WO2021194360A1

WO2021194360A1 - Extraction method for bio-active fractions

Info

Publication number: WO2021194360A1
Application number: PCT/NZ2021/050052
Authority: WO
Inventors: Craig Lachlan Mcintosh; Gary Neil Monk; Rau Makeri Pohatu
Original assignee: Pharmazen Limited
Priority date: 2020-03-27
Filing date: 2021-03-29
Publication date: 2021-09-30
Also published as: NZ793006A

Abstract

An extraction method for obtaining oil based bioactive fractions, from plant tissue and/or animal tissue ('the substrate'), especially Antarctic Krill ( euphasia superba), and New Zealand Green-Shelled Mussel (perna canaliculus ) tissue. The method including the steps of a) subjecting the substrate to a dehydration or drying step, to produce a dehydrated or dried sample having less than 5% water content; b) subjecting the dehydrated substrate to a 96% - 100% ethanol extraction step to produce an extract from the sample; c) collecting the extract from step b); d) repeating steps b) and c); e) combining all the extracts obtained from steps b) and c) following step d) and subjecting the extracts to a primary evaporation step to obtain the oil based bioactive fraction in a semi-concentrated form.

Description

EXTRACTION METHOD FOR BIO-ACTIVE FRACTIONS

FIELD OF INVENTION

The invention relates to an extraction method for extracting bio-active fractions.

In particular, although not exclusively, the invention relates to an efficient method for extracting and blending at least two separate oil based bioactive fractions obtained from separate sources.

BACKGROUND TO THE INVENTION

The manufacture of high potency natural nutritional ingredients sourced from plant and animal tissue requires new extraction techniques which can not only: effectively capture the bioactive fractions with high purity and minimal degradation; but also

- extract bioactive fractions in a form which enables them to be usefully combined with other bioactive fractions obtained from one or more different sources; to help formulate new products and deliver more health benefits.

One non-limiting example of a useful combination of differently sourced oil based bioactive fractions is Krill oil and greenshell mussel (GSM) oil.

However, a problem that exists currently is how to create a stable homogenous blend of these two bioactive oil fractions, at a desired ratio.

Presently, GSM oil and Krill oil are extremely hard to effectively homogenously blend together given the respective viscosities involved. Thus, current commercially available GSM oil and Krill oil blends are not that homogenous. This is problematic when looking to encapsulate a blend of GSM and Krill oils in gel capsules - as it is difficult to know with certainty what percentage of Krill oil or GSM oil is present in any given capsule.

It would also be useful to have greater control when blending two bioactive oil fractions so as to control what percentage of the respective fractions is present in the overall final blended product.

For a non-limiting example, of blending two bioactive oil fractions together with greater control than conventional blends of bioactive oil fractions, it would be useful to have a blend formulated to contain 1% - 99% Krill oil and 1%-99% GSM oil, which is stable and homogeneous enabling this new formulation to be put into capsules and commercialised as having a desired percentage ratio of Krill oil to GSM oil. It is an object of the invention to address the foregoing problems or at least provide the public with a useful choice.

DEFINITIONS

The term ‘bioactive fraction’ refers to a final form of an extract which has been isolated and purified from source plant or animal tissue. The bioactive fraction includes at least one biologically active substance therein, in a concentrated form compared to the source plant or animal tissue.

The term ‘extract’ refers to both the act of removing unwanted material from the tissue as well the resultant product after removal of said unwanted tissue or other matter which may include materials used in the extraction process. An extract may be in an initial or final form. A final form of an extract being a more purified or otherwise more refined form of the initial extract.

The term 'at least 96% ethanol’ as used herein refers to ethanol having minimum 96% v/v of C2H5OH at 20°C.

The terms Anhydrous ethanol’ and Pure ethanol’ and ‘Absolute ethanol’ and ‘Dehydrated ethanol’ as used herein refer to ethanol having minimum 99% v/v C2H5OH at 20°C.

The term ‘semi-concentrated form’ as used herein refers to an oil based bioactive fraction which is not fully evaporated and still retains more water/solvent therein than the final concentrated form. Generally, the semi-concentrated form of the bioactive fraction has a loss on drying (LoD) endpoint of 5% for Krill and 10% for GSM.

The term “parallel extracted blend” as used herein refers to a blend of two or more bioactive fractions wherein each fraction to be blended has undergone substantially the same key extraction process steps to arrive at the respective bioactive fractions that are to be blended.

Unless the context clearly requires otherwise, throughout the description and the claims, the words “comprise”, “comprising”, and the like, are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense, that is to say, in the sense of “including, but not limited to”.

SUMMARY OF THE INVENTION

According to a first aspect of the present invention, there is provided an extraction method for obtaining oil based bioactive fractions, from plant tissue and/or animal tissue (‘the substrate’) the method including the steps of: a) subjecting the substrate to a dehydration or drying step, to produce a dehydrated or dried sample having less than 5% water content; b) subjecting the dehydrated substrate to a 96% - 100% ethanol extraction step to produce an extract from the sample; c) collecting the extract from step b); d) repeating steps b) and c); e) combining all the extracts obtained from steps b) and c) following step d) and subjecting the extracts to a primary evaporation step to obtain the oil based bioactive fraction in a semi-concentrated form.

Generally step d) may be repeated two, three or four times.

In preferred embodiments step d) may be repeated until the refractometer reading of the extraction is no more than 21-22%.

According to a second aspect of the present invention, there is provided a method for creating a homogenous blend of at least two bioactive fractions wherein each bioactive fraction is obtained via the method substantially as described above including the further step of: f) blending the two bioactive fractions together whilst subjecting to a secondary evaporation step.

According to a third aspect of the present invention, there is provided a method substantially as described above wherein one bioactive fraction is GSM oil and the other bio-active fraction is Krill oil.

According to a fifth aspect of the present invention, there is provided a method for creating blend of at least two bioactive fractions sourced from plant and/or animal tissue, wherein:

- at least one bio-active fraction is obtained by the extraction method of the first aspect of the present invention, or a blend of two bioactive fractions as per the second aspect of the present invention, (herein individually or collectively referred to as the First Bioactive Fraction(s) (FBF)); and wherein

- at least one other bioactive fraction (alternative bioactive fraction(s) (ABF)) is in a powder form obtained via an alternative extraction technique; the method comprising the steps of: a) dissolving the ABF in ethanol; b) blending together - the ABF with the FBF obtained from a method substantially as described above in relation to the first to third aspects - whilst performing a further evaporation step.

According to a sixth aspect of the present invention, there is provided a method substantially as described above wherein the ABF is obtained from blackcurrant tissue.

According to a seventh aspect of the present invention, there is provided a method substantially as described above wherein the FBF is a blend of GSM oil and Krill oil.

The ratio of GSM oil to Krill oil in the blend may vary as may be desired. Some preferred but non-limiting examples are given below.

The blend may contain a ratio of 1% GSM oil to 99% Krill oil.

The blend may further contain a ratio of 5% GSM oil to 95% Krill oil;

The blend may further contain a ratio of 90% GSM oil to 10% Krill oil; or vice versa.

The blend may further contain a ratio of 80% GSM oil to 20% Krill oil; or vice versa.

The blend may further contain a ratio of 70% GSM oil to 30% Krill oil; or vice versa.

The blend may further contain a ratio of 60% GSM oil to 40% Krill oil; or vice versa.

The blend may further contain a ratio of 50% GSM oil to 50% Krill oil; or vice versa.

According to an eighth aspect of the present invention, there is provided a method substantially as described above wherein the FBF is GSM oil.

According to a ninth aspect of the present invention, there is provided a method substantially as described above wherein the FBF is Krill oil.

According to a 10^th aspect of the present invention, there is provided a method of obtaining a Krill oil bioactive fraction wherein the method comprises the step of freeze drying the Krill tissue to have less than 5% water content.

According to a 11^th aspect of the present invention, there is provided a method of obtaining a Krill oil bioactive fraction wherein the method comprises the step of additional mincing and freeze drying the Krill tissue to have less than 3% water content.

According to a 12^th aspect of the present invention, there is provided a method of obtaining a Krill oil bioactive fraction substantially as described above wherein the freeze dried Krill tissue is then subjected to a 96% - 100% ethanol extraction step to produce an extract from the sample. According to a 13^th aspect of the present invention, there is provided a method of obtaining a Krill oil bioactive fraction substantially as described above wherein the freeze dried Krill tissue is then subjected to a 99.9% ethanol extraction step to produce an extract from the sample.

According to a 14^th aspect of the present invention, there is provided a method of obtaining a Krill oil bioactive fraction substantially as described above wherein the freeze dried Krill tissue is then subjected to repeated steps b) and c), namely multiple ethanol extraction steps, to produce Krill oil yields of approximately 35-45%.

According to a 15^th aspect of the present invention, there is provided a method of obtaining a Krill oil bioactive fraction substantially as described above wherein the ethanol extracts are recirculated and pumped back through the Krill tissue to facilitate efficient extractions and maximise Krill oil yields.

According to a 16^th aspect of the present invention, there is provided a method of producing a blend of at least two bioactive oil fractions obtained from plant or animal tissue wherein the method comprises the step of:

• subjecting respective tissue fractions - from which the bioactive oil fractions are obtained - to substantially the same key extraction process steps - to achieve a parallel extracted blend.

BRIEF DESCRIPTION OF THE DRAWINGS

One or more embodiments of the invention will be described below by way of example only, and without intending to be limiting, with reference to the following drawings, in which:

Figure 1 shows a flow chart detailing the key extraction process steps of the present invention according to one preferred embodiment.

Figure 2 shows a flow chart detailing the key extraction process steps for GSM and Krill to achieve a parallel extracted blend.

Figure 3 shows a line graph of the correlation between the refractometer readings and Krill oil content of the extracted samples.

Further aspects of the invention, which should be considered in all its novel aspects, will become apparent to those skilled in the art upon reading of the following description which provides at least one example of a practical application of the invention.

BEST MODES With respect to the drawings Figure 1 shows the key extraction process steps of the present invention that are performed to obtain the oil based bioactive fractions. If two or more plant or animal tissues are being separately extracted to obtain bioactive oil fractions using the same key extraction steps outlined in Figure 1 the respective tissues are being extracted in parallel. Figure 2 shows the key extraction steps being performed in parallel for obtaining a blend of GSM oil and Krill oil.

In relation to the key extraction steps of Figures 1 and 2.

• The mincing of the raw material is an optional step in relation to Figure 1 as certain plant and animal tissue raw materials may not need mincing or other pre-preparation prior to being freeze dried.

• The freeze drying step importantly brings the water content down to less than 5%, and preferably less than 3%.

• The ethanol extraction step importantly uses 96 -100% ethanol, preferably 99.9% ethanol.

• The primary evaporation step importantly removes the ethanol and any remaining water to between substantially 5% — substantially 15%, and not less than 5%.

Figure 3 shows the correlation between refractometer readings and Krill oil content of the extracted samples. If the refractometer reading of an extracted sample is above 21-22%, an additional extraction step will be undertaken.

Example 1 The process steps used to perform the method of the present invention as detailed in the first aspect according to one preferred embodiment are detailed in Table 1.

Table 1 details the key parameters of process steps and extract composition of the extraction method/blending method of the first and second aspects of the present invention.

Table 1

1. Starting raw GSM received as raw deshelled Krill received as processed meal materials frozen mussel meat. (known as OlyMeg from Olympic

Seafoods). Mincing and freezing Minced to size 16-19 mm. Minced to size 1-5 mm. preparation

Processing temperatures Processing temperatures -18-10°C. -18-10°C.

Process times 2-4 hrs. Process times 2-4 hrs.

Blast frozen 12-24 hrs. Blast frozen 4-12 hrs. Freeze drying Cycle length 24-49 hrs. Cycle length 8-16 hrs.

Dryer temperatures 30-80°C. Dryer temperatures 30-70°C. Product temperatures -18-60°C. Product temperatures -18-45°C. Vacuum 20-300 pascals Vacuum 20-50 pascals Product water activity Aw 0.2-0.7 Product water activity Aw 0.2-0.3. Product moisture 1-5%

Product moisture 1-3% Freeze dried (FD) FD GSM composition (g/100g FD Krill Meal composition product quality Triglyceride (TG)) (g/1 OOg Triglyceride (TG))

Eicosapentaenoic acid (EPA) EPA 13.0 18.8

DHA 7.4

Docosahexaenoic acid (DHA)

Omega³27.5 13.8

Phospholipids 45.7

Omega³ 34.55

Astaxanthin (Asta) 255.0 mg/kg Phospholipids 34.4

Viscosity 654.3cP Viscosity lOOOOcP Ethanol Extraction Un-milled FD GSM FD Krill meal

Counter-current extraction or Counter current extraction or Cross current extraction. Cross-current extraction.

Ethanol: raw material (ETOH:RM) ETOHiRM ratio 6-8L ratio 8-12L

ETOH conc.% min 96%.

ETOH conc.% min 96%. Extraction time 45-75 minutes. Extraction time 45-75 minutes. Extraction temperatures 45-50°C.

Extraction temperatures 20-45°C.

Extraction endpoint

Extraction endpoint refractometer refractometer reading 21-22% reading 21-22% brix. brix.

LOD endpoint 2-5%.

LOD endpoint 2-5%. Primary evaporation Key operating parameters: Key operating parameters: in FFE (Falling film

1. Temp set point 40-60 °C 1. Temp set point 35-50 °C evaporator)

2. Product temp 20-35°C. 2. Product temp 20-45°C.

3. Evaporation rate 3. Evaporation rate 1000-1500 L/hr. 600-1500 L/hr.

4. Specific Gravity (SG) 4. SG concentration set concentration set point point 0.94-0.96 0.91-0.93

Blending and Key operating parameters for 1. Temp set point 65-70 °C secondary blending GSM bioactive oil

2. Product temp 45-60°C. evaporation in WFE fraction with Krill bioactive oil (Wiped film fraction: 3. Evaporation rate 70-100 evaporator). L/hr.

4. LOD endpoint 0.60-0.80%.

Process Step Product Parameters

Finished product Blended GSM/Krill Lipid quality See Tables 4(i) and (ii) below transferred to Parameters: storage tanks ready for final packaging Specifically, total lipids content of incoming Krill and GSM raw materials may be checked on Extraction System B-811 (BIJCHI Labortechnik AG) using the Soxhlet analytical method with ethanol as the solvent extractant. The Extraction System B-811 Operation Manual provides details on how to carry out the method. Krill and GSM raw materials may have dry weight lipid contents of 48-50%, which may be later compared against to determine actual production recoveries. Preferably, 80% product recovery may be aimed for.

In addition, residual lipids after extraction may be checked using the Soxhlet analytical method on occasions to determine that residual lipids in the extracted substrates are lower than 6-8%, as compared to 48-50% of the raw materials before extraction, thereby confirming the extraction efficiency on the particular substrate material. The extraction efficiency in each extraction step may also be monitored using a correlation between end point refractometer readings in the ethanol extracts and the oil content in the extracts.

An example of how this is achieved is now explained in relation to Krill oil but it will be understood that the same methodology in relation to refractometer readings may also be used for GSM oil .

The correlation is established by first collecting multiple extracted samples from the process of the present invention. The oil content of these extracted samples are then determined by identifying LoD on drying at 105°C for 1 hour. The refractometer readings of these samples are then measured on the refractometer in % brix. The correlation between refractometer readings in the ethanol extracts and the oil content in the extracts is then determined by the best fit as shown in Figure 3.

This correlation may be used to predict from refractometer readings what the oil content is in the Krill extracted samples. Specifically, the final extraction step must contain no more than a 21-22% refractometer reading which indicates that most of the Krill or GSM oil has been extracted. If the refractometer reading is above 21-22%, an additional extraction step will be undertaken.

Table 2 below details the yield percentage from the raw material (RM) achieved at process step 7 above for each of Krill oil and GSM oil.

Table 2

Table 3 below details the product parameters for each of the GSM bioactive oil and Krill bioactive oil obtained at process step 7 in Table 1 above.

Table 3

Tables 4(i) and 4(ii) detail the Krill/GSM Oil Blend Quality parameters that can be achieved at Process Step 8 above.

Table 4(i)

Table 4(ii)

Example 3 CASSIPURE™ powdered anthocyanin rich blackcurrant (BC) extract manufactured by Waitaki Biosciences was used for making the following blend with Krill Oil. The BC extract is measured to the desired amount and then dissolved in 99% ethanol. The dissolved extract is then blended with the desired amount of Krill Oil obtained from the process of the present invention during the secondary scraper evaporation stage (for producing the Krill Oil).

It should be noted if a BC extract is being obtained from raw material (e.g. New Zealand Blackcurrant berry ( ribes nigrum) whole berries or pomace sourced from growers and stored frozen) via an ethanol extraction technique or an enzyme/resin purification technique it may be possible to introduce the ethanol BC extract mix to the Krill Oil at step 6 of the present invention as detailed in Example 1.

Table 5 below details the yield percentage from the raw material (RM) achieved for each of Krill oil and BC extract.

Table 5

Tables 6(i) and 6(ii) detail the Krill oil and Black Currant extract Blend Quality parameters that can be achieved when blended as described above. Table 6(i)

Table 6(ii)

Example 4

A GSM blackcurrant blend can be made using the same method as detailed in Example 3 above.

Table 7 below details the yield percentage from the raw material (RM) achieved for each of GSM oil and BC extract. Table 7

Example 5

A GSM oil/Krill oil blend can also be further blended with blackcurrant using substantially the same method as detailed in Example 3. DETAILED DESCRIPTION OF THE INVENTION INCLUDING ALTERNATE WAYS TO

IMPLEMENT THE INVENTION

The plant or animal tissue sample may generally be subjected to one or more pre-treatments, prior to being subjected to, the process of the present invention and being subjected to the initial dehydration step. However, it will also be appreciated that in some cases, dependent on the tissue, no pre treatment may be necessary, prior to being subjected to, the process of the present invention and being subjected to the initial dehydration step.

In some preferred embodiments the plant or animal tissue may be minced/macerated or otherwise in a reduced, diced or chopped form, as necessary depending on the tissue, prior to being subjected to the dehydration step of the present invention.

In some further preferred embodiments, the minced tissue may be frozen prior to being subjected to the dehydration step of the present invention. Alternatively, the plant or animal tissue may be dissolved, or otherwise pre-processed in a suitable manner as would be well known to a person skilled in the art, so as to facilitate the extraction of the bioactive oil fraction.

The dehydration step may be performed in a number of ways without departing from the present invention.

In some embodiments the dehydration step may comprise exposing the plant or animal tissue to infrared radiation.

In preferred embodiments the dehydration step may comprise freeze dying the tissue sample in a freeze drier for a period sufficient for the water content of the tissue to drop to substantially 5% or less, preferably less than 3%.

In a further preferred embodiment, the freeze drier may include an NIR sensor.

The plant or animal tissue from which the bioactive oil fraction is to be extracted may determine the optimum technique for dehydration.

The water content of the tissue may preferably be monitored/measured using near infrared (NIR) testing. Other methods for testing the water content may be employed without departing from the scope of the present invention.

The 96%-100%, preferably 99.9% ethanol extraction step preferably uses of the purest ethanol available. That said the inventors have found the invention performs significantly better than conventional extraction techniques for bioactive oil fractions when the minimum threshold of at least 96% ethanol is used.

In some preferred embodiments the ethanol extraction step may use anhydrous ethanol (also known as “absolute ethanol” or “dehydrated alcohol”).

For some tissues a counter-current ethanol extraction technique may be used. One example of such a tissue is GSM.

For other tissues a cross-current ethanol extraction may be employed. An example of such a tissue is Krill.

The extract obtained from the method of the present invention once the 96% - 100% ethanol extraction has been performed is then subjected to an evaporation step.

In preferred embodiments the ethanol extraction occurs at temperatures substantially between 20°C - 50°C. In some preferred embodiments the ethanol extraction step may be performed by a high throughput pressurised liquid extraction apparatus.

Preferably, the ethanol extraction step may be carried out at atmospheric pressures using a liquid recirculation system through the substrate being extracted.

The primary evaporation step may be undertaken in a variety of different ways.

In a preferred embodiment the primary evaporation step may take place in a falling film evaporator (FFE).

In a further preferred embodiment, the FFE may include an NIR sensor.

The primary evaporation step once complete yields a semi-concentrated bioactive oil fraction.

However, the inventors have found that if you want to blend the bioactive oil fraction (say GSM oil) with another bioactive oil fraction (say Krill oil) it is advantageous to subject the two oil fractions to a combined blending and secondary evaporation step.

The blending and secondary evaporation step may be undertaken in a variety of different ways.

In a preferred embodiment the blending and evaporation step may take place in a wiped film evaporator (WFE).

However, other evaporators suitable for high viscosity substrates may also be employed.

In a further preferred embodiment, the WFE may include an NIR sensor.

Upon completion of the primary evaporation step a semi-concentrated oil based bioactive fraction is obtained.

The oil based bioactive fraction obtained may be in an initial form (if further processing/refinement is to be undertaken); or a final form extract (if the bioactive fraction is ready to be used/sold with no further processing steps other than packaging).

Without wishing to be bound by theory the inventors have found that having water content in oils too late in the extraction process is detrimental to producing a stable homogenous blend of two or more bioactive oil-based fractions. Typically, what is encountered when trying to blend together two oil based bioactive fractions using conventional techniques is that the evaporation step requires higher temperatures and/or longer times to remove water so as to blend the two bioactive oil fractions together. However, counter to achieving the desired blending is a concomitant increase in the viscosity of the two oil fractions which resists the dispersion of the two oil fractions to form a blend. The invention may also be said broadly to consist in the parts, elements and features referred to or indicated in the specification of the application, individually or collectively, in any or all combinations of two or more of said parts, elements or features.

It should be noted that various changes and modifications to the presently preferred embodiments described herein will be apparent to those skilled in the art. Such changes and modifications may be made without departing from the spirit and scope of the invention and without diminishing its attendant advantages. It is therefore intended that such changes and modifications be included within the present invention as claimed.

Claims

WHAT WE CLAIM IS:

1. An extraction method for obtaining oil based bioactive fractions, from plant tissue and/or animal tissue ('the substrate’) the method including the steps of: a) subjecting the substrate to a dehydration or drying step, to produce a dehydrated or dried sample having less than 5% water content; b) subjecting the dehydrated substrate to a 96% - 100% ethanol extraction step to produce an extract from the sample; c) collecting the extract from step b); d) repeating steps b) and c); e) combining all the extracts obtained from steps b) and c) following step d) and subjecting the extracts to a primary evaporation step to obtain the oil based bioactive fraction in a semi-concentrated form.

2. A method for creating a homogenous blend of at least two bioactive fractions with each bioactive fraction obtained via the method of claim 1 including the further step of: e) blending the two bioactive fractions together whilst subjecting to a secondary evaporation step.

3. A method as claimed in claim 1 wherein the dehydrated or dried sample has less than 3% water content.

4. A method as claimed in claim 1 wherein the dehydrated substrate is subjected to a 99.9% ethanol extraction step to produce an extract from the sample.

5. A method as claimed in claim 1 wherein steps b) and c) are repeated for multiple times before being subjected to step d).

6. A method as claimed in claim 2 wherein one bioactive fraction is GSM oil and the other bio-active fraction is Krill oil.

7. A method for creating a blend of at least two bioactive fractions sourced from plant and/or animal tissue, wherein:

- at least one bio-active fraction is obtained by the extraction method claimed in claim 1 , or is a blend of bioactive fractions as claimed in claim 2 or claim 6, (herein individually or collectively referred to as the First Bioactive Fraction(s) (FBF)); and wherein

- at least one other bioactive fraction (alternative bioactive fraction(s) (ABF)) is in a powder form obtained via an alternative extraction technique; the method comprising the steps of: a) Dissolving the ABF in ethanol; b) blending together the ABF with the FBF whilst performing a further evaporation step.

8. A method as claimed in claim 7 wherein the ABF is obtained from blackcurrant tissue.

9. A method as claimed in claim 8 wherein the FBF is a blend of GSM oil and Krill oil.

10. A method as claimed in claim 8 wherein the FBF is GSM oil.

11. A method as claimed in claim 8 wherein the FBF is Krill oil.

12. A method of obtaining a Krill oil bioactive fraction wherein the method comprises the step of freeze drying the Krill tissue to have less than 5% water content.

13. A method as claimed in claim 12 wherein the method comprises the step of additional mincing and freeze drying the Krill tissue to have less than 3% water content.

14. A method of obtaining a Krill oil bioactive fraction as claimed in claim 12 wherein the freeze-dried Krill tissue is then subjected to a 96% - 100% ethanol extraction step to produce an extract from the sample.

15. A method as claimed in claim 14 wherein the freeze-dried Krill tissue is subjected to a 99.9% ethanol extraction step to produce an extract from the sample.

16. A method of obtaining a Krill oil bioactive fraction as claimed in claim 14 wherein the ethanol extracts are recirculated and pumped back through the Krill tissue to facilitate efficient extractions and maximise Krill oil yields.

17. A method of producing a blend of at least two bioactive oil fractions obtained from plant or animal tissue wherein the method comprises the step of: a) subjecting respective tissue fractions from which the bioactive oil fractions are to be obtained to substantially the same key extraction process steps - to achieve a parallel extracted blend.