Method for preparing hydrogenated soybean phosphatidylcholine

Abstract

The present invention relates to the field of synthetic phospholipids, and in particular to a method for preparing hydrogenated soybean phosphatidylcholine. The method comprises the following steps: A. hydrolyzing hydrogenated soybean oil to obtain an intermediate mixture 1 and adjusting the pH of the intermediate mixture 1 to obtain an intermediate mixture 2; B. mixing the intermediate mixture 2 with glycerophosphorylcholine, a condensing agent, an acid-binding agent, and a reaction solvent, and carrying out an esterification reaction and solid-liquid separation to obtain intermediate mixture 3; C. mixing the intermediate mixture 3 with an adsorbent for adsorption, and then carrying out solid-liquid separation to obtain an intermediate mixture 4; and D. mixing the obtained intermediate mixture 4 with an organic solvent, and carrying out pulping, separation, solid-liquid separation, and drying to obtain a hydrogenated soybean phosphatidylcholine product. The present invention solves the problems of difficult impurity removal, potential safety hazards, high costs, etc. in existing techniques for preparing hydrogenated soybean phosphatidylcholine by means of a hydrogenation reaction.

Description

Preparation method of hydrogenated soybean phosphatidylcholine TECHNICAL FIELD

[0001] The present application relates to the field of natural phospholipid synthesis, in particular to a preparation method of hydrogenated soybean phosphatidylcholine. BACKGROUND

[0002] Soybean phospholipid is a phosphorus-containing lipid, which is a mixture of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI) and triglyceride, and can be used as an emulsifier and a liposome membrane material in pharmaceutical preparations. However, soybean phospholipid has problems such as peculiar smell, deep color, easy oxidation and instability in application, which limits its use. Some researchers use catalytic hydrogenation method to partially saturate the unsaturated bonds contained in phospholipid to form hydrogenated soybean phospholipid, which greatly improves the stability of soybean phospholipid and reduces or eliminates its color and odor.

[0003] Due to the presence of a small amount of substances with similar properties and structures and non-lipid substances in the raw material soybean lecithin, other impurities are easily produced during hydrogenation. The presence of these impurities will affect the quality of HSPC. In addition, hydrogenation reaction has great safety hazards. Hydrogenation reaction has high flammability and belongs to exothermic reaction. Hydrogen gas is in contact with metal equipment under high temperature and high pressure, which is easy to cause hydrogen embrittlement phenomenon, reduce the strength of metal equipment and increase the safety risk of production. In the activation and regeneration process of the catalyst, explosion accidents are easy to occur. The tail gas in the hydrogenation cracking reaction process contains unreacted hydrogen, which is easy to cause fire or explosion accidents when discharged. In addition, the current market price of soybean phospholipid is about 800 yuan / kg, and the cost of raw materials for preparing HSPC is high. These conditions limit the industrialization development of hydrogenated soybean phosphatidylcholine prepared by hydrogenation reaction of soybean phospholipid. SUMMARY

[0004] The purpose of the present application is to solve the problems of difficult removal of impurities, safety hazards and high cost in the existing hydrogenation reaction for preparing hydrogenated soybean phosphatidylcholine, and to design a technical route for preparing hydrogenated soybean phosphatidylcholine from hydrogenated soybean oil with high quality, low cost and safety.

[0005] In order to solve the above technical problems, the technical scheme adopted by the present application is as follows:

[0006] A preparation method of hydrogenated soybean phosphatidylcholine, comprising the following steps:

[0007] A. Hydrolyzing hydrogenated soybean oil (CAS No. 8016-70-4) to obtain an intermediate mixture 1, and adjusting the pH of the intermediate mixture 1 to obtain an intermediate mixture 2;

[0008] B, mixing the intermediate mixture 2 with glycerylphosphorylcholine, condensing agent, acid binding agent and reaction solvent to carry out esterification reaction, and after solid-liquid separation, obtaining intermediate mixture 3;

[0009] C, mixing and adsorbing the intermediate mixture 3 with adsorbent, and then carrying out solid-liquid separation to obtain intermediate mixture 4;

[0010] D, mixing the obtained intermediate mixture 4 with organic solvent, and after beating, precipitation, solid-liquid separation and drying, obtaining hydrogenated soybean phosphatidylcholine product.

[0011] Preferably, in step C, the adsorbent includes modified silica gel; the modified silica gel includes a mixture of silica gel and metal salt; the metal salt includes one or more of transition metal elements of IB-VIIIB; the mass ratio of the intermediate mixture 3 to the adsorbent includes 1:0.1-0.5; the temperature of the mixing and adsorbing includes 40-70℃; the time of the mixing and adsorbing includes 2-6 hours; the solid-liquid separation method includes filtration; and after the filtration, the intermediate mixture 4 is obtained by further concentration.

[0012] Preferably, the preparation of the modified silica gel includes: mixing the silica gel and the metal salt in aqueous solution, washing with water and drying to obtain the modified silica gel; the mixing method of the silica gel and the metal salt includes stirring; the stirring temperature includes 20-100℃; the stirring time includes 0.5-24 hours; the mass ratio of the silica gel to the metal salt includes 1:0.05-0.5; and the metal salt includes one or more of iron salt, copper salt, zinc salt, cobalt salt, nickel salt, manganese salt, chromium salt and ruthenium salt.

[0013] Preferably, the operation of step A includes one of the following:

[0014] A1, mixing the hydrogenated soybean oil with acid, sodium alkylsulfonate and water to carry out the hydrolysis reaction to generate the intermediate mixture 1, washing the intermediate mixture 1 with water until the pH is 6-7, then standing and separating to obtain the upper liquid to obtain the intermediate mixture 2;

[0015] or

[0016] A2, mixing the hydrogenated soybean oil with inorganic base, lower alcohol and water to carry out the hydrolysis reaction to generate the intermediate mixture 1, adding acid to adjust the pH to 6-7, then cooling and crystallizing, and centrifuging to obtain the intermediate mixture 2.

[0017] Preferably, in step A1, the mass ratio of the hydrogenated soybean oil, the acid, the sodium alkylsulfate, and the water comprises 1:0.02-0.1:0.02-0.2:0.3-0.8; the reaction temperature of the hydrolysis reaction comprises 90-115℃; the reaction time of the hydrolysis reaction comprises 8-24 hours; the acid comprises one or more of sulfuric acid, hydrochloric acid, nitric acid, acetic acid, and citric acid.

[0018] Preferably, in step A2, the mass ratio of the hydrogenated soybean oil, the inorganic base, the lower alcohol, and the water comprises 1:0.1-0.3:1-20:0.05-4; the reaction temperature of the hydrolysis reaction comprises 50-100℃; the reaction time of the hydrolysis reaction comprises 0.5-8 hours; the inorganic base comprises one or more of sodium hydroxide, potassium hydroxide, sodium carbonate, and sodium bicarbonate; the lower alcohol comprises one or more of methanol, ethanol, isopropyl alcohol, and n-butyl alcohol; the acid comprises one or more of sulfuric acid, hydrochloric acid, nitric acid, acetic acid, and citric acid.

[0019] Preferably, in step B, the weight ratio of the glycerophosphocholine, the intermediate mixture 2, the condensing agent, the acid-binding agent, and the reaction solvent comprises 1:2-3:2-3:2-3:1-90; the reaction time of the esterification reaction comprises 4-24 hours; the reaction temperature of the esterification reaction comprises 40-80℃; the solid-liquid separation method comprises filtration.

[0020] Preferably, the condensing agent comprises one or more of dicyclohexyl carbodiimide, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N,N'-diisopropyl carbodiimide, 1-hydroxybenzotriazole, O-benzotriazol-N,N,N',N'-tetramethyluronium tetrafluoroborate, 2-(7-azobenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate; the acid-binding agent comprises one or more of pyridine, 4-dimethylaminopyridine, triethylamine, N,N-diisopropyl ethylamine, and N-hydroxysuccinimide; the reaction solvent comprises one or more of n-hexane, dichloromethane, trichloromethane, dimethyl sulfoxide, and ethyl acetate.

[0021] Preferably, in step D, the mass ratio of the intermediate mixture 4 and the organic solvent comprises 1:1-20; the beating and precipitation is repeated 1-4 times; the organic solvent comprises one or more of n-hexane, octane, dichloromethane, trichloromethane, ethyl acetate, acetone, tetrahydrofuran, methyl tert-butyl ether, methanol, ethanol, and water; the solid-liquid separation method comprises filtration.

[0022] A hydrogenated soybean phosphatidylcholine product obtained by a method for preparing the hydrogenated soybean phosphatidylcholine described above.

[0023] Compared with the prior art, the present application has the following beneficial effects:

[0024] The present application overcomes the problems of high raw material cost, difficult impurity removal and poor safety in hydrogenation synthesis process in the prior art of preparing HSPC, and designs a new modified silica gel by using the structural differences of HSPC, condensing agent and acid-binding agent, thereby improving the separation degree of the product and impurities, and developing an efficient and widely applicable purification method to promote the industrial production of HSPC. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 is a liquid chromatogram of the intermediate mixture 3 of Example 1.

[0026] Figure 2 is a liquid chromatogram of the HSPC finished product of Example 1.

[0027] Figure 3 is a liquid chromatogram of the sample after hydrogenation reaction of Comparative Example 1.

[0028] Figure 4 is a liquid chromatogram of the HSPC finished product of Comparative Example 1.

[0029] Figure 5 is a liquid chromatogram of the HSPC finished product of Comparative Example 2. DETAILED DESCRIPTION

[0030] In order to make the purpose, technical scheme and advantages of the present application clearer, the present application will be further described in detail below with reference to the drawings.

[0031] Example 1

[0032] Take 200g of hydrogenated soybean oil (commercially available, purchased from Jiangxi Alpha High-tech Pharmaceutical Co., Ltd.), 10ml of sulfuric acid, 15g of sodium dodecyl sulfonate and 80g of water in a round-bottom flask, and stir at 105℃ for 16h to obtain intermediate mixture 1. Wash the intermediate mixture 1 with water until the pH is 6-7, then stand for separation to obtain 185g of intermediate mixture 2. Take 180g of DMAP, 85g of glycerylphosphorylcholine, 200g of DCC and 1.5L of ethyl acetate in a round-bottom flask, and react at 70℃ for 9h, then filter to obtain 573g of intermediate mixture 3. Dissolve 50g of iron sulfate in 200ml of pure water, add 200g of silica gel, stir at 60℃ for 4h, then filter and wash the residue with a small amount of water, and dry to obtain modified silica gel. Stir and adsorb the prepared modified silica gel with the intermediate mixture 3 at 55℃ for 2h, then filter and concentrate to obtain 348g of intermediate mixture 4. Stir and beat the solid in 3L of acetone for 1h, then filter to obtain white residue, repeat the beating step for 2 times, and dry under reduced pressure to obtain 213g of HSPC, with a purity of 99.23% detected by liquid chromatography, as shown in Figures 1 and 2.

[0033] Example 2

[0034] Take 200 g of hydrogenated soybean oil, 35 g of sodium hydroxide, 1.8 L of ethanol and 200 ml of water in a round-bottom flask, stir at 65°C for 2 h to obtain intermediate mixture 1, adjust the pH to 6-7 with citric acid, cool to 20°C to precipitate, centrifuge to obtain 162 g of intermediate mixture 2. Take 170 g of HOBT, 70 g of glycerophosphorylcholine, 160 g of EDCI and 3 L of chloroform in a round-bottom flask, react at 60°C for 16 h, filter to obtain 472 g of intermediate mixture 3. Dissolve 40 g of calcium chloride in 200 ml of pure water, add 150 g of silica gel to the mixture, stir at 80°C for 2 h, filter and wash the residue with a small amount of water, dry to obtain modified silica gel. Stir the modified silica gel with intermediate mixture 3 at 55°C for 3 h, filter and concentrate to obtain 302 g of intermediate mixture 4. Stir the solid in 2 L of ethyl acetate for 0.5 h, filter to obtain white residue, repeat the stirring step once, and dry under reduced pressure to obtain 196 g of HSPC with a purity of 98.85% detected by liquid chromatography.

[0035] Example 3

[0036] Take 200 g of hydrogenated soybean oil, 35 g of sodium hydroxide, 1.8 L of ethanol and 200 ml of water in a round-bottom flask, stir at 65°C for 2 h to obtain intermediate mixture 1, adjust the pH to 6-7 with citric acid, cool to 20°C to precipitate, centrifuge to obtain 162 g of intermediate mixture 2. Take 170 g of HOBT, 70 g of glycerophosphorylcholine, 160 g of EDCI and 3 L of chloroform in a round-bottom flask, react at 60°C for 16 h, filter to obtain 472 g of intermediate mixture 3. Dissolve 40 g of calcium chloride in 200 ml of pure water, add 150 g of silica gel to the mixture, stir at 80°C for 2 h, filter and wash the residue with a small amount of water, dry to obtain modified silica gel. Stir the modified silica gel with intermediate mixture 3 at 55°C for 3 h, filter and concentrate to obtain 302 g of intermediate mixture 4. Stir the solid in 2 L of ethyl acetate for 0.5 h, filter to obtain white residue, repeat the stirring step once, and dry under reduced pressure to obtain 196 g of HSPC with a purity of 98.85% detected by liquid chromatography.

[0037] Example 4

[0038] Take 50 kg of hydrogenated soybean oil, 10 kg of sodium hydroxide, 400 L of methanol and 150 L of water in the reaction tank, stir at 77°C for 6h to obtain intermediate mixture 1, adjust pH to 6-7 with hydrochloric acid, then cool to 20°C to crystallize, centrifuge to obtain 47 kg of intermediate mixture 2. Take 50 kg of HOBT, 20 kg of glycerophosphocholine, 50 kg of DCC and 1000 L of ethyl acetate in the reaction tank, react at 72°C for 10 hours, filter to obtain 128 kg of intermediate mixture 3. Dissolve 5 kg of zinc chloride in 100 L of pure water, add 50 kg of silica gel, stir at 65°C for 12h, then filter and wash the residue with a small amount of water, dry to obtain modified silica gel. Stir the prepared modified silica gel with intermediate mixture 3 at 62°C for 6h, filter and concentrate to obtain 69 kg of intermediate mixture 4. Add 310 L of acetone and stir for 0.5 hours, filter to obtain white residue, repeat the slurry step 3 times, and dry under reduced pressure to obtain 52 kg of HSPC, with a purity of 99.19% detected by liquid chromatography.

[0039] Comparative Example 1

[0040] Into the reaction kettle, add soybean phospholipid 125g, Pd / C (palladium on carbon, content 10%, water content about 60%) 50g, toluene 300ml, water 50ml and methanol 250ml, react under the condition of reaction temperature 50°C, pressure 0.8-1.3MPa. After 3 hours of reaction, discharge the material, filter the reaction solution to remove palladium on carbon, wash the filtrate with a mixture of 30ml toluene and 20ml methanol, then concentrate to dryness, add petroleum ether-acetone mixed solution 500ml to stir and wash again, filter, dry the filter cake to obtain 98g HSPC, with a purity of 95.62% detected by liquid chromatography, as shown in Figures 3 and 4.

[0041] Comparative Example 2

[0042] Add intermediate mixture 3 in Example 1 to silica gel without metal salt treatment, and the rest of the preparation process is consistent with Example 1. The final HSPC has a purity of 97.18% detected by liquid chromatography, as shown in Figure 5.

[0043] Effect Example 1

[0044] By comparing Example 1 and Comparative Example 1, it can be seen that the impurities after the reaction of Example 1 are unreacted fatty acids and LPC, which have a very different structure from HSPC and can be easily removed by simple slurry refining process. However, Comparative Example 1 produces some impurities that are very similar in structure to HSPC after hydrogenation, making subsequent refining difficult, with poor results and affecting the quality of HSPC. In addition, the raw material used in Example 1, hydrogenated soybean oil, is much cheaper than soybean phospholipid, and the preparation process does not require the use of hydrogen gas, which has safety hazards, making it more suitable for industrial production than Comparative Example 1.

[0045] By comparing Example 1 and Comparative Example 2, it can be seen that the modified silica gel after treatment of intermediate mixture 3 with metal salt adsorption is obviously better than direct adsorption with ordinary silica gel, and the purity of the finished product is increased by more than 2%.

[0046] The above disclosure is only the preferred embodiment of the present application, and of course cannot limit the scope of the patent rights of the present application, so the equivalent changes made according to the claims of the present application still fall within the scope of the present application.

Claims

1. A method for preparing hydrogenated soy phosphatidylcholine, characterized by, The method comprises the following steps: A. Hydrolyzing hydrogenated soybean oil to obtain intermediate mixture 1, and adjusting the pH of the intermediate mixture 1 to obtain intermediate mixture 2; B. Mixing the intermediate mixture 2 with glycerophosphocholine, a condensing agent, an acid-binding agent and a reaction solvent to perform an esterification reaction, and then performing solid-liquid separation to obtain intermediate mixture 3; C. Mixing and adsorbing the intermediate mixture 3 with an adsorbent, and then performing solid-liquid separation to obtain intermediate mixture 4; D. Mixing the intermediate mixture 4 with an organic solvent, performing beating and precipitation, and then performing solid-liquid separation and drying to obtain hydrogenated soybean phosphatidylcholine product.

2. The method for preparing hydrogenated soybean phosphatidylcholine according to claim 1, characterized in that, In step C, the adsorbent comprises modified silica gel; the modified silica gel comprises a mixture of silica gel and metal salt; the metal salt comprises one or more of transition metal elements of IB-VIIIB; the mass ratio of the intermediate mixture 3 to the adsorbent is 1:0.1-0.5; the temperature of the mixing and adsorbing is 40-70°C; the time of the mixing and adsorbing is 2-6 hours; the solid-liquid separation is performed by filtration; and the intermediate mixture 4 is obtained by further concentrating after the filtration.

3. The method for preparing hydrogenated soybean phosphatidylcholine according to claim 2, characterized in that, The preparation of the modified silica gel comprises mixing the silica gel and the metal salt in an aqueous solution, washing with water and drying to obtain the modified silica gel; the mixing of the silica gel and the metal salt is performed by stirring; the stirring temperature is 20-100°C; the stirring time is 0.5-24 hours; the mass ratio of the silica gel to the metal salt is 1:0.05-0.5; and the metal salt comprises one or more of iron salt, copper salt, zinc salt, cobalt salt, nickel salt, manganese salt, chromium salt and ruthenium salt.

4. The method for preparing hydrogenated soybean phosphatidylcholine according to claim 1, characterized in that, The operation of step A comprises one of the following: A1. Mixing the hydrogenated soybean oil with acid, sodium alkyl sulfonate and water to perform the hydrolysis reaction to generate the intermediate mixture 1, washing the intermediate mixture 1 with water to a pH of 6-7, and then standing and separating to obtain the intermediate mixture 2; Or A2. Mixing the hydrogenated soybean oil with inorganic base, lower alcohol and water to perform the hydrolysis reaction to generate the intermediate mixture 1, adding acid to adjust the pH to 6-7, and then cooling and crystallizing to obtain the intermediate mixture 2.

5. The method for preparing hydrogenated soybean phosphatidylcholine according to claim 4, characterized in that, In step A1, the mass ratio of the hydrogenated soybean oil to the acid, the sodium alkyl sulfonate, the water is 1:0.02-0.1:0.02-0.2:0.3-0.8; the reaction temperature of the hydrolysis reaction is 90-115°C; the reaction time of the hydrolysis reaction is 8-24 hours; and the acid comprises one or more of sulfuric acid, hydrochloric acid, nitric acid, acetic acid and citric acid.

6. The method for preparing hydrogenated soybean phosphatidylcholine according to claim 4, characterized in that, In step A2, the mass ratio of the hydrogenated soybean oil, the inorganic base, the lower alcohol, and the water includes 1:0.1-0.3:1-20:0.05-4; the reaction temperature of the hydrolysis reaction includes 50-100°C; the reaction time of the hydrolysis reaction includes 0.5-8 hours; the inorganic base includes one or more of sodium hydroxide, potassium hydroxide, sodium carbonate, and sodium bicarbonate; the lower alcohol includes one or more of methanol, ethanol, isopropanol, and n-butanol; and the acid includes one or more of sulfuric acid, hydrochloric acid, nitric acid, acetic acid, and citric acid.

7. The method for preparing hydrogenated soybean phosphatidylcholine according to claim 1, characterized in that, In step B, the weight ratio of the glycerophosphocholine, the intermediate mixture 2, the condensing agent, the acid binding agent, and the reaction solvent includes 1:2-3:2-3:2-3:1-90; the reaction time of the esterification reaction includes 4-24 hours; the reaction temperature of the esterification reaction includes 40-80°C; and the solid-liquid separation method includes filtration.

8. The method for preparing hydrogenated soybean phosphatidylcholine according to claim 7, characterized in that, The condensing agent includes one or more of dicyclohexyl carbodiimide, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N,N'-diisopropyl carbodiimide, 1-hydroxybenzotriazole, O-benzotriazol-N,N,N',N'-tetramethyluronium tetrafluoroborate, and 2-(7-azobenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate; the acid binding agent includes one or more of pyridine, 4-dimethylaminopyridine, triethylamine, N,N-diisopropylethylamine, and N-hydroxysuccinimide; and the reaction solvent includes one or more of n-hexane, dichloromethane, trichloromethane, dimethyl sulfoxide, and ethyl acetate.

9. The method for preparing hydrogenated soybean phosphatidylcholine according to claim 1, characterized in that, In step D, the mass ratio of the intermediate mixture 4 and the organic solvent includes 1:1-20; the beating and precipitation is repeated 1-4 times; the organic solvent includes one or more of n-hexane, octane, dichloromethane, trichloromethane, ethyl acetate, acetone, tetrahydrofuran, methyl tert-butyl ether, methanol, ethanol, and water; and the solid-liquid separation method includes filtration.

10. A hydrogenated soybean phosphatidylcholine product obtained by the method of claim 1.

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