HK1176866B - Liquid/liquid extraction - Google Patents

Description

Liquid/liquid extraction

Technical Field

The present invention relates to a process for extracting unsaponifiable parts (especially parts or whole parts) of vegetable oils, vegetable butters or oils derived from microorganisms.

Background

The unsaponifiable matter or unsaponifiable fraction of fat (fraction) is a part of the fat, remains insoluble in water after the continued action of the alkali and can be extracted with an organic solvent.

Most unsaponifiables of vegetable oils or butter contain several large families of substances. Which may include saturated or unsaturated hydrocarbons, fatty alcohols or triterpene alcohols, sterols, tocopherols, carotenoid pigments, lutein, and in the case of certain oils and butters, one or two specific and distinct families.

The standard method for obtaining unsaponifiables in vegetable oils and vegetable butters aims at extracting all or part of said large family of substances contained therein for the preparation of part or all of the unsaponifiable fraction.

In particular, part or all of the unsaponifiable fraction is studied for its pharmaceutical, cosmetic and nutritional properties.

The standard method for obtaining unsaponifiables in vegetable oils and vegetable butters comprises the steps of saponification of the fat and extraction of the desired product (the unsaponifiable) by means of an organic solvent.

The most commonly used solvents are oil-based solvents and include alkanes (hexane, heptane, etc.) and chlorinated solvents (1, 2-Dichloroethane (DCE), trichloroethane, 1-chlorobutane, carbon tetrachloride, etc.). Among the latter, in particular, DCE and 1-chlorobutane are the best candidates in view of extraction yield and selectivity.

However, from an industrial point of view, the toxicity of the solvents used and their chemical stability must be taken into account. For this reason, chlorinated solvents (in particular 1, 2-Dichloroethane (DCE) and 1-chlorobutane) have three major disadvantages: they can degrade in alkaline media (as in the case of saponification with soap solutions), they are classified as toxic solvents (DCE in particular is classified as CMR), and they have a negative impact on the environment.

Chlorinated solvents often have unacceptable toxicity and/or risk.

Furthermore, processes for obtaining unsaponifiable fractions may require the use of organic solvents in amounts that are not suitable for the viability of the process, may include too many extraction steps and may be slow, and/or have unacceptable phase separation (e.g., the production of unwanted emulsions), both from an economic and environmental standpoint. In particular, it may be necessary or beneficial to adjust the amount of fat used in the saponification process in order to optimize the proportion of solvent used.

Disclosure of Invention

It is therefore an object of the present invention to address all or part of the above problems. In particular, the object of the present invention is to provide a more economical, more direct, more environmentally friendly process which requires a smaller amount of organic solvent, is easier to implement, is faster, and which produces less toxic and/or less hazardous conditions, improves the phase separation, making it possible to obtain unsaponifiables with a yield, cost and/or selectivity at least comparable to existing processes.

In particular, it would be desirable to use solvents which are less hazardous, less toxic, in particular not classified as CMR, which are chemically more stable than 1, 2-dichloroethane and/or 1-chlorobutane, and/or which make it possible to extract unsaponifiables with yields and/or selectivities at least comparable to those obtained with 1, 2-dichloroethane and/or 1-chlorobutane.

So-called "solvents classified as CMR" may be those indicated in the list in the appendix of the 2009/2/CE directive on 1, 15 of 2009, and in particular may be selected fromhttp://eur-lex.europa.eu/LexUriServ/ LexUriServ.do?uri=OJ:L:2009:011:0006:0082:FR:PDFIs obtained herein and referred to as "EU 1 (French abbreviation UE 1) CMR List"; those listed under the European regulatory Classification of chemicals (la Classification europ ene ne ergaletaire products chimiques canaquee rog e nes, mutag des et toxiques point ur la reproduction-31e ATP, 2009) in ATP 31 th year, in particular, can be selected fromhttp://www.prc.cnrs-gif.fr/en telechargement/cmr31.pdfObtained from the address of (a), which second list is referred to herein as the "EU 2 (french abbreviation UE 2) CMR list"; and/or those solvents listed in the "California cosmetics safety Act of 2005" (California cosmetics Act 2005) document entitled "Chemicals known or suspected to cause cancer or reproductive toxicity" established by the "California public health department of public health, occupational health branch, California safety cosmetics project," date 2009, 9/1, the third list referred to herein as the "CMUS R List".

In the context of the present invention, the expression "EU CMR list" refers to the EU1CMR list and/or the EU2CMR list.

One of the more specific aims is to obtain specific unsaponifiable fractions, for example fractions with a high content of certain compounds and a low content of other compounds, in particular fractions comprising only one or certain families of the total unsaponifiable compounds, and/or fractions with a composition comparable to that obtained by processes using DCE and/or 1-chlorobutane.

The unsaponifiable matter may be composed of a variety of components, in particular comprising a large family of substances as defined above and/or a special family of substances. It is desirable to extract as completely as possible at least one of these families, in particular at least two, in particular at least three, more in particular at least four, or at least five, more in particular at least six, even more in particular all families of substances which make up unsaponifiables in a given oil or butter.

On the other hand, the process according to the invention is intended to provide a specific fraction of unsaponifiables (fraction of unsaponifiable matter) and in particular a content rich in at least one family of substances that make up said unsaponifiables, or to extract one or more specific compounds of said unsaponifiables, or to provide a total or nearly total fraction of unsaponifiables.

The invention therefore relates to a process for the extraction of an unsaponifiable fraction, in particular a partial or total fraction, contained in vegetable oils or butter, in oils derived from microorganisms, in concentrates of vegetable oils or butter or of oils derived from microorganisms, or in by-products (co-products) of the refining industry involving vegetable oils or oils derived from microorganisms, such as the deodorisation emissions (lees chapps de sorption), said process comprising at least:

A) comprising a step of converting said oil, said butter or said by-products of the refining industry involving vegetable oils or oils derived from microorganisms into a hydro-alcoholic solution, in particular by a step selected from saponification and esterification,

B) comprising the step of extracting said hydro-alcoholic solution by means of liquid/liquid extraction or distillation, wherein the fat fraction is separated from the unsaponifiable fraction,

C) optionally, a step selected from crystallization and liquid/liquid extraction comprising purification of the unsaponifiable matter,

wherein the process is characterized in that at least one of the liquid/liquid extraction step in step B, the crystallization step in step C and the liquid/liquid extraction step in step C is carried out with a first solvent system comprising a solvent in an amount of at least 50% by volume relative to the total volume of the solvent system, said solvent being selected from the group consisting of fluorinated aromatic solvents, in particular trifluorotoluene (BTF) and hexafluorobenzene (BHF), tert-butyl ethers, in particular 2-ethoxy-2-methylpropane, also known as ethyl tert-butyl ether (ETBE), solvents comprising at least one silicon atom, in particular Hexamethyldisiloxane (HMDS) and Tetramethylsilane (TMS), methyltetrahydrofuran (MeTHF) and mixtures thereof.

CAS numbers for these different solvents are as follows, BTF: 98-08-8; BHF: 392-56-3; ETBE: 37-92-3; MTBE: 1634-04-4; HMDS: 107-46-0; TMS: 75-76-3; and MeTHF: 96-47-9.

In the present invention, the expression "fraction" means that the fraction contains all the substances constituting the unsaponifiable matter present in a given vegetable oil, oil derived from a microorganism or vegetable butter.

In the present invention, the expression "fraction part" means that this part comprises at least one family of substances constituting said unsaponifiable matter present in a given vegetable oil or butter or in an oil derived from a microorganism.

According to a particular embodiment, the present invention relates to a process wherein step B) comprises or consists of liquid/liquid extraction with said first solvent system.

According to another particular embodiment, the present invention relates to a process wherein step C) comprises or consists of crystallization or liquid/liquid extraction with said first solvent system.

According to an even more particular embodiment, the process comprises or consists of a liquid/liquid extraction with the first solvent system and step B) comprising or consisting of a liquid/liquid extraction with the first solvent system, the step C) comprising or consisting of a crystallization and/or a liquid/liquid extraction with the first solvent system, the first solvent system being the same as or different from the one used in step a).

In particular, step C) may comprise the purification of an unsaponifiable fraction enriched in one or more families of substances that make up the unsaponifiable, present in a given vegetable oil, oil from a microorganism or margarine. In particular, this step may further comprise the isolation of a specific fraction of the soybean unsaponifiable matter (such as sterol compounds) or of the avocado unsaponifiable matter (such as furan and/or sterol compounds).

The first solvent system may comprise a solvent selected from fluorinated aromatic solvents, in particular Benzotrifluoride (BTF) and hexafluorobenzene (BHF), tert-butyl ether, in particular 2-ethoxy-2-methylpropane, also known as ethyl tert-butyl ether (ETBE), solvents comprising at least one silicon atom, in particular Hexamethyldisiloxane (HMDS) and Tetramethylsilane (TMS), methyltetrahydrofuran (MeTHF) and mixtures thereof, in a content of at least 60%, in particular at least 75%, in particular at least 90%, more in particular at least 95%, even more in particular at least 99% by volume relative to the total volume of the first solvent system.

In particular, the first solvent system consists of fluorinated aromatic solvents, in particular trifluorotoluene (BTF) and hexafluorobenzene (BHF), tert-butyl ether, in particular 2-ethoxy-2-methylpropane, also known as ethyl tert-butyl ether (ETBE), solvents comprising at least one silicon atom, in particular Hexamethyldisiloxane (HMDS) and Tetramethylsilane (TMS), methyltetrahydrofuran (MeTHF) or mixtures thereof.

The first solvent system may comprise a solvent selected from fluorinated aromatic solvents, in particular Benzotrifluoride (BTF) and hexafluorobenzene (BHF), tert-butyl ether, in particular 2-ethoxy-2-methylpropane, also known as ethyl tert-butyl ether (ETBE), solvents comprising at least one silicon atom, in particular Hexamethyldisiloxane (HMDS) and Tetramethylsilane (TMS), and methyltetrahydrofuran (MeTHF), in a content of at least 50% by volume, in particular at least 75% by volume, in particular at least 90% by volume, more in particular at least 95% by volume, even more in particular at least 99% by volume, relative to the total volume of the first solvent system.

According to one variant, the first solvent system consists of a fluorinated aromatic solvent, in particular trifluorotoluene (BTF) and hexafluorobenzene (BHF), a tert-butyl ether, in particular 2-ethoxy-2-methylpropane, also known as ethyl tert-butyl ether (ETBE), a solvent comprising at least one silicon atom, in particular Hexamethyldisiloxane (HMDS), Tetramethylsilane (TMS), or methyltetrahydrofuran (MeTHF).

According to a second variant, the first solvent system consists of a first solvent chosen from fluorinated aromatic solvents, in particular trifluorotoluene (BTF) and hexafluorobenzene (BHF), tert-butyl ether, in particular 2-ethoxy-2-methylpropane, also known as ethyl tert-butyl ether (ETBE), solvents comprising at least one silicon atom, in particular Hexamethyldisiloxane (HMDS) and Tetramethylsilane (TMS), methyltetrahydrofuran (MeTHF), and a second solvent, different from the first solvent, chosen from fluorinated aromatic solvents, in particular trifluorotoluene (BTF) and hexafluorobenzene (BHF), tert-butyl ether, in particular 2-ethoxy-2-methylpropane, also known as ethyl tert-butyl ether (ETBE), and 2-methoxy-2-methylpropane, also known as methyl tert-butyl ether (MTBE)), a solvent comprising at least one silicon atom, in particular Hexamethyldisiloxane (HMDS) and Tetramethylsilane (TMS), methyltetrahydrofuran (MeTHF).

According to a third variant, the first solvent system consists of one or more solvents selected from HMDS, HFB and BTF and one or more solvents selected from MeTHF, MTBE, ETBE and TMS.

According to a fourth variant, the first solvent system consists of HMDS and MeTHF, in particular in a ratio range from 50/50 to 90/90 (volume/volume).

Furthermore, the proportion of HMDS used in the first solvent system may play an important role in the consumption of solvent or wash water and/or in the extraction time. It may also facilitate decantation, which in turn leads to the formation of low quality emulsions and/or faster phase separation during the extraction and/or washing steps.

According to a fifth variant, the first solvent system consists of ETBE.

According to a particular embodiment, the first solvent system has a CMR solvent (in particular a solvent present in the EU1, EU2 and/or US CMR list) in a content lower than or equal to 10% by volume, in particular lower than or equal to 5% by volume, in particular lower than or equal to 2% by volume, more in particular lower than or equal to 1% by volume, even more in particular lower than or equal to 0.5% by volume, even lower than or equal to 0.1% by volume, relative to the total volume of the first solvent system.

Even more particularly, the first solvent system is free of solvents present in the EU1, EU2, and/or US CMR list.

The solvent used in the first solvent system has a purity of at least 90%, in particular at least 95%, in particular at least 98%, more in particular at least 99%, even at least 99.5%.

In particular, step A) comprising the conversion of said by-products of the oil, butter or vegetable oil refining industry into a hydro-alcoholic solution is carried out in a second solvent systemIn particular by a step selected from saponification and esterification, said second solvent system comprising a solvent in an amount of at least 50% by volume relative to the total volume of said second solvent system, said solvent being selected from C₂-C₄Alcohols (especially ethanol, n-propanol, isopropanol, butanol, especially n-butanol), MeTHF and mixtures thereof.

The second solvent system may comprise a solvent in an amount of at least 60 vol.%, in particular at least 75 vol.%, in particular at least 90 vol.%, more in particular at least 95 vol.%, even more in particular at least 99 vol.%, relative to the total volume of the second solvent system, said solvent being selected from C₂-C₄Alcohols (especially ethanol, n-propanol, isopropanol, butanol, especially n-butanol), MeTHF and mixtures thereof.

In particular, the second solvent system consists of ethanol, n-propanol, isopropanol, butanol, MeTHF or mixtures thereof.

The second solvent system may comprise a solvent in an amount of at least 50 vol.%, in particular at least 75 vol.%, in particular at least 90 vol.%, more in particular at least 95 vol.%, even more in particular at least 99 vol.%, relative to the total volume of the second solvent system, said solvent being selected from C₂-C₄Alcohols (especially ethanol, n-propanol, isopropanol, butanol, especially n-butanol) and MeTHF.

According to a particular embodiment, the second solvent system comprises solvents present in the EU1, EU2 and/or US CMR list in a content lower than or equal to 10% by volume, in particular lower than or equal to 5% by volume, in particular lower than or equal to 2% by volume, more in particular lower than or equal to 1% by volume, even more in particular lower than or equal to 0.5% by volume, even lower than or equal to 0.1% by volume, relative to the total volume of the second solvent system.

Even more particularly, the second solvent system is free of solvents present in the EU1, EU2, and/or US CMR list.

The solvent used in the second solvent system has a purity of at least 90%, in particular at least 95%, in particular at least 98%, more in particular at least 99%, even at least 99.5%.

In the context of the present invention, the expression "diluted aqueous-alcoholic solution" ("DAS" abbreviated by the french term "SHD") refers to a diluted saponification reaction medium, in particular comprising water and one or more apolar solvents, in particular chosen from alcohols (for example C)₂-C₄Alcohol) and MeTHF.

The extracted diluted aqueous-alcoholic solution (DAS) may have a water content higher than or equal to 50 vol%, in particular higher than or equal to 60 vol%, in particular higher than or equal to 65 vol%, more in particular higher than or equal to 70 vol%, even higher than or equal to 72 vol%, relative to the volume of said aqueous-alcoholic solution.

The diluted aqueous-alcoholic solution (DAS) to be extracted may have a water content lower than or equal to 95% by volume, in particular lower than or equal to 90% by volume, in particular lower than or equal to 85% by volume, more in particular lower than or equal to 80% by volume, even lower than or equal to 75% by volume, with respect to the volume of said aqueous-alcoholic solution.

The ratio (volume/volume) of DAS to solvent system to be extracted may range from 0.1 to 10, especially from 0.25 to 5, especially from 0.5 to 2.

The process according to the invention provides an unsaponifiable fraction which is practically identical to that obtained by the "classical" process described in example 1 below using 1, 2-dichloroethane or 1-chlorobutane.

In the context of the present invention, the expression "substantially identical unsaponifiable fraction" means an unsaponifiable fraction having a similar chromatogram (profile chromatographique) and/or composition as the reference fraction obtained by extraction in DCE or 1-chlorobutane, in particular by classical extraction in DCE or 1-chlorobutane.

In the context of the present invention, the expression "similar chromatogram" means a chromatogram which is characterized in that it comprises the chromatograms of all the families of unsaponifiables observed in the reference fraction obtained by classical methods according to extraction in 1, 2-dichloroethane or 1-chlorobutane.

In the context of the present invention, the expression "similar composition" means a composition characterized in that the content of compounds of the various families of the unsaponifiable extract is of the same order of magnitude as the content observed in the reference fraction obtained by classical means according to extraction in 1, 2-dichloroethane or 1-chlorobutane, and more particularly in relation to the reference fraction, the composition corresponds to the following contents (% by weight with respect to the weight of the reference fraction):

in this case, "+/-Y%" means that, if the reference value is X%, the content may range from (X-Y)% to (X + Y)%. If X is 70% and Y is 15%, the content may range from 55% to 85%.

In particular, the "unidentified" moiety is not within the ranges specified above.

In the context of the present invention, the expression "classical extraction in 1, 2-dichloroethane or 1-chlorobutane" refers to extraction in 1, 2-dichloroethane or 1-chlorobutane according to the method defined in example 1, starting from the same starting materials as those used in the extraction of unsaponifiables with another solvent system.

In the context of the present invention, the expression "reference fraction" refers to the unsaponifiable matter extracted in 1, 2-dichloroethane or 1-chlorobutane, starting from the same raw materials as those used in the extraction of the unsaponifiable matter with another solvent system.

For example, according to the protocol described in example 1 of patent EP1246633B 1.

The invention further relates to a process for the extraction of the fraction (in particular part or all) of unsaponifiables contained in vegetable oils, oils derived from microorganisms, plant butters or by-products of the refining industry relating to vegetable oils (in particular avocado and/or soybean oils) or oils derived from microorganisms, said process comprising at least:

A) a saponification step in which the oil, the butter or the by-products of the refining industry involving vegetable oils or oils derived from microorganisms are converted into a hydro-alcoholic solution,

B) comprising the step of extracting the aqueous-alcoholic solution by means of a first solvent system as defined above.

More particularly, the method of extraction of the fraction of unsaponifiable matter of the soybean oil according to the invention is such that: liquid-liquid extraction is carried out by bringing the DAS together with a first DAS solvent system, in particular using counter-current extraction by means of a first solvent system as defined above, in particular consisting of BTF, wherein the ratio of solvent system to DAS ranges (volume/volume) from 0.1 to 10, in particular from 0.25 to 5, in particular from 0.5 to 2.

More particularly, the extraction process of the fraction of unsaponifiables of the avocado oil according to the invention is such that: liquid-liquid extraction is carried out by bringing the DAS together with a first DAS solvent system, in particular using counter-current extraction by means of a solvent system, in particular consisting of a mixture of ETBE, wherein the ratio of solvent system to DAS (volume/volume) ranges from 0.1 to 10, in particular from 0.25 to 5, in particular from 0.5 to 2.

The vegetable oil or oil derived from microorganisms used in the present process may be selected from the group consisting of soybean, quinoa, rapeseed, corn, sunflower, sesame, lupin, cotton, coconut, olive, palm leaf, wheat germ, alfalfa, avocado, palm kernel, groundnut, copra, flax, castor bean, grape seed, squash seed, blackcurrant seed, melon seed, tomato seed, pumpkin seed, almond, hazelnut, walnut, evening primrose, borage, safflower, linseed, poppy seed, macroalgae, microalgae, such as chlorella (chlorella), and/or oils of microorganisms (especially saline, freshwater or groundwater microorganisms, especially yeasts, molds, more especially bacteria), and mixtures thereof.

The margarine may be selected from cocoa butter, indian red iron butter, shea butter, and mixtures thereof.

Comparison of the content of unsaponifiables in the various vegetable oils (soybean, cotton, coconut, olive and avocado) shows that the avocado oil obtained by extraction according to the various known methods has a particularly high level of unsaponifiables.

Typically, the fraction of unsaponifiable matter obtained ranges from 2% to 10% in avocado oil, about 0.5% in coconut oil, about 1% in soybean oil and about 1% in olive oil.

Avocado unsaponifiables can be prepared by extraction from avocado oil.

The extraction of the unsaponifiable avocado oil can be carried out as follows.

According to methods known to the person skilled in the art:

-alternatively, the fresh pulp is pressed in a cage press (press shell) in the presence of third water-absorbent fibrous bodies (tier colors fibrous adsorbent l' eau), such as coffee shell, and the oil and water emulsions obtained are separated by decantation and/or centrifugation;

alternatively, the fresh pulp is comminuted and brought together with a suitable organic solvent (e.g. a methanol-chloroform mixture) and the oil is recovered by evaporating the solvent. Several methods for extracting the unsaponifiable fraction from vegetable oils have been described in the prior art.

Mention may in particular be made of the process for preparing the unsaponifiable avocado oil described and claimed in patent FR2678632, owned by laboratories pharmacy. In contrast to the classical process for the preparation of avocado oil unsaponifiables, the process provides avocado oil unsaponifiables rich in furan moieties, also known as fraction H.

Thus, as described in patent FR2678632, the avocado oil unsaponifiable used according to the invention can be obtained from fresh fruit before the extraction and saponification of the oil, but it is preferred to prepare said avocado unsaponifiable from previously heat-treated fruit.

The heat treatment consists of a controlled drying of the fruit, which is preferably fresh, carried out at a temperature of preferably at least about 80 ℃ and preferably between about 80 ℃ and about 120 ℃ for at least 4 hours, advantageously at least 10 hours, preferably between about 24 and about 48 hours.

Also mentioned is a process for preparing a soybean oil unsaponifiable matter obtained from a soybean oil unsaponifiable matter concentrate.

The unsaponifiable concentrate can be prepared by means of molecular distillation according to the process described in patent application FR2762512 for lupin oil, but suitable for soybean oil.

In this process, the soybean oil is distilled in a centrifugal or wiped film molecular still, the distillation being carried out at a temperature between about 210 ℃ and 250 ℃ and under a high vacuum in the range between 0.01 and 0.001mm Hg (i.e. 0.13 to 1.3 Pa).

The distillate obtained has an unsaponifiable content of between 5% and 40% by volume and therefore constitutes a soybean oil unsaponifiable concentrate.

The concentrate is then saponified with a base (such as plant ash or soda) in a polar medium (especially an alcoholic medium, preferably ethanol, n-propanol, isopropanol, butanol (especially n-butanol), MeTHF, or a mixture thereof) and then subjected to one or more extractions with the first solvent system.

The obtained extraction solution is then preferably centrifuged, filtered and washed with water in order to remove residual traces of base.

The extraction solvent is carefully evaporated to facilitate recovery of the unsaponifiable matter.

Finally, the oil can be enriched in unsaponifiables by separating the main component of the unsaponifiables recovered in the concentrate before its saponification. Various methods may be used, including: cold crystallization, liquid/liquid extraction and molecular distillation.

The previous concentration of unsaponifiables in the oil makes it possible to reduce the volume of oil to be saponified.

Molecular distillation is particularly preferred, preferably at a temperature between about 180 ℃ and about 230 ℃ and at a pressure of between 10^-3And 10^-2mm Hg, preferably between 10^-3In the order of mm Hg.

The concentration of unsaponifiables in the distillate can be up to 60% by weight, relative to the total weight.

In particular, the present invention relates to a process as described above, wherein the unsaponifiable matter obtained is selected from the group consisting of soybean unsaponifiables, avocado unsaponifiables, in particular avocado unsaponifiables rich in furan fractions and/or avocado unsaponifiables rich in sterol fractions, more in particular mixtures of Avocado and Soybean Unsaponifiables (ASU).

The process according to the invention makes it possible to extract the fraction of unsaponifiables contained in vegetable oils, oils derived from microorganisms or margarines. It may also make it possible to extract the unsaponifiable fraction coming from by-products of the refining industry (generated during the refining of vegetable oils or oils derived from microorganisms) involving vegetable oils or oils derived from microorganisms, such as deodorizing emissions, also known as deodorizers (d).

Indeed, the fatty acids and partial glycerides present in the deodorizer can be saponified or esterified with lower alcohols with the aim of separating the fatty fraction from the unsaponifiable fraction either by liquid/liquid extraction or by vacuum distillation.

Finally, the purification of the unsaponifiable or isolated active fractions, most commonly tocopherols (including vitamin E) and sterols, involves in particular a step of crystallization or liquid/liquid extraction in an organic solvent.

The invention also relates to a process for the extraction of the fraction of unsaponifiable matter of a by-product of the refining industry involving vegetable oils or oils derived from microorganisms, wherein said by-product is a deodorizer of vegetable oils or oils derived from microorganisms, said process comprising at least:

-a step comprising saponification, wherein the deodorizer is converted into a water-alcohol solution,

-a step comprising a counter-current extraction of the water-alcohol solution by means of the first solvent system,

-a step comprising crystallization of a sterol and/or a triterpene alcohol,

a step comprising the isolation of active compounds such as tocopherols, tocotrienols, squalene and carotene, wherein the isolation step is selected from the group consisting of extraction (in particular by means of the first solvent system) and distillation.

In particular, crystallization of sterols and/or triterpene alcohols may be performed in the first solvent system.

The invention further relates to an unsaponifiable fraction, in particular a partial or complete fraction, which is free of solvents classified in the EU1, EU2 and/or US CMR list, wherein in particular said fraction is obtained by the extraction process according to the invention.

The invention further relates to the use of said fraction for the preparation of a composition, in particular a pharmaceutical, edible and/or cosmetic composition, or an edible supplement.

The invention further relates to a composition, in particular an edible, cosmetic or pharmaceutical composition, or an edible supplement, comprising at least one unsaponifiable fraction of at least one vegetable oil or oil derived from a microorganism, said fraction being free of solvents classified in the EU1, EU2 and/or US CMR list and/or being obtainable or directly obtainable according to the process of the invention, and optionally comprising excipients, in particular cosmetically, dietetically or pharmaceutically acceptable excipients.

According to a particular embodiment, the present invention relates to a composition, in particular a pharmaceutical, edible or cosmetic composition, or an edible supplement, comprising at least one unsaponifiable matter, in particular a soybean unsaponifiable matter, an avocado unsaponifiable matter, more particularly an avocado unsaponifiable matter rich in furan fraction and/or an avocado unsaponifiable matter rich in sterol fraction, even more particularly a mixture of Avocado and Soybean Unsaponifiables (ASU), obtainable or directly obtained by the process of the invention.

The pharmaceutical composition may be intended for the prevention and/or treatment of connective tissue disorders, in particular arthrosis, periodontal disease and/or skin ageing.

The edible composition or the edible supplement may be intended for the prevention and/or treatment of connective tissue disorders, in particular arthrosis, periodontal disease and ageing and/or inflammation of the skin.

The cosmetic composition may be intended for the prevention and/or treatment of epidermal, dermal and/or subcutaneous skin disorders.

In the context of the present invention, the expression "free of solvents classified in the EU1, EU2 and/or US CMR list" means that the total content of solvents classified in the EU1, EU2 and/or US CMR list is less than 10ppm, in particular less than 5ppm, in particular less than 2ppm, or less than 1 ppm.

The invention further relates to a cosmetic treatment method in which a cosmetic composition according to the invention is applied topically, and to the use of the vegetable oil, oil derived from microorganisms, or unsaponifiables of vegetable butter obtained according to the invention for the preparation of a medicament, in particular intended for the treatment or prevention of connective tissue disorders, in particular arthrosis.

Of course, various features described in this specification may be combined together.

The following experiments were conducted as examples to explain the present invention.

Detailed Description

Example 1: extraction of avocado unsaponifiable matter by DCE (ref 1)

The first step consists of the saponification of the concentrate prepared by molecular distillation of avocado oil.

To this end, a given weight of avocado oil concentrate (45.6 g) was added continuously to a 100ml round bottom flask equipped with a condenser, followed by ethanol (36.6 g), 50% plant ash (5.2 g) and several pumice pellets.

The system was then refluxed for a total of 3.5 hours and, after cooling, diluted with demineralised water (60 ml).

After saponification, a water-alcohol solution is obtained, said solution containing the unsaponifiable matter (or unsaponifiable fraction). The unsaponifiable is then extracted using a first solvent system, specifically DCE.

Several successive extractions (5X 70 ml) were carried out; the collected organic phases were then combined and washed to neutrality with tap water (5 × 70 ml) (phenolphthalein test).

The solvent phase obtained is then dried over anhydrous sodium sulfate and filtered; the unsaponifiable is then recovered by evaporation of the solvent in a rotary evaporator. The unsaponifiable thus extracted is weighed and stored in a pill making machine (pilulier) under an inert gas atmosphere.

The composition of the extracted unsaponifiable matter is then measured by gas chromatography (GPC, CPG for french abbreviation). The results are shown in table 1 below.

Example 2: extraction of avocado unsaponifiables using HMDS

The extraction was carried out according to the procedure of example 1. After optimization to adapt it to the new solvent system, the extraction procedure consisted of an extraction step with 4 × 70ml of HDMS (instead of DCE) and a step comprising washing the organic phase with 3 × 70ml of water. Measurements were made as in example 1 and the results are shown in table 1 below.

Example 3: extraction of avocado unsaponifiables using HMDS-MeTHF mixture (90/10, by volume)

The extraction was carried out according to the procedure of example 1. After optimization to adapt it to the new solvent system, the extraction procedure consisted of an extraction step with 4 × 70ml of HDMS-MeTHF mixture (90/10 by volume) (instead of DCE) and a step comprising washing the organic phase with 2 × 70ml of water. Measurements were made as in example 1 and the results are shown in table 1 below.

Example 4: extraction of avocado unsaponifiables using HMDS-MeTHF mixture (50/50, by volume)

The extraction was carried out according to the procedure of example 1. After optimization to adapt it to the new solvent system, the extraction procedure consisted of an extraction step with 6 × 70ml of HDMS-MeTHF mixture (50/50 by volume) (instead of DCE) and a step comprising washing the organic phase with 8 × 70ml of water. Measurements were made as in example 1 and the results are shown in table 1 below.

TABLE 1

And (3) analysis: chromatograms and compositions of the unsaponifiables extracted in examples 1 to 4

Weight yield relative to concentrate =100 × (weight of extracted unsaponifiable/weight of concentrate used)

The mixture tested resulted in extraction of the unsaponifiable matter. The unsaponifiables obtained all had chromatograms similar to those observed with the reference extracted with DCE.

However, their composition, taking into account the components of the extracted unsaponifiable matter, illustrates the influence of the polar character of the first solvent system on the potential selectivity. The most polar compounds are extracted in greater proportion using the most polar mixture.

The use of HMDS alone enables selective extraction of non-polar compounds.

Thus, the HMDS/MeTHF mixture makes it possible to extract unsaponifiables with chromatograms similar to the reference unsaponifiable. The composition of the extracted unsaponifiable matter is directly related to the composition of the solvent system, more particularly to the polarity of the solvent system: the more polar the solvent system, the more abundant the polar compounds (e.g. trihydric alcohols) in the unsaponifiable matter are. Conversely, the less polar the solvent system, the better the selectivity of the extraction and the more abundant the non-polar compounds (e.g., furans) in the unsaponifiable material.

Furthermore, the composition of the first solvent system makes it possible, due to its weak polarity, to improve the extraction and washing steps by limiting the amount of solvent and/or water used and by promoting phase separation.

Example 5: extraction of avocado unsaponifiables using ETBE

The extraction was carried out according to the procedure of example 1. After optimization to adapt it to the new solvent system, the extraction procedure consisted of an extraction step with 4 × 60ml of ETBE (instead of DCE) and a step comprising washing the organic phase with 4 × 70ml of water. Measurements were made as in example 1 and the results are shown in table 2 below.

TABLE 2

And (3) analysis: chromatograms and compositions of the unsaponifiables extracted in examples 1 and 5

The first solvent system tested resulted in an extraction of the unsaponifiable with a weight yield similar to that obtained in a classical extraction process with DCE. The unsaponifiable matter obtained has a chromatogram and a composition similar to those observed for the reference extracted with DCE.

Furthermore, ETBE makes it possible to reduce the number of extractions, and thus the amount of solvent used and the number of washes, and thus the amount of water used, in order to be applied in the unsaponifiable fraction of the extracted avocado concentrate.

In summary, ETBE is a good alternative to the use of DCE as the first solvent system in a process for extracting avocado unsaponifiables.

Example 6: extraction of avocado unsaponifiables using 1-chlorobutane(Ref.2)

The extraction and washing was carried out according to the procedure of example 1 using 1-chlorobutane (instead of DCE). Measurements were made as in example 1 and the results are shown in table 3 below.

Example 7: extraction of avocado unsaponifiables using BTF

Saponification was carried out according to the procedure of example 1 using isopropanol (36.6 ml) instead of ethanol. The extraction was carried out according to the procedure of example 1. After optimization to adapt it to the new solvent system, the extraction process consisted of an extraction step with 4 × 35ml of BTF (instead of DCE) and a step comprising washing the organic phase with 4 × 35ml of water. Measurements were made as in example 1 and the results are shown in table 1 below.

Example 8: extraction of avocado unsaponifiables using HMDS

Saponification was carried out according to the procedure of example 1 using MeTHF (36.6 ml) instead of ethanol. The extraction was carried out according to the procedure of example 1. After optimization to adapt it to the new solvent system, the extraction process consisted of an extraction step with 3 × 70ml HMDS (instead of DCE) and a step comprising washing the organic phase with 3 × 70ml water. Measurements were made as in example 1 and the results are shown in table 3 below.

TABLE 3

And (3) analysis: chromatograms and compositions of the unsaponifiables extracted in examples 6 to 8

The use of MeTHF in the second solvent system in combination with the use of HMDS in the first solvent system results in the extraction of unsaponifiables. The unsaponifiable matter obtained has a chromatogram and a composition similar to those observed for the reference extracted with 1-chlorobutane.

Similarly, the use of isopropanol in the second solvent system in combination with the use of BTF in the first solvent system results in the extraction of unsaponifiables. The unsaponifiable matter obtained has a chromatogram and a composition similar to those observed for the reference extracted with 1-chlorobutane.

The use of MeTHF in the second solvent system in combination with the use of HMDS in the first solvent system and the use of isopropanol in the second solvent system in combination with the use of BTF in the first solvent system reduces the number of extractions and the amount of solvent used as well as the number of washes and the amount of water used.

In summary, the combination of MeTHF/HMDS and isopropanol/BTF is an excellent alternative to the use of ethanol/1-chlorobutane, as indicated by enabling extraction of unsaponifiables with chromatograms and compositions similar to the reference, but with reduced amounts of solvent and water used.

Example 9: extraction of soybean unsaponifiable matter using DCE

The first step consists of saponification of the soy concentrate. To this end, a 100ml round bottom flask equipped with a condenser was charged successively with a given weight of soybean oil concentrate (10.0 g), followed by ethanol (23.3 ml), 50% plant ash (1.7 ml) and several pumice pellets.

The system was then refluxed for a total of 3 hours and further diluted with demineralised water (60 ml).

After saponification, a water-alcohol solution is obtained, said solution containing the unsaponifiable matter (or unsaponifiable fraction). The unsaponifiable is then extracted using a first solvent system, specifically DCE.

Several successive extractions (5X 43 ml) were carried out; the collected organic phases were then combined and washed with water (7 × 43 ml) to neutrality (phenolphthalein test).

The solvent phase obtained is then dried over anhydrous sodium sulfate and filtered; the unsaponifiable is then recovered by evaporation of the solvent in a rotary evaporator. The unsaponifiable thus extracted is weighed and stored in a pill-making machine under an atmosphere of inert gas.

The composition of the extracted unsaponifiable matter is then measured by Gas Phase Chromatography (GPC). The results are shown in table 4 below.

Example 10: extraction of soybean unsaponifiable matter using BTF

Saponification was performed according to the procedure of example 9 using increasing volumes of ethanol (50.0 ml). The extraction was carried out according to the procedure of example 9. After optimization to adapt it to the new solvent system, the extraction process consisted of an extraction step with 7 × 35ml of BTF (instead of DCE) and a step comprising washing the organic phase with 10 × 35ml of water. Measurements were made as in example 9 and the results are shown in table 4 below.

Example 11: extraction of Soybean unsaponifiables Using HMDS-MeTHF mixture (50/50, by volume)

The extraction was carried out according to the procedure of example 9. After optimization to adapt it to the new solvent system, the extraction procedure consisted of an extraction step with 5 × 43ml of HDMS-MeTHF mixture (50/50 by volume) (instead of DCE) and a step comprising washing the organic phase with 4 × 43ml of water. Measurements were made as in example 9 and the results are shown in table 4 below.

TABLE 4

And (3) analysis: chromatograms and compositions of the unsaponifiables extracted in examples 9 to 11

	Example 9	Example 10	Example 11
				Hydrocarbons	0.1	0.2	0.1
Squalene	4.4	4.5	3.9
				Tocopherol	25.8	23.4	19.8
Sterols	44.3	43.7	41.0
				Others	6.0	6.1	4.6
Weight yield relative to concentrate	45%	38%	49%

The first solvent system tested resulted in the extraction of the unsaponifiable matter. The unsaponifiable matter obtained has a chromatogram and a composition similar to those observed for the reference extracted with DCE.

In summary, BTF and a 50% by volume HMDS/MeTHF mixture appear to be excellent alternatives to the use of DCE.

Example 12: extraction of unsaponifiable matter of rapeseed by BTF

Saponification was carried out according to the procedure of example 10, using rapeseed concentrate as concentrate. Extraction was carried out according to the procedure of example 10. Measurements were made as in example 10 and the results are shown in table 5 below.

Example 13: extraction of corn unsaponifiables using BTF

Saponification was carried out according to the procedure of example 10, using corn concentrate as the concentrate. Extraction was carried out according to the procedure of example 10. Measurements were made as in example 10 and the results are shown in table 5 below.

Example 14: extraction of sunflower unsaponifiable matter by BTF

Saponification was carried out according to the procedure of example 10, using sunflower concentrate as concentrate. Extraction was carried out according to the procedure of example 10. Measurements were made as in example 10 and the results are shown in table 5 below.

TABLE 5

And (3) analysis: chromatograms and compositions of the unsaponifiables extracted in examples 10 and 12 to 14

	Example 10	Example 12	Example 13	Example 14
					Tocopherol (%)	23.4	6.4	7.9	9.7
Sterol (%)	43.7	63.1	55.3	58.7
					Weight yield relative to concentrate	38%	7.2%	9.0%	4.0%
Weight yield relative to unsaponifiable matter	79%	76%	83%	78%

BTF applied to various plant substances gives equivalent results, considering the weight yield relative to the unsaponifiable matter.

Example 15: extraction of unsaponifiable sesame seeds by using ETBE

Saponification was carried out according to the procedure of example 5, using sesame concentrate as the concentrate. The extraction was carried out according to the procedure of example 5. Measurements were made as in example 5 and the results are shown in table 6 below.

Example 16: extraction of sesame unsaponifiables with HMDS-MeTHF mixture (50/50, by volume)

Saponification was carried out according to the procedure of example 4, using sesame concentrate as the concentrate. The extraction was carried out according to the procedure of example 4. Measurements were made as in example 4 and the results are shown in table 6 below.

TABLE 6

And (3) analysis: chromatograms and compositions of the unsaponifiables extracted in examples 15 and 16

	Example 15	Example 16
			Sterol (%)	17.4	20.0
Sesamin (%)	28.0	31.4
			Sesame Lin element (%)	13.3	15.7
Weight yield relative to concentrate	12%	15%
			Weight yield relative to unsaponifiable matter	83%	98%

For sesame unsaponifiables, both ETBE and HMDS-MeTHF mixtures (50/50 by volume) were good extraction solvents. The HMDS-MeTHF mixture had excellent extraction capacity with an unsaponifiable extraction yield of 98 wt%.

Example 17: extraction of palm unsaponifiable matter by BTF

Saponification was carried out according to the procedure of example 10, using palm concentrate as the concentrate. Extraction was carried out according to the procedure of example 10. Measurements were made as in example 10 and the results are shown in table 7 below.

Example 18: extraction of palm unsaponifiable Material with HMDS-MeTHF mixture (50/50, by volume)

Saponification was carried out according to the procedure of example 11, using palm concentrate as the concentrate. Extraction was carried out according to the procedure of example 11. Measurements were made as in example 11 and the results are shown in table 7 below.

TABLE 7

And (3) analysis: chromatograms and compositions of the unsaponifiables extracted in examples 17 and 18

	Example 17	Example 18
			Sterol (%)	23.0	25.0
Tocopherol (%)	3.7	3.2
			Fertility trinenol (%)	11.4	8.9
Carotene	0.6	0.5
			Weight yield relative to concentrate	2%	3%
Weight yield relative to unsaponifiable matter	75%	99%

For palm unsaponifiables, both BTF and HMDS-MeTHF mixtures (50/50 by volume) were good extraction solvents. The HMDS-MeTHF mixture had excellent extraction capacity with 99 wt.% extraction yield of unsaponifiables.

Claims (17)

1. A method for the extraction of unsaponifiable matter fractions contained in vegetable oils, oils derived from microorganisms or vegetable butters, or in by-products of the refining industry involving vegetable oils or oils derived from microorganisms, said method comprising at least:

A) comprising a step of converting said oil, said butter or said by-products of the refining industry involving vegetable oils or oils derived from microorganisms into a hydro-alcoholic solution by a step selected from saponification and esterification,

B) comprising a step of extracting said hydro-alcoholic solution by means of liquid/liquid extraction, wherein a fat fraction is separated from an unsaponifiable fraction,

wherein the process is characterized in that the liquid/liquid extraction step in step B is carried out with a first solvent system comprising a solvent in an amount of at least 50% by volume relative to the total volume of the solvent system, the solvent being ethyl tert-butyl ether (ETBE) or a mixture of ethyl tert-butyl ether (ETBE) and: a fluorinated aromatic solvent, a solvent comprising at least one silicon atom, methyltetrahydrofuran (MeTHF), or a mixture thereof.

2. The process according to claim 1, further comprising a step C selected from crystallization and liquid/liquid extraction to purify the unsaponifiable fraction.

3. The process according to claim 2, wherein the crystallization step in step C and the liquid/liquid extraction step in step C are carried out with a first solvent system comprising a solvent in an amount of at least 50% by volume relative to the total volume of the solvent system, said solvent being ethyl tert-butyl ether (ETBE) or a mixture of ethyl tert-butyl ether (ETBE) and: a fluorinated aromatic solvent, a solvent comprising at least one silicon atom, methyltetrahydrofuran (MeTHF), or a mixture thereof.

4. The method of claim 1 or claim 2, wherein the fluorinated aromatic solvent is trifluorotoluene (BTF) or hexafluorobenzene (BHF) and the solvent comprising at least one silicon atom is Hexamethyldisiloxane (HMDS) or Tetramethylsilane (TMS).

5. The process of claim 1 or claim 2, wherein the first solvent system comprises a solvent in an amount of at least 75% by volume relative to the total volume of the first solvent system, the solvent being ethyl tert-butyl ether (ETBE) or a mixture of ethyl tert-butyl ether and: a fluorinated aromatic solvent, a solvent comprising at least one silicon atom, methyltetrahydrofuran (MeTHF), or a mixture thereof.

6. The process of claim 1 or claim 2, wherein the first solvent system consists of ethyl tert-butyl ether (ETBE) or a mixture of ethyl tert-butyl ether (ETBE) and a solvent of: a fluorinated aromatic solvent, a solvent comprising at least one silicon atom, methyltetrahydrofuran (MeTHF), or a mixture thereof.

7. The process according to claim 1 or claim 2, wherein the first solvent system comprises a solvent in an amount of at least 90% by volume relative to the total volume of the first solvent system, the solvent being selected from ethyl tert-butyl ether (ETBE) or a mixture of ethyl tert-butyl ether (ETBE) and: a fluorinated aromatic solvent, a solvent comprising at least one silicon atom, methyltetrahydrofuran (MeTHF), or a mixture thereof.

8. The process according to claim 1, wherein step a) comprising the conversion of the oil or butter into a hydro-alcoholic solution by a step selected from saponification and esterification is carried out in a second solvent system comprising a solvent in an amount of at least 50% by volume with respect to the total volume of the solvent system, said solvent being selected from C₂-C₄Alcohols, MeTHF and mixtures thereof.

9. The method of claim 8, wherein the second solvent system comprises a solvent in an amount of at least 60 volume percent relative to the total volume of the second solvent system, the solvent selected from C₂-C₄Alcohols, MeTHF and mixtures thereof.

10. The method of claim 8, wherein the second solvent system consists of C₂-C₄Alcohol, MeTHF or mixtures thereof.

11. The method of claim 8, wherein the second solvent system comprises a solvent in an amount of at least 75 vol% for a total volume of the second solvent system, the solvent selected from C₂-C₄Alcohol or MeTHF.

12. The method of claim 1, wherein the vegetable oil, oil derived from a microorganism, or oleo is selected from the group consisting of soybean, quinoa, rapeseed, corn, sunflower, sesame, lupin, cotton, coconut, olive, palm leaf, wheat germ, alfalfa, avocado, palm kernel, groundnut, copra, flax, castor bean, grape seed, squash seed, blackcurrant seed, melon seed, tomato seed, pumpkin seed, almond, hazelnut, walnut, evening primrose, borage, safflower, poppy seed, macroalgae, and/or microbial oils and mixtures thereof, and cocoa butter, indian red iron tree butter, shea butter, and mixtures thereof.

13. The method of claim 12, wherein the flax is flax seed.

14. The method of claim 12, wherein the microorganism is a microalgae, yeast, mold, or bacteria.

15. The method of claim 1 or claim 2, wherein the first solvent system consists of ethyl tert-butyl ether (ETBE).

16. The process according to claim 1 or claim 2, wherein the first solvent system consists of ethyl tert-butyl ether (ETBE) and a second solvent, different from the first solvent, selected from fluorinated aromatic solvents, solvents comprising at least one silicon atom and methyl tetrahydrofuran (MeTHF).

17. A process according to claim 1 or claim 2, wherein the unsaponifiable obtained is selected from the group consisting of soybean unsaponifiables, avocado unsaponifiables rich in furan fractions and/or avocado unsaponifiables rich in sterol fractions, or mixtures of Avocado and Soybean Unsaponifiables (ASU).