Algal plant extracts rich in eicosapentaenoic acid as triglyceride / diglyceride conjugates

Abstract

Provided is a plant extract comprising an ethanol extract of Nannochloropsis algae having 10 wt% to 50 wt% polar lipids and 45 wt% to 60 wt% fatty acids, the fatty acids including greater than 10 wt% eicosapentaenoic acid (EPA) as triglyceride conjugates and / or diglyceride conjugates, and less than 15 wt% EPA as free fatty acids. EPA levels can reach up to 90 wt% and the plant extract can be flowable at room temperature. Growth conditions are set, adjusted, and monitored to promote EPA formation in the cell vacuole, resulting in a high proportion of EPA as triglyceride / diglyceride conjugates and a high concentration of Ω-3 fatty acids, which requires only a gentle extraction procedure to reach the final product, thereby preserving the biochemical properties of the compound without excessive modification.

Description

[Technical Field]

[0001] 1.Technical Field The present invention relates to the field of foods and supplements rich in Omega 3 fatty acids, and more particularly to algal plant extracts rich in EPA as triglyceride and / or diglyceride conjugates. [Background technology]

[0002] 2. Description of Related Technology U.S. Pat. Nos. 9,629,820, 10,123,986, and 10,039,734, as well as U.S. Patent Application Publication No. 20140179781, the entireties of which are incorporated herein by reference, describe eicosapentaenoic acid (EPA) compositions and their product and use indications. The present invention discloses a composition comprising about 15 wt% to about 90 wt% EPA and about 10 wt% to about 70 wt% polar lipids, the polar lipids including phospholipid conjugates and glycolipid conjugates, about 3 wt% to about 50 wt% of the EPA in the composition is phospholipid conjugates, about 5 wt% to about 50 wt% of the EPA in the composition is glycolipid conjugates, and about 0 wt% to about 10 wt% of the EPA is triglyceride conjugates or diglyceride conjugates, the composition comprises about 0.1 wt% to about 3.0 wt% mannitol, the composition does not comprise docosahexaenoic acid (DHA), fatty acid methyl esters, or fatty acid ethyl esters, and the composition is suitable for human consumption. Note that eicosapentaenoic acid (EPA) refers to cis-5,8,11,14,17-eicosapentaenoic acid (20:5,n-3) and docosahexaenoic acid (DHA) refers to cis-4,7,10,13,16,19-docosahexaenoic acid (22:6(n-3)). Summary of the Invention

[0003] The following is a simplified summary to provide an initial understanding of the invention. This summary is not intended to necessarily identify key elements or to delineate the scope of the invention, but rather merely to serve as an introduction to the description that follows.

[0004] One aspect of the invention provides a plant extract comprising an ethanol extract of Nannochloropsis algae having 10 wt% to 20 wt% polar lipids and 45 wt% to 60 wt% fatty acids, wherein the fatty acids comprise more than 10 wt% eicosapentaenoic acid (EPA) as triglyceride conjugates and / or diglyceride conjugates, and less than 15 wt% EPA as free fatty acids.

[0005] These additional and / or other aspects and / or advantages of the present invention will be set forth in the detailed description that follows, and in some cases may be inferred from the detailed description and / or may be learned by practice of the invention. [Brief explanation of the drawings]

[0006] For a better understanding of embodiments of the present invention and to show how the same may be carried into effect, reference will now be made, by way of example only, to the accompanying drawings in which like numerals designate corresponding elements or parts throughout and in which:

[0007] [Figure 1] FIG. 1 is a high-level schematic diagram of a bioreactor for extracting plant extracts, according to some embodiments of the present invention.

[0008] It will be appreciated that for simplicity and clarity of illustration, elements shown in the figures have not necessarily been drawn to scale. For example, the dimensions of some elements may be exaggerated relative to other elements for clarity. Further, where considered appropriate, reference numerals may be repeated among the figures to indicate corresponding or analogous elements. DETAILED DESCRIPTION OF THE INVENTION

[0009] In the following description, various aspects of the present invention will be described. For purposes of explanation, specific configurations and details are set forth in order to provide a thorough understanding of the present invention. However, it will also be apparent to one skilled in the art that the present invention may be practiced without the specific details presented herein. Furthermore, well-known features may be omitted or simplified in order not to obscure the present invention. With particular reference to the drawings, it is emphasized that the particulars shown are by way of example and for the purpose of exemplary explanation of the invention only, and are presented for the purpose of providing what is believed to be the most useful and readily understood explanation of aspects of the principles and concepts of the invention. In this regard, no attempt has been made to show the structural details of the invention in more detail than is necessary for a fundamental understanding of the invention, and by describing the invention in conjunction with the drawings, it will be apparent to those skilled in the art how several forms of the invention may be embodied in practice.

[0010] Before describing at least one embodiment of the present invention in detail, it is to be understood that the invention is not limited in its application to the details of construction and the arrangement of components set forth in the following description or illustrated in the drawings. The invention is applicable not only to combinations of the disclosed embodiments but also to other embodiments that can be practiced or carried out in various ways. Also, it is to be understood that the phraseology and terminology employed herein is for the purpose of description and should not be regarded as limiting.

[0011] U.S. Patent Application Publication Nos. 2019 / 0345427, 2020 / 0231925, and 2022 / 0256884, each of which is incorporated by reference in its entirety, disclose systems and methods for growing algae and extracting an Omega-3 rich extract therefrom.

[0012] Embodiments of the present invention provide an efficient and economical method for growing algae and extracting therefrom an algal plant extract that is rich in EPA as triglyceride conjugates and / or diglyceride conjugates, thereby improving the art of algal Omega 3-enriched plant extracts, as well as foods and supplements containing the extracts. Some embodiments provide a plant extract comprising an ethanol extract of Nannochloropsis algae, having 10 wt% to 20 wt% polar lipids and 45 wt% to 60 wt% fatty acids, the fatty acids comprising more than 10 wt% eicosapentaenoic acid (EPA) as triglyceride and / or diglyceride conjugates, and less than 15 wt% EPA as free fatty acids. EPA levels can reach up to 90 wt% and the plant extract can be flowable at room temperature. Growth conditions are set, adjusted, and monitored to promote EPA formation in the cell vacuole, resulting in a high proportion of EPA as triglyceride / diglyceride conjugates and a high concentration of omega-3 fatty acids, which requires only a gentle extraction procedure to reach the final product, thereby preserving the biochemical properties of the compound without excessive modification.

[0013] FIG. 1 is a high-level schematic diagram of a bioreactor 100 for extracting a plant extract 140, according to some embodiments of the present invention. The bioreactor 100 includes one or more tanks filled with water and Nannochloropsis algae 90 (e.g., N. oculata, N. australis, N. gaditana, N. granulate, N. limnetica, N. oceanica, N. salina, Nitzschia paleacea, Phaeodactylum tricornutum, Pavlova lutheri, Rebecca salina, any strain or combination thereof, or equivalent algae), and further includes (i) an irradiation system 120 and (ii) a bubbling system 110.

[0014] The illumination system 120 may comprise a plurality of powerful light sources 125, e.g., light emitting diodes (LEDs, e.g., between 400-700 nm, subranges thereof, e.g., 400-500 nm, 500-600 nm, 600-700 nm, and / or specific wavelengths, e.g., 650 nm), which may be arranged in one or more horizontal and / or vertical panels (shown schematically and only partially). The illumination system 120 directly illuminates the algae with photosynthetically active radiation in the red and blue wavelength ranges (rather than the sunlight or white light spectrum as in the prior art), and the intensity of the photosynthetically active radiation in these wavelength ranges is greater than or equal to 750 μmol / s / m 2 It may be configured to reach over

[0015] A bubbling system 110 associated with a gas source (e.g., CO2-enriched air or nitrogen, containing a CO2 concentration of 30% or greater) includes multiple spargers of at least two types, i.e., a bubbling system that rapidly (e.g., 100 / min) pumps gas through the algae culture. This allows for a cumulative bubble volume relative to the vessel volume. The algal culture includes sparger(s) 110A having large nozzles (e.g., greater than 1 mm in diameter, possibly in the range of 1-5 mm or a sub-range thereof) to generate large bubbles 115A that move slowly (e.g., 5 / min, which represents a cumulative bubble volume relative to the vessel volume, possibly in the range of ±30%, 2-30 / min or a sub-range thereof) through the algal culture and mix it, and sparger(s) 110B having small nozzles (e.g., less than 1 mm in diameter, possibly in the range of 0.1-1 mm or a sub-range thereof) to generate small bubbles 115B that move slowly (e.g., 5 / min, which represents a cumulative bubble volume relative to the vessel volume, possibly in the range of ±30%, 2-30 / min or a sub-range thereof) through the algal culture, allowing CO to diffuse into the algal cells. In some cases, more than two types of nozzles may be used to control mixing of the algae culture and delivery of sufficient concentrations of CO2 to the algae. Two or more types of spargers (each with multiple nozzles) may be distributed in one or more locations and configured to generate turbulent mixing of the algae within the culture vessel and provide CO2 to the algae to an extent that maximizes or optimizes its growth and / or its Omega-3 fatty acid content and / or composition, e.g., maximizes its EPA (and / or DHA) content and / or association with triglycerides / diglycerides within the cell vacuoles.

[0016] As shown schematically in the enlarged area of ​​Figure 1, a very dense algal culture (e.g., having an algal density of at least 5 g / l, at least 10 g / l, and possibly in the range of 5-15 g / l or any subrange thereof) and a localized, powerful light source 125 create illuminated and dark zones within the bioreactor 100, and powerful agitation of the algal culture by large bubbles 115B continuously mixes the liquid and moves the algal cells 90 between the dark and illuminated zones.

[0017] In a non-limiting example, the light source 125 may be, for example, 4 m long with a light path of about 2.5 cm. 2 Use at least 24 LEDs over an area of ​​at least 700 µmol m² (defining the reach of the illumination zone). -2 s -1 , at least 1000 µmol m -2 s -1 , at least 1200 µmol m -2 s -1 , or any intermediate value of the illumination intensity of each. It has also been found that the intense heterogeneous illumination, dark zone periods, and high levels of CO2 supplied to the algal cells 90 by small bubbles 115A result in faster algal growth rates and alter the biosynthetic pathways utilized by the algae to form organic compounds. For example, in high-density conditions, the illuminated zones may extend up to several millimeters (e.g., 1-5 mm) from the point light source 125, while the dark zones between illuminated zones may extend over tens of millimeters (e.g., 20-30 mm) between successive illuminated zones, and thus individual algal cells spend periods in the dark zones to assimilate CO2 using the light energy absorbed in the illuminated zones.

[0018] Growth conditions may be monitored to maintain optimal growth; for example, temperature may be kept constant (e.g., 15°C, 20°C, 25°C, 27°C, 12-17°C, 15-20°C, 20-25°C, 25-30°C, 20-40°C, 20-50°C, or any subrange thereof) or may be varied within a specific range; and chemical conditions such as pH, O2 and CO2 content, and / or the content of various ions or compounds may be kept constant or varied within a specific range (e.g., a pH value of 6.7-7.2, or any subrange thereof). For example, the organic carbon content may be maintained at or about 20 wt%. Alternatively or complementary, the CO2 concentration may be kept below 40% and / or the pH may be monitored to indicate and regulate the CO2 concentration. Similarly, the algae density may be kept constant or may be varied within a particular range. The flow rate of small bubbles 115B from the sparger(s) 110B with small nozzles may be adjusted to increase or decrease CO2 levels (e.g., increased to greater than 5 / min, e.g., 7-10 / min to increase CO2 levels, or decreased to 2 / min to decrease CO2 levels). In certain embodiments, the levels of various nutrients may be monitored and additional nutrients may be provided via one or both bubble streams; for example, phosphorus may be added to the gas supply if low P levels are detected or if growth is inhibited.

[0019] The rapid cycling of ultra-high density algal cultures between highly illuminated and dark regions, provided by intensive mixing and intensive CO2 supply, are exceptional growth conditions that result in the unexpected EPA composition being extracted from the algae by ethanol wet extraction, as described herein. In particular, the inventors surprisingly found that these growth conditions cause the algae to form more EPA in the cell vacuoles (in the form of triglyceride- and / or diglyceride-conjugates) and less EPA in the cell membrane (in the form of polar lipids). Furthermore, the harsh growth conditions also resulted in relatively high concentrations of EPA in the algal biomass.

[0020] The algae slurry is periodically or continuously removed from the bioreactor 100 (e.g., to balance biomass growth) and mixed with ethanol (step 130), and the plant extract 140 is extracted therefrom by phase separation of one or more of the separated phases (step 132) and thermal separation(s) (step 134). Non-limiting examples of separation process parameters include a volume ratio of at least 5:1 (or optionally in the range of 3.5:1 to 7:1 or intermediate ranges) between ethanol and algae slurry (which mixture may be stirred for 0.5 to 4 hours, e.g., 2 hours), the use of centrifugation for phase separation, optionally under vacuum (e.g., at a pressure of 75 to 125 mbar), and a temperature range of 55°C to 65°C for thermal separation, which may be sufficient to reach the required concentration while minimizing thermal and chemical modification of the plant extract due to the initial high concentration of Omega-3 fatty acids such as EPA. Thus, the plant extract 140 reaches a concentration of 10 wt% to 20 wt% polar lipids and 45 wt% to 65 wt% fatty acids. In certain embodiments, up to 90% of the algae mass can be removed per day. In certain embodiments, the yield from the bioreactor 100 can reach 2.8 gr / l resulting in a yield of 190 mg / l of omega-3 fatty acids per day.

[0021] In certain embodiments, the plant extract 140 reaches a concentration of 10 wt%-50 wt% polar lipids (e.g., glycolipids and phospholipids) and 15 wt%-90 wt% EPA, which can be partitioned into 3-25 wt% phospholipids, 3-25 wt% glycolipids, and 15-35 wt% (non-polar) triglycerides / diglycerides. It should be noted that the viscosity and very dark color of the plant extract 140 limits the analysis of its constituents by common profiling methods, such as optical and spectroscopic methods, as well as gas chromatography-mass spectrometry (GC-MS).

[0022] In particular, the inventors have surprisingly found that the extract 140 obtained by the separation steps 132, 134 is already highly concentrated (due to the special growth conditions) and does not require further purification and isolation of EPA to reach higher EPA concentrations. Thus, the plant extract 140 is kept in a relatively pure form, avoiding high temperatures and chemical modification. Furthermore, due to the different distribution of EPA within the algal cells, the resulting plant extract 140 contains more than 10 wt% EPA as triglyceride and / or diglyceride conjugates (due to the higher EPA concentration in the cell vacuoles) and less than 15 wt% EPA as free fatty acids (due to the lower EPA concentration in the polar lipids of the cell membrane). In certain embodiments, the plant extract 140 comprises greater than 15 wt% EPA as triglyceride and / or diglyceride conjugates and less than 10 wt% EPA as free fatty acids. In certain embodiments, the plant extract 140 comprises about 18 wt% EPA as triglyceride and / or diglyceride conjugates and about 7 wt% EPA as free fatty acids.

[0023] In certain embodiments, the resulting plant extract 140 contains more than 10 wt% EPA and / or DHA as triglyceride conjugates and / or diglyceride conjugates (due to higher EPA and / or DHA concentrations in the cell vacuoles) and less than 15 wt% EPA and / or DHA as free fatty acids (due to lower EPA and / or DHA concentrations in the polar lipids of the cell membrane). In certain embodiments, the plant extract 140 comprises greater than 15 wt% EPA and / or DHA as triglyceride conjugates and / or diglyceride conjugates and less than 10 wt% EPA and / or DHA as free fatty acids. In certain embodiments, the plant extract 140 comprises about 18 wt% EPA and / or DHA as triglyceride conjugates and / or diglyceride conjugates, and about 7 wt% EPA and / or DHA as free fatty acids.

[0024] Various embodiments include food products and / or dietary supplements that include the botanical extract 140. In certain embodiments, foods and / or dietary supplements containing the botanical extract 140 may be used to aid in antidepressant treatment and / or cholesterol management.

[0025] Advantageously, compared to the prior art, such as U.S. Pat. Nos. 9,629,820, 10,123,986, and 10,039,734, and U.S. Patent Application Publication No. 20140179781, which cultivate algae in open pond systems and result in different compositions, some of the disclosed embodiments provide EPA compositions having 15 wt% to 90 wt% EPA and 10 wt% to 50 wt% polar lipids, wherein the polar lipids include phospholipid conjugates and glycolipid conjugates, and wherein 3 wt% to 25 wt% of the EPA in the composition is phospholipid conjugates, 3 wt% to 25 wt% of the EPA in the composition is glycolipid conjugates, and 15 wt% to 35 wt% of the EPA in the composition is triglyceride conjugates or diglyceride conjugates. In some embodiments, less than 10 wt% of the EPA in the composition is in the form of free fatty acids. In some embodiments, the EPA composition is docosahexaenoic acid (DHA)-free and suitable for human consumption.

[0026] For example, Table 1 shows non-limiting examples of biomass analyses used to extract the disclosed compositions. Table 1 shows the EPA concentration, polar lipid (including glycolipids and phospholipids), and non-polar lipid (triglyceride and diglyceride) concentrations on an ash-free dry weight (AFDW) basis. Data are obtained from the analysis of 25 batches over a period of approximately 3 years and are compared to biomass analysis of N. oculata grown in open ponds (e.g., as taught by U.S. Patent Application Publication No. 20140179781).

[0027] Table 1: Compositional analysis of N. oculata biomass (controlled growth) compared to the composition of the prior art (open pond). Data in wt%. [Table 1]

[0028] For example, compared to prior art biomass of Nannochloropsis oculata cultured in open pond systems, algal biomass grown under the controlled conditions disclosed herein has twice the average EPA content (9.7 wt% vs. 5.4 wt%) and twice the average triglyceride (non-polar) content (28.5 wt% vs. 15 wt%). The disclosed plant extract 140 obtained from aqueous ethanol extraction of the biomass was 39-53% of the algal AFDW biomass with an EPA yield of over 90%. Considering the % extraction rate and % yield, the data show that the concentration of EPA in the extract (which corresponds to the algal biomass composition) is greater than 15 wt%, the concentration of polar lipids (glycolipids and phospholipids) is greater than 13 wt%, the concentration of triglycerides is greater than 50 wt% (e.g., 59 wt%), and the concentration of EPA-triglyceride conjugates is greater than 15 wt% (e.g., 28 wt%), all in contrast to the prior art such as U.S. Patent Application Publication No. 20140179781. A further difference is that the disclosed extracts have an unusually low free fatty acid composition of less than 10 wt% (eg, 7 wt%).

[0029] In another example, Table 2 shows a non-limiting example of an analysis of EPA in the triglyceride and free fatty acid forms in the disclosed extract compositions.

[0030] Table 2: NMR analysis of the forms of EPA in the extract (%). [Table 2]

[0031] As shown in Table 2, in the disclosed extracts, a low percentage (less than 5 wt%) of EPA is found in the free fatty acid form (FFA), while a large percentage (greater than 15 wt%) of EPA is in the triglyceride form (SN2 indicates a central position of EPA on the glycerol backbone, while SN1 / 3 indicates a terminal position of EPA on the glycerol backbone), distinguishing the disclosed extracts from prior art extracts.

[0032] Additionally, whereas prior art extracts, likely due to their high levels of polar lipids and free fatty acids and low levels of glyceride-lipid conjugates, have a waxy viscosity and cannot be handled at temperatures below 90°C (see U.S. Patent Application Publication No. 20140179781), the disclosed extract 140 is typically liquid or semi-liquid at about 40°C, likely due to their low levels of free fatty acids and high levels of glyceride-lipid conjugates. In certain embodiments, the plant extract 140 has different rheological properties than prior art extracts such as those disclosed in U.S. Pat. Nos. 9,629,820, 10,123,986, and 10,039,734, and U.S. Patent Application Publication No. 20140179781, and in particular has better flow properties, possibly due to different growing and extraction conditions resulting in different extracted components that improve the flowability of the extract 140. For example, different growth and extraction conditions may result in different profiles of algal polysaccharides, including a large proportion of ethanol-soluble polysaccharides (e.g., algaenans, e.g., Scholz et al. 2014, Ultrastructure and composition of the Nannochloropsis gaditana cell wall, Eukaryotic Cell 13(11):1450-1464), thereby making the plant extract more fluid (e.g., Bernaerts 2019, The role of structural biopolymers in the functionalization of microalgae for food processing. Dissertation, KU Leuven, Belgium; Zailer 2019, Holistic control of fats and oils by NMR spectroscopy, Encyclopedia of Food Chemistry, vol. 2, 168-181 and Audo et al. 2012, Relationship between microalgae lipid extracts composition and rheological properties, 2nd International Symposium on (See Asphalt Pavements et Environnement, Transportation Research Board, of The National Academies, France). The inventors suggest that the growth and extraction conditions disclosed herein result in an extract 140 with modified and improved rheological properties over the prior art, for example, by modifying the profile of algal polysaccharides.

[0033] It should be noted that bioreactor 100, as briefly described above, may further include additional piping and valves, pumping and filtering elements, and control, sensing and regulating elements for regulating material flow (e.g., water, gas, slurry and extraction processes), as described, for example, in WIPO Publication Nos. 2018 / 134818, 2018 / 134819, 2018 / 134820, 2018 / 154565 and 2020 / 240551, the entireties of which are incorporated herein by reference. It is further noted that values ​​modified by the term "about" are understood to encompass ±10% of the value.

[0034] In the above description, one embodiment is an example or implementation of the present invention. The various appearances of "one embodiment," "one embodiment," "particular embodiments," or "some embodiments" do not necessarily all refer to the same embodiments. Although various features of the invention may be described in the context of a single embodiment, the features may also be provided separately or in any suitable combination. Conversely, although the invention may be described herein in the context of separate embodiments for clarity, the invention may also be practiced in a single embodiment. Certain embodiments of the present invention may include features from different embodiments disclosed above, and certain embodiments may incorporate elements from other embodiments disclosed above. The disclosure of elements of the invention in the context of a particular embodiment is not to be construed as limiting them to use only in that particular embodiment. Furthermore, it is to be understood that the present invention can be practiced or carried out in various ways and that the present invention can be embodied in specific embodiments other than those outlined in the description above.

[0035] The present invention is not limited to the above drawings or the corresponding descriptions. For example, the flow need not move through each box or state shown, nor need it move in the exact same order as shown and described. Unless otherwise defined, the meanings of technical and scientific terms used herein are those commonly understood by one of ordinary skill in the art to which this invention belongs. While the present invention has been described with respect to a limited number of embodiments, these should not be construed as limiting the scope of the invention, but rather as illustrative of some of the preferred embodiments. Other possible variations, modifications, and applications are also within the scope of the present invention. Accordingly, the scope of the present invention should not be limited by what has been described above, but rather by the appended claims and their legal equivalents.

Claims

1. 1. An EPA (eicosapentaenoic acid) composition comprising 15 wt% to 90 wt% EPA and 10 wt% to 50 wt% polar lipids, the polar lipids include phospholipid conjugates and glycolipid conjugates; 3 wt% to 25 wt% of the EPA in the composition is phospholipid conjugate; 3 wt% to 25 wt% of the EPA in the composition is a glycolipid conjugate; 15 wt% to 35 wt% of the EPA in the composition is triglyceride conjugate or diglyceride conjugate; The EPA composition is free of docosahexaenoic acid (DHA) and is suitable for human consumption.

2. 10. The composition of claim 1, wherein less than 10 wt% of the EPA in the composition is in the form of free fatty acids.

3. 3. A food or dietary supplement comprising the EPA composition of claim 1 or 2.

4. 1. A plant extract comprising an ethanol extract of Nannochloropsis algae having 10 wt% to 50 wt% polar lipids and 45 wt% to 60 wt% fatty acids, wherein the fatty acids are: greater than 10 wt. % eicosapentaenoic acid (EPA) as triglyceride conjugates and / or diglyceride conjugates, and Less than 15 wt% EPA as free fatty acids A plant extract comprising:

5. 5. The plant extract of claim 4, wherein the fatty acids comprise more than 15 wt. % EPA as triglyceride conjugates and / or diglyceride conjugates and less than 10 wt. % EPA as free fatty acids.

6. 6. The plant extract of claim 4, wherein the fatty acids comprise 18 wt. % EPA as triglyceride conjugates and / or diglyceride conjugates and 7 wt. % EPA as free fatty acids.

7. 7. The plant extract of claim 4, wherein the Nannochloropsis algae includes N. oculata.

8. The Nannochloropsis algae have a density of at least 700 micromol m －２ s －１ The cells are grown at a high density of at least 5 g / l under high illumination intensity of 1000 kJ / L and continuously bubbling and CO ２ The plant extract according to any one of claims 4 to 7, which has been concentrated.

9. A food or nutritional supplement comprising the plant extract of any one of claims 4 to 8.

10. 1. A plant extract comprising an ethanol extract of Nannochloropsis algae having 10 wt% to 50 wt% polar lipids and 45 wt% to 60 wt% fatty acids, wherein the fatty acids are: More than 10 wt % eicosapentaenoic acid (EPA) and / or docosahexaenoic acid (DHA) as triglyceride conjugates and / or diglyceride conjugates, and Less than 15 wt% EPA and / or DHA as free fatty acids A plant extract comprising:

11. 11. The plant extract of claim 10, wherein the fatty acids comprise more than 15 wt. % EPA and / or DHA as triglyceride conjugates and / or diglyceride conjugates and less than 10 wt. % EPA and / or DHA as free fatty acids.

12. 12. The plant extract of claim 10 or 11, wherein the fatty acids comprise 18 wt. % EPA and / or DHA as triglyceride conjugates and / or diglyceride conjugates and 7 wt. % EPA and / or DHA as free fatty acids.

13. 13. The plant extract of any one of claims 10 to 12, wherein the Nannochloropsis algae includes N. oculata.

14. The Nannochloropsis algae have a density of at least 700 micromol m －２ s －１ The cells are grown at a high density of at least 5 g / l under high illumination intensity of 1000 kJ / L and continuously bubbling and CO ２ The plant extract according to any one of claims 10 to 13, which has been concentrated.

15. A food or nutritional supplement comprising the plant extract of any one of claims 10 to 14.

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