ratio is 1: 1-1: 10, the column passing pressure is 0.3-1.0 mPa, and the column passing temperature is 20-40 ℃.
Preferably, in the step C, the percentage content of water in the short-chain alcohol aqueous solution a is 5% to 30%; the short-chain alcohol comprises one or more of methanol, ethanol, n-butanol and isopropanol.
Preferably, in step C, the deoiled yolk powder extract is completely dissolved by using short-chain alcohol, wherein the short-chain alcohol comprises one or more of methanol, ethanol, n-butanol and isopropanol; the reverse filler comprises one or more of C18 bonded silica gel, C8 bonded silica gel and C4 bonded silica gel; the deoiled yolk powder extract and the reverse filler are mixed according to the mass ratio of 1: 1-10 column packing, wherein the diameter-height ratio is 1: 1-1: 10.
preferably, in the step C, after the crude lysophosphatidylcholine product is obtained, the aqueous solution b of the short-chain alcohol is used as an eluent to perform secondary elution on the chromatographic column filled with the reverse filler, collect the eluent, concentrate the eluent, decolor the eluent with activated carbon, filter, concentrate, refine and freeze-dry the eluent to obtain an egg yolk lecithin product; the content of yolk lecithin in the yolk lecithin product is more than 70 percent; in the short-chain alcohol aqueous solution b, the percentage content of water is 0-10%; the short-chain alcohol comprises one or more of methanol, ethanol, n-butanol and isopropanol.
The main purpose of the step is to elute the residual PC in the reverse column, the short-chain alcohol with lower water content can obtain higher elution efficiency and less introduced impurities, the best water content proportion is 0-5%, but LPC is completely eluted at the moment, and the product with the PC content of more than 70% can also be obtained with the water content of 0-10%.
From the viewpoint of solvent recovery, the short chain alcohol used for dissolving the deoiled yolk powder extract in the step C should be kept consistent with the short chain alcohol in the aqueous solution a of the short chain alcohol and the aqueous solution b of the short chain alcohol.
Preferably, in the step B, the deoiled yolk powder alcohol extract is completely dissolved by 95% ethanol, and the mass ratio of the deoiled yolk powder alcohol extract to alumina is 1: 1-5 fillers with a diameter-height ratio of 1: 1-1: eluting with 95% ethanol at-0.1 to 3.0mPa and 10 to 60 ℃.
The main purpose of the step is to separate most PE (phosphatidylethanolamine) in the deoiled yolk powder alcohol extract, so the step adopts open column operation at normal temperature, and 95% ethanol is used as a solvent and an eluent, thereby not only reducing the volatilization risk of the solvent and the recovery difficulty of the solvent, but also realizing the most convenient PE separation. And detecting by using a thin-layer chromatography spot plate, and determining that the deoiled yolk powder is completely eluted.
Preferably, in step a, the alcohol used for alcohol extraction includes one or more of methanol, ethanol and aqueous solution thereof; the mass part ratio of the deoiled yolk powder to the alcohol is 1: 5-15 times of extraction, 2-6 times of extraction, 1-8 h of extraction time and 15-50 ℃ of extraction temperature.
Preferably, in step a, the de-oiled egg yolk powder is obtained by de-oiling egg yolk powder, wherein the de-oiling comprises one of acetone de-oiling method and supercritical method; when the acetone deoiling method is applied, the mass ratio of the egg yolk powder to the acetone is 1: 1-10 times of deoiling, wherein the deoiling temperature is 20-45 ℃, and after deoiling, combining acetone insoluble substances, and drying to obtain the deoiled yolk powder; when the supercritical method is applied, the egg yolk powder is placed in an extraction kettle for continuous extraction, the temperature is kept at 25-45 ℃, the CO2 flow rate is controlled at 50-120 kg/h, the pressure of the extraction kettle is 10-25 MPa, the pressure of a separation kettle 1 is 5-20 MPa, the pressure of a separation kettle 2 is 5-15 MPa, and the continuous extraction lasts for 4-10 h, so that the de-oiled egg yolk powder is obtained.
A lysophosphatidylcholine product obtained by the method for extracting high-purity lysophosphatidylcholine from egg yolk powder, wherein the purity of the lysophosphatidylcholine product is higher than 98%.
Compared with the prior art, the implementation of the invention has the following beneficial effects:
firstly, the technical scheme of the invention can obtain the lysophosphatidylcholine with the purity of more than 98 percent, and the attribute of the natural source of the lysophosphatidylcholine is ensured.
Compared with the prior art, the technical scheme of the invention has obviously higher elution efficiency, and does not adopt organic solvents with higher toxicity, such as halogenated alkane and the like.
The technical scheme of the invention realizes the full utilization of the yolk powder, greatly reduces the acquisition cost of natural lysophosphatidylcholine, and can obtain the medicinal yolk lecithin with the PC content of more than 70 percent and meeting pharmacopoeia standards while obtaining the lysophosphatidylcholine with the purity of more than 98 percent.
Drawings
FIG. 1 is a schematic diagram of the extraction process of lysophosphatidylcholine according to the technical embodiment of the present invention;
FIG. 2 is a diagram showing the water content examination of the LPC elution process.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention clearer, the present invention will be further described in detail with reference to the accompanying drawings so that those skilled in the art can better understand the present invention and can implement the present invention, but the present invention is not limited by the illustrated examples. Modifications or substitutions to methods, procedures, or conditions of the invention may be made without departing from the spirit and scope of the invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art.
Example 1
As shown in FIG. 1, 1kg of egg yolk powder was deoiled by adding 5L of acetone, deoiled at 25 deg.C for 3 times, filtered, and dried to obtain 726g of deoiled egg yolk powder. And then 3630ml of 95% ethanol is used for carrying out ethanol extraction, the extraction temperature is 20 ℃, the extraction time is 2 hours, filter residues are filtered, repeated extraction is carried out for 4 times, filtrates are combined, and the vacuum concentration and drying are carried out, thus obtaining 471g of the deoiled yolk powder ethanol extract. Performing alumina column chromatography with 95% ethanol as eluent until yolk lecithin is completely eluted, concentrating, drying, adding methanol to dissolve to obtain raw material solution, filling 2.35kgC18 bonded silica gel for column chromatography (diameter-height ratio is 1: 6), column temperature is 25 deg.C, eluting with methanol: eluting with water (85: 15) under column pressure of 2.7mPa for 8 column volumes, mixing eluates, concentrating and drying to obtain crude lysophosphatidylcholine product 10.16g with purity of 96.3%, eluting with pure methanol as eluent for the second time under column pressure of 1.4mPa for 10 column volumes, and mixing eluates containing egg yolk lecithin to obtain crude egg yolk lecithin product. Dissolving crude lysophosphatidylcholine in chloroform to prepare a raw material solution, filling 12g of silica gel (the diameter-height ratio is 1: 2), keeping the column temperature at 25 ℃, and dissolving the crude lysophosphatidylcholine in chloroform to prepare a raw material solution, wherein the volume ratio of the silica gel to the chloroform: methanol (1: 1) is used as eluent, the column pressure in the elution process is 0.3mPa, 10 column volumes are eluted, the eluates are combined, concentrated and dried to obtain lysophosphatidylcholine 9.65g with the purity of 99.8%.
Example 2
As shown in figure 1, 2kg of egg yolk powder was taken, 2L of acetone was added for deoiling, deoiled at 35 deg.C for 5 times, filtered, and dried to obtain 1.48kg of deoiled egg yolk powder. And then carrying out alcohol extraction by using 14.8L 95% ethanol at the extraction temperature of 30 ℃ for 2h, filtering filter residues, repeatedly extracting for 2 times, combining filtrates, and concentrating under reduced pressure to obtain 973g of the deoiled yolk powder alcohol extract. Performing alumina column chromatography with 95% ethanol as eluent until yolk lecithin is completely eluted, concentrating, drying, adding ethanol, dissolving to obtain raw material solution, filling 7kgC18 bonded silica gel, performing column chromatography (diameter-height ratio of 1: 8), at 35 deg.C, eluting with ethanol: eluting with water (75: 25) under column pressure of 2.4mPa for 8 column volumes, mixing eluates, concentrating, drying to obtain crude lysophosphatidylcholine product 23.68g with purity of 91.6%, eluting with ethanol: eluting with water (98: 2) under column pressure of 1.2mPa for 10 column volumes, and mixing the eluates containing egg yolk lecithin to obtain crude egg yolk lecithin. Dissolving crude lysophosphatidylcholine in chloroform to prepare a raw material solution, filling 46g of silica gel (the diameter-height ratio is 1: 3), carrying out column temperature at 25 ℃, and mixing the raw material solution with chloroform: methanol (1: 1) is used as eluent, the column pressure in the elution process is 0.4mPa, 10 column volumes are eluted, the eluates are combined, concentrated and dried to obtain 21.08g of lysophosphatidylcholine with the purity of 99.4%.
Example 3
As shown in FIG. 1, egg yolk powder (500 g) was deoiled by adding 5L of acetone, deoiled at 40 deg.C for 1 time, filtered, and dried to obtain deoiled egg yolk powder (355 g). Extracting with 5.32L 95% ethanol at 35 deg.C for 2 hr, filtering to remove residue, mixing filtrates, and concentrating under reduced pressure to obtain 231.3g deoiled yolk powder ethanol extract. Performing alumina column chromatography with 95% ethanol as eluent until yolk lecithin is completely eluted, concentrating, drying, adding isopropanol, dissolving to obtain raw material solution, filling 1.2kgC8 bonded silica gel for column chromatography (diameter-height ratio of 1: 6), column temperature of 35 deg.C, eluting with isopropanol: eluting with water (70: 30) under column pressure of 2.2mPa for 8 column volumes, mixing eluates, concentrating, and drying to obtain crude lysophosphatidylcholine product 6.93g with purity of 85.2%, eluting with isopropanol: eluting with water (80: 20) under column pressure of 1.1mPa for 10 column volumes, and mixing the eluates containing egg yolk lecithin to obtain crude egg yolk lecithin. Dissolving crude lysophosphatidylcholine in chloroform to prepare a raw material solution, filling 25g of silica gel (the diameter-height ratio is 1: 4), carrying out column temperature at 40 ℃, and mixing the raw material solution with chloroform: isopropanol (1: 1) is used as eluent, the column pressure in the elution process is 1.0mPa, 10 column volumes are eluted, the eluates are combined, concentrated and dried to obtain 5.64g of lysophosphatidylcholine with the purity of 99.1%.
Example 4
As shown in figure 1, yolk powder 5kg is added into supercritical extraction kettle at 25 deg.C and CO for continuous extraction2The flow rate is 60kg/h, the pressure of the extraction kettle is 15MPa, the pressure of the separation kettle 1 is 10MPa, the pressure of the separation kettle 2 is 8MPa, and the process lasts for 6 hours to obtain 3.56kg of the deoiled yolk powder. Extracting with 21.3L 95% ethanol at 25 deg.C for 8 hr, filtering, extracting the residue for 3 times, concentrating, and drying to obtain 2.02kg of deoiled yolk powder ethanol extract. Performing alumina column chromatography with 95% ethanol as eluent until yolk lecithin is completely eluted, concentrating, drying, adding ethanol, dissolving to obtain stock solution, and filling 12kg of ethanolColumn chromatography (1: 5 diameter to height ratio) was performed on silica gel C4 at 30 ℃ with the first eluent methanol: eluting with water (90: 10) under column pressure of 2.9mPa for 8 column volumes, mixing eluates, concentrating, and drying to obtain crude lysophosphatidylcholine 56.96g with purity of 88.9%, eluting with ethanol: eluting with water (90: 10) under column pressure of 1.1mPa for 10 column volumes, and mixing the eluates containing egg yolk lecithin to obtain crude egg yolk lecithin. Dissolving crude lysophosphatidylcholine in chloroform to prepare a raw material solution, filling 170g of silica gel (the diameter-height ratio is 1: 4), carrying out column temperature at 20 ℃, and mixing the raw material solution with dichloromethane: ethanol (1: 1) is used as eluent, the column pressure in the elution process is 0.5mPa, 10 column volumes are eluted, the eluates are combined, concentrated and dried to obtain 50.34g of lysophosphatidylcholine with the purity of 98.9%.
Example 5
As shown in figure 1, egg yolk powder 10kg is added into supercritical extraction kettle at 35 deg.C and CO for continuous extraction2The flow rate is 100kg/h, the pressure of the extraction kettle is 20MPa, the pressure of the separation kettle 1 is 15MPa, the pressure of the separation kettle 2 is 12MPa, and the process lasts for 4 hours to obtain 6.98kg of the deoiled yolk powder. Extracting with 34.9L 95% ethanol at 35 deg.C for 4 hr, filtering, extracting the residue for 2 times, concentrating, and drying to obtain 4.48kg deoiled yolk powder ethanol extract. Performing alumina column chromatography with 95% ethanol as eluent until yolk lecithin is completely eluted, concentrating, drying, adding methanol to dissolve to obtain raw material solution, filling 31kgC18 bonded silica gel for column chromatography (diameter-height ratio is 1: 6), column temperature is 30 deg.C, eluting with methanol: eluting with water (95: 5) under column pressure of 3mPa for 8 column volumes, mixing eluates, concentrating, and drying to obtain crude lysophosphatidylcholine 144.39g with purity of 72.6%, eluting with isopropanol: eluting with water (90: 10) under column pressure of 1.1mPa for 10 column volumes, and mixing the eluates containing egg yolk lecithin to obtain crude egg yolk lecithin. Dissolving crude lysophosphatidylcholine in chloroform to prepare a raw material solution, filling 650g of neutral alumina for column chromatography (the diameter-height ratio is 1: 8), carrying out column chromatography at the temperature of 30 ℃, and adding chloroform: eluting with methanol (1: 1) as eluentEluting 10 column volumes under 1mPa, mixing eluates, concentrating and drying to obtain 106.08g lysophosphatidylcholine with purity of 98.7%.
Example 6
As shown in figure 1, 20kg yolk powder is added into supercritical extraction kettle at 40 deg.C and CO for continuous extraction2The flow rate is 110kg/h, the pressure of the extraction kettle is 23MPa, the pressure of the separation kettle 1 is 17MPa, the pressure of the separation kettle 2 is 12MPa, and the operation lasts for 3 hours to obtain 13.4kg of the deoiled yolk powder. And then 160L of 95 percent ethanol is used for carrying out ethanol extraction at the extraction temperature of 45 ℃ for 4h, filter residues are filtered, concentrated and dried, and 9.37kg of deoiled yolk powder ethanol extract is obtained. Performing alumina column chromatography with 95% ethanol as eluent until yolk lecithin is completely eluted, concentrating, drying, adding methanol to dissolve to obtain raw material solution, filling 90kgC8 bonded silica gel to perform column chromatography (diameter-height ratio is 1: 5), at 35 deg.C, eluting with ethanol: eluting with water (75: 25) under column pressure of 2.7mPa for 8 column volumes, mixing eluates, concentrating, and drying to obtain crude product 335g of lysophosphatidylcholine with purity of 70.4%, eluting with methanol: eluting with water (90: 10) under column pressure of 1.1mPa for 10 column volumes, and mixing the eluates containing egg yolk lecithin to obtain crude egg yolk lecithin. Dissolving the crude lysophosphatidylcholine product in chloroform to prepare a raw material solution, filling 1.6kg of neutral alumina for column chromatography (the diameter-height ratio is 1: 6), keeping the column temperature at 40 ℃, and dissolving the crude lysophosphatidylcholine product in chloroform: methanol (1: 1) is used as eluent, the column pressure in the elution process is 0.8mPa, 10 column volumes are eluted, the eluates are combined, concentrated and dried to obtain 235.44g of lysophosphatidylcholine with the purity of 98.3%.
Comparative example 1
To fully evaluate the effectiveness of the technique of the present invention, the key step of lysophosphatidylcholine elution was examined. In the process of adopting yolk powder as a starting material to extract lysophosphatidylcholine, a reversed-phase chromatographic column is adopted to extract crude LPC, because the polarities of LPC and PC are not very different, if pure short-chain alcohol is adopted as an eluent, components such as PC, LPC, SM and the like can be eluted simultaneously, the LPC is eluted firstly along with the increase of the polarity of the eluent, the LPC can be obtained in one step by utilizing the characteristic, but the LPC, PC and SM which can be eluted are less and less along with the increase of the polarity, so that the LPC, PC and SM which are not eluted are remained in yolk lecithin, and the content of LPC in the yolk lecithin is finally over standard. Therefore, the comparison example fully considers the proportion of water in the reverse-direction column elution process, and the investigation result shows that when the water content is between 5% and 30%, high-purity LPC can be obtained and the yield is high, and when the water content exceeds 30%, the elution proportion of LPC, PC and SM is low, and the quality of the subsequent egg yolk lecithin is influenced. As can be seen from figure 1, the purity of more than 98% can be realized, and the content of crude lysophosphatidylcholine is at least more than 80%, so that the water content of 5-30% is the key point for meeting the effect of the technical scheme in consideration of the quality of the product. The results are shown in FIG. 2.
The above disclosure is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the scope of the present invention, therefore, the present invention is not limited by the appended claims.
Claims (10)
1. A method for extracting high-purity lysophosphatidylcholine from egg yolk powder is characterized by comprising the following steps:
A. extracting deoiled yolk powder with alcohol, filtering to remove residue, mixing filtrates, concentrating, and drying to obtain deoiled yolk powder alcohol extract;
B. completely dissolving the deoiled yolk powder alcohol extract, separating by alumina column chromatography, collecting the filtrate obtained by separation, concentrating and drying to obtain the deoiled yolk powder extract;
C. completely dissolving the deoiled yolk powder extract, adopting a short-chain alcohol water solution a as an eluent, eluting through a chromatographic column filled with reverse packing, combining the eluents, concentrating and drying to obtain a crude product of lysophosphatidylcholine;
D. and completely dissolving the crude lysophosphatidylcholine product, performing secondary column chromatography, eluting by adopting an alkane-short chain alcohol system, collecting eluent, concentrating and freeze-drying to obtain a lysophosphatidylcholine product.
2. The method for extracting lysophosphatidylcholine with high purity from egg yolk powder according to claim 1, wherein in step D, said alkane-short chain alcohol system comprises: alkane and short-chain alcohol are mixed according to the volume ratio of 1: 1, the alkane comprises one or more of chloroform and dichloromethane, and the short-chain alcohol comprises one or more of methanol, ethanol, isopropanol and the like.
3. The method for extracting high-purity lysophosphatidylcholine from egg yolk powder according to claim 1, wherein in step D, the crude lysophosphatidylcholine is completely dissolved by chloroform, the packing material of the secondary column chromatography comprises one or more of silica gel and alumina, and the mass ratio of the crude lysophosphatidylcholine to the packing material of the secondary column chromatography is 1: 1-5 filling columns, wherein the diameter-height ratio is 1: 1-1: 10, the column passing pressure is 0.3-1.0 mPa, and the column passing temperature is 20-40 ℃.
4. The method for extracting lysophosphatidylcholine with high purity from egg yolk powder according to claim 1, wherein in the step C, the water content in the aqueous solution a of short chain alcohol is 5% to 30%; the short-chain alcohol comprises one or more of methanol, ethanol, n-butanol and isopropanol.
5. The method for extracting highly pure lysophosphatidylcholine from egg yolk powder according to claim 1, wherein in step C, said deoiled egg yolk powder extract is completely dissolved with short chain alcohols, said short chain alcohols comprising one or more of methanol, ethanol, n-butanol, isopropanol; the reverse filler comprises one or more of C18 bonded silica gel, C8 bonded silica gel and C4 bonded silica gel; the mass ratio of the deoiled yolk powder extract to the reverse filler is 1: 1-10 columns are filled, and the diameter-height ratio is 1: 1-1: 10.
6. the method for extracting high-purity lysophosphatidylcholine from egg yolk powder according to claim 1, wherein in the step C, after the crude lysophosphatidylcholine is obtained, the chromatographic column filled with the reverse packing is subjected to secondary elution by using the short-chain alcohol aqueous solution b as an eluent, and the eluent is collected, concentrated, decolored by activated carbon, filtered, concentrated, refined and freeze-dried to obtain an egg yolk lecithin product; the content of yolk lecithin in the yolk lecithin product is more than 70 percent; in the short-chain alcohol aqueous solution b, the percentage content of water is 0-10%; the short-chain alcohol comprises one or more of methanol, ethanol, n-butanol and isopropanol.
7. The method for extracting highly pure lysophosphatidylcholine from egg yolk powder according to claim 1, wherein in step B, said deoiled egg yolk powder alcohol extract is completely dissolved with 95% ethanol, and the mass ratio of said deoiled egg yolk powder alcohol extract to alumina is 1: 1-5 fillers with a diameter-height ratio of 1: 1-1: 10, elution was performed with 95% ethanol.
8. The method for extracting highly pure lysophosphatidylcholine from egg yolk powder according to claim 1, wherein in step a, the alcohol used for the alcohol extraction comprises one or more of methanol, ethanol and aqueous solution thereof; the mass part ratio of the deoiled yolk powder to the alcohol is 1: 5-15 times of extraction, 2-6 times of extraction, 1-8 h of extraction time and 15-50 ℃ of extraction temperature.
9. The method for extracting highly pure lysophosphatidylcholine from egg yolk powder according to claim 1, wherein in step a, the de-oiled egg yolk powder is obtained by de-oiling egg yolk powder, wherein said de-oiling comprises one of acetone de-oiling and supercritical de-oiling; when the acetone deoiling method is applied, the mass ratio of the egg yolk powder to the acetone is 1: 1-10 times of deoiling, wherein the deoiling temperature is 20-45 ℃, and after deoiling, combining acetone insoluble substances, and drying to obtain the deoiled yolk powder; when the supercritical method is applied, the yolk powder is placed in an extraction kettle for continuous extraction, the temperature is kept at 25-45 ℃, and CO is kept2The flow rate is controlled to be 50-120 kg/h, and the pressure of the extraction kettle10-25 Mpa, 5-20 Mpa of the pressure of the separation kettle 1, 5-15 Mpa of the pressure of the separation kettle 2, and lasting for 4-10 hours to obtain the de-oiled yolk powder.
10. A lysophosphatidylcholine product obtained by the method for extracting high purity lysophosphatidylcholine from egg yolk powder according to claim 1, wherein said lysophosphatidylcholine product has a purity of greater than 98%.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4849137A (en) * | 1987-04-09 | 1989-07-18 | Kewpie Kabushiki Kaisha | Process for producing lysophospholipids containing substantially no lysophospholipids except LPC |
| JPH02113891A (en) * | 1988-10-22 | 1990-04-26 | Toyama Chem Co Ltd | Novel process for production of lysophosphatidyl-choline |
| JPH06228171A (en) * | 1993-02-05 | 1994-08-16 | Taiyo Kagaku Co Ltd | Method for purifying sphingomyelin |
| CN104558021A (en) * | 2014-12-30 | 2015-04-29 | 广州白云山汉方现代药业有限公司 | Preparation method of natural LPC (lysophosphatidylcholine) |
| CN109912645A (en) * | 2019-03-16 | 2019-06-21 | 广州隽沐生物科技股份有限公司 | The preparation method of egg yolk lecithin |
| CN110144369A (en) * | 2019-05-22 | 2019-08-20 | 杨利平 | A method of preparing high-purity lysophosphatidyl choline |
-
2020
- 2020-12-21 CN CN202011516027.8A patent/CN114644649B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4849137A (en) * | 1987-04-09 | 1989-07-18 | Kewpie Kabushiki Kaisha | Process for producing lysophospholipids containing substantially no lysophospholipids except LPC |
| JPH02113891A (en) * | 1988-10-22 | 1990-04-26 | Toyama Chem Co Ltd | Novel process for production of lysophosphatidyl-choline |
| JPH06228171A (en) * | 1993-02-05 | 1994-08-16 | Taiyo Kagaku Co Ltd | Method for purifying sphingomyelin |
| CN104558021A (en) * | 2014-12-30 | 2015-04-29 | 广州白云山汉方现代药业有限公司 | Preparation method of natural LPC (lysophosphatidylcholine) |
| CN109912645A (en) * | 2019-03-16 | 2019-06-21 | 广州隽沐生物科技股份有限公司 | The preparation method of egg yolk lecithin |
| CN110144369A (en) * | 2019-05-22 | 2019-08-20 | 杨利平 | A method of preparing high-purity lysophosphatidyl choline |
Non-Patent Citations (1)
| Title |
|---|
| 李招等: "柱色谱纯化溶血磷脂酰胆碱的方法研究" * |
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