CN110642711B - Method for separating alpha-linolenic acid from peony seed oil - Google Patents
Method for separating alpha-linolenic acid from peony seed oil Download PDFInfo
- Publication number
- CN110642711B CN110642711B CN201910831360.9A CN201910831360A CN110642711B CN 110642711 B CN110642711 B CN 110642711B CN 201910831360 A CN201910831360 A CN 201910831360A CN 110642711 B CN110642711 B CN 110642711B
- Authority
- CN
- China
- Prior art keywords
- temperature
- seed oil
- linolenic acid
- peony seed
- layer
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11C—FATTY ACIDS FROM FATS, OILS OR WAXES; CANDLES; FATS, OILS OR FATTY ACIDS BY CHEMICAL MODIFICATION OF FATS, OILS, OR FATTY ACIDS OBTAINED THEREFROM
- C11C1/00—Preparation of fatty acids from fats, fatty oils, or waxes; Refining the fatty acids
- C11C1/02—Preparation of fatty acids from fats, fatty oils, or waxes; Refining the fatty acids from fats or fatty oils
- C11C1/025—Preparation of fatty acids from fats, fatty oils, or waxes; Refining the fatty acids from fats or fatty oils by saponification and release of fatty acids
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
- C07C51/42—Separation; Purification; Stabilisation; Use of additives
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
- C07C51/42—Separation; Purification; Stabilisation; Use of additives
- C07C51/43—Separation; Purification; Stabilisation; Use of additives by change of the physical state, e.g. crystallisation
Abstract
The invention discloses a method for separating alpha-linolenic acid from peony seed oil. The method for separating alpha-linolenic acid from peony seed oil provided by the invention comprises the following steps: (1) primarily purifying the peony seed oil through saponification reaction to obtain a saponification reactant; (2) adding a solvent into the saponification reactant obtained in the step (1) to obtain a mixed solution; putting the mixed solution into a melting layer crystallizer with a double-layer jacket, and respectively controlling the temperature of the inner layer and the outer layer of the jacket, the cooling time and the maintaining time to crystallize the melting layer; (3) collecting mother liquor crystallized in the melting layer, and removing the solvent to obtain the alpha-linolenic acid. According to the method, unsaturated fatty acid with high solubility at low temperature is separated by utilizing a melting layer crystallization coupling mode according to different solubilities of mixed fatty acid in a solvent, so that the effect of separating alpha-linolenic acid in the mixed fatty acid is achieved.
Description
Technical Field
The invention relates to a preparation method of linolenic acid, in particular to a method for separating alpha-linolenic acid from peony seed oil.
Background
The peony seed oil is also called peony oil, and is woody nut plant oil extracted from peony seeds, the peony seed oil contains more than 100 kinds of nutrient substances with physiological activity, such as linolenic acid, oleic acid, linoleic acid, palmitic acid, stearic acid, paeonol, saponin, polysaccharide, squalene, VA, VE, carotene and the like, and the peony seed oil is rich in unsaturated fatty acid, wherein the content of alpha-linolenic acid reaches about 40%. Alpha-linolenic acid is an omega-3 type fatty acid containing 3 double bonds, can not be synthesized in a human body and is a necessary polyunsaturated fatty acid which needs to be taken from the outside, the alpha-linolenic acid can be converted into a series of metabolites including EPA and DHA by the human body, the EPA and the DHA have obvious physiological activity, are not only main components of brain tissues, but also have important effects on enhancing intelligence, improving immunity, protecting vision, reducing 'three highs', preventing cardiovascular and cerebrovascular diseases and the like, and are extremely important to the health of the human body.
The separation and purification method of alpha-linolenic acid is a hot point concerned by the food and medicine field, and the existing separation and purification methods of unsaturated fatty acid comprise a low-temperature crystallization method, a urea inclusion method, a silver ion complexation method, a column chromatography method and a molecular distillation method. The low temperature crystallization method is also called solvent fractionation method, and separation and purification are carried out according to different solubilities and freezing points of fatty acid or fatty acid ester in organic solvent at low temperature. In general, the solubility of fatty acids in organic solvents decreases with increasing carbon chain length, increases with increasing number of double bonds, and becomes more pronounced with decreasing temperature. Therefore, the mixed fatty acid is dissolved in the organic solvent under the low temperature state, and a large amount of undissolved saturated fatty acid and partial monounsaturated fatty acid are removed by filtration, thereby achieving the purpose of enriching and purifying linolenic acid. The alpha linolenic acid has a large number of double bonds and high solubility in organic solvents. The low-temperature crystallization method has simple process and convenient operation, and the active ingredients are not easy to generate denaturation reactions such as fusion, oxidation, isomerization and the like, so the mixed fatty acid is separated by adopting the cooling method more commonly. At present, the mixed fatty acid after low-temperature crystallization is separated by commonly using a filtering method and a centrifuging method, but the requirements of the filtering method and the centrifuging method on operation conditions are very strict, crystal melting possibly exists in the operation process, the operation temperature needs to be strictly controlled, otherwise, the yield of unsaturated fatty acid is influenced, and the separation efficiency is low.
The layer crystallization is a crystallization mode of forming a crystallization layer by slowly solidifying a crystallization material directly on a cooling interface, and is also called a gradual freezing method or a directional crystallization method.
Disclosure of Invention
In view of the above, the main objective of the present invention is to provide a method for separating α -linolenic acid from peony seed oil, which is helpful for separating unsaturated fatty acids with high solubility at low temperature, so as to achieve the effect of separating α -linolenic acid from mixed fatty acids.
In order to achieve the above object, according to one aspect of the present invention, a method for separating alpha-linolenic acid from peony seed oil is provided.
The method for separating alpha-linolenic acid from peony seed oil provided by the invention comprises the following steps:
(1) primarily purifying the peony seed oil through saponification reaction to obtain a saponification reactant;
(2) adding a solvent into the saponification reactant obtained in the step (1) to obtain a mixed solution; putting the mixed solution into a melting layer crystallizer with a double-layer jacket, and respectively controlling the temperature of the inner layer and the outer layer of the jacket, the cooling time and the maintaining time to crystallize the melting layer;
(3) collecting mother liquor crystallized in the melting layer, and removing the solvent to obtain the alpha-linolenic acid.
The saponification reaction is carried out by mixing peony seed oil with NaOH ethanol water solution.
The method further comprises the step of removing ethanol from the saponification reactant by distillation under reduced pressure before the step (2), and then acidifying and separating the oil phase.
The mass percentage concentration of NaOH in the NaOH ethanol water solution is 2.5-4.0%, and the volume percentage of the solvent ethanol water is 90-100% of ethanol water solution;
the volume ratio of the peony seed oil to the NaOH ethanol aqueous solution is 1:4 to 6.
The saponification reaction in the step (1) is carried out in a reaction tank capable of accurately controlling the temperature, the set temperature is 40-60 ℃ within 10-20 min, and the reaction is carried out for 1-3 h.
In the step (2), the temperature, the cooling time and the maintaining time of the inner layer and the outer layer of the control jacket are as follows: the temperature of the outer layer jacket is controlled to be reduced to-12 ℃ to-30 ℃ within 120min to 180min, the temperature of the inner layer jacket is controlled to be reduced to-15 ℃ to-30 ℃ within 120min to 180min, and the temperature is maintained for 4h to 5 h.
The dosage of the solvent in the step (2) is that the mass ratio of the mixed fatty acid to the solvent is 1: 0.5-3; the solvent is ethanol or acetone.
The step of acidifying and separating the oil phase comprises the steps of distilling the saponification reactant obtained in the step (1) under reduced pressure to remove ethanol, and then adding H2SO4Mixing and stirring, separating oil phase, washing with water to neutrality, and drying with anhydrous sodium sulfate to obtain mixed fatty acid;
said H2SO4Is H with the mass percentage concentration of 5-10 percent2SO4An aqueous solution; said saponification reactant with said H2SO4The volume ratio of the aqueous solution is 1: 2.5-7.0.
The solvent in the step (3) is removed by distilling the mother liquor under reduced pressure to remove the solvent;
the reduced pressure distillation conditions are as follows: distilling under reduced pressure for 10min to 30min under the conditions that the vacuum degree is-0.05 MPa to-0.1 MPa and the temperature is 45 ℃ to 55 ℃.
The invention also provides a melting layer crystallizer with a double-layer jacket.
The melting layer crystallizer with the double-layer jacket comprises a main crystallizer 3, wherein an inner-layer temperature control pipeline 6 is arranged inside the main crystallizer 3, an outer-layer temperature control pipeline 4 is arranged outside the main crystallizer 3, and a layer crystallization pipe 5 with a frosted outer surface is arranged on the outer wall of an inner crystallization pipe of the inner-layer temperature control pipeline 6; a mother liquor outlet 7 is arranged at the bottom of the crystallizer 3, the outer temperature control pipeline 4 is connected with the first temperature control water bath 1 through a pipeline, and the inner temperature control pipeline 6 is connected with the second temperature control water bath 2 through a pipeline.
Preferably, in the ethanol aqueous solution of NaOH in the step (1), the concentration of NaOH is 2.5-3.75% by mass percent, and the ethanol aqueous solution is 95% by volume percent.
Preferably, the saponification reaction in the step (1) is carried out at the temperature of 40-60 ℃ for 1-2 h, and the volume ratio of the peony seed oil to the NaOH ethanol aqueous solution is 1: 4.
mixing peony seed oil and an ethanol aqueous solution of sodium hydroxide for saponification reaction, removing ethanol through reduced pressure distillation, acidifying, separating liquid, washing and drying to obtain mixed fatty acid, adding the mixed fatty acid into an ethanol solvent with the dilution multiple of 0.5-3 for dilution, putting the diluted mixed fatty acid into a crystallization tank of a crystallization device with a double-layer jacket, performing melt layer crystallization, controlling the temperature of an outer-layer jacket to be reduced to-12 to-30 ℃ within 120-180 min, reducing the temperature of an inner-layer jacket to-15 to-30 ℃ within 120-180 min, and maintaining for 4h to enable the outer surface of the inner-layer jacket to grow a crystal layer; and (3) after collecting the mother liquor, removing the solvent in the mother liquor through reduced pressure distillation to obtain the high-concentration alpha-linolenic acid.
According to the method, unsaturated fatty acid with high solubility at low temperature is separated by utilizing a melting layer crystallization coupling mode according to different solubilities of mixed fatty acid in a solvent, so that the effect of separating alpha-linolenic acid in the mixed fatty acid is achieved. The invention simultaneously utilizes the melting point difference and solubility difference of the alpha linolenic acid and other mixed fatty acids, namely the alpha linolenic acid has the highest solubility in a target solvent and is not easy to separate out when being cooled, and simultaneously utilizes the lowest melting point of the alpha linolenic acid and is not easy to separate out when being cooled. The technology utilizes the principle that fatty acid crystals with low solubility and high melting point are directly solidified into a crystallization layer on a cooling interface by layer crystallization, avoids the phenomenon that the crystals are melted when mixed fatty acid is separated under the freezing condition, and greatly improves the yield of alpha-linolenic acid. Meanwhile, the melting layer crystallizer with a special structure is utilized to separate out and collect fatty acid with low solubility and high melting point, and the mother liquor enriches alpha linolenic acid to realize the separation of the alpha linolenic acid.
Drawings
For purposes of illustration and not limitation, the present invention will now be described in accordance with its preferred embodiments, particularly with reference to the accompanying drawings, in which:
FIG. 1 is a schematic view of a crystallization tank of a double-jacketed melt-layer crystallizer;
wherein, the method comprises the following steps of 1-first temperature control water bath, 2-second temperature control water bath, 3-main crystallizer, 4-outer layer temperature control pipeline and 5-layer crystallization pipe with frosted outer surface; 6-inner layer temperature control pipeline, 7-mother liquor outlet.
Detailed Description
Example 1 isolation of alpha-linolenic acid from peony seed oil
The method for separating alpha-linolenic acid from peony seed oil comprises the following steps:
(1) saponification: placing the peony seed oil in a reaction tank capable of accurately controlling the temperature, wherein the volume ratio of the peony seed oil to the NaOH ethanol water solution is 1:4, adding 2.5 percent of NaOH ethanol aqueous solution (ethanol water is 95 percent of ethanol solution by volume percent) by mass percentage, fully stirring and mixing, controlling the stirring speed at 150rpm, reaching the set temperature of 40 ℃ within 10min, and reacting for 3h to obtain the saponification reactant.
(2) Distilling the saponification reactant obtained in the step (1) under reduced pressure at a vacuum degree of-0.05 MPa and a temperature of 55 ℃ for 30min, removing ethanol, and adding 5 times of volume of H with a mass percent concentration of 5%2SO4Mixing the water solution, stirring, separating oil phase, washing with water for 3 times to neutrality, and drying with anhydrous sodium sulfate to obtain mixed fatty acid.
(3) Adding solvent ethanol with the volume 0.5 time that of the mixed fatty acid into the mixed fatty acid obtained in the step (2) to obtain mixed solution; putting the mixed solution into a crystallization tank of a melting layer crystallizer with a double-layer jacket, reducing the temperature of the jacket of the outer layer to-12 ℃ within 120min, reducing the temperature of the jacket of the inner layer to-15 ℃ within 120min, and maintaining for 5 h. Collecting mother liquor, wherein the mother liquor collection is completed once, and the collected mother liquor is subjected to reduced pressure distillation under the reduced pressure distillation conditions: vacuum distilling at 45 deg.C under-0.05 MPa for 30min, and removing ethanol to obtain 61 wt% alpha-linolenic acid.
As shown in fig. 1, the melt layer crystallizer with a double-layer jacket comprises a main crystallizer 3, an inner-layer temperature control pipeline 6 is arranged inside the main crystallizer 3, an outer-layer temperature control pipeline 4 is arranged outside the main crystallizer 3, and a layer crystallization pipe 5 with a frosted outer surface is arranged on the outer wall of a crystallization pipe in the inner-layer temperature control pipeline 6; a mother liquor outlet 7 is arranged at the bottom of the crystallizer 3, the outer temperature control pipeline 4 is connected with the first temperature control water bath 1 through a pipeline, and the inner temperature control pipeline 6 is connected with the second temperature control water bath 2 through a pipeline. And (3) putting the mixed solution obtained in the step (2) into a main crystallizer to crystallize a molten layer. Along with the temperature reduction of the outer layer pipeline and the inner layer temperature control pipeline, when the temperature is reduced to the crystallization temperature of the component with high melting point and low solubility, the component is firstly separated out and attached to the outer wall of the inner crystallization pipe of the inner layer temperature control pipeline 6 due to the temperature difference of the inner crystallization pipe and the outer crystallization pipe. The concentration and separation of the linolenic acid in the mixed solution are realized by controlling the temperature reduction rate and stopping the temperature difference between the inner pipe and the outer pipe.
And (3) measuring the content of alpha-linolenic acid: the alpha-linolenic acid content of the collected mother liquor was determined by gas chromatography-mass spectrometry (GC-MS).
The alpha-linolenic acid content determination method comprises the following specific steps:
methyl esterification of fatty acids:
1) the test tube was filled with 1.0mL of a methanol sulfate solution (5% sulfuric acid + 95% methanol), and sealed after being filled with nitrogen. 90 ℃ water
And bathing for 1 h.
2) After cooling at room temperature, 1.5mL of n-pentane and 1.0mL of water were added to the tube, and the mixture was shaken and mixed to terminate the methylation reaction.
3) After shaking at 2500rpm, the mixture was centrifuged for 10min, and 1.0mL of the supernatant was collected, and 5.0mL of redistilled n-hexane was added after concentration with nitrogen.
4) Adding 20 μ L of internal standard (heptadecanoic acid) into 980 μ L of extractive solution sample, rapidly mixing, filtering with 0.22um organic phase microporous filter, and bottling for GC-MS analysis.
Preparation of a standard substance: the standard included 5 major components of mixed fatty acids, palmitic acid, stearic acid, oleic acid, linoleic acid and alpha-linolenic acid (all available from Sigma-Aldrich, Shanghai, China, usa) with heptadecanoic acid (available from Sigma-Aldrich, Shanghai, China, usa) not present in peony seed oil as an internal standard. Preparing a series of standard solutions with different concentrations by using the standard substance, wherein the concentration gradient comprises 0.015625mg/mL, 0.03125mg/mL, 0.0625mg/mL, 0.125mg/mL, 0.25mg/mL, 0.5mg/mL and 1.0mg/mL, taking 980 mu L of the standard solutions with different concentrations, adding 20 mu L of the internal standard substance respectively, sealing in a sample bottle, and storing at-20 ℃ for later use.
GC-MS analysis of fatty acid methyl esters:
the analysis system for testing the standard substance and analyzing the sample is Agilent GC-MS, and the chromatographic column is HP-88 silicone
Column for chromatography of an alkane polymer, 30 m.times.0.25 mm, 0.20 μm, (Agilent, USA). The temperature ramp program used was as follows: keeping the temperature at 100 ℃ for 2min, then increasing the temperature to 230 ℃ at 15 ℃/min, keeping the temperature for 5min, feeding the sample at the temperature of a sample inlet of 250 ℃, shunting and feeding the sample, wherein the shunting ratio is 10:1, the sample feeding volume is 1 mu L, the carrier gas is high-purity helium gas, and the flow rate is 1.0 mL/min; the ionization mode is EI, the electron energy is 70eV, the transmission line temperature is 280 ℃, the ion source temperature is 230 ℃, the quadrupole rod temperature is 150 ℃, and the mass scanning range (m/z): 30-450 u, and performing searching and comparison and qualitative determination with a standard substance by using a NIST05 Library.
Quantitative analysis of fatty acids:
linear regression quantification was performed using an internal standard curve method. Firstly, a series of standard products added with internal standards are respectively tested, the concentration of the standard products is taken as the abscissa (x), the peak area ratio of the corresponding standard products to the internal standards is taken as the ordinate (y), linear regression is carried out, and a standard curve is made. And then measuring the ratio of the peak areas of the sample to be detected and the internal standard substance under the same chromatographic condition, and finding out the concentration of the component to be detected in the sample on a standard curve according to the ratio so as to calculate the content of the alpha linolenic acid in the sample of the component to be detected.
Example 2 isolation of alpha-linolenic acid from peony seed oil
The method for separating alpha-linolenic acid from peony seed oil comprises the following steps:
(1) saponification: placing the peony seed oil in a reaction tank capable of accurately controlling the temperature, wherein the volume ratio of the peony seed oil to the NaOH ethanol water solution is 1: and 5, adding a NaOH ethanol aqueous solution with the mass percentage concentration of 3% (the ethanol water is a 90% ethanol solution by volume percentage), fully stirring and mixing, controlling the stirring speed at 200rpm, reaching the set temperature of 60 ℃ within 15min, and carrying out saponification reaction for 1h to obtain a saponification reactant.
(2) Distilling the saponification reactant obtained in the step (1) under reduced pressure at the vacuum degree of-0.1 MPa and the temperature of 45 ℃ for 30min, removing ethanol, and adding 10% H with the mass percentage concentration of 3 times of the volume2SO4Mixing the aqueous solution, stirring, separating oil phase, washing with water for 3 times to neutral, and adding anhydrous sulfuric acidSodium is dried to obtain the mixed fatty acid.
(3) Adding solvent ethanol with the volume 1.5 times that of the mixed fatty acid into the mixed fatty acid obtained in the step (2) to obtain mixed solution; putting the mixed solution into a crystallization tank of a melting layer crystallizer with a double-layer jacket, reducing the temperature of the jacket at the outer layer to-18 ℃ within 140min, reducing the temperature of the jacket at the inner layer to-21 ℃ within 140min, and maintaining for 5 h. Collecting mother liquor, wherein the mother liquor collection is completed once, and the collected mother liquor is subjected to reduced pressure distillation under the reduced pressure distillation conditions: and (3) carrying out reduced pressure distillation for 20min under the conditions that the vacuum degree is-0.1 MPa and the temperature is 50 ℃, and removing ethanol to obtain the alpha-linolenic acid with the mass percentage content of 63%.
The crystallization tank of the double-jacketed melt layer crystallizer was the same as in example 1.
The measurement method of the content of alpha-linolenic acid was the same as in example 1.
Example 3 isolation of alpha-linolenic acid from peony seed oil
The method for separating alpha-linolenic acid from peony seed oil comprises the following steps:
(1) saponification: placing the peony seed oil in a reaction tank capable of accurately controlling the temperature, wherein the volume ratio of the peony seed oil to the NaOH ethanol water solution is 1: 6, adding a NaOH ethanol aqueous solution with the mass percentage concentration of 3.5% (the ethanol water is a 100% ethanol solution by volume percentage), fully stirring and mixing, controlling the stirring speed at 250rpm, reaching the set temperature of 50 ℃ within 20min, and reacting for 1.5h to obtain a saponification reactant.
(2) Distilling the saponification reactant obtained in the step (1) under reduced pressure at a vacuum degree of-0.1 MPa and a temperature of 55 ℃ for 30min to remove ethanol, and adding 10% H with a mass percentage concentration of 3 times of the volume of the saponification reactant2SO4Mixing the water solution, stirring, separating oil phase, washing with water for 3 times to neutrality, and drying with anhydrous sodium sulfate to obtain mixed fatty acid.
(3) Adding acetone which is a solvent and is 2 times of the volume of the mixed fatty acid into the mixed fatty acid obtained in the step (2) to obtain a mixed solution; putting the mixed solution into a crystallization tank of a melting layer crystallizer with a double-layer jacket, reducing the temperature of the jacket of the outer layer to-23 ℃ within 160min, reducing the temperature of the jacket of the inner layer to-26 ℃ within 160min, and maintaining for 4 h. Collecting mother liquor, wherein the mother liquor collection is completed once, and the collected mother liquor is subjected to reduced pressure distillation under the reduced pressure distillation conditions: and (3) carrying out reduced pressure distillation for 10min under the conditions that the vacuum degree is-0.05 MPa and the temperature is 45 ℃, and removing ethanol to obtain the alpha-linolenic acid with the mass percentage content of 62%.
The crystallization tank of the double-jacketed melt layer crystallizer was the same as in example 1.
The measurement method of the content of alpha-linolenic acid was the same as in example 1.
Example 4 isolation of alpha-linolenic acid from peony seed oil
The method for separating alpha-linolenic acid from peony seed oil comprises the following steps:
(1) saponification: placing the peony seed oil in a reaction tank capable of accurately controlling the temperature, wherein the volume ratio of the peony seed oil to the NaOH ethanol water solution is 1:4, adding a 3.75 percent NaOH ethanol water solution (ethanol water is a 95 percent ethanol solution by volume), fully stirring and mixing, controlling the stirring speed at 200rpm, reaching the set temperature of 50 ℃ within 15min, and reacting for 1h to obtain a saponification reactant.
(2) Distilling the saponification reactant obtained in the step (1) under reduced pressure at the vacuum degree of-0.1 MPa and the temperature of 45 ℃ for 10min, removing ethanol, and adding 2.5 times of H with the mass percentage concentration of 10%2SO4Mixing the water solution, stirring, separating oil phase, washing with water for 3 times to neutrality, and drying with anhydrous sodium sulfate to obtain mixed fatty acid.
(3) Adding solvent ethanol with the volume 3 times that of the mixed fatty acid into the mixed fatty acid obtained in the step (2) to obtain mixed solution; putting the mixed solution into a crystallization tank of a melting layer crystallizer with a double-layer jacket, reducing the temperature of the jacket at the outer layer to-30 ℃ within 180min, reducing the temperature of the jacket at the inner layer to-30 ℃ within 180min, and maintaining for 4 h. Collecting mother liquor, wherein the mother liquor collection is completed once, and the collected mother liquor is subjected to reduced pressure distillation under the reduced pressure distillation conditions: distilling under reduced pressure at 45 deg.C under-0.1 MPa for 10min, and removing ethanol to obtain 65% alpha-linolenic acid.
The crystallization tank of the double-jacketed melt layer crystallizer was the same as in example 1.
The measurement method of the content of alpha-linolenic acid was the same as in example 1.
Example 5 isolation of alpha-linolenic acid from peony seed oil
The method for separating alpha-linolenic acid from peony seed oil comprises the following steps:
(1) saponification: placing the peony seed oil in a reaction tank capable of accurately controlling the temperature, wherein the volume ratio of the peony seed oil to the NaOH ethanol water solution is 1:4, adding a 3.5 percent NaOH ethanol water solution (ethanol water is a 95 percent ethanol solution by volume), fully stirring and mixing, controlling the stirring speed at 250rpm, reaching the set temperature of 60 ℃ within 20min, and reacting for 1h to obtain a saponification reactant.
(2) Distilling the saponification reactant obtained in the step (1) under reduced pressure at 45 ℃ and vacuum degree of-0.05 MPa for 10min to remove ethanol, and adding 7 times of H with the mass percent concentration of 5%2SO4Mixing the water solution, stirring, separating oil phase, washing with water for 3 times to neutrality, and drying with anhydrous sodium sulfate to obtain mixed fatty acid.
(3) Adding solvent ethanol with the volume 3 times that of the mixed fatty acid into the mixed fatty acid obtained in the step (2) to obtain mixed solution; putting the mixed solution into a crystallization tank of a melting layer crystallizer with a double-layer jacket, reducing the temperature of the jacket at the outer layer to-27 ℃ within 180min, reducing the temperature of the jacket at the inner layer to-30 ℃ within 180min, and maintaining for 4 h. Collecting mother liquor, wherein the mother liquor collection is completed once, and the collected mother liquor is subjected to reduced pressure distillation under the reduced pressure distillation conditions: and (3) carrying out reduced pressure distillation for 10min under the conditions that the vacuum degree is-0.05 MPa and the temperature is 45 ℃, and removing ethanol to obtain the alpha-linolenic acid with the mass percentage content of 65%.
The crystallization tank of the double-jacketed melt layer crystallizer was the same as in example 1.
The measurement method of the content of alpha-linolenic acid was the same as in example 1.
The above-described embodiments should not be construed as limiting the scope of the invention. Those skilled in the art will appreciate that various modifications, combinations, sub-combinations, and substitutions can occur, depending on design requirements and other factors. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (5)
1. The method for separating alpha-linolenic acid from peony seed oil comprises the following steps:
(1) primarily purifying the peony seed oil through saponification reaction to obtain a saponification reactant; the saponification reaction is carried out in a reaction tank capable of accurately controlling the temperature, the set temperature is 40-60 ℃ within 10-20 min, and the reaction is carried out for 1-3 h;
(2) removing ethanol in the saponification reactant by reduced pressure distillation, and then adding H with the mass percent concentration of 5-10%2SO4Mixing and stirring the aqueous solution, separating an oil phase, washing the oil phase to be neutral, and drying the oil phase by using anhydrous sodium sulfate to obtain mixed fatty acid; said saponification reactant with said H2SO4The volume ratio of the aqueous solution is 1: 2.5-7.0;
(3) adding a solvent into the mixed fatty acid obtained in the step (2) to obtain a mixed solution; putting the mixed solution into a melting layer crystallizer with a double-layer jacket, and respectively controlling the temperature of the inner layer and the outer layer of the jacket, the cooling time and the maintaining time to crystallize the melting layer; the temperature, the cooling time and the maintaining time of the inner layer and the outer layer of the control jacket are as follows: the temperature of the outer layer jacket is controlled to be reduced to-12 ℃ to-30 ℃ within 120min to 180min, the temperature of the inner layer jacket is reduced to-15 ℃ to-30 ℃ within 120min to 180min, and the temperature is maintained for 4h to 5 h;
(4) collecting mother liquor crystallized in the melting layer, and removing the solvent to obtain the alpha-linolenic acid.
2. The method for separating alpha-linolenic acid from peony seed oil as claimed in claim 1, wherein: the saponification reaction is carried out by mixing peony seed oil with NaOH ethanol water solution.
3. The method for separating alpha-linolenic acid from peony seed oil as claimed in claim 2, wherein:
the mass percentage concentration of NaOH in the NaOH ethanol water solution is 2.5-4.0%, and the volume percentage of the solvent ethanol water is 90-100% of ethanol water solution;
the volume ratio of the peony seed oil to the NaOH ethanol water solution is 1: 4-6.
4. The method for separating alpha-linolenic acid from peony seed oil as claimed in claim 1, wherein: the dosage of the solvent in the step (3) is that the mass ratio of the mixed fatty acid to the solvent is 1: 0.5-3; the solvent is ethanol or acetone.
5. The method for separating alpha-linolenic acid from peony seed oil as claimed in claim 1, wherein:
the solvent in the step (4) is removed by distilling the mother liquor under reduced pressure to remove the solvent;
the reduced pressure distillation conditions are as follows: distilling under reduced pressure for 10min to 30min under the conditions that the vacuum degree is-0.05 MPa to-0.1 MPa and the temperature is 45 ℃ to 55 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910831360.9A CN110642711B (en) | 2019-09-03 | 2019-09-03 | Method for separating alpha-linolenic acid from peony seed oil |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910831360.9A CN110642711B (en) | 2019-09-03 | 2019-09-03 | Method for separating alpha-linolenic acid from peony seed oil |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110642711A CN110642711A (en) | 2020-01-03 |
| CN110642711B true CN110642711B (en) | 2020-09-22 |
Family
ID=68991521
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910831360.9A Active CN110642711B (en) | 2019-09-03 | 2019-09-03 | Method for separating alpha-linolenic acid from peony seed oil |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110642711B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112255328A (en) * | 2020-09-21 | 2021-01-22 | 河南师范大学 | Optimized method for measuring fatty acids in different tissues of fish body |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102311882A (en) * | 2011-08-11 | 2012-01-11 | 青岛农业大学 | Extraction method for unsaturated fatty acid |
| CN103771374A (en) * | 2013-12-30 | 2014-05-07 | 广西明利化工有限公司 | Electronic grade phosphoric acid clapboard crystallizing device |
| CN104212636A (en) * | 2014-09-02 | 2014-12-17 | 天津大学 | Method for separating milk fat based on melting layer crystallizing technology |
| CN104418726A (en) * | 2013-08-20 | 2015-03-18 | 陕西中鼎华盛农业发展有限公司 | Extraction method of [alpha]-linolenic acid |
| CN110054560A (en) * | 2019-05-23 | 2019-07-26 | 河南农业大学 | A kind of method of purification and its application of pharmaceutical woody alpha-linolenic acid |
-
2019
- 2019-09-03 CN CN201910831360.9A patent/CN110642711B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102311882A (en) * | 2011-08-11 | 2012-01-11 | 青岛农业大学 | Extraction method for unsaturated fatty acid |
| CN104418726A (en) * | 2013-08-20 | 2015-03-18 | 陕西中鼎华盛农业发展有限公司 | Extraction method of [alpha]-linolenic acid |
| CN103771374A (en) * | 2013-12-30 | 2014-05-07 | 广西明利化工有限公司 | Electronic grade phosphoric acid clapboard crystallizing device |
| CN104212636A (en) * | 2014-09-02 | 2014-12-17 | 天津大学 | Method for separating milk fat based on melting layer crystallizing technology |
| CN110054560A (en) * | 2019-05-23 | 2019-07-26 | 河南农业大学 | A kind of method of purification and its application of pharmaceutical woody alpha-linolenic acid |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110642711A (en) | 2020-01-03 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP1211304B1 (en) | Method for isolating high-purified unsaturated fatty acids using crystallization | |
| JP6465938B2 (en) | Method for producing highly unsaturated fatty acid alkyl ester-containing composition | |
| CN105462687B (en) | Method for purifying olive oil by adopting mixed adsorbent | |
| KR100972703B1 (en) | Improvements in or relating to separation technology | |
| KR20140113921A (en) | Compositions comprising 20-carbon fatty acids and methods of making and using same | |
| CN110642711B (en) | Method for separating alpha-linolenic acid from peony seed oil | |
| WO2016041506A1 (en) | Method for preparing high-content conjugated linoleic acid through purification of vegetable oil as raw material | |
| CN103483305B (en) | Method for gathering/recovering VE (Vitamins E), squalene and polyunsaturated fatty acids from deodorized distillate of plant oil | |
| CN105132189B (en) | A kind of fine separation method of C18 series and C20~C22 series fatty acid methyl esters | |
| Gunstone et al. | Improved procedures for the isolation of pure oleic, linoleic, and linolenic acids or their methyl esters from natural sources | |
| KR20160096110A (en) | Method for chromatographic purification of a fatty acid | |
| CN102030629B (en) | Alpha linolenic acid cryogenic separation and purification method | |
| CN105753689A (en) | Method of extracting purified Alpha-linolenic acid from linseed oil | |
| CN103467432A (en) | Method for extracting vitamin E from deodorizer distillate of idesia polycarpa oil | |
| EP3208257B1 (en) | Method for recycling urea in urea adduct process | |
| CN108164415B (en) | Method for completely separating EPA and DHA from fish oil | |
| CN110590545A (en) | Method for completely separating oleic acid and linoleic acid | |
| US11535583B1 (en) | Preparation method of eicosapentaenoic acid ethyl ester | |
| CN106892791A (en) | Extract method and the medicine containing squalene of plant source spiny dogfish ene compositions and its preparation method and application | |
| CN104557542B (en) | A kind of Supercritical fluid chromatography prepares the method for high purity EPA ester and DHA ester monomer | |
| CN108191638B (en) | Method for purifying GLA in algae oil by using urea | |
| CN107056605B (en) | Method for separating unsaturated fatty acid and phytosterol from gardner seed oil | |
| CN106316851B (en) | A kind of new method of methyl chloroacetate purification | |
| CN110845300A (en) | Method for separating high-purity α -linolenic acid from peony seed oil by utilizing column chromatography | |
| CN106890200A (en) | Extract method and the medicine containing squalene of plant source spiny dogfish ene compositions and its preparation method and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20201217 Address after: 100093 Beijing City, Haidian District Xiangshan Nanxin Village No. 20 Patentee after: Institute of Botany of the Chinese Academy of Sciences Patentee after: Shandong Sijiyuan peony Biotechnology Co.,Ltd. Address before: 100093 Beijing City, Haidian District Xiangshan Nanxin Village No. 20 Patentee before: Institute of Botany of the Chinese Academy of Sciences |