CN110790626B - Method for extracting squalene from rapeseed oil - Google Patents

Abstract

The invention relates to the technical field of squalene extraction, in particular to a method for extracting squalene from rapeseed oil, comprising the following steps: (1) adding an aqueous solution containing compound enzymes to the rapeseed oil, and then placing it in a pulsed direct current electric field (2) adding a microcrystalline fiber solution to the resultant of step (1) and mixing, then adding a crystallizing agent and mixing under the condition of 50-55° C., heat preservation treatment, filtering, and removing the filtrate; (3) adding a crystallizing agent to step ( 2) add ethanol in the resultant, add L-vitamin C simultaneously, be placed in a constant temperature water bath at 60-65 ℃ and reflux, cooled, washed with water until the washing solution is neutral, and distilled under vacuum conditions; squalene in the present invention The extraction rate is high, the reagent residue is less, the purity of squalene is improved, and the extraction cost is reduced.

Description

A kind of extraction method of squalene in rapeseed oil

technical field

The invention relates to the technical field of squalene extraction, in particular to a method for extracting squalene from rapeseed oil.

Background technique

Rapeseed oil, also known as rapeseed oil or coriander oil, is also known as clear oil in parts of Hunan. It is widely used in people's daily life because of its golden color and fragrant smell. It is one of the main edible oils at home and abroad. The content of unsaturated fatty acids in rapeseed oil is as high as 85%, and it is rich in oleic acid, linoleic acid, linolenic acid and phospholipids. The digestion and absorption rate of rapeseed oil is as high as 99%. Rapeseed oil is rich in vitamin E and polyphenols. , squalene, sterols and other biologically active ingredients.

Squalene is a highly unsaturated hydrocarbon compound, a lipid unsaponifiable, its chemical name is 2,6,10,15,19,23-hexamethyl-2,6,10,14,18 ,22-Tetracosahexaene, is an open-chain triterpenoid compound, which can improve the activity of superoxide dismutase (SOD) in vivo, enhance immunity, improve sexual function, anti-aging, anti-fatigue, anti-tumor It is widely used in medicine, food, cosmetics, chemical industry and other fields.

For example, patent document CN201510956808.1 discloses a method for extracting a plant-derived squalene composition, a preparation method for a squalene-containing medicine, a squalene-containing medicine prepared by the method, and applications thereof. The method for extracting a plant-derived squalene composition includes: sequentially performing pretreatment and enrichment separation on a squalene-rich raw material, wherein the conditions for the enrichment and separation are controlled so that the obtained plant-derived squalene composition contains Containing 35-80 wt% squalene, the squalene-rich feedstock contains 5-30 wt% squalene and 10-40 wt% sterol.

Another example is the method for extracting squalene with Camellia oleifera seeds as a raw material disclosed in patent document CN201110027119.4. The technological process of its extraction method is: material preparation, pulverization, extraction, concentration, methyl esterification reaction, complexing agent-borax reaction, Macroporous resin chromatography separation and purification, concentration, extraction, washing, concentration, obtaining crude squalene oil, supercritical carbon dioxide extraction, namely obtaining squalene essential oil with squalene content ≥ 80%; its tea oil squalene soft capsule It is made from the following raw materials by weight percentage: squalene essential oil 87-90 with squalene content ≥80%, polyethylene glycol 4003-5, beeswax 2-3 and phospholipid 2-5.

In the document "Effect of Refining on the Quality of Rapeseed Oil" (published by Ma Qibing et al. in 2018), it is also mentioned that "the acid value, peroxide value, color and phosphorus content of refined rapeseed oil all decreased, VE, sterol , polyphenols, and squalene content were significantly reduced; trans fatty acid and 3-chloropropanol content did not change much", so this application, in order to prevent the reduction of squalene content and the reduction of squalene utilization value, squalene was carried out. extract.

SUMMARY OF THE INVENTION

In order to solve the above technical problems, the present invention provides a method for extracting squalene from rapeseed oil.

Specifically, this is achieved through the following technical solutions:

A method for extracting squalene in rapeseed oil, comprising the steps:

(1) Add an aqueous solution containing a composite enzyme to the rapeseed oil to a water-oil ratio of 1: (0.2-0.5), and then place it under a pulsed DC electric field for 10-20min;

(2) adding the microcrystalline fiber solution to the resultant of step (1) and mixing for 10-15min, then adding the crystallizing agent and mixing for 3-5min under the condition of 50-55 ℃, then heat preservation for 5-10min, filter, and remove the filtrate;

(3) in step (2) gained, add ethanol to alcohol oil molar ratio is (5-7): 1, add L-vitamin C simultaneously to it accounts for 2-6ppm of system total mass fraction, place 60-65 ℃ Under the condition of constant temperature water bath, reflux for 20-30min, cool, wash with water until the washing liquid is neutral, then distill under 70-90Pa vacuum condition.

The working conditions of the pulsed DC electric field: peak voltage 30-35V, pulse frequency 1-2kHz, duty ratio 0.1-0.5.

The composite enzyme content in the composite enzyme-containing aqueous solution is 30-50 ppm.

The composite enzyme is composed of phospholipase, protease and cellulase in an inoculum ratio of 1:(0.5-1):(0.2-0.4).

The temperature of the distillation is 220-230°C.

The mass concentration of the microcrystalline fiber solution is 20-25%.

The added amount of the microcrystalline fiber solution is 4-10% of the mass of the rapeseed oil.

The crystallizing agent is a citric acid solution containing catechin, and its mass fraction is: catechin 0.01-0.05%, citric acid 10-15%, and the rest is water.

The added amount of the crystallizing agent is 15-25 ppm of the filtrate mass.

Beneficial effects:

In the present invention, the extraction rate of squalene is high, the residue of reagents is less, the purity of squalene is improved, and the extraction cost is reduced, and the details are as follows:

First, the present invention utilizes an aqueous solution containing compound enzymes and a pulsed direct current electric field to promote the demulsification of rapeseed oil and provide conditions for fully releasing effective substances.

Secondly, the present invention utilizes the action of the microcrystalline fiber and the cellulase in the composite enzyme, so that the crystal structure of the microcrystalline fiber is damaged, and the fatty acid and impurity elements are coated in the mixing process, and the crystal structure is improved by using a crystallizing agent, so that the angle Squalene can be effectively separated from fatty acids and impurity elements.

Then, in the present invention, the rapeseed oil is esterified, and the addition of L-vitamin C during the esterification treatment can prevent oxidation, thereby improving the yield of squalene.

Finally, the present invention utilizes vacuum distillation to effectively negatively negative squalene, reduce the residual amount of reagents, and improve the purity and yield of squalene.

Detailed ways

The specific embodiments of the present invention are described in further detail below, but the present invention is not limited to these embodiments, and any improvement or substitution in the basic spirit of the present embodiment still belongs to the scope of protection of the claims of the present invention.

Example 1

A method for extracting squalene in rapeseed oil, comprising the steps:

(1) Add an aqueous solution containing a composite enzyme to the rapeseed oil to a water-oil ratio of 1:0.5, and then place it under a pulsed DC electric field for 20 minutes;

(2) adding the microcrystalline fiber solution to the resultant of step (1) and mixing for 15min, then adding the crystallizer and mixing for 5min under the condition of 55 ℃, then heat preservation for 10min, filter, and remove the filtrate;

(3) in step (2) gained, add ethanol to alcohol oil mol ratio to be 7:1, add L-vitamin C to 6ppm that it accounts for the total mass fraction of the system simultaneously, be placed in the constant temperature water bath under 65 ℃ condition and reflux 30min, Cool, wash with water until the washing liquid is neutral, then distill under 90Pa vacuum for 1h, and collect the distillate;

The working conditions of the pulsed DC electric field: peak voltage 35V, pulse frequency 2kHz, duty cycle 0.5;

The composite enzyme content in the described composite enzyme-containing aqueous solution is 50 ppm;

The composite enzyme is composed of phospholipase, protease, and cellulase in an inoculum ratio of 1:1:0.4;

The temperature of the distillation is 230°C;

The mass concentration of the microcrystalline fiber solution is 25%;

The added amount of the microcrystalline fiber solution is 10% of the rapeseed oil mass;

The crystallizing agent is a citric acid solution containing catechin, and its mass fraction is: 0.05% catechin, 15% citric acid, and the rest is water;

The added amount of the crystallizing agent is 25 ppm of the filtrate mass.

Example 2

A method for extracting squalene in rapeseed oil, comprising the steps:

(1) Add an aqueous solution containing a composite enzyme to the rapeseed oil to a water-oil ratio of 1:0.2, and then place it under a pulsed DC electric field for 10 minutes;

(2) adding a microcrystalline fiber solution to the resultant of step (1) and mixing for 10 minutes, then adding a crystallizer and mixing for 3 minutes at 50° C., then maintaining the temperature for 5 minutes, filtering, and removing the filtrate;

(3) adding ethanol to the alcohol-oil molar ratio of step (2) gained is 5:1, adding L-vitamin C to 2ppm of the total mass fraction of the system, and placing it in a constant temperature water bath at 60°C for 20min reflux, Cool, wash with water until the washing liquid is neutral, then distill under 70Pa vacuum for 1h, and collect the distillate;

The working conditions of the pulsed DC electric field: peak voltage 30V, pulse frequency 1kHz, duty cycle 0.1;

The composite enzyme content in the described composite enzyme-containing aqueous solution is 30 ppm;

The composite enzyme is composed of phospholipase, protease, and cellulase in an inoculum ratio of 1:0.5:0.2;

The temperature of the distillation is 220°C;

The mass concentration of the microcrystalline fiber solution is 20%;

The added amount of the microcrystalline fiber solution is 4% of the rapeseed oil mass;

The crystallizing agent is a citric acid solution containing catechin, and its mass fraction is: 0.01% catechin, 10% citric acid, and the rest is water;

The added amount of the crystallizer was 15 ppm of the filtrate mass.

Example 3

A method for extracting squalene in rapeseed oil, comprising the steps:

(1) Add an aqueous solution containing a composite enzyme to the rapeseed oil to a water-oil ratio of 1:0.3, and then place it under a pulsed DC electric field for 15 minutes;

(2) adding the microcrystalline fiber solution to the resultant of the step (1) and mixing for 12 minutes, then adding the crystallizer and mixing for 4 minutes at 53° C., holding the temperature for 8 minutes, filtering, and removing the filtrate;

(3) adding ethanol to the alcohol-oil molar ratio of step (2) gained is 6:1, adding L-vitamin C simultaneously to 4ppm of the total mass fraction of the system, and placing it in a constant temperature water bath at 63 ° C for refluxing for 25min, Cool, wash with water until the washing liquid is neutral, then distill under 80Pa vacuum for 1h, and collect the distillate;

The working conditions of the pulsed DC electric field: peak voltage 32V, pulse frequency 1.5kHz, duty cycle 0.3;

The composite enzyme content in the described composite enzyme-containing aqueous solution is 40 ppm;

The composite enzyme is composed of phospholipase, protease, and cellulase in an inoculum ratio of 1:0.8:0.3;

The temperature of the distillation is 225°C;

The mass concentration of the microcrystalline fiber solution is 22%;

The added amount of the microcrystalline fiber solution is 7% of the rapeseed oil mass;

The crystallizing agent is a citric acid solution containing catechin, and its mass fraction is: catechin 0.03%, citric acid 12%, and the rest is water;

The added amount of the crystallizer is 20 ppm of the filtrate mass.

Comparative Example 1

The difference from Example 4 is that the pulsed DC electric field treatment is replaced by mixing.

Comparative Example 2

The difference from Example 4 is that cellulase is not added to the composite enzyme.

Comparative Example 3

The difference from Example 4 is that no crystallizing agent is added.

Comparative Example 4

The difference from Example 4 is that L-vitamin C is not added.

Comparative Example 5

The difference from Example 4 is that step (2) is not taken.

The rapeseed oil in the present embodiment and the comparative example selects the same product. According to the method of "Determination of Squalene Content in Deodorized Distillate by Gas Chromatography" (Published by Li Xiangqing et al. in 2017), the squalene in the rapeseed oil before extraction is analyzed. The content of alkene was measured, and the result was 12.5%, and the method of embodiment and comparative example was used to extract squalene, and the same document "Gas Chromatography Determination of the Content of Squalene in Deodorized Distillate" (Li Xiangqing was equal to published in 2017) ) method carries out squalene content determination, and its yield result is as shown in Table 1:

Table 1

Meanwhile, the inventors respectively weighed 0.075g of the distillate of the embodiment and the comparative example, used 200-mesh silica gel to further separate and purify the squalene, and used dry method for sample loading, and the eluent was n-hexane: ethyl acetate= 100:1 for elution, each 10 mL is collected as a tube, the gas phase is detected, and the squalene content of each tube is calculated according to the standard curve;

The preparation method of the standard song is as follows: Accurately weigh 20 mg of squalene standard substance, dilute to 10 mL with n-hexane, and obtain 2 mg/mL squalene standard stock solution; take 0.05, 0.25, 0.50, 1.00, 7.50 mL of stock solution respectively , dilute to 10 mL with n-hexane to obtain each concentration. Taking the peak area as the ordinate (y) and each mass concentration as the abscissa (x), draw a standard curve.

The squalene purity results are shown in Table 2:

Table 2

project Example 1 Example 2 Example 3 Comparative Example 1 purity/% 95.2 94.6 97.8 72.3 project Comparative Example 2 Comparative Example 3 Comparative Example 4 Comparative Example 5 purity/% 81.9 77.5 73.7 85.9

Claims (7)

1. an extraction method of squalene in rapeseed oil, is characterized in that, comprises the steps: (1) Add an aqueous solution containing a composite enzyme to the rapeseed oil to a water-oil ratio of 1: (0.2-0.5), and then place it under a pulsed DC electric field for 10-20min; (2) adding the microcrystalline fiber solution to the resultant of step (1) and mixing for 10-15min, then adding the crystallizing agent and mixing for 3-5min under the condition of 50-55 ℃, then heat preservation for 5-10min, filter, and remove the filtrate; (3) in step (2) gained, add ethanol to alcohol oil molar ratio is (5-7): 1, add L-vitamin C simultaneously to it accounts for 2-6ppm of system total mass fraction, place 60-65 ℃ Under the condition of constant temperature water bath, reflux for 20-30min, cool, wash with water until the washing liquid is neutral, then distill under 70-90Pa vacuum condition; The composite enzyme is composed of phospholipase, protease, and cellulase in an inoculum ratio of 1:(0.5-1):(0.2-0.4); The crystallizing agent is a citric acid solution containing catechin, and its mass fraction is: 0.01-0.05% of catechin, 10-15% of citric acid, and the rest is water. 2. the extraction method of squalene in rapeseed oil as claimed in claim 1, is characterized in that, the working condition of described pulsed direct current electric field: peak voltage 30-35V, pulse frequency 1-2kHz, duty ratio 0.1-0.5 . 3. the extraction method of squalene in rapeseed oil as claimed in claim 1, is characterized in that, the compound enzyme content in the described aqueous solution containing compound enzyme is 30-50ppm. 4. the extraction method of squalene in rapeseed oil as claimed in claim 1, is characterized in that, the mass concentration of described microcrystalline fiber solution is 20-25%. 5. the extraction method of squalene in rapeseed oil as claimed in claim 1, is characterized in that, the addition of described microcrystalline fiber solution is 4-10% of rapeseed oil quality. 6. the extraction method of squalene in rapeseed oil as claimed in claim 1, is characterized in that, the add-on of described crystallizing agent is 15-25ppm of filtrate quality. 7. the extraction method of squalene in rapeseed oil as claimed in claim 1, is characterized in that, the temperature of described distillation is 220-230 ℃.