CN114478622A

CN114478622A - Preparation method of phosphatidylcholine

Info

Publication number: CN114478622A
Application number: CN202210263401.0A
Authority: CN
Inventors: 王维; 于凯; 邵英禄; 闫奇; 惠翠; 王伟; 杨浩然; 任伟民
Original assignee: Shenyang Gold Jyouki Technology Co ltd
Current assignee: Shenyang Gold Jyouki Technology Co ltd
Priority date: 2022-03-17
Filing date: 2022-03-17
Publication date: 2022-05-13

Abstract

The invention belongs to the technical field of organic synthesis, and particularly relates to a preparation method of phosphatidylcholine. The preparation method of the phosphatidylcholine provided by the invention comprises the following steps: dissolving (R) -glycidol and a first acylation reagent in a first organic solvent, and carrying out a first acylation reaction to obtain (S) -glycidol fatty acid ester; and (S) -glycidyl fatty acid ester, phosphorylcholine and a second organic solvent are mixed for ring-opening reaction to obtain the lysophosphatidylcholine. And dissolving lysophosphatidylcholine and a second acylation reagent in a third organic solvent, and carrying out a second acylation reaction to obtain phosphatidylcholine. The method has the advantages of simple process, few reaction steps, mild reaction conditions and more convenient operation; in addition, the post-treatment method is simple and convenient, the discharge of experimental waste is reduced, and the purity of the prepared phosphatidylcholine is more than 98%, so that the method is suitable for industrial preparation.

Description

Preparation method of phosphatidylcholine

Technical Field

The invention belongs to the technical field of organic synthesis, and particularly relates to a preparation method of phosphatidylcholine.

Background

Phosphatidylcholine is widely present in animal and plant bodies, is a basic substance for life activities, has important physiological functions and unique emulsifying property, and has important application in the industries of food, health care products, medicines and the like. The medicine prepared from phosphatidylcholine can be used for preparing high-energy parenteral fluid-fat emulsion, carriers of various anticancer drugs and liposome. For example, distearoylphosphatidylcholine is one of the important excipients for the preparation of liposomes, erucylphosphatidylcholine is used in the slow-release polycarbivacaine liposomes known under the name of Exparel, dioleoylphosphatidylcholine is used in the product known under the name of Depocyt (cytarabine) for the treatment of lymphoma meningitis. Extraction from animal and plant carriers rich in phospholipids is a traditional way of preparing phosphatidylcholine, but the substances obtained in the traditional way are mixtures and do not fully meet the market demand.

In order to obtain high-purity phosphatidylcholine with a single and definite structure-activity relationship, researchers synthesize and prepare phosphatidylcholine by a chemical method. For example, in the literature (Synthesis of a Small Library of Mixed-Acid Phospholipids from D-Mannitols as a Homochiral structural Material chem.pharm.Bull.1999,47(11) 1659) -1663), a benzyl-protected chiral diol is used as a starting Material, two identical or different acyl groups are attached according to the purpose, different acyl groups can be introduced by using different activities of primary and secondary hydroxyl groups or by using selective protecting groups, and finally the protecting groups are removed and coupled with phospholipid head groups; the introduction of the acyl group may utilize the corresponding fatty acid, fatty acid anhydride or more reactive acid chloride. For example, in the Master thesis of Chenguang at northwest university, "study on total synthesis of phosphatidylcholine, phosphatidylethanolamine and phosphatidylglycerol", glycerol is used as a precursor compound to design a total synthesis route of glycerophospholipid containing the same carbon chain; ketal, 3, 4-dimethoxybenzyl are respectively used as dihydroxy and monohydroxy protecting groups of a glycerol skeleton, and acetic acid aqueous solution and 2, 3-dichloro-5, 6-dicyanobenzoquinone (DDQ) are used as corresponding deprotection reagents; the distearoyl phosphatidylcholine is prepared by the steps of reduction, chlorination, condensation, protection and deprotection of dihydroxyl and monohydroxy, acylation, phosphorylation, head group protection, coupling, deprotection and the like.

However, the existing route for preparing phosphatidylcholine has long steps, complex reaction and high requirements on equipment conditions, and is not suitable for scale-up preparation.

Disclosure of Invention

In view of the above, the invention provides a preparation method of phosphatidylcholine, and the preparation method provided by the invention is simple in process, convenient to operate and easy for industrial production.

In order to solve the technical problem, the invention provides a preparation method of phosphatidylcholine, which comprises the following steps:

dissolving (R) -glycidol and a first acylation reagent in a first organic solvent, and carrying out a first acylation reaction to obtain (S) -glycidol fatty acid ester;

mixing the (S) -glycidyl fatty acid ester, phosphorylcholine and a second organic solvent, and carrying out a ring-opening reaction to obtain lysophosphatidylcholine;

and dissolving lysophosphatidylcholine and a second acylation reagent in a third organic solvent, and carrying out a second acylation reaction to obtain phosphatidylcholine.

Preferably, the first acylating agent and the second acylating agent independently comprise a carboxylic acid, an acid chloride, or an acid anhydride.

Preferably, the number of carbon atoms in the carboxylic acid, the acyl chloride and the anhydride is 8-27 independently.

Preferably, the carboxylic acid comprises n-octanoic acid, lauric acid, palmitic acid, stearic acid, arachidic acid, oleic acid or arachidonic acid;

the acyl chloride comprises fatty acyl chloride, myristoyl chloride, palmitoyl chloride or stearoyl chloride;

the acid anhydride includes fatty acid anhydride, lauric acid anhydride, stearic acid anhydride or oleic acid anhydride.

Preferably, the molar ratio of the (R) -glycidol to the first acylating agent is 1:1 to 1.8;

the temperature of the first acylation reaction is-15-50 ℃; the time of the first acylation reaction is 2-8 h.

Preferably, the molar ratio of the lysophosphatidylcholine to the second acylating agent is 1: 1-1.8;

the temperature of the second acylation reaction is 50-150 ℃; the time of the second acylation reaction is 8-24 h.

Preferably, the molar ratio of the phosphorylcholine to the (S) -glycidyl fatty acid ester is 1: 1-1.5.

Preferably, the ring-opening reaction is carried out under the condition of a catalyst;

the mol ratio of the phosphorylcholine to the catalyst is 1: 0.1-1.

Preferably, the catalyst comprises one or more of transition metal oxide, sulfuric acid, nitric acid, hydrochloric acid, cationic resin and solid super acid.

Preferably, the temperature of the ring-opening reaction is 60-150 ℃; the ring-opening reaction time is 12-48 h.

The invention provides a preparation method of phosphatidylcholine, which comprises the following steps: dissolving (R) -glycidol and a first acylation reagent in a first organic solvent, and carrying out a first acylation reaction to obtain (S) -glycidol fatty acid ester; and (S) -glycidyl fatty acid ester, phosphorylcholine and a second organic solvent are mixed for ring-opening reaction to obtain the lysophosphatidylcholine. And dissolving lysophosphatidylcholine and a second acylation reagent in a third organic solvent, and carrying out a second acylation reaction to obtain phosphatidylcholine. The preparation method provided by the invention has the advantages of simple process, few reaction steps and convenience in operation, and is suitable for industrial production. The preparation method provided by the invention has the following advantages:

(1) the preparation method provided by the invention can prepare the phosphatidylcholine with a single and definite structure, and can prepare the high-purity phosphatidylcholine containing symmetrical or asymmetrical, saturated or unsaturated, substituted or unsubstituted double fatty acid chains according to the structure-activity relationship.

(2) The invention takes common (R) -glycidol, carboxylic acid, acyl chloride and acid anhydride as starting raw materials, the conventional chemical phosphorylcholine is used in the intermediate step, the solvents are common industrial reagents, no further treatment is needed, and the invention has the advantage of easily obtained raw materials.

(3) The method has the advantages of simple process, few reaction steps, mild reaction conditions and more convenient operation; in addition, the post-treatment method is simple and convenient, the discharge of experimental waste is reduced, and the purity of the prepared phosphatidylcholine is more than 98%, so that the method is suitable for industrial preparation.

Drawings

FIG. 1 shows lauroyl lysophosphatidylated substance prepared in example 2¹H NMR spectrum;

FIG. 2 is a liquid chromatogram of lauroyl lysophosphatidylcholine prepared in example 2;

FIG. 3 shows the preparation of dilauroylphosphatidylcholine in example 2¹H NMR spectrum;

FIG. 4 is a liquid chromatogram of dilauroyl phosphatidylcholine prepared in example 2;

FIG. 5 shows the preparation of myristoyl lysophosphatidylcholine in example 3¹H NMR spectrum;

FIG. 6 is a liquid chromatogram of myristoyl lysophosphatidylcholine prepared in example 3;

FIG. 7 shows dimyristoyl phosphatidylcholine prepared in example 3¹H NMR spectrum;

FIG. 8 is a liquid chromatogram of dimyristoyl phosphatidylcholine prepared in example 3;

FIG. 9 shows the preparation of palmitoyl lysophosphatidylcholine in example 4¹H NMR spectrum;

FIG. 10 is a liquid chromatogram of palmitoyl lysophosphatidylcholine prepared in example 4;

FIG. 11 shows dipalmitoylphosphatidylcholine prepared in example 4¹H NMR spectrum;

FIG. 12 is a liquid chromatogram of dipalmitoylphosphatidylcholine prepared in example 4;

FIG. 13 shows stearoyl lysophosphatidylcholine prepared in example 5¹H NMR chart;

FIG. 14 is a liquid chromatogram of stearoyl lysophosphatidylcholine prepared in example 5;

FIG. 15 shows the preparation of distearoylphosphatidylcholine in example 5¹H NMR spectrum;

FIG. 16 is a liquid chromatogram of distearoylphosphatidylcholine prepared in example 5;

FIG. 17 shows the preparation of dioleoylphosphatidylcholine prepared in example 7¹H NMR spectrum;

FIG. 18 is a liquid chromatogram of dioleoylphosphatidylcholine prepared in example 7;

FIG. 19 shows the preparation of 1-stearoyl-2-palmitoyl phosphatidylcholine from example 10¹H NMR spectrum;

FIG. 20 is a liquid chromatogram of 1-stearoyl-2-palmitoyl phosphatidylcholine prepared in example 10.

Detailed Description

The invention provides a preparation method of phosphatidylcholine, which comprises the following steps:

and (S) -glycidyl fatty acid ester, phosphorylcholine and a second organic solvent are mixed for ring-opening reaction to obtain the lysophosphatidylcholine.

The present invention provides a method for producing (S) -glycidol fatty acid ester by dissolving (R) -glycidol and a first acylating agent in a first organic solvent to carry out a first acylation reaction. In the present invention, the first acylating agent preferably comprises a carboxylic acid, an acid chloride or an acid anhydride, more preferably a carboxylic acid or an acid anhydride. In the invention, the number of carbon atoms in the carboxylic acid, the acyl chloride and the anhydride is preferably 8-27, and more preferably 10-26. In the present invention, the carboxylic acid preferably has a structure represented by the formula a-1; the acid chloride preferably has a structure represented by a-2; the acid anhydride preferably has a structure represented by formula a-3;

wherein R is¹Preferably C7-C26 alkyl or substituted alkyl, more preferably C7-C26 alkyl. In the present invention, the carboxylic acid preferably includes fatty acid, n-octanoic acid, lauric acid, palmitic acid, stearic acid, arachidic acid, oleic acid, or arachidonic acid, and more preferably, arachidic acid, oleic acid, or arachidonic acid. In the present invention, the acid chloride preferably includes fatty acid chloride, myristoyl chloride, palmitoyl chloride, or stearoyl chloride, and more preferably myristoyl chloride or stearoyl chloride. In the present invention, the acid anhydride preferably includes fatty acid anhydride, lauric acid anhydride, stearic acid anhydride or oleic acid anhydride, and more preferably lauric acid anhydride or oleic acid anhydride.

In the present invention, the first organic solvent preferably includes one or more of acetonitrile, 1, 4-dioxane, N-dimethylformamide, N-dimethylacetamide, ethylene glycol dimethyl ether, dimethyl sulfoxide, sulfolane, dichloromethane, dichloroethane, toluene and xylene, more preferably 1, 4-dioxane, dichloromethane or toluene. In the present invention, when the first organic solvent is two or more of the above specific substances, the ratio of the specific substances in the present invention is not particularly limited, and any ratio may be used.

In the present invention, the molar ratio of the (R) -glycidol to the first acylating agent is preferably 1:1 to 1.8, and more preferably 1:1 to 1.5.

In the present invention, the ratio of the mass of the (R) -glycidol to the volume of the first organic solvent is preferably 19.6g:200 to 500mL, more preferably 19.6g:320 to 450 mL.

In the present invention, the (R) -glycidol, the first acylating agent, and the mixed solution in which the first acylating agent is dissolved in the first organic solvent preferably further include a dehydrating agent, and the dehydrating agent preferably includes N, N-dicyclohexylcarbodiimide, triethylamine, or 4-dimethylaminopyridine. In the present invention, the mass ratio of the (R) -glycidol to the dehydrating agent is preferably 19.6:0.32 to 55, more preferably 19.6:28 to 54.5. In the present invention, it is preferable that the reaction solution obtained by dissolving (R) -glycidol and the first acylating agent in the first organic solvent is heated to a temperature required for the first acylation reaction, and then the dehydrating agent is added.

The invention has no special requirements on the dissolving mode as long as the dissolving mode can be completely dissolved.

In the invention, the temperature of the first acylation reaction is preferably-15-50 ℃; the time of the first acylation reaction is preferably 2-8 h. In the invention, when the first acylating agent is carboxylic acid, the temperature of the first acylation reaction is preferably 45-50 ℃, and the time of the first acylation reaction is preferably 7-8 h; when the first acylation reagent is acid anhydride, the temperature of the first acylation reaction is preferably 25-30 ℃, and the time of the first acylation reaction is preferably 4-5 h; when the first acylation reagent is acyl chloride, the temperature of the first acylation reaction is preferably-15 to-10 ℃, and the time of the first acylation reaction is preferably 2 to 3 hours. In the present invention, the first acylation reaction is preferably accompanied by stirring. In the present invention, the stirring is not particularly limited as long as the reaction can be sufficiently performed.

In the present invention, the equation of the first acylation reaction is shown in formula 1:

in the present invention, it is preferable that the first acylation reaction further comprises: and (2) cooling the system after the first acylation reaction, filtering, concentrating the filtrate obtained by filtering, and recrystallizing to obtain the (S) -glycidyl fatty acid ester.

In the invention, the temperature of the cooled system is preferably room temperature, and the temperature of the room temperature is preferably 20-35 ℃, and more preferably 25-30 ℃. The cooling mode is not particularly limited, and the required temperature can be achieved. The invention has no special requirements on the filtration, and can adopt the conventional filtration mode in the field. In the present invention, the concentration is preferably concentration under reduced pressure. The present invention does not require any particular step for the concentration under reduced pressure, as long as most of the solvent in the system can be removed. In the present invention, the solvent for recrystallization preferably includes a mixed solution of dichloromethane and methanol, a mixed solution of toluene and isopropanol, a mixed solution of ethyl acetate and n-butanol, or a mixed solution of acetone and n-hexane. In the invention, the mass ratio of the dichloromethane and the methanol in the mixed solution of the dichloromethane and the methanol is preferably 1: 2.8-3.2, and more preferably 1:3. In the present invention, the mass ratio of toluene to isopropyl alcohol in the mixed solution of toluene and isopropyl alcohol is preferably 1:2.8 to 3.2, and more preferably 1:3. In the invention, the mass ratio of ethyl acetate to n-butanol in the mixed solution of ethyl acetate and n-butanol is preferably 1: 2.8-3.2, and more preferably 1:3. In the invention, the mass ratio of acetone to n-hexane in the mixed solution of acetone and n-hexane is preferably 1: 2.8-3.2, and more preferably 1:3. In the invention, the mass ratio of the crude product obtained after concentration to the solvent for recrystallization is preferably 1: 3.8-4.2, and more preferably 1: 4.

After obtaining the (S) -glycidyl fatty acid ester, the invention mixes the (S) -glycidyl fatty acid ester, the phosphorylcholine and a second organic solvent, and carries out ring-opening reaction to obtain the lysophosphatidylcholine. In the present invention, the second organic solvent preferably includes one or more of toluene, N-dimethylformamide, N-dimethylacetamide, methanol, ethanol, isopropanol, N-butanol, t-butanol, ethylene glycol methyl ether, ethylene glycol dimethyl ether, dimethyl sulfoxide and sulfolane, and more preferably ethylene glycol methyl ether, sulfolane or N, N-dimethylformamide. In the present invention, when the second organic solvent includes two or more of the above-mentioned specific substances, the ratio of the specific substances in the present invention is not particularly limited, and any ratio may be used.

In the present invention, the mass ratio of the (S) -glycidyl fatty acid ester to the second organic solvent is preferably 32.8 to 91g:400 to 450mL, and more preferably 32.8 to 90.5g:400 to 420 mL. In the present invention, the molar ratio of the phosphorylcholine to the (S) -glycidyl fatty acid ester is preferably 1:1 to 1.5, and more preferably 1:1.2 to 1.4.

In the present invention, the ring-opening reaction is preferably carried out under catalyst conditions; the catalyst preferably comprises one or more of transition metal oxide, sulfuric acid, nitric acid, hydrochloric acid, cationic resin and solid super acid, and more preferably transition metal oxide, sulfuric acid or solid super acid. In the present invention, the transition metal oxide preferably includes acidic alumina. In the present invention, the solid super acid preferably includes a zirconia-sulfuric acid solid super acid. In the present invention, when the catalyst includes two or more of the above-mentioned specific substances, the ratio of the specific substances in the present invention is not particularly limited, and any ratio may be used.

In the present invention, the molar ratio of the phosphorylcholine to the catalyst is preferably 1:0.1 to 1, and more preferably 1:0.2 to 0.8.

The invention has no special requirement on the mixing as long as the uniform mixing can be realized. In the invention, the temperature of the ring-opening reaction is preferably 60-150 ℃, and more preferably 70-140 ℃; the time of the ring-opening reaction is preferably 12-48 h, and more preferably 15-40 h.

In the present invention, the ring-opening reaction is preferably accompanied by stirring, and the stirring is not particularly required in the present invention as long as the reaction can be sufficiently performed.

In the present invention, the formula of the ring-opening reaction is shown in formula 2:

in the present invention, it is preferable that the ring-opening reaction further includes: and (3) cooling the system after the ring-opening reaction, filtering, concentrating the filtrate obtained by filtering, and recrystallizing to obtain the lysophosphatidylcholine.

In the invention, the temperature of the cooled system is preferably room temperature, and the temperature of the room temperature is preferably 20-35 ℃, and more preferably 25-30 ℃. The cooling mode is not particularly limited, and the required temperature can be achieved. The invention has no special requirements on the filtration, and can adopt the conventional filtration mode in the field. In the present invention, the concentration is preferably concentration under reduced pressure. The present invention does not require any particular step for the concentration under reduced pressure, as long as most of the solvent in the system can be removed. In the present invention, the solvent for recrystallization preferably includes a mixture of ethyl acetate and n-hexane, a mixture of chloroform and diethyl ether, or a mixture of acetonitrile and methyl t-butyl ether, and more preferably a mixture of ethyl acetate and n-hexane or a mixture of chloroform and diethyl ether. In the present invention, when the recrystallization solvent is a mixed solution of ethyl acetate and n-hexane, the mass ratio of ethyl acetate to n-hexane is preferably 0.8 to 1.2:1, and more preferably 1: 1; the recrystallization temperature is preferably-1 to 1 ℃, and more preferably 0 ℃. In the invention, when the solvent for recrystallization is a mixed solution of chloroform and diethyl ether, the mass ratio of the chloroform to the diethyl ether is preferably 2.8-3.2: 5, and more preferably 3: 5; the recrystallization temperature is preferably 38-42 ℃, and more preferably 40 ℃. In the invention, when the solvent for recrystallization is a mixed solution of acetonitrile and methyl tert-butyl ether, the mass ratio of the acetonitrile to the methyl tert-butyl ether is preferably 1.8-2.2: 6, and more preferably 2: 6; the temperature of the recrystallization is preferably-42 to-38 ℃, more preferably-40 ℃. In the invention, the mass ratio of the crude product obtained after concentration to the solvent for recrystallization is preferably 1: 7.8-8.2, and more preferably 1: 8. In the present invention, the number of recrystallization is preferably 1 to 5, and more preferably 2 to 4.

After obtaining the lysophosphatidylcholine, the lysophosphatidylcholine and a second acylation reagent are dissolved in a third organic solvent to carry out a second acylation reaction, so that the phosphatidylcholine is obtained. In the present invention, the second acylating agent preferably comprises a carboxylic acid, an acid chloride or an acid anhydride, more preferably a carboxylic acid or an acid anhydride. In the invention, the number of carbon atoms in the carboxylic acid, the acyl chloride and the anhydride is preferably 8-27 independently, and more preferably 10-16 independently. In the present invention, the carboxylic acid preferably has a structure represented by the formula b-1; the acid chloride preferably has a structure represented by b-2; the acid anhydride preferably has a structure represented by the formula b-3;

wherein R is²Preferably C7-C26 alkyl or substituted alkyl, more preferably C7-C26 alkyl. In the present invention, the carboxylic acid preferably includes fatty acid, n-octanoic acid, lauric acid, palmitic acid, stearic acid, arachidic acid, oleic acid, or arachidonic acid, and more preferably arachidic acid, oleic acid, or arachidonic acid. In the present invention, the acid chloride preferably includes fatty acid chloride, myristoyl chloride, palmitoyl chloride, or stearoyl chloride, and more preferably myristoyl chloride or stearoyl chloride. In the present invention, the acid anhydride preferably includes fatty acid anhydride, lauric acid anhydride, stearic acid anhydride or oleic acid anhydride, and more preferably lauric acid anhydride or oleic acid anhydride.

In the present invention, the acyl groups provided by the first and second acylating agents are preferably not identical, i.e. R¹And R²Are not identical groups.

In the present invention, the third organic solvent preferably includes one or more of acetonitrile, 1, 4-dioxane, N-dimethylformamide, N-dimethylacetamide, ethylene glycol dimethyl ether, dimethyl sulfoxide, sulfolane, dichloromethane, dichloroethane, and toluene and xylene, more preferably toluene, sulfolane or dimethyl sulfoxide, and still more preferably toluene or dimethyl sulfoxide.

In the present invention, the molar ratio of the lysophosphatidylcholine to the second acylating agent is preferably 1:1 to 1.8, and more preferably 1:1.2 to 1.6.

In the invention, the volume ratio of the mass of the phosphatidylcholine to the third organic solvent is preferably 115-150 g:500mL, and more preferably 120-145 g:500 mL.

In the present invention, the mixed solution of the lysophosphatidylcholine and the second acylating agent dissolved in the third organic solvent preferably further includes a dehydrating agent, and the dehydrating agent preferably includes N, N-Dicyclohexylcarbodiimide (DCC), triethylamine, or 4-Dimethylaminopyridine (DMAP). In the invention, the mass ratio of the lysophosphatidylcholine to the dehydrating agent is preferably 115-150: 0.32-55, and more preferably 120-145: 28-55. In the invention, the reaction solution obtained by dissolving lysophosphatidylcholine and a second acylation reagent in a third organic solvent is preferably heated to the temperature required by the second acylation reaction, and then the dehydrating agent is added.

In the invention, the temperature of the second acylation reaction is preferably 50-150 ℃; the time of the acylation reaction is preferably 8-24 h. In the invention, when the second acylating agent is carboxylic acid, the temperature of the second acylation reaction is preferably 145-150 ℃, and the time of the second acylation reaction is preferably 22-24 h; when the second acylation reagent is acid anhydride, the temperature of the second acylation reaction is preferably 100-110 ℃, and the time of the second acylation reaction is preferably 14-16 h; when the second acylation reagent is acyl chloride, the temperature of the second acylation reaction is preferably 50-60 ℃, and the time of the second acylation reaction is preferably 8-10 h. In the present invention, the second acylation reaction is preferably accompanied by stirring. In the present invention, the stirring is not particularly limited as long as the reaction can be sufficiently performed.

In the present invention, the equation of the second acylation reaction is shown in formula 3:

in the present invention, it is preferable that the second acylation reaction further comprises: and cooling the system after the second acylation reaction, filtering, concentrating the filtrate obtained by filtering, and recrystallizing to obtain the phosphatidylcholine.

In the invention, the temperature of the cooled system is preferably room temperature, and the temperature of the room temperature is preferably 20-35 ℃, and more preferably 25-30 ℃. The cooling mode is not particularly limited, and the required temperature can be achieved. The invention has no special requirements on the filtration, and can adopt the conventional filtration mode in the field. In the present invention, the concentration is preferably concentration under reduced pressure. The present invention does not require any particular step for the concentration under reduced pressure, as long as most of the solvent in the system can be removed. In the present invention, the solvent for recrystallization preferably includes a good solvent for phosphatidylcholine and a poor solvent for phosphatidylcholine. In the present invention, the good solvent of phosphatidylcholine preferably includes one or more of acetonitrile, ethyl acetate, dichloromethane, dichloroethane, toluene, xylene, chloroform, methanol, ethanol, n-butanol, isopropanol and tert-butanol, more preferably toluene, methanol or acetonitrile. In the invention, the mass ratio of the concentrated solution to the good solvent is preferably 2-4: 1, and more preferably 3: 1. In the present invention, the poor solvent of phosphatidylcholine preferably comprises one or more of n-hexane, cyclohexane, acetone, butanone, diethyl ether and methyl tert-butyl ether, more preferably n-hexane, butanone or methyl tert-butyl ether. In the invention, the mass ratio of the concentrated solution to the poor solvent is preferably 4-6: 1, and more preferably 5: 1. In the examples of the present invention, the solvent for recrystallization is specifically a mixture of toluene and methyl ethyl ketone at a mass ratio of 3:5, a mixture of acetonitrile and methyl tert-butyl ether at a mass ratio of 2:6, or a mixture of methanol and n-hexane at a mass ratio of 1:1. In the examples of the present invention, the mass ratio of the concentrated solution to the recrystallization solvent is specifically 1: 8.

In the invention, the recrystallization temperature is preferably-40 ℃, and more preferably 0-20 ℃. In the present invention, the number of recrystallization is preferably 1 to 5, and more preferably 2 to 4.

The invention adopts the reactant with definite structure and single carbon chain length to ensure that the prepared phosphatidylcholine has definite structure, and the asymmetric phosphatidylcholine can be obtained only by adjusting the types of the first acylation reagent and the second acylation reagent.

In order to further illustrate the present invention, the following embodiments are described in detail, but they should not be construed as limiting the scope of the present invention.

Example 1

Dissolving (R) -glycidol (19.6g, 0.264mol) and N-octanoic acid (68.5g, 0.475mol) in 320mL of 1, 4-dioxane, heating to 50 ℃, adding a dehydrating agent N, N-Dicyclohexylcarbodiimide (DCC) (54.5g, 0.264mol) into the dissolved solution, carrying out a first acylation reaction (accompanied by stirring) for 8h, cooling to 25 ℃, filtering, collecting filtrate, and concentrating under reduced pressure to obtain a crude product; recrystallizing the obtained crude product (the mass ratio of the crude product to the mixed solvent is 1:4) by using the mixed solvent of toluene and isopropanol with the mass ratio of 1:3 to obtain 49.6g of (S) -glycidol n-caprylate;

mixing phosphorylcholine (30.2g, 0.164mol), (S) -glycidol n-caprylate (32.8g, 0.164mol) and 400mL of ethylene glycol monomethyl ether, heating to 60 ℃, carrying out ring-opening reaction (with stirring) for 12h, cooling to room temperature, filtering, collecting filtrate, and concentrating under reduced pressure to obtain a crude product; recrystallizing the crude product with a mixed solution of ethyl acetate and n-hexane in a mass ratio of 1:1 at 0 deg.C for 3 times (the mass ratio of the crude product to the mixed solution is 1:8) to obtain 38.3g of n-octanoyl lysophosphatidylcholine;

dissolving n-octanoyl lysophosphatidylcholine (101.1g, 0.264mol) and n-octanoic anhydride (52.3g, 0.370mol) in toluene (500mL), heating to 100 ℃, adding 4-dimethylaminopyridine (DMAP, 0.32g, 2.64mmol) into the dissolved solution to perform a second acylation reaction (with stirring) for 14h, cooling to 25 ℃, filtering, collecting filtrate, concentrating under reduced pressure to obtain a crude product, and recrystallizing the crude product with a mixed solution of toluene and butanone at a mass ratio of 3:5 at 0 ℃ (the mass ratio of the crude product to the mixed solution is 1:8) for 3 times to obtain 116.9g of di-n-octanoyl phosphatidylcholine.

Example 2

Dissolving (R) -glycidol (19.6g, 0.264mol) and lauric acid (95.1g, 0.475mol) in 320mL of 1, 4-dioxane, heating to 50 ℃, adding a dehydrating agent N, N-Dicyclohexylcarbodiimide (DCC) (54.5g, 0.264mol) into the dissolved solution, carrying out a first acylation reaction (accompanied by stirring) for 8h, cooling to 25 ℃, filtering, collecting filtrate, and concentrating under reduced pressure to obtain a crude product; recrystallizing the obtained crude product (the mass ratio of the crude product to the mixed solvent is 1:4) by using the mixed solvent of toluene and isopropanol with the mass ratio of 1:3 to obtain 63.5g of (S) -glycidyl laurate;

mixing phosphorylcholine (30.2g, 0.164mol), (S) -glycidyllaurate (42.0g, 0.164mol), sulfuric acid (1.6g, 16.4mmol) and ethylene glycol monomethyl ether (400mL), heating to 60 ℃, carrying out ring-opening reaction (with stirring) for 12h, cooling to 25 ℃, filtering, collecting filtrate, carrying out reduced pressure concentration to obtain a crude product, and recrystallizing the crude product (the mass ratio of the crude product to the mixed liquid is 1:8) for 3 times by using the mixed liquid of ethyl acetate and n-hexane with the mass ratio of 1:1 at the temperature of 0 ℃ to obtain 46.8g of lauroyl lysophosphatidylcholine;

lauroyl lysophosphatidylcholine (115.9g, 0.264mol) and lauric anhydride (141.4g, 0370mol) are dissolved in toluene (500mL), after the temperature is raised to 100 ℃, 4-dimethylaminopyridine (DMAP, 0.32g, 2.64mmol) is added into the dissolved solution to carry out a second acylation reaction (with stirring) for 14h, the cooled temperature is 25 ℃, the filtered solution is filtered, the filtered solution is collected and concentrated under reduced pressure to obtain a crude product, and the crude product is recrystallized by using a mixed solution of toluene and butanone in a mass ratio of 3:5 at the temperature of 0 ℃ (the mass ratio of the crude product to the mixed solution is 1:8) for 3 times to obtain 142.6g of dilauroyl phosphatidylcholine.

Example 3

Mixing (R) -glycidol (19.6g, 0.264mol) and dichloromethane (320mL), cooling to-15 ℃, adding triethylamine (28.1g, 0.278mol) into the mixed solution, adding myristoyl chloride (65.2g, 0.264mol) into the mixed solution of (R) -glycidol, dichloromethane and triethylamine through a constant pressure dropping funnel at a constant speed within 1 hour (with stirring), continuing to react for 1 hour (with stirring), filtering, collecting filtrate, and concentrating under reduced pressure to obtain a crude product; recrystallizing the obtained crude product by using a mixed solvent of ethyl acetate and n-butanol in a mass ratio of 1:3 to obtain 68.2g of (S) -glycidylmyristate;

mixing phosphorylcholine (30.2g, 0.164mol), (S) -glycidylmyristate (69.9g, 0.246mol), acidic alumina (16.6g, 0.164mol) and sulfolane (420mL), heating to 150 ℃, carrying out ring-opening reaction (with stirring) for 48h, cooling to 25 ℃, filtering, collecting filtrate, concentrating under reduced pressure to obtain a crude product, and reacting at 40 ℃ in a mass ratio of 3:5, recrystallizing the crude product for 1 time (the mass ratio of the crude product to the mixed solution is 1:8) by using a mixed solution of chloroform and ether to obtain 50.5g of myristoyl lysophosphatidylcholine;

myristoyl lysophosphatidylcholine (123.3g, 0.264mol), myristic anhydride (162.3g, 0.370mol) and toluene (500mL) are mixed, the temperature is raised to 100 ℃, 4-dimethylaminopyridine (DMAP, 0.32g, 2.64mmol) is added to the mixed solution to carry out a second acylation reaction (with stirring) for 14h, the mixed solution is cooled to 25 ℃, the filtration is carried out, the filtrate is collected and concentrated under reduced pressure to obtain a crude product, and the crude product is recrystallized by using a mixed solvent of toluene and butanone in a mass ratio of 3:5 at the temperature of 0 ℃ (the mass ratio of the crude product to the mixed solution is 1:8) for 3 times to obtain 148.3g of dimyristoyl phosphatidylcholine.

Example 4

Dissolving (R) -glycidol (19.6g, 0.264mol) and palmitic acid (121.8g, 0.475mol) in 320mL of 1, 4-dioxane, heating to 50 ℃, adding a dehydrating agent N, N-Dicyclohexylcarbodiimide (DCC) (54.5g, 0.264mol) into the dissolved solution, carrying out a first acylation reaction (accompanied by stirring) for 8h, cooling to 25 ℃, filtering, collecting filtrate, and concentrating under reduced pressure to obtain a crude product; recrystallizing the obtained crude product (the mass ratio of the crude product to the mixed solvent is 1:4) by using the mixed solvent of toluene and isopropanol with the mass ratio of 1:3 to obtain 73.3g of (S) -glycidol palmitate;

mixing phosphorylcholine (30.2g, 0.164mol), (S) -glycidol palmitate (76.8g, 0.246mol), acidic alumina (16.6g, 0.164mol) and sulfolane (420mL), heating to 150 ℃, carrying out ring-opening reaction (with stirring) for 48h, cooling to 25 ℃, filtering, collecting filtrate, concentrating under reduced pressure to obtain a crude product, and reacting at 40 ℃ in a mass ratio of 3:5, recrystallizing the crude product for 1 time by using a mixed solution of chloroform and diethyl ether (the mass ratio of the crude product to the mixed solution is 1:8) to obtain 56.8g of palmitoyl lysophosphatidylcholine;

mixing palmitoyl lysophosphatidylcholine (123.3g, 0.264mol), palmitic anhydride (183.1g, 0.370mol) and toluene (500mL), heating to 100 ℃, adding 4-dimethylaminopyridine (DMAP, 0.32g, 2.64mmol) to the mixed solution to perform a second acylation reaction (with stirring) for 14h, cooling to 25 ℃, filtering, collecting the filtrate, concentrating under reduced pressure to obtain a crude product, and recrystallizing the crude product (the mass ratio of the crude product to the mixed solution is 1:8)1 time at 40 ℃ by using a mixed solvent of toluene and butanone in a mass ratio of 3:5 to obtain 164.5g dipalmitoyl phosphatidylcholine.

Example 5

Dissolving (R) -glycidol (19.6g, 0.264mol) and triethylamine (28.1g, 0.278mol) in 320mL of dichloromethane, cooling to-15 ℃, dropwise adding stearoyl chloride (80.0g, 0.264mol) within 1h at a constant speed, reacting for 1h while keeping the temperature, filtering, collecting filtrate, and concentrating under reduced pressure to obtain a crude product; recrystallizing the obtained crude product (the mass ratio of the crude product to the mixed solvent is 1:4) by using the mixed solvent of methanol and dichloromethane with the mass ratio of 1:3 to obtain 82.5g of (S) -glycidol stearate;

mixing phosphorylcholine (30.2g, 0.164mol), (S) -glycide stearate (83.6g, 0.246mol), acidic alumina (16.6g, 0.164mol) and sulfolane (420mL), heating to 150 ℃, carrying out cyclization reaction (stirring) for 48h, cooling to 25 ℃, filtering, collecting filtrate, carrying out reduced pressure concentration to obtain a crude product, and recrystallizing the crude product (the mass ratio of the crude product to the mixed solution is 1:8) by using a mixed solution of chloroform and diethyl ether with the mass ratio of 3:5 at the temperature of 40 ℃ to obtain 59.2g stearoyl lysophosphatidylcholine;

stearoyl lysophosphatidylcholine (138.1g, 0.264mol), stearic anhydride (183.1g, 0.370mol) and toluene (500mL) are mixed, heated to 100 ℃, 4-dimethylaminopyridine (DMAP, 0.32g, 2.64mmol) is added to the mixed solution to carry out a second acylation reaction (with stirring) for 14h, cooled to 25 ℃, filtered, the filtrate is collected and concentrated under reduced pressure to obtain a crude product, and the crude product is recrystallized (the mass ratio of the crude product to the mixed solution is 1:8)1 time by using a mixed solvent of toluene and butanone in a mass ratio of 3:5 at the temperature of 40 ℃ to obtain 183.3g distearoyl phosphatidylcholine.

Example 6

Dissolving (R) -glycidol (19.6g, 0.264mol) and triethylamine (28.1g, 0.278mol) in 320mL of dichloromethane, cooling to-15 ℃, dropwise adding arachidylchloride (87.4g, 0.264mol) within 1h at a constant speed, reacting for 1h while keeping the temperature, filtering, collecting filtrate, and concentrating under reduced pressure to obtain a crude product; recrystallizing the obtained crude product (the mass ratio of the crude product to the mixed solvent is 1:4) by using the mixed solvent of methanol and dichloromethane with the mass ratio of 1:3 to obtain 92.3g of (S) -glycidic arachidic acid ester;

mixing phosphorylcholine (30.2g, 0.164mol), (S) -glycide arachidic acid ester (90.5g, 0.246mol), acidic alumina (16.6g, 0.164mol) and sulfolane (420mL), heating to 150 ℃, carrying out ring-opening reaction (with stirring) for 48h, cooling to 25 ℃, filtering, collecting filtrate, carrying out reduced pressure concentration to obtain a crude product, and recrystallizing the crude product (the mass ratio of the crude product to the mixed solution is 1:8)1 time by using a mixed solution of chloroform and diethyl ether with the mass ratio of 3:5 at the temperature of 40 ℃ to obtain 63.3g of arachidoyl lysophosphatidylcholine;

mixing arachidylic lysophosphatidylcholine (145.5g, 0.264mol), arachidic acid (148.4g, 0.475mol) and sulfolane (500mL), heating to 150 ℃, adding N, N-Dicyclohexylcarbodiimide (DCC), 54.5g, 0.264mmol) into the mixed solution, carrying out a second acylation reaction (with stirring) for 24h, cooling to 25 ℃, filtering, collecting filtrate, concentrating under reduced pressure to obtain a crude product, and recrystallizing the crude product (the mass ratio of the crude product to the mixed solution is 1:8) by using a mixed solvent of toluene and butanone with a mass ratio of 3:5 at 40 ℃ for 1 time to obtain 200.8g of arachidylic acylphosphatidylcholine.

Example 7

Dissolving (R) -glycidol (19.6g, 0.264mol) and oleic anhydride (202.2g, 0.370mol) in toluene (450mL), heating to 30 ℃, adding 4-dimethylaminopyridine (DMAP, 0.32g, 2.64mmol) into the dissolved solution to perform a first acylation reaction (with stirring) for 5h, cooling to 25 ℃, filtering, collecting filtrate, concentrating under reduced pressure to obtain a crude product of (S) -glycidol oleate, and recrystallizing the crude product at (-5 ℃) by using a mixed solvent of acetone and n-hexane with the mass ratio of 1:3 to obtain 77.7g of (S) -glycidol oleate;

mixing phosphorylcholine (30.2g, 0.164mol), (S) -glycidol oleate (66.6g, 0.197mol), zirconia-sulfuric acid solid super acid (10.1g, 0.082mol) and N, N-dimethylformamide (420mL), heating to 100 ℃, carrying out ring-opening reaction (with stirring) for 30h, cooling to 25 ℃, filtering, collecting filtrate, concentrating under reduced pressure to obtain a crude product, and recrystallizing the crude product with a mixed solution of acetonitrile and methyl tert-butyl ether in a mass ratio of 2:6 (the mass ratio of the crude product to the mixed solution is 1:8) for 5 times at-40 ℃ to obtain 47g of oleoyl lysophosphatidylcholine;

oleoyl lysophosphatidylcholine (137.5g, 0.264mol), oleic acid (134.2g, 0.475mol) and toluene (500mL) are mixed, heated to 150 ℃, 4-dimethylaminopyridine (DMAP, 0.32g, 2.64mmol) is added to the mixed solution to carry out a second acylation reaction (with stirring) for 14h, cooled to 25 ℃, filtered, the filtrate is collected and concentrated under reduced pressure to obtain a crude product, and the crude product is recrystallized (the mass ratio of the crude product to the mixed solution is 1:8)4 times by using a mixed solvent of acetonitrile and methyl tert-butyl ether with the mass ratio of 2:6 at the temperature of-40 ℃ to obtain 149.2g of dioleoyl phosphatidylcholine.

Example 8

Dissolving (R) -glycidol (19.6g, 0.264mol) and arachidonic acid anhydride (218.3g, 0.370mol) in 450mL of toluene, heating to 30 ℃, adding 4-Dimethylaminopyridine (DMAP) (0.32g, 2.64mmol) into a reaction system, preserving heat for reaction for 5h, filtering, collecting filtrate, and concentrating under reduced pressure to obtain a crude product; recrystallizing the obtained crude product (the mass ratio of the crude product to the mixed solvent is 1:4) by using the mixed solvent of acetone and n-hexane with the mass ratio of 1:3 to obtain 76.2g of (S) -glycidyl arachidonate;

mixing phosphorylcholine (30.2g, 0.164mol), (S) -glycidol arachidonate (70.9g, 0.197mol), zirconia-sulfuric acid solid super acid (10.1g, 0.082mol) and N, N-dimethylformamide (400mL), heating to 100 ℃, carrying out ring opening reaction (with stirring) for 30h, cooling to 25 ℃, filtering, collecting filtrate, carrying out reduced pressure concentration to obtain a crude product, and recrystallizing the crude product (the mass ratio of the crude product to the mixed solution is 1:8) for 5 times by using a mixed solution of acetonitrile and methyl tert-butyl ether with the mass ratio of 2:6 at the temperature of-40 ℃ to obtain 38.3g of arachidonoyl lysophosphatidylcholine;

mixing arachidonic acid lysophosphatidylcholine (143.4g, 0.264mol), arachidonic acid (144.7g, 0.475mol) and sulfolane (500mL), heating to 150 ℃, adding N, N-dicyclohexylcarbodiimide (DCC, 54.5g, 0.264mmol) into the mixed solution, carrying out a second acylation reaction (with stirring) for 24h, cooling to 25 ℃, filtering, collecting filtrate, concentrating under reduced pressure to obtain a crude product, and recrystallizing the crude product (the mass ratio of the crude product to the mixed solution is 1:8)5 times by using a mixed solvent of acetonitrile and methyl tert-butyl ether with the mass ratio of 2:6 at the temperature of-40 ℃ to obtain 142.4g of the eicosatetraylphosphatidylcholine.

Example 9

Stearoyl lysophosphatidylcholine was prepared according to the method of example 5;

mixing stearoyl lysophosphatidylcholine (138.1g, 0.264mol), oleic acid (134.2g, 0.475mol) and sulfolane (500mL), heating to 150 ℃, adding a dehydrating agent N, N-dicyclohexylcarbodiimide (DCC, 54.5g, 0.264mol) into the mixed solution to perform a second acylation reaction (with stirring) for 24h, cooling to 25 ℃, filtering, collecting filtrate, concentrating under reduced pressure to obtain a crude product, and recrystallizing the crude product (the mass ratio of the crude product to the mixed solution is 1:8) for 3 times by using a mixed solvent of acetonitrile and methyl tert-butyl ether with the mass ratio of 2:6 at the temperature of-40 ℃ to obtain 153.7g of 1-stearoyl-2-oleoyl phosphatidylcholine.

Example 10

mixing stearoyl lysophosphatidylcholine (138.1g, 0.264mol) and dimethyl sulfoxide (500mL), heating to 50 ℃, adding triethylamine (28.1g, 0.278mol) into the mixed solution, adding palmitoyl chloride (72.5g, 0.264mol) into the mixed solution of stearoyl lysophosphatidylcholine, dimethyl sulfoxide (500mL) and triethylamine through a constant-pressure dropping funnel within 7 hours, continuing to keep the temperature and stir for 1 hour, cooling to 25 ℃, filtering, collecting filtrate, concentrating under reduced pressure to obtain a crude product, and recrystallizing the crude product (the mass ratio of the crude product to the mixed solution is 1:8) for 1 time by using a mixed solvent of methanol and n-hexane with the mass ratio of 1:1 at the temperature of 40 ℃ to obtain 186.8g of 1-stearoyl-2-palmitoyl phosphatidylcholine.

Example 11

Palmitoyl lysophosphatidylcholine was prepared according to the method of example 4;

mixing palmitoyl lysophosphatidylcholine (130.7g, 0.264mol) and dimethyl sulfoxide (500mL), heating to 50 ℃, adding triethylamine (28.1g, 0.278mol) into the mixed solution, adding stearoyl chloride (82.5g, 0.264mol) into the mixed solution of palmitoyl lysophosphatidylcholine, dimethyl sulfoxide and triethylamine through a constant-pressure dropping funnel within 7 hours, continuing to keep the temperature and stir for 1h, cooling to 25 ℃, filtering, collecting filtrate, concentrating under reduced pressure to obtain a crude product, and recrystallizing the crude product (the mass ratio of the crude product to the mixed solution is 1:8)1 time by using a mixed solvent of methanol and n-hexane with the mass ratio of 1:1 at the temperature of 40 ℃ to obtain 186.8g of 1-palmitoyl-2-stearoyl phosphatidylcholine.

The calculated yields of (S) -glycidyl fatty acid ester, lysophosphatidylcholine and phosphatidylcholine in examples 1 to 11 are shown in Table 1; the purity of lysophosphatidylcholine and phosphatidylcholine in examples 1 to 11 measured by liquid chromatography is shown in table 1.

TABLE 1 yield and purity of intermediate and final products in examples 1-11

Nuclear magnetic detection is carried out on lauroyl lysophosphatidylcholine prepared in example 2 to obtain¹The H NMR spectrum is shown in FIG. 1; performing liquid chromatography detection on lauroyl lysophosphatidylcholine prepared in example 2 to obtain a liquid chromatogram shown in FIG. 2; performing nuclear magnetic resonance detection on the dilauroyl phosphatidylcholine prepared in example 2 to obtain¹The H NMR spectrum is shown in FIG. 3; the dilauroyl phosphatidylcholine prepared in example 2 was subjected to liquid chromatography, and the liquid chromatogram obtained is shown in fig. 4.

Nuclear magnetic assay of myristoyl lysophosphatidylcholine prepared in example 3 to obtain myristoyl lysophosphatidylcholine¹The H NMR spectrum is shown in FIG. 5; performing liquid chromatography detection on myristoyl lysophosphatidylcholine prepared in example 3 to obtain a liquid chromatogram shown in fig. 6; nuclear magnetic resonance detection was performed on dimyristoyl phosphatidylcholine obtained in example 3 to obtain dimyristoyl phosphatidylcholine¹The H NMR spectrum is shown in FIG. 7; dimyristoyl phosphatidyl compound prepared in example 3Liquid chromatogram obtained by liquid chromatography detection of choline is shown in FIG. 8.

Subjecting palmitoyl lysophosphatidylcholine obtained in example 4 to nuclear magnetic detection to obtain¹The H NMR spectrum is shown in FIG. 9; performing liquid chromatography detection on palmitoyl lysophosphatidylcholine prepared in example 4 to obtain a liquid chromatogram shown in fig. 10; subjecting the dipalmitoylphosphatidylcholine prepared in example 4 to nuclear magnetic resonance detection to obtain¹The H NMR spectrum is shown in FIG. 11; the dipalmitoylphosphatidylcholine prepared in example 4 was subjected to liquid chromatography, and the liquid chromatogram obtained was shown in FIG. 12.

Performing nuclear magnetic detection on stearoyl lysophosphatidylcholine prepared in example 5 to obtain stearoyl lysophosphatidylcholine¹The H NMR spectrum is shown in FIG. 13; detecting stearoyl lysophosphatidylcholine prepared in example 5 by liquid chromatography, and obtaining a liquid chromatogram as shown in fig. 14; performing nuclear magnetic detection on the distearoyl phosphatidylcholine prepared in example 5 to obtain¹The H NMR spectrum is shown in FIG. 15; the distearoyl phosphatidylcholine prepared in example 5 was subjected to liquid chromatography detection, and the liquid chromatogram obtained was shown in fig. 16.

Subjecting the dioleoylphosphatidylcholine prepared in example 7 to nuclear magnetic resonance to obtain¹The H NMR spectrum is shown in FIG. 17; the dioleoylphosphatidylcholine prepared in example 7 was subjected to liquid chromatography, and the liquid chromatogram obtained was shown in FIG. 18.

Subjecting the 1-stearoyl-2-palmitoyl phosphatidylcholine obtained in example 10 to nuclear magnetic resonance to obtain¹The H NMR spectrum is shown in FIG. 19; the liquid chromatogram obtained by performing liquid chromatography detection on 1-stearoyl-2-palmitoyl phosphatidylcholine prepared in example 10 is shown in fig. 20.

The invention is based on¹H NMR spectra are assigned to hydrogen in different chemical environments, so that the structure of the product can be determined, the intermediate product prepared in the embodiment is lysophosphatidylcholine, and the intermediate product is phosphatidylcholine, such as nuclear magnetic resonance hydrogen spectra provided by the invention; meanwhile, according to the retention time of different substances in the high performance liquid chromatogramThe difference in (3) can also be confirmed by comparing with a standard sample to confirm the structure and purity.

Although the present invention has been described in detail with reference to the above embodiments, it is only a part of the embodiments of the present invention, not all of the embodiments, and other embodiments can be obtained without inventive step according to the embodiments, and the embodiments are within the scope of the present invention.

Claims

1. A preparation method of phosphatidylcholine comprises the following steps:

2. The method of claim 1, wherein the first and second acylating reagents independently comprise a carboxylic acid, an acid chloride, or an acid anhydride.

3. The method according to claim 2, wherein the number of carbon atoms in the carboxylic acid, the acid chloride and the acid anhydride is 8 to 27 independently.

4. The production method according to claim 3, wherein the carboxylic acid comprises n-octanoic acid, lauric acid, palmitic acid, stearic acid, arachidic acid, oleic acid or arachidonic acid;

5. The production method according to any one of claims 1 to 4, wherein the molar ratio of the (R) -glycidol to the first acylating agent is 1:1 to 1.8;

6. The method according to any one of claims 1 to 4, wherein the molar ratio of the lysophosphatidylcholine to the second acylating agent is 1:1 to 1.8;

7. The method according to claim 1, wherein the molar ratio of the phosphorylcholine to the (S) -glycidyl fatty acid ester is 1:1 to 1.5.

8. The production method according to claim 1, wherein the ring-opening reaction is carried out under a catalyst condition;

the mol ratio of the phosphorylcholine to the catalyst is 1: 0.1-1.

9. The method of claim 8, wherein the catalyst comprises one or more of transition metal oxide, sulfuric acid, nitric acid, hydrochloric acid, cationic resin, and solid super acid.

10. The method according to any one of claims 4 and 7 to 9, wherein the ring-opening reaction is carried out at a temperature of 60 to 150 ℃; the ring-opening reaction time is 12-48 h.