CN107384585B - Method for preparing antarctic krill oil in grading manner - Google Patents
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Abstract
The invention provides a method for preparing antarctic krill oil in a grading manner, which comprises the steps of carrying out first-stage extraction on antarctic krill powder by using acetone to obtain first-stage extracted antarctic krill oil and first-stage extracted antarctic krill powder; performing secondary extraction on the first-stage extracted antarctic krill powder by using an alkane solvent to obtain secondary extracted antarctic krill oil and secondary extracted antarctic krill powder; and performing third-stage extraction on the second-stage extracted antarctic krill powder by using an alcohol solvent to obtain third-stage extracted antarctic krill oil. The krill meal is extracted in three steps by using three solvents with different polarities, so that the lipid is finely classified, and the lipid in the krill meal can be maximally extracted.
Description
Technical Field
The invention belongs to the field of marine organism deep processing, and particularly relates to a method for preparing antarctic krill oil in a grading manner.
Background
Antarctic krill (Euphausia superba), commonly known as Antarctic krill, is an important marine organism that lives in the southern ocean, with resource volumes of approximately 3.79 million tons. The protein content of the fresh antarctic krill is about 11.9-15.4%, the lipid content is 0.5-3.6%, the ash content is 3%, the sugar content is 2%, and the balance is water. The lipids extracted from antarctic krill are called antarctic krill oil, and in recent years, the antarctic krill oil has been receiving increasing attention due to its unique structure and function.
The main component of common edible oil is triglyceride (95%), while the composition of antarctic krill oil is more complex. Research reports that the triglyceride content in the antarctic krill lipid is only 22.1-38.4%, and the polar lipid content is as high as 56.4-72.0%. The polar lipid is mainly phospholipid, diglyceride, monoglyceride, etc. Antarctic krill are fed to the algae and are therefore rich in EPA (10-20%) and DHA (8-18%) in the oil and present in combination with phospholipids. In addition, krill oil is rich in trace components such as astaxanthin, tocopherol, vitamin a, sterols, and the like.
At present, most of extraction technologies are 'one-pot' extensive extraction, but the 'one-pot' extensive extraction technology only focuses on the extraction efficiency of lipid, but the composition of the extracted krill oil is limited by krill raw materials, the consistency is difficult to maintain, and especially the contents of phospholipid and trace components in the krill oil have large differences. Therefore, there is a need for a new and more refined extraction technique for extracting krill lipids in a graded manner to produce krill oils with different phospholipid contents and trace components, so as to enrich the types of krill oils and produce krill oils with different functional characteristics, and to blend krill oils with a desired composition ratio as required to maintain the quality of the final krill oil product.
Disclosure of Invention
This section is for the purpose of summarizing some aspects of embodiments of the invention and to briefly introduce some preferred embodiments. In this section, as well as in the abstract and the title of the invention of this application, simplifications or omissions may be made to avoid obscuring the purpose of the section, the abstract and the title, and such simplifications or omissions are not intended to limit the scope of the invention.
The present invention is proposed in view of the above and/or the technical blank of the existing method for preparing antarctic krill oil by classification.
Therefore, the invention aims to provide a method for preparing the antarctic krill oil in a grading way.
In order to solve the technical problems, the invention provides the following technical scheme: a method for preparing Antarctic krill oil in a grading manner comprises the steps of carrying out first-stage extraction on Antarctic krill powder by using acetone to obtain first-stage extracted Antarctic krill oil and first-stage extracted Antarctic krill powder; performing secondary extraction on the first-stage extracted antarctic krill powder by using an alkane solvent to obtain secondary extracted antarctic krill oil and secondary extracted antarctic krill powder; and performing third-stage extraction on the second-stage extracted antarctic krill powder by using an alcohol solvent to obtain third-stage extracted antarctic krill oil.
As a preferred embodiment of the method for preparing antarctic krill oil by classification according to the present invention, wherein: the method comprises the following steps of performing first-stage extraction on antarctic krill powder by using acetone, wherein the first-stage extraction is performed by mixing the antarctic krill powder with the acetone according to the mass-to-volume ratio of 1: 1-10, uniformly mixing, stirring for reaction, immediately filtering after stirring is finished, removing a solvent from filtrate to obtain first-stage extracted antarctic krill oil, and vacuum-drying filter residues to obtain first-stage extracted antarctic krill powder.
As a preferred embodiment of the method for preparing antarctic krill oil by classification according to the present invention, wherein: the temperature of the acetone is-10 ℃.
As a preferred embodiment of the method for preparing antarctic krill oil by classification according to the present invention, wherein: the stirring reaction is carried out at-10 ℃ and 100-1000 rpm for 5-40 min.
As a preferred embodiment of the method for preparing antarctic krill oil by classification according to the present invention, wherein: and performing second-stage extraction on the first-stage extracted antarctic krill powder by using an alkane solvent, uniformly mixing the first-stage extracted antarctic krill powder with the alkane solvent, stirring at 20-40 ℃ and 100-1000 rpm for 5-40 min, immediately filtering after stirring, removing the solvent from filtrate to obtain second-stage extracted antarctic krill oil, and performing vacuum drying on filter residues to obtain second-stage extracted antarctic krill powder.
As a preferred embodiment of the method for preparing antarctic krill oil by classification according to the present invention, wherein: uniformly mixing the first-stage extracted antarctic krill powder with an alkane solvent, wherein the mass-volume ratio of the first-stage extracted antarctic krill powder to the alkane solvent is 1: 1-10, and mixing uniformly.
As a preferred embodiment of the method for preparing antarctic krill oil by classification according to the present invention, wherein: the alkane solvent comprises n-hexane or isohexane.
As a preferred embodiment of the method for preparing antarctic krill oil by classification according to the present invention, wherein: and carrying out third-stage extraction on the second-stage extracted antarctic krill powder by using an alcohol solvent, uniformly mixing the second-stage extracted antarctic krill powder with the alcohol solvent, stirring at 30-50 ℃ and 100-1000 rpm for 5-40 min, immediately filtering after stirring, and removing the solvent from the filtrate to obtain third-stage extracted antarctic krill oil.
As a preferred embodiment of the method for preparing antarctic krill oil by classification according to the present invention, wherein: and uniformly mixing the second-stage extracted antarctic krill powder with an alcohol solvent, wherein the mass-volume ratio of the second-stage extracted antarctic krill powder to the alcohol solvent is 1: 1-10, and mixing uniformly.
As a preferred embodiment of the method for preparing antarctic krill oil by classification according to the present invention, wherein: the alcohol solvent comprises ethanol or isopropanol.
The invention has the following beneficial effects:
(1) according to the invention, three solvents with different polarities are adopted to carry out three-step extraction on the krill meal, so that not only is the refined grading carried out on the lipid, but also the lipid in the krill meal can be maximally extracted, and the prepared antarctic krill oil has excellent oxidation resistance.
(2) The three kinds of shrimp sauce prepared by the invention have unique composition characteristics and are beneficial to developing shrimp sauce products with different functions.
(3) The three types of shrimp sauce prepared by the method can be prepared differently according to requirements, so that the composition of a constant krill product is facilitated, and different consumption requirements are met.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without inventive exercise. Wherein:
FIG. 1 is a graph showing the results of the DPPH radical scavenging experiment in example 7.
Detailed Description
In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with examples are described in detail below.
In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, but the present invention may be practiced in other ways than those specifically described and will be readily apparent to those of ordinary skill in the art without departing from the spirit of the present invention, and therefore the present invention is not limited to the specific embodiments disclosed below.
Furthermore, reference herein to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one implementation of the invention. The appearances of the phrase "in one embodiment" in various places in the specification are not necessarily all referring to the same embodiment, nor are separate or alternative embodiments mutually exclusive of other embodiments.
Example 1
(1) Taking 100g of antarctic krill powder, and performing first-stage extraction: mixing krill powder with acetone (precooled for 6h at 4 ℃) according to a ratio of 1:2(w/v), stirring for 15min at 4 ℃, wherein the stirring speed is 300r/min, filtering by using a Buchner funnel after stirring is finished, desolventizing and drying filtrate to obtain first-stage extracted antarctic krill oil, weighing, calculating the oil yield and determining the chemical composition. Drying and desolventizing the euphausia superba powder filtered after primary extraction at the temperature of 60 ℃ and under the pressure of-0.1 Mpa, and weighing for later use;
(2) and (3) second-stage extraction: mixing the shrimp meal obtained by the extraction in the step (1) with n-hexane according to a ratio of 1:2(w/v), stirring for 15min at 30 ℃, wherein the stirring speed is 300r/min, filtering by using a Buchner funnel after the stirring is finished, desolventizing the filtrate, drying to obtain the second-stage extracted antarctic krill oil, weighing, calculating the oil yield and determining the chemical composition. Drying and desolventizing the euphausia superba powder filtered after the secondary extraction at the temperature of 60 ℃ and under the pressure of-0.1 Mpa, and weighing for later use;
(3) and (3) third-stage extraction: mixing the shrimp meal obtained after the extraction in the step (2) with ethanol according to a ratio of 1:3(w/v), stirring for 15min at 40 ℃, wherein the stirring speed is 300r/min, filtering by adopting a Buchner funnel after the stirring is finished, desolventizing the filtrate, drying to obtain the Antarctic krill oil extracted in three stages, weighing, calculating the oil yield and determining the chemical composition.
Example 1 the yields, main composition and percentage of each component in the total extracted components of the krill oil obtained by fractionation are shown in the table below.
| First grade krill oil | Second grade krill oil | Tertiary krill oil | |
| Oil extraction rate (% shrimp meal) | 5.08 | 4.80 | 9.11 |
| Phospholipid (g/100g shrimp sauce) | 2.32 | 39.00 | 59.52 |
| (g100g shrimp meal) | 0.12 | 1.88 | 5.42 |
| (% total extracted phospholipid) | 1.62 | 25.34 | 73.05 |
| Astaxanthin (mg/kg shrimp sauce) | 505.00 | 34.92 | 7.48 |
| (mg/kg shrimp meal) | 25.56 | 1.66 | 0.68 |
| (% Total extract astaxanthin) | 91.61 | 5.95 | 2.44 |
| Tocopherol (mg/100g shrimp sauce) | 29.39 | 12.98 | 3.65 |
| (mg/100g shrimp meal) | 1.48 | 0.62 | 0.33 |
| (% total extracted tocopherol) | 60.91 | 25.51 | 13.58 |
| Vitamin A (mg/100g shrimp sauce) | 34.32 | 12.33 | 2.27 |
| (mg/100g shrimp meal) | 1.74 | 0.59 | 0.21 |
| (% Total extracted vitamin A) | 68.50 | 23.23 | 8.27 |
| Sterol (mg/g shrimp sauce)) | 27.95 | 17.72 | 9.97 |
| (mg/g shrimp meal) | 1.42 | 0.85 | 0.91 |
| (% Total extract sterol) | 44.65 | 26.73 | 28.62 |
Example 2
(1) Taking 200g of antarctic krill powder, and performing first-stage extraction: mixing krill powder with acetone (precooled for 6h at 10 ℃) according to a ratio of 1:3(w/v), stirring for 10min at 10 ℃, wherein the stirring speed is 200r/min, filtering by using a Buchner funnel after stirring is finished, desolventizing the filtrate, drying to obtain first-stage extracted antarctic krill oil, weighing, calculating the oil yield and determining the chemical composition. Drying and desolventizing the euphausia superba powder filtered after primary extraction at the temperature of 60 ℃ and under the pressure of-0.1 Mpa, and weighing for later use;
(2) and (3) second-stage extraction: mixing the shrimp meal obtained by the extraction in the step (1) with isohexane according to a ratio of 1:3(w/v), stirring at 30 ℃ for 15min at a stirring speed of 300r/min, filtering by using a Buchner funnel after the stirring is finished, desolventizing the filtrate, drying to obtain the second-stage extracted euphausia superba oil, weighing, calculating the oil yield and determining the chemical composition. Drying and desolventizing the euphausia superba powder filtered after the secondary extraction at the temperature of 60 ℃ and under the pressure of-0.1 Mpa, and weighing for later use;
(3) and (3) third-stage extraction: mixing the shrimp meal obtained after the extraction in the step (2) with ethanol according to a ratio of 1:4(w/v), stirring at 35 ℃ for 25min at a stirring speed of 300r/min, filtering by using a Buchner funnel after the stirring is finished, desolventizing the filtrate, drying to obtain the Antarctic krill oil extracted in three stages, weighing, calculating the oil yield, and determining the chemical composition.
Example 2 the yields, main composition and percentage of each component in the total extracted components of the krill oil obtained by fractionation are shown in the table below.
Example 3
(1) Taking 250g of antarctic krill powder, and performing first-stage extraction: mixing krill powder with acetone (precooled for 6h at 0 ℃) according to a ratio of 1:3(w/v), stirring for 20min at 0 ℃, wherein the stirring speed is 100r/min, filtering by using a Buchner funnel after stirring is finished, desolventizing the filtrate, drying to obtain first-stage extracted antarctic krill oil, weighing, calculating the oil yield and determining the chemical composition. Drying and desolventizing the euphausia superba powder filtered after primary extraction at the temperature of 60 ℃ and under the pressure of-0.1 Mpa, and weighing for later use;
(2) and (3) second-stage extraction: mixing the shrimp meal obtained by the extraction in the step (1) with n-hexane according to a ratio of 1:3(w/v), stirring for 20min at 25 ℃, wherein the stirring speed is 200r/min, filtering by using a Buchner funnel after the stirring is finished, desolventizing the filtrate, drying to obtain the second-stage extracted antarctic krill oil, weighing, calculating the oil yield and determining the chemical composition. Drying and desolventizing the euphausia superba powder filtered after the secondary extraction at the temperature of 60 ℃ and under the pressure of-0.1 Mpa, and weighing for later use;
(3) and (3) third-stage extraction: mixing the shrimp meal obtained after the extraction in the step (2) with isopropanol according to a ratio of 1:4(w/v), stirring for 20min at 45 ℃, wherein the stirring speed is 200r/min, filtering by using a Buchner funnel after the stirring is finished, desolventizing the filtrate, drying to obtain the Antarctic krill oil extracted in three stages, weighing, calculating the oil yield and determining the chemical composition.
Example 3 the yields, main composition and percentage of each component in the total extracted components of the krill oil obtained by fractionation are shown in the table below.
| First grade krill oil | Second grade krill oil | Tertiary krill oil | |
| Oil extraction rate (% shrimp meal) | 5.69 | 6.74 | 9.77 |
| Phospholipid (g/100g shrimp sauce) | 3.18 | 31.24 | 61.43 |
| (g100g shrimp meal | 0.18 | 2.11 | 6.00 |
| (% total extracted phospholipid) | 2.18 | 25.40 | 72.41 |
| Astaxanthin (mg/kg shrimp sauce) | 568.10 | 26.74 | 5.13 |
| (mg/kg shrimp meal) | 32.32 | 1.80 | 0.50 |
| (% Total extract astaxanthin) | 93.35 | 5.20 | 1.45 |
| Tocopherol (mg/100g shrimp sauce) | 35.13 | 10.63 | 4.92 |
| (mg/100g shrimp meal) | 2.00 | 0.72 | 0.48 |
| (% total extracted tocopherol) | 62.54 | 22.42 | 15.04 |
| Vitamin A (mg/100g shrimp sauce) | 40.14 | 11.36 | 1.67 |
| (mg/100g shrimp meal) | 2.28 | 0.77 | 0.16 |
Example 4
To obtain a krill oil product with a phospholipid content of 40%, the three krill oils of example 1 were formulated in different ratios: 34% of first-stage extracted krill oil and 66% of third-stage extracted krill oil; or 22% of the first-stage extracted krill oil, 35% of the second-stage extracted krill oil and 43% of the third-stage extracted krill oil.
Example 5
To obtain a krill oil product with 35% phospholipid and 200ppm astaxanthin content, the three krill oils of example 1 were blended at different ratios: 38% of first-stage extracted krill oil, 14% of second-stage extracted krill oil and 48% of third-stage extracted krill oil.
Example 6
To obtain a krill oil product with a phospholipid content of 30% and a tocopherol content of 18mg/100g, the three krill oils of example 2 were formulated in different proportions: 40% of first-stage extracted krill oil, 20% of second-stage extracted krill oil and 40% of third-stage extracted krill oil.
Example 7
By adopting a DPPH free radical scavenging experiment, the in-vitro antioxidant capacity of the three kinds of the shrimp oil is evaluated by the Trolox equivalent antioxidant capacity by comparing the DPPH free radical scavenging inhibition rate of the shrimp oil with the free radical scavenging inhibition rate of a standard water-soluble tocopherol (Trolox). The results prove that the three types of shrimp sauce have significant difference in oxidation resistance.
The experimental method comprises the following steps: 0.5g of each of the shrimp oil of example 1 was dissolved in 100mL of ethyl acetate, and 2mL of each solution and 2mL of a DPPH solution having a concentration of 0.15mmoL/L were mixed uniformly, reacted in the dark for 2 hours, and the absorbance of the sample was measured at a wavelength of 517 nm. The free radical clearance rate is calculated according to the absorbance, and then the inhibition rate is calculated
The calculation method of the inhibition rate comprises the following steps: k% [1- (A) ]1-A2)/A3]×100％
Wherein A is1: absorbance of 2mL of PPH solution and 2mL of solution to be measured
A2: absorbance of 2mL of the solution to be measured and 2mL of the solvent,
A3: absorbance of 2mL DPPH solution +2mL solvent.
Meanwhile, taking Trolox as a standard substance, drawing a standard curve of Trolox free radical clearance rate under different concentrations, comparing the free radical clearance rate of the shrimp oil with the standard curve, and calculating the Trolox equivalent oxidation resistance of the shrimp oil.
The experimental result is shown in figure 1 of the attached drawing of the specification, and the result shows that the three kinds of the shrimp oil have strong antioxidant capacity, and particularly the shrimp oil extracted in three stages has the strongest antioxidant capacity.
Therefore, the three solvents with different polarities are adopted to carry out three-step extraction on the krill meal, so that not only is the refined classification carried out on the lipid, but also the lipid in the krill meal can be extracted to the maximum extent, and the prepared antarctic krill oil has excellent oxidation resistance; the three kinds of shrimp sauce prepared by the invention have unique composition characteristics, and are beneficial to developing shrimp sauce products with different functions; the three types of shrimp sauce prepared by the method can be prepared differently according to requirements, so that the composition of a constant krill product is facilitated, and different consumption requirements are met.
It should be noted that the above-mentioned embodiments are only for illustrating the technical solutions of the present invention and not for limiting, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions may be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, which should be covered by the claims of the present invention.
Claims (7)
1. A method for preparing antarctic krill oil in a grading manner is characterized by comprising the following steps: comprises the steps of (a) preparing a mixture of a plurality of raw materials,
performing first-stage extraction on the antarctic krill powder by using acetone to obtain first-stage extraction antarctic krill oil and first-stage extraction antarctic krill powder, performing first-stage extraction on the antarctic krill powder by using the acetone, wherein the first-stage extraction is performed on the antarctic krill powder by using the acetone according to the mass-to-volume ratio of 1: 1-10, uniformly mixing, stirring for reaction, immediately filtering after stirring is finished, removing a solvent from filtrate to obtain first-stage extracted antarctic krill oil, and vacuum-drying filter residues to obtain first-stage extracted antarctic krill powder;
performing second-stage extraction on the first-stage extracted antarctic krill powder by using an alkane solvent to obtain second-stage extracted antarctic krill oil and second-stage extracted antarctic krill powder, performing second-stage extraction on the first-stage extracted antarctic krill powder by using an alkane solvent, uniformly mixing the first-stage extracted antarctic krill powder with the alkane solvent, stirring at the temperature of 20-40 ℃ and at the speed of 100-1000 rpm for 5-40 min, immediately filtering after stirring, removing the solvent from filtrate to obtain second-stage extracted antarctic krill oil, and performing vacuum drying on filter residues to obtain second-stage extracted antarctic krill powder;
and carrying out third-stage extraction on the second-stage extracted antarctic krill powder by using an alcohol solvent to obtain third-stage extracted antarctic krill oil, and carrying out third-stage extraction on the second-stage extracted antarctic krill powder by using the alcohol solvent, wherein the second-stage extracted antarctic krill powder and the alcohol solvent are uniformly mixed, stirring for 5-40 min at the temperature of 30-50 ℃ and the rpm of 100-1000, immediately filtering after stirring is finished, and removing the solvent from the filtrate to obtain the third-stage extracted antarctic krill oil.
2. The method for preparing antarctic krill oil by classification as claimed in claim 1, wherein: the temperature of the acetone is-10 ℃.
3. The method for preparing antarctic krill oil by classification as claimed in claim 2, wherein: the stirring reaction is carried out at-10 ℃ and 100-1000 rpm for 5-40 min.
4. The method for preparing antarctic krill oil by classification as claimed in claim 1, wherein: uniformly mixing the first-stage extracted antarctic krill powder with an alkane solvent, wherein the mass-volume ratio of the first-stage extracted antarctic krill powder to the alkane solvent is 1: 1-10, and mixing uniformly.
5. The method for preparing antarctic krill oil by classification as claimed in claim 4, wherein: the alkane solvent comprises n-hexane or isohexane.
6. The method for preparing antarctic krill oil by classification as claimed in claim 1, wherein: and uniformly mixing the second-stage extracted antarctic krill powder with an alcohol solvent, wherein the mass-volume ratio of the second-stage extracted antarctic krill powder to the alcohol solvent is 1: 1-10, and mixing uniformly.
7. The method for preparing antarctic krill oil by classification as claimed in claim 6, wherein: the alcohol solvent comprises ethanol or isopropanol.
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