CN109880858B - Method for reducing content of free fatty acid in marine phospholipid - Google Patents
Method for reducing content of free fatty acid in marine phospholipid Download PDFInfo
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Abstract
The invention discloses a method for reducing the content of free fatty acid in marine phospholipid. The method for reducing the content of free fatty acid in marine phospholipid comprises the step of converting free polyunsaturated fatty acid containing three or more double bonds in crude marine phospholipid into phospholipid. The method for reducing the content of free fatty acid in the marine phospholipid improves the content of phospholipid while reducing the acid value, ensures the content of polyunsaturated fatty acid such as EPA, DHA and the like, reduces the loss of components such as fat-soluble vitamins and the like, has high separation degree, does not damage the structure of phospholipid, and improves the quality of products.
Description
Technical Field
The invention relates to a method for reducing the content of free fatty acid in marine phospholipid.
Background
A large number of researches at home and abroad prove that the marine phospholipid has safe and reliable effects in the aspects of adjuvant therapy of cardiovascular and cerebrovascular diseases and diabetes, reduction of arthritis symptoms, alleviation of menstrual period syndromes and dysmenorrheal, antioxidation and the like, and has good development prospects in the aspect of development of functional nutritional food industries. Research shows that the marine phospholipid contains rich n-3 polyunsaturated fatty acids such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), and rich fat-soluble components such as vitamin A, vitamin E, astaxanthin and the like, wherein the EPA, the DHA and the like are combined with the phospholipid through chemical bonds. In view of the influence of the freshness of raw materials on the quality of products, the content of free fatty acid in the crude marine phospholipid is generally high, the acid value is more than 10mg/g (even as high as 60mg/g), and the free fatty acid needs to be further removed to be used as a food raw material.
Currently, in the grease industry, the methods for removing free fatty acid mainly include the following methods: (1) and (3) neutralization deacidification method: the method can cause saponification of phospholipid and glyceride in marine phospholipid while reducing acid value by using sodium hydroxide to neutralize free fatty acid in grease, and the sodium fatty acid generated by neutralization and saponification can be used as an emulsifier to emulsify a reaction system, so that oil-water separation is difficult. (2) Molecular distillation method: the method realizes substance separation by means of the difference of the mean free path of motion of different substance molecules, but the molecular distillation temperature is usually higher (more than 160 ℃), the structure of functional phospholipid is easy to be damaged by continuous high temperature, and the method is not suitable for deacidification of marine phospholipid. (3) The supercritical fluid extraction deacidification method comprises the following steps: the method has certain effect, but has large equipment investment and high process cost, and can remove fat-soluble vitamins, astaxanthin, triglyceride and other components in the marine phospholipid while removing free fatty acid. (4) The Zhoudaiyong, etc. of Dalian Industrial university has developed a method for deacidifying acetone solvent, which can effectively reduce the content of free fatty acid in marine phospholipid, but can also remove fat-soluble vitamins, astaxanthin and triglyceride in marine phospholipid, resulting in poor fluidity of marine phospholipid and loss of nutrient components. In addition, since free fatty acids of marine phospholipids also contain a certain amount of EPA and DHA, the above-mentioned various deacidification methods also result in the loss of n-3 polyunsaturated fatty acids.
Disclosure of Invention
In order to make up for the defects of the prior art, the invention provides a method for reducing the content of free fatty acid in marine phospholipid, which comprises the steps of firstly adopting a urea inclusion method to primarily remove free saturated fatty acid and low unsaturated fatty acid in crude marine phospholipid, and then catalyzing the free polyunsaturated fatty acid and carrying out ester synthesis reaction by phospholipase or lipase to convert the free polyunsaturated fatty acid into phospholipid, so that the acid value is reduced, the phospholipid content is increased, the product quality is improved, and the loss rate of the product is reduced.
The invention is realized by the following technical scheme:
a method of reducing the free fatty acid content of marine phospholipids, said method comprising the step of converting free polyunsaturated fatty acids containing three and more double bonds in marine phospholipids into phospholipids.
The polyunsaturated fatty acid is converted into phospholipid through reaction with glycerol phosphoryl substances, and the glycerol phosphoryl substances are any one or more of glycerol phosphorylcholine, glycerol phosphorylinositol, glycerol phosphorylethanolamine or glycerol phosphorylserine.
The reaction comprises the following operation steps: firstly, dissolving crude marine phospholipid in a first solvent, then adding a proper amount of glycerophosphorylcholine and an enzyme preparation catalyst, uniformly mixing, reacting, and removing the first solvent and the enzyme preparation catalyst after the reaction is finished to obtain a marine phospholipid product.
Preferably, the first solvent is any one or more of n-hexane, chloroform, dichloromethane, tert-butanol, diethyl ether, ethyl acetate or petroleum ether.
Preferably, the enzyme preparation catalyst is a phospholipase or a lipase, including but not limited to phospholipase A1, phospholipase A2, and lipases from Candida antarctica, Rhizomucor miehei, Rhizopus arrhizus.
Preferably, the addition amount of the glycerophosphoryl substance is 10-50% of the mass of the marine phospholipid, the addition amount of the enzyme preparation catalyst is 5-20% of the mass of the marine phospholipid, the addition amount of the first solvent is 1-10 times of the mass of the marine phospholipid, the reaction time is 2-12h, and the reaction temperature is 30-70 ℃.
In order to avoid the decrease of the content of EPA and DHA caused by the reaction of saturated fatty acid and low unsaturated fatty acid in the crude marine phospholipid with glycerophosphorylcholine and the like, the method for decreasing the content of free fatty acid in the marine phospholipid preferably further comprises the step of deacidifying the marine phospholipid before the ester synthesis reaction, wherein the deacidification is mainly used for removing free saturated fatty acid and low unsaturated fatty acid containing one or two double bonds in the marine phospholipid.
The deacidification method is a urea inclusion method and comprises the following specific steps:
dissolving urea and marine phospholipid in a second solvent according to a certain proportion at a first temperature to obtain a first mixed solution, cooling the first mixed solution to a second temperature to precipitate urea crystals, separating the urea crystals to obtain a second mixed solution, and removing the second solvent from the second mixed solution to obtain primarily deacidified marine phospholipid;
in order to obtain purified marine phospholipid as much as possible, after the second solvent in the second mixed solution is removed, the obtained marine phospholipid is concentrated to further separate out urea crystals, and then the urea crystals are separated out, and the second solvent is removed to obtain the second marine phospholipid which is deacidified again.
Preferably, the mass ratio of the second solvent to the urea and the marine phospholipid is 10 (0.1-2) to (0.1-4), the second solvent is methanol or ethanol with the volume concentration of 70-100%, the first temperature is 70-80 ℃, and the second temperature is below 20 ℃.
The invention has the beneficial technical effects that:
according to the method for reducing the content of free fatty acid in marine phospholipid provided by the invention, the free polyunsaturated fatty acid is catalyzed by phospholipase or lipase to perform ester synthesis reaction with glycerophosphorylcholine, oleophosphorylinositol, glycerophosphorylethanolamine or glycerophosphorylserine to convert the free polyunsaturated fatty acid into phospholipid, so that the acid value of the product is reduced, and the content of phospholipid is increased.
In order to avoid the reduction of the content of EPA and DHA caused by the reaction of saturated fatty acid, low unsaturated fatty acid and glycerophosphorylcholine in the crude marine phospholipid, a urea inclusion method is adopted to preliminarily remove free saturated fatty acid and low unsaturated fatty acid in the crude marine phospholipid before the ester synthesis reaction, in the urea inclusion process, the linear chain or saturated fatty acid and low unsaturated fatty acid which are close to the linear chain easily enter a urea crystal pipeline to form a urea inclusion compound to be crystallized out of a solution, and the polyunsaturated fatty acid molecules with more double bonds have larger volume and are difficult to enter the crystal pipeline, so that the saturated fatty acid and the low unsaturated fatty acid can be removed.
The method of the invention reduces the acid value, improves the content of phospholipid, ensures the content of polyunsaturated fatty acid such as EPA, DHA and the like, reduces the loss of components such as fat-soluble vitamin, has high separation degree, does not damage the structure of phospholipid, and improves the quality of products.
The specific implementation mode is as follows:
the present invention will be further illustrated with reference to the following examples, but the present invention is not limited thereto, and the preparation methods in the examples are all conventional ones and will not be described in detail.
Example 1:
weighing 40g of herring lecithin (acid value 42.1mg/g, phospholipid content 70.1%), 100g of 70% ethanol and 20g of urea, mixing, filling nitrogen, sealing, heating in a water bath at 70 ℃, magnetically stirring, and completely dissolving to obtain a uniform and transparent mixed solution; placing the mixed solution in a cold water bath at 5 ℃ to reduce the temperature of the mixed solution to below 20 ℃ and separate out urea crystals, and then filtering or centrifuging to obtain a clear marine phospholipid ethanol solution; concentrating the marine phospholipid ethanol solution to 30% of the original volume under vacuum condition, filtering or centrifuging again to obtain clear marine phospholipid ethanol concentrated solution, removing ethanol to obtain marine phospholipid with primarily reduced acid value, and detecting that the acid value is 12.4 mg/g.
Dissolving 10g of marine phospholipid with the primarily reduced acid value in 50g of tert-butyl alcohol, preheating to 60 ℃, adding 1.0g of candida antarctica lipase B and 1.0g of glycerophosphorylcholine, filling nitrogen for sealing reaction for 4 hours, centrifuging to remove the lipase, and removing the tert-butyl alcohol by vacuum distillation of a tert-butyl alcohol solution to finally obtain a marine phospholipid product with the acid value of 1.2mg/g, wherein the phospholipid content is 98.42% by detection.
Example 2:
weighing 1g of herring lecithin (acid value 42.1mg/g, phospholipid content 70.1%), 100g of 100% ethanol and 1g of urea, mixing, introducing nitrogen gas, sealing, heating in water bath at 80 deg.C, and magnetically stirring for dissolving completely to obtain uniform and transparent mixed solution; placing the mixed solution in a cold water bath at 5 ℃ to reduce the temperature of the mixed solution to below 20 ℃ and separate out urea crystals, and then filtering or centrifuging to obtain a clear marine phospholipid ethanol solution; concentrating the marine phospholipid ethanol solution to 30% of the original volume under vacuum condition, filtering or centrifuging again to obtain clear marine phospholipid ethanol concentrated solution, and removing ethanol to obtain marine phospholipid with primarily reduced acid value, wherein the acid value is 16.5mg/g by detection.
Dissolving 10g of marine phospholipid with the primarily reduced acid value in 100g of tertiary butanol, preheating to 70 ℃, adding 12.0 g of phospholipase A and 2.0g of glycerophosphoryl inositol, filling nitrogen, sealing, reacting for 12h, centrifuging to remove lipase, distilling the tertiary butanol solution under reduced pressure to remove the tertiary butanol, and finally obtaining a marine phospholipid product with the acid value of 4.9mg/g, wherein the phospholipid content is 95.33% by detection.
Example 3:
weighing 20g of herring lecithin (acid value 42.1mg/g, phospholipid content 70.1%), 100g of 95% ethanol and 5g of urea, mixing, introducing nitrogen gas, sealing, heating in 80 deg.C water bath, and magnetically stirring for dissolving completely to obtain uniform and transparent mixed solution; placing the mixed solution in a cold water bath at 5 ℃ to reduce the temperature of the mixed solution to below 20 ℃ and separate out urea crystals, and then filtering or centrifuging to obtain a clear marine phospholipid ethanol solution; concentrating the marine phospholipid ethanol solution to 30% of the original volume under vacuum condition, filtering or centrifuging again to obtain clear marine phospholipid ethanol concentrated solution, removing ethanol to obtain marine phospholipid with primarily reduced acid value, and detecting that the acid value is 7.4 mg/g.
Dissolving 10g of marine phospholipid with the primarily reduced acid value in 100g of n-hexane, preheating to 50 ℃, adding 20.5 g of phospholipase A and 2.0g of glycerol phosphoethanolamine, filling nitrogen, sealing, reacting for 12h, centrifuging to remove lipase, distilling the tertiary butanol solution under reduced pressure to remove the tertiary butanol, and finally obtaining a marine phospholipid product with the acid value of 2.2mg/g, wherein the phospholipid content is 97.81% by detection.
Example 4:
weighing 30g of herring lecithin (acid value 42.1mg/g, phospholipid content 70.1%), 100g of 95% ethanol and 10g of urea, mixing, introducing nitrogen gas, sealing, heating in 80 deg.C water bath, and magnetically stirring for dissolving completely to obtain uniform and transparent mixed solution; placing the mixed solution in a cold water bath at 5 ℃ to reduce the temperature of the mixed solution to below 20 ℃ and separate out urea crystals, and then filtering or centrifuging to obtain a clear marine phospholipid ethanol solution; concentrating the marine phospholipid ethanol solution to 30% of the original volume under vacuum condition, filtering or centrifuging again to obtain clear marine phospholipid ethanol concentrated solution, removing ethanol to obtain marine phospholipid product with primarily reduced acid value, and detecting that the acid value is 9.2 mg/g.
Dissolving 10g of marine phospholipid with the primarily reduced acid value in 100g of marine phospholipid, preheating to 50 ℃, adding 22.0g of phospholipase A and 2.0g of glycerophosphorylserine, filling nitrogen, sealing, reacting for 12h, centrifuging to remove lipase, and distilling the tertiary butanol solution under reduced pressure to remove the tertiary butanol, thereby finally obtaining a marine phospholipid product with the acid value of 2.2mg/g, wherein the phospholipid content is 98.01% by detection.
Finally, it should be noted that the embodiments are only preferred embodiments of the present invention, and are not intended to limit the present invention, and although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications and equivalents may be made to the technical solutions described in the foregoing embodiments, or some technical features may be substituted. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (8)
1. A method for reducing the content of free fatty acid in marine phospholipid is characterized in that the marine phospholipid is deacidified by a urea inclusion method to obtain deacidified marine phospholipid; the deacidified marine phospholipid is converted into phospholipid through reaction with glycerol phosphoryl substances under the action of a catalyst, wherein the glycerol phosphoryl substances are any one or more of glycerol phosphorylcholine, glycerol phosphorylinositol, glycerol phosphorylethanolamine or glycerol phosphorylserine, and the catalyst is phospholipase or lipase.
2. The method for reducing the content of free fatty acids in marine phospholipids as claimed in claim 1, characterized in that the reaction is carried out by the following steps: firstly, dissolving crude marine phospholipid in a first solvent, then adding a proper amount of glycerophosphoryl substance and an enzyme preparation catalyst, uniformly mixing, reacting, and removing the first solvent and the enzyme preparation catalyst after the reaction is finished to obtain a marine phospholipid product.
3. The method for reducing the content of free fatty acids in marine phospholipids according to claim 2, characterized in that the first solvent is any one or more of n-hexane, chloroform, dichloromethane, tert-butanol, diethyl ether, ethyl acetate or petroleum ether.
4. The method of claim 1, wherein the phospholipase or lipase comprises phospholipase A1, phospholipase A2, and lipase derived from Candida antarctica, Rhizomucor miehei, Rhizopus arrhizus.
5. The method for reducing the content of free fatty acids in marine phospholipids as claimed in claim 2, wherein the addition amount of glycerophosphoryl substances is 10% -50% of the mass of marine phospholipids, the addition amount of enzyme preparation catalysts is 5% -20% of the mass of marine phospholipids, the addition amount of first solvents is 1-10 times of the mass of marine phospholipids, the reaction time is 2-12h, and the reaction temperature is 30-70 ℃.
6. The method for reducing the content of free fatty acids in marine phospholipids as claimed in claim 1, wherein the deacidification method is a urea-coating method, comprising the following steps:
dissolving urea and marine phospholipid in a second solvent according to a certain proportion at a first temperature to obtain a first mixed solution, cooling the first mixed solution to a second temperature to precipitate urea crystals, separating the urea crystals to obtain a second mixed solution, and removing the second solvent from the second mixed solution to obtain the deacidified marine phospholipid.
7. The method of claim 6, wherein the marine phospholipid obtained after removing the second solvent from the second mixed solution is concentrated to separate out urea crystals, and then the urea crystals are separated and the second solvent is removed to obtain the second marine phospholipid which is deacidified again.
8. The method of claim 6, wherein the mass ratio of the second solvent to urea to marine phospholipids is 10 (0.1-2) to (0.1-4), the second solvent is 70-100% methanol or ethanol by volume, the first temperature is 70-80 ℃, and the second temperature is 20 ℃ or lower.
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| CN101701229A (en) * | 2009-11-17 | 2010-05-05 | 广州海莎生物科技有限公司 | Method for preparing texture phospholipid and lecithin |
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