CA2073051A1 - Process for extracting minor fatty compounds which occur in a substance of biological origin - Google Patents
Process for extracting minor fatty compounds which occur in a substance of biological originInfo
- Publication number
- CA2073051A1 CA2073051A1 CA002073051A CA2073051A CA2073051A1 CA 2073051 A1 CA2073051 A1 CA 2073051A1 CA 002073051 A CA002073051 A CA 002073051A CA 2073051 A CA2073051 A CA 2073051A CA 2073051 A1 CA2073051 A1 CA 2073051A1
- Authority
- CA
- Canada
- Prior art keywords
- cyclodextrin
- substance
- fatty
- treated
- water
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B3/00—Refining fats or fatty oils
- C11B3/006—Refining fats or fatty oils by extraction
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C15/00—Butter; Butter preparations; Making thereof
- A23C15/12—Butter preparations
- A23C15/14—Butter powder; Butter oil, i.e. melted butter, e.g. ghee ; Anhydrous butter
- A23C15/145—Removal of steroids, e.g. cholesterol or free acids; Fractionation of anhydrous milkfat by extraction with solvents other than solvent crystallisation or with supercritical gases or by distillation
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/27—Removal of unwanted matter, e.g. deodorisation or detoxification by chemical treatment, by adsorption or by absorption
Abstract
ABSTRACT
A method for extracting minor fatty compounds, particularly steroids, from biological matter by means of cyclodextrin. According to the method, the cyclodextrin is stirred into said biological matter in a water-containing reaction medium to allow the formation of inclusion complexes between the cyclodextrin and the minor fatty compounds, whereafter said complexes are separated from said matter, the cyclodextrin being stirred in at a temperature such that the glycerides in said matter are in a congealed state.
A method for extracting minor fatty compounds, particularly steroids, from biological matter by means of cyclodextrin. According to the method, the cyclodextrin is stirred into said biological matter in a water-containing reaction medium to allow the formation of inclusion complexes between the cyclodextrin and the minor fatty compounds, whereafter said complexes are separated from said matter, the cyclodextrin being stirred in at a temperature such that the glycerides in said matter are in a congealed state.
Description
wO 92/04430 - 1 - PCT/FR91/00707 Process for extractlnq mlnor fatty comp~ounds which occur in a substance of bioloqical_oriqin The present invention relates to a process for extracting minor fatty compounds, especially steroids, and more particularly cholesterol, which occur in a substance Gf biological origin.
"Minor fatty compouTIds'' designates all non-glyceride fatty constituents which occur in natural fatty substances. They may be steroids, especially sterols, fatty alcohols, fat-soluble vitamins, pigments, hydro-carbons, free fatty acids, abietic and pimaric acids, flavouring compounds and the like.
For more detail, reference can be made to the definition given in "Manuel d'analyse des corps gras (Manual of the analysis of fatty acids~" JP WOLF - Ed Azoulay - Paris 1969, pages 11 and 25.
Within the context of the present invention, the term "steroids" designates all compounds containing a more or less hydrogenated cyclopentanophenanthrene skeleton. Many of these compound~ are alcohols and are termed sterols. They may occur in the form of esters especially of fatty acids, commonly designated by the term "cerides".
Cholesterol belongs to the sterol family. It ha~
been known for some time that it is the main constituent of biliary calculi. Its reputation has recently been strengthened by its involvement in circulatory di~orders, and more particularly the hardening of the arteries.
Nonesterified cholesterol is the main constituent of fatty substances of animal origin. These animal fats, which are present in most of our foodstuffs, constitute an important source of cholesterol which, in the event of an excess, may be the cause of serious cardiovascular diseases including myocardial infarction in particular.
Fatty substances of plant origin contain not cholesterol but phytosterols whose structure is very similar to it, such as for example: ~-sito~terol, campesterol, stigmasterol, brassica~terol, 207305~
~-7-stigmasterol, ~-5-campesterol, ~-5-avenasterol, Q-7-avenasterol, ~-7-9-sigmatadienol, fucosterol or ergosterol.
The process according to the invention also applies to the oxidised derivatives of steroids:
sterones. These compounds occur especially in frying oils used repeatedly. Their presence in food products is not desirable as they are not devoid of toxic effect.
In addition, steroids are product~ of high added-value both economically and technically. Indeed, theyhave the potential of constituting high grade raw materials for the preparation of vitamin compounds (ergosterol: vitamin D2) or hormonal compounds (equilenin, oestrone, progesterone, testosterone or cortisone and the like). In addition to their pharma-ceutical applications, steroids, and more particularly sterols and their fatty acid esters (cerides), may be used in the cosmetics industry as emulsifiers.
As far as the other minor fatty compounds are concerned, there may be distinguished, on the one hand, compounds which are undesirable from the food or techno-logical point of view such as free fatty acids which promote oxidation, degradation compounds such as polymerised acids, foul-smelling compounds, colouring substances and savoury substances and the like; and on the other hand, compound~ of high food or technological value such aB fat-soluble vitamins, es~ential fatty acids (linoleic acid, ~-linoleic acid, ~-linoleic acid, arachidonic acid, DHA and EPA), rare fatty acids (ricinoleic and abietic acids), and flavouring compound~.
There are thus several justifications for all the technological developments designed to extract the minor fatty compounds from fatty biological substances. This term designates in particular, on the one hand, animal fats, what they are storage fats such a~ fish oils or animal oils (tallow, chicken fat, lard and the like), structural fats such a~ those present in muscular, placenteral or nerve tissue (meat, egg, spinal cord, 2073~1 brain and the like) or fats from glandular secretion such as milk fats, or fats from skin ~ecretion such as cerides (lanolin); and, on the other hand, vegetable fats.
Finally, the fats may be a mixture of animal fats and vegetable fats such as margarines.
Various methods for extractinq minor fatty compounds from fat-containing biological substances have thus already been proposed.
Primarily, techniques exist for extracting steroids, especially cholesterol.
One of them consists in placing fatty substance of animal origin in contact with digitonin (digitogenin glucodise), which has the property of reacting with cholesterol to give an insoluble precipitate. The efficiency and the re~ults of this method are not satisfactory due to the difficulty of separating the precipitate from the mixture. And in any case, this method is inapplicable industrially, especially for food products.
Cholesterol may also be extracted from fatty substance3 by entrainment by means o$ a solvent. The main disadvantage of this process is that the solvents generally used are toxic and that traces of them always remain in the fatty substances considered.
Processes of microdistillation, whi¢h are inap-plicable at the industrial level, or alternatively of ab~orption on columns such a~ tho~e descxibed for example in European Patent Applications No. 0,174,848 and No. 0,318,326, are al~o known. These applications describe a process according to which the fatty substance, maintained in a liquid Ytate, passes through an absorption column, in this case activated charcoal. It is evident that such a process i~ very hard to implement and, furthermore, the extraction which is achieved by it is not very selective.
Another physicochemical process for extracting cholesterol from fats i9 disclosed by Japanese Patent Application No. 59/140,299. It consists in placing a 2073~51 substance charged with cholesterol, such as milk powder, in contact with supercritical CO2 at a temperature of between 35 and 45C and at a pre~sure of between 130 and 200 atm. Obtaining these phy~ical conditions requires the use of complex and expensive equipment. Control of the process is thus very intricate as a result. Furthermore, as specified in the patent application, other lipid compounds are entrained by the supercritical CO2. This process is not therefore selective.
A process for the biodegradation of the said sterols, which is de~cribed by European Patent Appli-cation No. 0,278,794, has also been conceived for removing sterols from fats. This process uses bacteria which, when placed in contact with the fatty substance, are capable of metabolising the sterols contained therein. Like all processes involving fermentations, this biodegradation process is very intricate to manage because of the inherent variability of living matter.
Furthermore, the equipment used and the relatively long duration are, inter alia, elements which make such a process expensive. Finally, the catabolites produced during these fermentations remain up until now completely unknown with respect to their nature and their toxicity and are, in any case, present in the fatty substance thus treated.
A proces~ for removing the cholesterol pre~ent in a fatty substance of animal origin is also disclosed by European Patent Application No. 0,256,911. It is based on the property, known since 1958, which is exhibited by cyclodextrin3 (cyclic polyglucose compound~ of tubular frustoconical conformation containing 6, 7 or 8 glucose units and respectively designated by alpha-, beta- or gamma-cyclodextrin) whereby they accept sterol molecules, and especially chole~terol molecules, into their hydrophobic cavity to form inclusion complexes which are soluble in water. According to this process, the fatty substance, which is maintained fluid, is placed in contact, in an aqueous medium, with a cyclodextrin, with 20730Sl stirring and under a nonoxidising a~mosphere for 30 minutes to 10 hours so as to allow the formation of complexes~ Separation of the latter is then carried out by entrainment with water which solubili~es these complexes. The aqueouq solution thu~ obtained is then collected after de~antation. The lipid phase obtained must be washed with water several times so as to remove the traces of complexes. The conditions for carrying out this process turn out to be relatively exacting. One of the main constraints to be met is the maintenance of a nitrogen-based nonoxidi~ing atmosphere over the entire period during which the compounds are placed in contact.
Thi~ period i~ moreover very long in practice since, if reference i9 made to the examples, it can be observed that it i8 always greater than or equal to 3 hours.
The yield of cholesterol extraction by this process is not very high. Indeed, in the best case, it is only 41%, and this after three successive extractions as indicated in Example 3 of the de~cription of this European patent application.
The amount of water required for the placing in contact before separation corresponds to not less than 1000% by weight relative to the fatty substance to be treated.
Still on the same principle of using cyclo-dextrins for the extraction of steroids, European Patent Application No. 0,326,469 de~cribe~ a procefls for removing steroidal compound~ present in a substance of biological origin. According to this process, the substance of biological origin, which is made fluid if it i~ a solid fat at normal temperature, is placed in contact with a cyclodextrin in aqueous medium. The contacting iB carried out with stirring for 5 hours at a temperature of the order of 40C. A separation of the cyclodextrin/ steroid complexes i~ then carried out. Here again, it should be noted that the actual duration of the plaGing in contact, which is given in all the examples, is very long: minimum of 5 hours.
"Minor fatty compouTIds'' designates all non-glyceride fatty constituents which occur in natural fatty substances. They may be steroids, especially sterols, fatty alcohols, fat-soluble vitamins, pigments, hydro-carbons, free fatty acids, abietic and pimaric acids, flavouring compounds and the like.
For more detail, reference can be made to the definition given in "Manuel d'analyse des corps gras (Manual of the analysis of fatty acids~" JP WOLF - Ed Azoulay - Paris 1969, pages 11 and 25.
Within the context of the present invention, the term "steroids" designates all compounds containing a more or less hydrogenated cyclopentanophenanthrene skeleton. Many of these compound~ are alcohols and are termed sterols. They may occur in the form of esters especially of fatty acids, commonly designated by the term "cerides".
Cholesterol belongs to the sterol family. It ha~
been known for some time that it is the main constituent of biliary calculi. Its reputation has recently been strengthened by its involvement in circulatory di~orders, and more particularly the hardening of the arteries.
Nonesterified cholesterol is the main constituent of fatty substances of animal origin. These animal fats, which are present in most of our foodstuffs, constitute an important source of cholesterol which, in the event of an excess, may be the cause of serious cardiovascular diseases including myocardial infarction in particular.
Fatty substances of plant origin contain not cholesterol but phytosterols whose structure is very similar to it, such as for example: ~-sito~terol, campesterol, stigmasterol, brassica~terol, 207305~
~-7-stigmasterol, ~-5-campesterol, ~-5-avenasterol, Q-7-avenasterol, ~-7-9-sigmatadienol, fucosterol or ergosterol.
The process according to the invention also applies to the oxidised derivatives of steroids:
sterones. These compounds occur especially in frying oils used repeatedly. Their presence in food products is not desirable as they are not devoid of toxic effect.
In addition, steroids are product~ of high added-value both economically and technically. Indeed, theyhave the potential of constituting high grade raw materials for the preparation of vitamin compounds (ergosterol: vitamin D2) or hormonal compounds (equilenin, oestrone, progesterone, testosterone or cortisone and the like). In addition to their pharma-ceutical applications, steroids, and more particularly sterols and their fatty acid esters (cerides), may be used in the cosmetics industry as emulsifiers.
As far as the other minor fatty compounds are concerned, there may be distinguished, on the one hand, compounds which are undesirable from the food or techno-logical point of view such as free fatty acids which promote oxidation, degradation compounds such as polymerised acids, foul-smelling compounds, colouring substances and savoury substances and the like; and on the other hand, compound~ of high food or technological value such aB fat-soluble vitamins, es~ential fatty acids (linoleic acid, ~-linoleic acid, ~-linoleic acid, arachidonic acid, DHA and EPA), rare fatty acids (ricinoleic and abietic acids), and flavouring compound~.
There are thus several justifications for all the technological developments designed to extract the minor fatty compounds from fatty biological substances. This term designates in particular, on the one hand, animal fats, what they are storage fats such a~ fish oils or animal oils (tallow, chicken fat, lard and the like), structural fats such a~ those present in muscular, placenteral or nerve tissue (meat, egg, spinal cord, 2073~1 brain and the like) or fats from glandular secretion such as milk fats, or fats from skin ~ecretion such as cerides (lanolin); and, on the other hand, vegetable fats.
Finally, the fats may be a mixture of animal fats and vegetable fats such as margarines.
Various methods for extractinq minor fatty compounds from fat-containing biological substances have thus already been proposed.
Primarily, techniques exist for extracting steroids, especially cholesterol.
One of them consists in placing fatty substance of animal origin in contact with digitonin (digitogenin glucodise), which has the property of reacting with cholesterol to give an insoluble precipitate. The efficiency and the re~ults of this method are not satisfactory due to the difficulty of separating the precipitate from the mixture. And in any case, this method is inapplicable industrially, especially for food products.
Cholesterol may also be extracted from fatty substance3 by entrainment by means o$ a solvent. The main disadvantage of this process is that the solvents generally used are toxic and that traces of them always remain in the fatty substances considered.
Processes of microdistillation, whi¢h are inap-plicable at the industrial level, or alternatively of ab~orption on columns such a~ tho~e descxibed for example in European Patent Applications No. 0,174,848 and No. 0,318,326, are al~o known. These applications describe a process according to which the fatty substance, maintained in a liquid Ytate, passes through an absorption column, in this case activated charcoal. It is evident that such a process i~ very hard to implement and, furthermore, the extraction which is achieved by it is not very selective.
Another physicochemical process for extracting cholesterol from fats i9 disclosed by Japanese Patent Application No. 59/140,299. It consists in placing a 2073~51 substance charged with cholesterol, such as milk powder, in contact with supercritical CO2 at a temperature of between 35 and 45C and at a pre~sure of between 130 and 200 atm. Obtaining these phy~ical conditions requires the use of complex and expensive equipment. Control of the process is thus very intricate as a result. Furthermore, as specified in the patent application, other lipid compounds are entrained by the supercritical CO2. This process is not therefore selective.
A process for the biodegradation of the said sterols, which is de~cribed by European Patent Appli-cation No. 0,278,794, has also been conceived for removing sterols from fats. This process uses bacteria which, when placed in contact with the fatty substance, are capable of metabolising the sterols contained therein. Like all processes involving fermentations, this biodegradation process is very intricate to manage because of the inherent variability of living matter.
Furthermore, the equipment used and the relatively long duration are, inter alia, elements which make such a process expensive. Finally, the catabolites produced during these fermentations remain up until now completely unknown with respect to their nature and their toxicity and are, in any case, present in the fatty substance thus treated.
A proces~ for removing the cholesterol pre~ent in a fatty substance of animal origin is also disclosed by European Patent Application No. 0,256,911. It is based on the property, known since 1958, which is exhibited by cyclodextrin3 (cyclic polyglucose compound~ of tubular frustoconical conformation containing 6, 7 or 8 glucose units and respectively designated by alpha-, beta- or gamma-cyclodextrin) whereby they accept sterol molecules, and especially chole~terol molecules, into their hydrophobic cavity to form inclusion complexes which are soluble in water. According to this process, the fatty substance, which is maintained fluid, is placed in contact, in an aqueous medium, with a cyclodextrin, with 20730Sl stirring and under a nonoxidising a~mosphere for 30 minutes to 10 hours so as to allow the formation of complexes~ Separation of the latter is then carried out by entrainment with water which solubili~es these complexes. The aqueouq solution thu~ obtained is then collected after de~antation. The lipid phase obtained must be washed with water several times so as to remove the traces of complexes. The conditions for carrying out this process turn out to be relatively exacting. One of the main constraints to be met is the maintenance of a nitrogen-based nonoxidi~ing atmosphere over the entire period during which the compounds are placed in contact.
Thi~ period i~ moreover very long in practice since, if reference i9 made to the examples, it can be observed that it i8 always greater than or equal to 3 hours.
The yield of cholesterol extraction by this process is not very high. Indeed, in the best case, it is only 41%, and this after three successive extractions as indicated in Example 3 of the de~cription of this European patent application.
The amount of water required for the placing in contact before separation corresponds to not less than 1000% by weight relative to the fatty substance to be treated.
Still on the same principle of using cyclo-dextrins for the extraction of steroids, European Patent Application No. 0,326,469 de~cribe~ a procefls for removing steroidal compound~ present in a substance of biological origin. According to this process, the substance of biological origin, which is made fluid if it i~ a solid fat at normal temperature, is placed in contact with a cyclodextrin in aqueous medium. The contacting iB carried out with stirring for 5 hours at a temperature of the order of 40C. A separation of the cyclodextrin/ steroid complexes i~ then carried out. Here again, it should be noted that the actual duration of the plaGing in contact, which is given in all the examples, is very long: minimum of 5 hours.
2~73051 The low yields of sterol extraction given in the example~ are evidence that this procass lacks efficiency.
During the placing in contact before separation, the reaction medium contains more than 100% by weight of water relative to the substance to be treated expre~sed on a dry basis.
Secondly, proces~es for extracting minor fatty compounds other than steroids have been considered.
Among these processes, the one described in US Patent No. 3,491,132 and de~igned to reduce the free fatty acid content of glyceridic oils uYing cyclodextrin may be mentioned. In this case, the cyclodextrin i8 placed in contact with the fatty medium in the presence of substantial amounts of water ranging from 200 to 1000%
relative to the fatty medium treated.
These last three processes, which are described in European Patent Applications Nos. 0,256,911 and 0,326,469 and in US Patent No. 3,491,132, have at least one major disadvantage in common, namely that they take place in aqueous medium and require very large amounts of water for the placing in contact.
It is evident that the use of ~uch amounts of water constituteQ a handicap from the industrial point of view since it makes it necessary to handle large reaction volumes and requires equipment of large aapacity which is therefore bul~y and more expenqive. Moreo~er, in addition to the hlgh direct cost which this entails, the treatment of the effluents obtained after the separation should also be taken into account; this treatment repre-senting a burden which increases as a function of theamount of effluent.
In view of the foregoing, it ~hould be noted that the techniques for extracting minor fatty compounds using cyclodextrin, which are the most satisfactory neverthe-less, especially with respect to the quality of theproducts obtained, have not been optimised from the point of view of economic and technical profitability.
Anxieu~ to promote technical development in this field, the applicant company carried out long and numerous studie~ at the end of which they succeeded, to their credit, in finding that it was preferable and advantageous to carry out the placing in contact at a temperature at which the biological substance to be treated/cyclodextrin/water mixture is in a semisolid state, that is to say in a state where not all the substances present in the said substance are liquid.
Accordingly, the present invention relates to a process for extracting minor fatty compounds, especially steroids, and more particularly cholesterol, which occur in a sub~tance of biological origin, using cyclodextrin, in which the cyclodextrin i8 placed in contact, with stirring and in a reaction medium containing water, with the substance to be treated 80 as to allow the formation of inclusion complexes between the cyclodextrin and the minor fatty compounds, these complexes being subsequently separated from the said substance, the process being characteri~ed in that the placing in contact of cyclodextrin/substance to be treated is carried out at a temperature which is chosen such that the fatty substances present in the said substance are in a congealed~state.
Congealed state, within the context of the present invention, is understood to mean a state in which the level of solid or crystalline fatty substan¢es is greater than or equal to 5% by weight.
This solid fat level is a common variable in the field of lipid technology. It is measured by low resolu-tion Nuclear Magnetic Resonance, using pulsed minispectrometers.
Pulsed NMR, which is based on the fact that fat molecules in the liquid phase have a degree of mobility which is greater than that of the molecules of the solid phase, enables the percentage of solid or liquid fat to be determined. The principle and the applications of this method are described for example in the journal R.I.B.C.
2~730~
of Januar~-February 1985 No. 80, pages 23 to 26, or in the journal I-A-A of May 1988, pages 463 to 471 or of June 1988, pages 463 to 470.
Contrary to what is recommended in the prior art and especially in European Patent Applications Nos. 0,256,911 and 0,326,469 and US Patent No. 3,491,132 which are mentioned above, the applicant company has demonstrated, surprisingly and unexpectedly, that the congealed state of the fatty ~ubstances which constitute the substance to be treated corresponds to an advantageous state for complexing minor fatty compounds, especially steroids, with cyclodextrin at the stage of placing in contact with stirring.
Preferably, the operating conditions are chosen such that the level of solid fats is greater than or equal to 10~ and still more preferably greater than or equal to 30%.
In conformity with an advantageous arrangement of the present invention, the amount of water present in the reaction medium, during the placing in contact and before separation, i8 less than 100% by weight, preferably between 10 and 90%, and still more preferably between 20 and 60%, by weight relative to the substance to be treated expressed on a dry basis.
By thus limiting the amounts of water used, all the costs o~ implementing the extraction proce~ are sub~tantially reduced while making the use more flexible and easier.
Furthermore, the extraction yields are improved for much shorter durations of placing in contact, and the amounts of effluents collected, and therefore to be retreated, are substantially reduced. These effluents are in concentxated form, which keeps down the costs of transportation and retreatment.
The cyclodextrin used may be of the alpha, beta or gamma type, preferably of the beta type, which is substituted or unsubstituted. The mono- or poly-sub-stituent group~ of the cyclodextrin may be in particular 2~7305~
alkyl groups such as the hydroxypropyl or ethyl group, or glycoside or maltosyl type saccharides and the like.
Cyclodextrin rendered insoluble in water, for example by polymerisation, may also be chosen.
Advantageously, the cyclodextrin is used in the process according to the invention in pulverulent form.
A cyclodextrin having a water content of less than or equal to 11% by weight, preferably le ~ than or equal to 7% by weight, and still more preferably less than or equal to 5~ by weight is preferably used. It may however be u~ed after rehydration up to a water content of less than or equal to 15% by weight.
It is worthwhile to note that the placing in contact takes place under an ambient atmosphere. Indeed, the process conditions are not harsh and do not carry the risk of bringing about degradations, such as oxidation, of the sensitive fatty compounds (unsaturated fatty acids). Better still, cyclodextrin makes it possible, in addition, to remove, at least partially, the undesirable prooxidant compound~ which may be present.
According to a preferred embodiment, the substance to be treated, water - if the said substance to be treated does not contain it or does not contain enough of it - and then cyclodextrin, preferably in pulverulent form, are placed together in a mixer e~uipped with kneading means. The sequence in which the products are introduced i9 preferably the one indicated above but it is also possible, among all the pos3ible variants, for example to mix the substance to be treated and water beforehand.
Advantageously, the mixture is provided in a pasty form which is subjected, during the entirz period of placing in contact, to a kneading operation during which the formation of cyclodextrin/minor fatty compound complexes occurs.
For the separation of the~e complexes, and this is also another novelty of the use of the process according to the invention, the kneaded paste is heated 20~30~1 to a temperature which is chosen such that the fatty substances constituting the ~ubstance to be treated are in a molten state, that is to say in a state charac-terised by a level of solid or cry3talline fatty substances which is iess than 1% by weight.
This conversion to the molten state makes it possible to obtain, on the one hand, a phase containing the fatty substances and with a reduced content of minor fatty compounds and, on the other hand, a mixed water-lipid phase containing the cyclodextrin/minor fattycompound inclusion complexes.
These two phase~ separate from each other very well and it is therefore easy to collect the fatty phase free of the minor fatty compounds, which may then be used in various applications. The mixed phase, for its part, may be retreated to extract therefrom the minor fatty compounds which may be upgraded, especially in the pharmaceutical or cosmetics field.
When the substance to be treated is an animal and/or vegetable substance with a high lipid content such as for example butter which is concentrated or unconcentrated, tallow, lard, chicken fat3, lanolins, hydrogenated ve~etable oils, palm or copra fats, margarines and the like, it is introduced into a dough type kneading device provided with mean~ for regulating the temperature.
Water i~ then optionally incorporated therein insofar as the substance of biological oriqin to be treated does not contain it in a sufficient amount for the implementation of the process in conformity with the invention. The amount of water required is preferably between 20 and 60% by weight relative to the substance of biological origin to be treated, expressed on a dry basis.
Cyclodextrin i9 then added, preferably in pulverulent form and in sufficient amount to form inclusion complexes with the minor fatty compounds to be extracted.
20~0~1 The determination of the appropriate amount of cyclodextrin to be used is a function of the amount of minor fatty compounds to be extracted, and of their affinities for cyclodextrin. Generally, the amount of cyclodextrin is 0.5% to 15% relative to the substance of biological origin to be treated, expressed on a dry basis, preferably from 1 to 10% and more preferably from 3 to 8%.
The kneading of the paste formed by the mixing of these constituents is carried out at a temperature which i~ maintained constant by regulating means and chosen such that the fatty substance is in the congealed state with a level of solid or crystalline fats above 5%, preferably above 10% and more preferably between 30 and 50% by weight. For concentrated butter for example, the temperature is preferably chosen between about 10 and 25C.
After a variable time, generally between 10 and 120 min, the kneading is stopped and the paste is heated sufficiently so that the level of solids in the fatty substance is less than 1~ by weight. A water-lipid pellet containing the complexes and an oily liquid supernatant consisting of the fatty substance with a reduced minor fatty compound content axe thereby formed.
As far as this embodiment ~or the process according to the invention i~ concerned, it should be noted that this proces~ can be perfectly integrated within the framework of the manufacture of butter from cream. This manufacture essentially comprises the following stage~:
- biological or physical maturation (NIZO
process) of the cream;
- continuous or batch churning with emulsion reversal and separation of buttermilk;
- optional kneading with biological maturation (NIZO process), - optional washing with water.
2~73051 In conformity with the invention, cyclodextrin may be added after separation of the buttermilk, during the knea~ing. At the end of a placing in contact which is carried out at the c~urning temperature (les~ than 15C), S the separation of the inclusion complexes formed i~
carried out b~ reverting to the molten state.
The process according to the invention enables steroids and in particular sterols to be extracted with yields as high as about 90% in a single treatment. These yields may be improved by increasing the number of treatments. These extraction efficiencies are obtained economically and easily in a relatively short time.
Another advantage of the process conforming to the invention i9 that it can permit the successive extraction of a certain number of minor fatty compounds from a substance of biological origin by controlling in a defined order one or more essential process parameters, namely in particular the mixing or kneading temperature, the amount of water present at the time of the complexing stage, the temperature for the change of phase, the pH of the medium.
The process according to the invention may be applied to natural fatty substances which are essentially composed of triglycerides but low in total phospholipids:
that is to say having a total phospholipid content of less than 10% by weight relative to the total lipids.
This process permits, in particular, the extraction of steroids such as cholesterol from numerous biological substancea. They may be animal fats and/or vegetable fats among which there may be mentioned:
- milk fats such a~ concentrated or nonconcen-trated butter;
- animal fats such as tallow, lard, horse fat, chicken fats, fish oils, lanolin;
- vegetable fats such as hydrogenated vegetable fats, palm oil, copra oil, cacao butter;
- and mixtures thereof such as margarines.
2073~5~
In ad~ition to removing these compounds from foodstuffs, the process conforming to the invention offers a useful source of steroids which are capable of being used in the preparation of pharmaceutical ingredients, especially hormonal and vitamin ingredients, or in the cosmetics industry as emulsi~iers.
Naturally, the invention is not limited to the embodiments described above but, on the contrary, it encompasses all the variants thereof.
The following examples will make it possible to understand more clearly the process according to the present invention by demonstrating all the advantages offered by it.
EXAMPLE I
Extraction of sterols~_ _d more particularly cholesterol, which are present in a concentrated butter in conformity with the process according to the invention.
The concentrated butter used has a conce~tration of fatty 6ubstances of the order of 99.9%. The standard concentrated butter marketed by CORMAN (Belgium) is chosen for example.
The cyclodextrin is a beta-cyclodextrin of the type marketed by the applicant company under the registered trade mark KLEPTOSE and provided in the form of a white powder containing about 12% moi~ture.
1. ~lacing in contact 300 g of concentrated butter are introduced into the container of a dough mixer of the type marketed by KUSTNER. The container of the mixer i8 provided with a double wall in which a temperature-regulating fluid may be circulated. The mixer is started and the temperature is maintained at a mean value of about 14C. At this temperature, the concentrated butter has a level of solid fats of about 28%.
150 g of water and 5.70 g of KLEPTOSE cyclo-dextrin are then added successively. The placing in 20730~
-- lq ~
contact then proceeds for 30 minutes.
2. Separatlon of the inclusion complexes The pasty concentrated butter/water/cyclodextrin mixture removed from the mixer is heated to a temperature of the order of 37C (level of solid fats of the con-centrated butter less than 1~). A change of phase occurs spontaneously and a pasty pellet is obtained containing in particular the inclusion complexes and water. This pellet is covered by an oily phase formed by the dechole-sterolised concentrated butter. More than 94% by weightof the concentrated butter used can then be recovered.
The cholesterol is assayed by a colorimetric method after oxidation to cholestenone using cholesterol oxidase, according to the method of, and using the lS reagents with the reference no. 139 OS0 from, BOEHRINGER
Mannheim.
The initial cholesterol content of the concen-trated butter is 0.270~ by weight.
The final content is 0.038%.
The extraction yield therefore amounts to 86%.
A second extraction, carried out under the same conditions, provides a yield of 74%, which gives an overall yield of 96%.
It can be observed that the efficiency of the process according to the invention i8 quite remarkable both from the point of view of the amount and that of the duration.
The amount of water used is quite small and poses no problem of handling.
The pasty pellet containing water which is used repre~ents only 173 g. This small and concentrated amount poses no problem with respect to its retreatment.
EXAMPLE_II
Extractions of sterols, and more particularly cholesterol, which are present in butter in conformity with the invention.
In this example, 9 tests for extracting cholesterols from concentrated butter were carried out using the same equipment and by following the same procedure as in Example I. Certain experimental parameters were simply changed. The data and re~ults of these tests are presented in Table I below:
20730~1 O
...
t~
O ~
o 3 u a o u ~
~ ~ o o o o o u~ o o o H C~ ~1 Ei ~ ~ ~ ~ IY) ~ _~
O u~
~/ ~ 1 ~\
Cl O N ~ ,CI
u a~ _u~ u~ IJ~ Il~ I~ IJ~ Il'~ ~ ~ '~
$ $
h l-l 4 c) ~ O O
o o o o o o o o o XP
~1 ~ ~ o u~
~ ~-1~ l O
3\ _o o o O o o o o o h Ll In In ~a_I ~ ~ ~ u~ ~o 1` OD a~ u~ _ ~ - ~ ~
E~
REPLACEMENT PAGE
207305~
Comments on Table I
By comparing the results of tests 1 to 4 which differ only in the amount of water used, it can be observed that the cholasterol extraction yield, expressed in %, is maximum for water contents of between 30 and 50%
relative to concentrated butter. The increase in the water content brings about a drop in the extraction yield.
~y comparing the results of tests 3 and 5 for which only the KLEPTOSE cyclodextrin concentration is different (5 and 7% respectively relative to concentrated butter), it can be observed that it i5 possible to increase the extraction yield by 9% purely by increasing the cyclodextrin concentration by 2%.
15In test 6, the duration of kneading was reduced to 15 minutes. The yield obtained iæ 75%, which con-stitutes a very satisfactory result.
For test 7, the temperature of placing in contact was increased to 26.6C. By reducing the solids level in the concentrated butter, the cholesterol extraction yield i9 reduced (65% against 84% for test 3).
~ y using 3% cyclodextrin relative to concentrated butter ~tests 8 and 9), it can be observed that beyond 30 min, prolonging the kneading does not enable the yields to be increased.
EXAMPLE III
Extraction of sterols, and more particularl~
cholesterol which are Present in butter in_conformity with the ~ysc~L~ Log-to the invention.
30The butter used in this example is a commercial butter containing 82% of fatty substances.
The cyclodextrin, the equipment as well as the procedure are the same as for the preceding examples, the only difference being that no water is added: it is the butter itself which provides the water required for complex formation.
20730~1 The data and results of this example are given in Table II below:
- - 19- 207305~
~1 ~ .~, ~1.'4 :
~V~ ~
_ ~ 8 ~, 2073~
It is shown here that the process according to the invention is also effective on a substance to be treated initially containing water in an amount conform-ing to the characteristics of the present invention.
EXAMPLE IV
Extraction of sterols, and more particularly cholesterol, which are present in tallow, in conformity with the process accordina to the invention.
200 g of tallow containing 99% of fatty sub-stances, of the type marketed by ROUSSELOT, are treatedin the same manner as in the preceding examples.
The initial cholesterol content of the tallow is 0.12% by weight.
Table III below shows the data and the results of the present example.
~ 21 - 20730~1 . . . ._ .~
X ~ 04 ~ ' ~ ~ o ~. ~
~ ,~ ~o D~
~ U~
~o~ ~
E~ _~
20730Sl _mments on Table III
It appears that an amount of water of 25~ by weight relative to the tallow is qufficient to extract 86% of the initial cholesterol, and this with only 3% by weight of cyclodextrin relative to the tallow.
An increase in the amount of water and/or the duration of placing in contact does not enable the extraction yield to be improved.
In this example, the variations in the acid values were also measured. The acid value i8 defined as the number of mg of potas~ium hydroxide required to neutralise the acidity of 1 g of tallow. The acid value of tallow before extraction is 0.85.
Table IV below presents the results obtained.
TABLE IV
............. .
Water, Tem- CD Kleptose, Duration Final Z reduc-X rela- perature % relative of plac- acid tion in tive to C to tallow ing in value the tallow contact value 20Tallow S0 26 3 30 0.65 30 200 g 25 16 3 30 0.12 86 It is shown here that the decrease in acid value, and therefore of the free fatty acid content, of the tallow i~ very pronounced with 25~ of water.
During the placing in contact before separation, the reaction medium contains more than 100% by weight of water relative to the substance to be treated expre~sed on a dry basis.
Secondly, proces~es for extracting minor fatty compounds other than steroids have been considered.
Among these processes, the one described in US Patent No. 3,491,132 and de~igned to reduce the free fatty acid content of glyceridic oils uYing cyclodextrin may be mentioned. In this case, the cyclodextrin i8 placed in contact with the fatty medium in the presence of substantial amounts of water ranging from 200 to 1000%
relative to the fatty medium treated.
These last three processes, which are described in European Patent Applications Nos. 0,256,911 and 0,326,469 and in US Patent No. 3,491,132, have at least one major disadvantage in common, namely that they take place in aqueous medium and require very large amounts of water for the placing in contact.
It is evident that the use of ~uch amounts of water constituteQ a handicap from the industrial point of view since it makes it necessary to handle large reaction volumes and requires equipment of large aapacity which is therefore bul~y and more expenqive. Moreo~er, in addition to the hlgh direct cost which this entails, the treatment of the effluents obtained after the separation should also be taken into account; this treatment repre-senting a burden which increases as a function of theamount of effluent.
In view of the foregoing, it ~hould be noted that the techniques for extracting minor fatty compounds using cyclodextrin, which are the most satisfactory neverthe-less, especially with respect to the quality of theproducts obtained, have not been optimised from the point of view of economic and technical profitability.
Anxieu~ to promote technical development in this field, the applicant company carried out long and numerous studie~ at the end of which they succeeded, to their credit, in finding that it was preferable and advantageous to carry out the placing in contact at a temperature at which the biological substance to be treated/cyclodextrin/water mixture is in a semisolid state, that is to say in a state where not all the substances present in the said substance are liquid.
Accordingly, the present invention relates to a process for extracting minor fatty compounds, especially steroids, and more particularly cholesterol, which occur in a sub~tance of biological origin, using cyclodextrin, in which the cyclodextrin i8 placed in contact, with stirring and in a reaction medium containing water, with the substance to be treated 80 as to allow the formation of inclusion complexes between the cyclodextrin and the minor fatty compounds, these complexes being subsequently separated from the said substance, the process being characteri~ed in that the placing in contact of cyclodextrin/substance to be treated is carried out at a temperature which is chosen such that the fatty substances present in the said substance are in a congealed~state.
Congealed state, within the context of the present invention, is understood to mean a state in which the level of solid or crystalline fatty substan¢es is greater than or equal to 5% by weight.
This solid fat level is a common variable in the field of lipid technology. It is measured by low resolu-tion Nuclear Magnetic Resonance, using pulsed minispectrometers.
Pulsed NMR, which is based on the fact that fat molecules in the liquid phase have a degree of mobility which is greater than that of the molecules of the solid phase, enables the percentage of solid or liquid fat to be determined. The principle and the applications of this method are described for example in the journal R.I.B.C.
2~730~
of Januar~-February 1985 No. 80, pages 23 to 26, or in the journal I-A-A of May 1988, pages 463 to 471 or of June 1988, pages 463 to 470.
Contrary to what is recommended in the prior art and especially in European Patent Applications Nos. 0,256,911 and 0,326,469 and US Patent No. 3,491,132 which are mentioned above, the applicant company has demonstrated, surprisingly and unexpectedly, that the congealed state of the fatty ~ubstances which constitute the substance to be treated corresponds to an advantageous state for complexing minor fatty compounds, especially steroids, with cyclodextrin at the stage of placing in contact with stirring.
Preferably, the operating conditions are chosen such that the level of solid fats is greater than or equal to 10~ and still more preferably greater than or equal to 30%.
In conformity with an advantageous arrangement of the present invention, the amount of water present in the reaction medium, during the placing in contact and before separation, i8 less than 100% by weight, preferably between 10 and 90%, and still more preferably between 20 and 60%, by weight relative to the substance to be treated expressed on a dry basis.
By thus limiting the amounts of water used, all the costs o~ implementing the extraction proce~ are sub~tantially reduced while making the use more flexible and easier.
Furthermore, the extraction yields are improved for much shorter durations of placing in contact, and the amounts of effluents collected, and therefore to be retreated, are substantially reduced. These effluents are in concentxated form, which keeps down the costs of transportation and retreatment.
The cyclodextrin used may be of the alpha, beta or gamma type, preferably of the beta type, which is substituted or unsubstituted. The mono- or poly-sub-stituent group~ of the cyclodextrin may be in particular 2~7305~
alkyl groups such as the hydroxypropyl or ethyl group, or glycoside or maltosyl type saccharides and the like.
Cyclodextrin rendered insoluble in water, for example by polymerisation, may also be chosen.
Advantageously, the cyclodextrin is used in the process according to the invention in pulverulent form.
A cyclodextrin having a water content of less than or equal to 11% by weight, preferably le ~ than or equal to 7% by weight, and still more preferably less than or equal to 5~ by weight is preferably used. It may however be u~ed after rehydration up to a water content of less than or equal to 15% by weight.
It is worthwhile to note that the placing in contact takes place under an ambient atmosphere. Indeed, the process conditions are not harsh and do not carry the risk of bringing about degradations, such as oxidation, of the sensitive fatty compounds (unsaturated fatty acids). Better still, cyclodextrin makes it possible, in addition, to remove, at least partially, the undesirable prooxidant compound~ which may be present.
According to a preferred embodiment, the substance to be treated, water - if the said substance to be treated does not contain it or does not contain enough of it - and then cyclodextrin, preferably in pulverulent form, are placed together in a mixer e~uipped with kneading means. The sequence in which the products are introduced i9 preferably the one indicated above but it is also possible, among all the pos3ible variants, for example to mix the substance to be treated and water beforehand.
Advantageously, the mixture is provided in a pasty form which is subjected, during the entirz period of placing in contact, to a kneading operation during which the formation of cyclodextrin/minor fatty compound complexes occurs.
For the separation of the~e complexes, and this is also another novelty of the use of the process according to the invention, the kneaded paste is heated 20~30~1 to a temperature which is chosen such that the fatty substances constituting the ~ubstance to be treated are in a molten state, that is to say in a state charac-terised by a level of solid or cry3talline fatty substances which is iess than 1% by weight.
This conversion to the molten state makes it possible to obtain, on the one hand, a phase containing the fatty substances and with a reduced content of minor fatty compounds and, on the other hand, a mixed water-lipid phase containing the cyclodextrin/minor fattycompound inclusion complexes.
These two phase~ separate from each other very well and it is therefore easy to collect the fatty phase free of the minor fatty compounds, which may then be used in various applications. The mixed phase, for its part, may be retreated to extract therefrom the minor fatty compounds which may be upgraded, especially in the pharmaceutical or cosmetics field.
When the substance to be treated is an animal and/or vegetable substance with a high lipid content such as for example butter which is concentrated or unconcentrated, tallow, lard, chicken fat3, lanolins, hydrogenated ve~etable oils, palm or copra fats, margarines and the like, it is introduced into a dough type kneading device provided with mean~ for regulating the temperature.
Water i~ then optionally incorporated therein insofar as the substance of biological oriqin to be treated does not contain it in a sufficient amount for the implementation of the process in conformity with the invention. The amount of water required is preferably between 20 and 60% by weight relative to the substance of biological origin to be treated, expressed on a dry basis.
Cyclodextrin i9 then added, preferably in pulverulent form and in sufficient amount to form inclusion complexes with the minor fatty compounds to be extracted.
20~0~1 The determination of the appropriate amount of cyclodextrin to be used is a function of the amount of minor fatty compounds to be extracted, and of their affinities for cyclodextrin. Generally, the amount of cyclodextrin is 0.5% to 15% relative to the substance of biological origin to be treated, expressed on a dry basis, preferably from 1 to 10% and more preferably from 3 to 8%.
The kneading of the paste formed by the mixing of these constituents is carried out at a temperature which i~ maintained constant by regulating means and chosen such that the fatty substance is in the congealed state with a level of solid or crystalline fats above 5%, preferably above 10% and more preferably between 30 and 50% by weight. For concentrated butter for example, the temperature is preferably chosen between about 10 and 25C.
After a variable time, generally between 10 and 120 min, the kneading is stopped and the paste is heated sufficiently so that the level of solids in the fatty substance is less than 1~ by weight. A water-lipid pellet containing the complexes and an oily liquid supernatant consisting of the fatty substance with a reduced minor fatty compound content axe thereby formed.
As far as this embodiment ~or the process according to the invention i~ concerned, it should be noted that this proces~ can be perfectly integrated within the framework of the manufacture of butter from cream. This manufacture essentially comprises the following stage~:
- biological or physical maturation (NIZO
process) of the cream;
- continuous or batch churning with emulsion reversal and separation of buttermilk;
- optional kneading with biological maturation (NIZO process), - optional washing with water.
2~73051 In conformity with the invention, cyclodextrin may be added after separation of the buttermilk, during the knea~ing. At the end of a placing in contact which is carried out at the c~urning temperature (les~ than 15C), S the separation of the inclusion complexes formed i~
carried out b~ reverting to the molten state.
The process according to the invention enables steroids and in particular sterols to be extracted with yields as high as about 90% in a single treatment. These yields may be improved by increasing the number of treatments. These extraction efficiencies are obtained economically and easily in a relatively short time.
Another advantage of the process conforming to the invention i9 that it can permit the successive extraction of a certain number of minor fatty compounds from a substance of biological origin by controlling in a defined order one or more essential process parameters, namely in particular the mixing or kneading temperature, the amount of water present at the time of the complexing stage, the temperature for the change of phase, the pH of the medium.
The process according to the invention may be applied to natural fatty substances which are essentially composed of triglycerides but low in total phospholipids:
that is to say having a total phospholipid content of less than 10% by weight relative to the total lipids.
This process permits, in particular, the extraction of steroids such as cholesterol from numerous biological substancea. They may be animal fats and/or vegetable fats among which there may be mentioned:
- milk fats such a~ concentrated or nonconcen-trated butter;
- animal fats such as tallow, lard, horse fat, chicken fats, fish oils, lanolin;
- vegetable fats such as hydrogenated vegetable fats, palm oil, copra oil, cacao butter;
- and mixtures thereof such as margarines.
2073~5~
In ad~ition to removing these compounds from foodstuffs, the process conforming to the invention offers a useful source of steroids which are capable of being used in the preparation of pharmaceutical ingredients, especially hormonal and vitamin ingredients, or in the cosmetics industry as emulsi~iers.
Naturally, the invention is not limited to the embodiments described above but, on the contrary, it encompasses all the variants thereof.
The following examples will make it possible to understand more clearly the process according to the present invention by demonstrating all the advantages offered by it.
EXAMPLE I
Extraction of sterols~_ _d more particularly cholesterol, which are present in a concentrated butter in conformity with the process according to the invention.
The concentrated butter used has a conce~tration of fatty 6ubstances of the order of 99.9%. The standard concentrated butter marketed by CORMAN (Belgium) is chosen for example.
The cyclodextrin is a beta-cyclodextrin of the type marketed by the applicant company under the registered trade mark KLEPTOSE and provided in the form of a white powder containing about 12% moi~ture.
1. ~lacing in contact 300 g of concentrated butter are introduced into the container of a dough mixer of the type marketed by KUSTNER. The container of the mixer i8 provided with a double wall in which a temperature-regulating fluid may be circulated. The mixer is started and the temperature is maintained at a mean value of about 14C. At this temperature, the concentrated butter has a level of solid fats of about 28%.
150 g of water and 5.70 g of KLEPTOSE cyclo-dextrin are then added successively. The placing in 20730~
-- lq ~
contact then proceeds for 30 minutes.
2. Separatlon of the inclusion complexes The pasty concentrated butter/water/cyclodextrin mixture removed from the mixer is heated to a temperature of the order of 37C (level of solid fats of the con-centrated butter less than 1~). A change of phase occurs spontaneously and a pasty pellet is obtained containing in particular the inclusion complexes and water. This pellet is covered by an oily phase formed by the dechole-sterolised concentrated butter. More than 94% by weightof the concentrated butter used can then be recovered.
The cholesterol is assayed by a colorimetric method after oxidation to cholestenone using cholesterol oxidase, according to the method of, and using the lS reagents with the reference no. 139 OS0 from, BOEHRINGER
Mannheim.
The initial cholesterol content of the concen-trated butter is 0.270~ by weight.
The final content is 0.038%.
The extraction yield therefore amounts to 86%.
A second extraction, carried out under the same conditions, provides a yield of 74%, which gives an overall yield of 96%.
It can be observed that the efficiency of the process according to the invention i8 quite remarkable both from the point of view of the amount and that of the duration.
The amount of water used is quite small and poses no problem of handling.
The pasty pellet containing water which is used repre~ents only 173 g. This small and concentrated amount poses no problem with respect to its retreatment.
EXAMPLE_II
Extractions of sterols, and more particularly cholesterol, which are present in butter in conformity with the invention.
In this example, 9 tests for extracting cholesterols from concentrated butter were carried out using the same equipment and by following the same procedure as in Example I. Certain experimental parameters were simply changed. The data and re~ults of these tests are presented in Table I below:
20730~1 O
...
t~
O ~
o 3 u a o u ~
~ ~ o o o o o u~ o o o H C~ ~1 Ei ~ ~ ~ ~ IY) ~ _~
O u~
~/ ~ 1 ~\
Cl O N ~ ,CI
u a~ _u~ u~ IJ~ Il~ I~ IJ~ Il'~ ~ ~ '~
$ $
h l-l 4 c) ~ O O
o o o o o o o o o XP
~1 ~ ~ o u~
~ ~-1~ l O
3\ _o o o O o o o o o h Ll In In ~a_I ~ ~ ~ u~ ~o 1` OD a~ u~ _ ~ - ~ ~
E~
REPLACEMENT PAGE
207305~
Comments on Table I
By comparing the results of tests 1 to 4 which differ only in the amount of water used, it can be observed that the cholasterol extraction yield, expressed in %, is maximum for water contents of between 30 and 50%
relative to concentrated butter. The increase in the water content brings about a drop in the extraction yield.
~y comparing the results of tests 3 and 5 for which only the KLEPTOSE cyclodextrin concentration is different (5 and 7% respectively relative to concentrated butter), it can be observed that it i5 possible to increase the extraction yield by 9% purely by increasing the cyclodextrin concentration by 2%.
15In test 6, the duration of kneading was reduced to 15 minutes. The yield obtained iæ 75%, which con-stitutes a very satisfactory result.
For test 7, the temperature of placing in contact was increased to 26.6C. By reducing the solids level in the concentrated butter, the cholesterol extraction yield i9 reduced (65% against 84% for test 3).
~ y using 3% cyclodextrin relative to concentrated butter ~tests 8 and 9), it can be observed that beyond 30 min, prolonging the kneading does not enable the yields to be increased.
EXAMPLE III
Extraction of sterols, and more particularl~
cholesterol which are Present in butter in_conformity with the ~ysc~L~ Log-to the invention.
30The butter used in this example is a commercial butter containing 82% of fatty substances.
The cyclodextrin, the equipment as well as the procedure are the same as for the preceding examples, the only difference being that no water is added: it is the butter itself which provides the water required for complex formation.
20730~1 The data and results of this example are given in Table II below:
- - 19- 207305~
~1 ~ .~, ~1.'4 :
~V~ ~
_ ~ 8 ~, 2073~
It is shown here that the process according to the invention is also effective on a substance to be treated initially containing water in an amount conform-ing to the characteristics of the present invention.
EXAMPLE IV
Extraction of sterols, and more particularly cholesterol, which are present in tallow, in conformity with the process accordina to the invention.
200 g of tallow containing 99% of fatty sub-stances, of the type marketed by ROUSSELOT, are treatedin the same manner as in the preceding examples.
The initial cholesterol content of the tallow is 0.12% by weight.
Table III below shows the data and the results of the present example.
~ 21 - 20730~1 . . . ._ .~
X ~ 04 ~ ' ~ ~ o ~. ~
~ ,~ ~o D~
~ U~
~o~ ~
E~ _~
20730Sl _mments on Table III
It appears that an amount of water of 25~ by weight relative to the tallow is qufficient to extract 86% of the initial cholesterol, and this with only 3% by weight of cyclodextrin relative to the tallow.
An increase in the amount of water and/or the duration of placing in contact does not enable the extraction yield to be improved.
In this example, the variations in the acid values were also measured. The acid value i8 defined as the number of mg of potas~ium hydroxide required to neutralise the acidity of 1 g of tallow. The acid value of tallow before extraction is 0.85.
Table IV below presents the results obtained.
TABLE IV
............. .
Water, Tem- CD Kleptose, Duration Final Z reduc-X rela- perature % relative of plac- acid tion in tive to C to tallow ing in value the tallow contact value 20Tallow S0 26 3 30 0.65 30 200 g 25 16 3 30 0.12 86 It is shown here that the decrease in acid value, and therefore of the free fatty acid content, of the tallow i~ very pronounced with 25~ of water.
Claims (8)
1. Process for extracting minor fatty compounds, especially steroids, which occur in a substance of biological origin, using cyclodextrin, in which the cyclodextrin is placed in contact, with stirring and in a reaction medium containing water, with the substance to be treated so as to allow the formation of inclusion complexes between the cyclodextrin and the minor fatty compounds, these complexes being subsequently separated from the said substance, characterised in that the placing in contact of cyclodextrin/substance to be treated is carried out at a temperature which is chosen such that the fatty substances present in the said substance are in a congealed state that is to say a state in which the level of solid or crystalline fatty substances is greater than or equal to 5% by weight.
2. Process according to Claim 1, characterised in that the temperature is chosen such that the level of solid or crystalline fatty substances is preferably greater than or equal to 10%, and still more preferably greater than or equal to 30%.
3. Process according to Claim 1 or Claim 2, characterised in that the amount of water present in the reaction medium is less than 100%, preferably between 10 and 90%, and still more preferably between 20 and 60%, by weight relative to the amount of substance of biological origin to be treated, expressed on a dry basis.
4. Process according to any one of Claims 1 to 3, characterised in that the cyclodextrin is a beta cyclodextrin which is substituted or unsubstituted.
5. Process according to any one of Claims 1 to 4, characterised in that the cyclodextrin is used in pulverulent form and in that it has a water content of less than or equal to 11% by weight, preferably less than or equal to 7% by weight, and still more preferably less than or equal to 5% by weight.
6. Process according to any one of Claims 1 to 5, characterised in that the substance to be treated/water/
cyclodextrin mixture is provided in a pasty form which is subjected, during the period of placing in contact, to a kneading operation during which cyclodextrin/minor fatty compound complexes are formed.
cyclodextrin mixture is provided in a pasty form which is subjected, during the period of placing in contact, to a kneading operation during which cyclodextrin/minor fatty compound complexes are formed.
7. Process according to any one of claims 1 to 6, characterised in that the separation of the cyclo-dextrin/minor fatty compound inclusion complexes is carried out by heating the mixture to a temperature which is chosen such that the fatty substances present in the substance of biological origin to be treated are in a molten state, that is to say in a state wherein the level of solid or crystalline fatty substances is less than 1%
by weight, by virtue of which a supernatant fatty phase with a reduced content of minor fatty compound is separated from a mixed water-lipid phase containing the cyclodextrin/ minor fatty compound inclusion complexes.
by weight, by virtue of which a supernatant fatty phase with a reduced content of minor fatty compound is separated from a mixed water-lipid phase containing the cyclodextrin/ minor fatty compound inclusion complexes.
8. Process according to any one of Claims 1 to 7, characterised in that the amount of cyclodextrin used is between 0.5 and 15%, preferably between 1 and 10%, and still more preferably between 3 and 8% relative to the amount of substance of biological origin to be treated expressed on a dry basis.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| FR909010969A FR2666345B1 (en) | 1990-09-04 | 1990-09-04 | PROCESS FOR THE EXTRACTION OF MINOR FATTY COMPOUNDS CONTAINED IN MATERIAL OF ORGANIC ORIGIN. |
| FR9010969 | 1990-09-04 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2073051A1 true CA2073051A1 (en) | 1992-03-05 |
Family
ID=9400052
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002073051A Abandoned CA2073051A1 (en) | 1990-09-04 | 1991-09-04 | Process for extracting minor fatty compounds which occur in a substance of biological origin |
| CA002073014A Abandoned CA2073014A1 (en) | 1990-09-04 | 1991-09-04 | Process for extracting minor fatty compounds from a substance of biological origin |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002073014A Abandoned CA2073014A1 (en) | 1990-09-04 | 1991-09-04 | Process for extracting minor fatty compounds from a substance of biological origin |
Country Status (13)
| Country | Link |
|---|---|
| US (2) | US5412127A (en) |
| EP (2) | EP0498869B1 (en) |
| JP (2) | JPH05505849A (en) |
| AT (2) | ATE124716T1 (en) |
| AU (2) | AU652182B2 (en) |
| CA (2) | CA2073051A1 (en) |
| DE (2) | DE69110856T2 (en) |
| DK (2) | DK0498869T3 (en) |
| ES (2) | ES2073770T3 (en) |
| FI (1) | FI914132A (en) |
| FR (1) | FR2666345B1 (en) |
| NO (1) | NO180494C (en) |
| WO (2) | WO1992004430A1 (en) |
Families Citing this family (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5382442A (en) * | 1992-05-15 | 1995-01-17 | Brandeis University | Modified fat blends |
| US5624703A (en) * | 1992-05-15 | 1997-04-29 | Brandeis University | Modified fat blends |
| ES2109188B1 (en) * | 1996-03-07 | 1998-07-16 | Quesos Boffard S A | PROCEDURE FOR THE PREPARATION OF FRESH CREAM WITH LOW CHOLESTEROL CONTENT AND LOW CHOLESTEROL CONTENT CHEESES. |
| FI105274B (en) * | 1996-03-15 | 2000-07-14 | Upm Kymmene Oy | Method for hydrogenation of non-hormone-containing unsaturated plant steroids and regeneration of spent catalyst |
| NZ335308A (en) * | 1997-01-10 | 2000-02-28 | Gerald Thomas Hinton | Detergent composition containing a granulated percarbonate |
| AU5258701A (en) * | 2000-04-28 | 2001-11-12 | Takeda Chemical Industries Ltd. | Method of searching for arteriosclerosis inhibitors and shrinkers |
| AU2001265937A1 (en) * | 2000-06-30 | 2002-01-14 | Societe Des Produits Nestle S.A. | Flavouring compositions |
| US6462568B1 (en) * | 2000-08-31 | 2002-10-08 | Micron Technology, Inc. | Conductive polymer contact system and test method for semiconductor components |
| KR100817934B1 (en) | 2006-11-29 | 2008-03-31 | 한국과학기술연구원 | SELECTIVE EXTRACTION METHOD OF STEROID HORMONES USING POLYMERIZED beta;-CYCLODEXTRIN POWDERS |
| US8110235B2 (en) * | 2008-05-06 | 2012-02-07 | Alliance Enterprises Of S.E. Wisconsin, Llc | Methods for reducing cholesterol in a milk product |
| CN102281773A (en) * | 2008-12-24 | 2011-12-14 | 好侍食品株式会社 | Method for preventing decomposition/deterioration of lipophilic component in the presence of water |
| US10564076B2 (en) | 2015-06-16 | 2020-02-18 | Agilent Technologies, Inc. | Compositions and methods for analytical sample preparation |
| EP3320241A4 (en) * | 2015-07-09 | 2018-07-18 | Vistadeltek, LLC | Control plate in a valve |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3491132A (en) * | 1967-05-01 | 1970-01-20 | Corn Products Co | Glyceride oil treatment |
| NZ209508A (en) * | 1984-09-11 | 1988-03-30 | New Zealand Dairy Res Inst | Removing steroids and/or pharmaceutically active compounds from fats and oils |
| FR2601959B1 (en) * | 1986-07-24 | 1988-12-02 | Monserbio Gie | PROCESS FOR REMOVAL OF CHOLESTEROL FROM ANIMAL FATTY MATERIAL AND DEPLETED CHOLESTEROL FATTY MATERIAL OBTAINED |
| NZ222728A (en) * | 1987-11-27 | 1991-08-27 | New Zealand Dairy Res Inst | Method of removing sterols and or sterol oxides from edible fats and oils |
| FR2626145B1 (en) * | 1988-01-22 | 1990-07-06 | Monserbio | PROCESS FOR THE REMOVAL OF STEROID COMPOUNDS CONTAINED IN A SUBSTANCE OF BIOLOGICAL ORIGIN |
| BE1003488A3 (en) * | 1989-03-14 | 1992-04-07 | Corman N Sa | PROCESS FOR REDUCING THE CONTENT OF CHOLESTEROL AND FREE FATTY ACIDS IN FAT MATERIAL OF ANIMAL ORIGIN AND FAT MATERIAL THUS OBTAINED. |
| AU633084B2 (en) * | 1989-05-10 | 1993-01-21 | Commonwealth Scientific And Industrial Research Organisation | Cholesterol removal |
| AU630446B2 (en) * | 1989-05-19 | 1992-10-29 | Commonwealth Scientific And Industrial Research Organisation | Cholesterol removal from eggs, dairy products and other aqueous emulsions |
| FR2649589B1 (en) * | 1989-07-12 | 1991-10-31 | Roquette Freres | PROCESS FOR THE PREPARATION OF DAIRY PRODUCTS WITH REDUCED STEROLS, IN PARTICULAR CHOLESTEROL |
| DE4013367A1 (en) * | 1990-04-26 | 1991-10-31 | Sueddeutsche Kalkstickstoff | METHOD FOR REMOVING CHOLESTERIN OR CHOLESTERINE STARS FROM EGG YELLOW |
-
1990
- 1990-09-04 FR FR909010969A patent/FR2666345B1/en not_active Expired - Fee Related
-
1991
- 1991-09-03 FI FI914132A patent/FI914132A/en not_active Application Discontinuation
- 1991-09-03 NO NO913453A patent/NO180494C/en unknown
- 1991-09-04 DE DE69110856T patent/DE69110856T2/en not_active Expired - Fee Related
- 1991-09-04 US US07/854,662 patent/US5412127A/en not_active Expired - Fee Related
- 1991-09-04 AT AT91916173T patent/ATE124716T1/en not_active IP Right Cessation
- 1991-09-04 JP JP91514949A patent/JPH05505849A/en active Pending
- 1991-09-04 ES ES91916173T patent/ES2073770T3/en not_active Expired - Lifetime
- 1991-09-04 ES ES91916049T patent/ES2073769T3/en not_active Expired - Lifetime
- 1991-09-04 WO PCT/FR1991/000707 patent/WO1992004430A1/en active IP Right Grant
- 1991-09-04 AU AU85170/91A patent/AU652182B2/en not_active Ceased
- 1991-09-04 JP JP3514950A patent/JPH05502910A/en active Pending
- 1991-09-04 DK DK91916049.9T patent/DK0498869T3/en active
- 1991-09-04 EP EP91916049A patent/EP0498869B1/en not_active Expired - Lifetime
- 1991-09-04 WO PCT/FR1991/000708 patent/WO1992004431A1/en active IP Right Grant
- 1991-09-04 EP EP91916173A patent/EP0498870B1/en not_active Expired - Lifetime
- 1991-09-04 AU AU85113/91A patent/AU652180B2/en not_active Ceased
- 1991-09-04 US US07/852,149 patent/US5371255A/en not_active Expired - Fee Related
- 1991-09-04 DE DE69111048T patent/DE69111048T2/en not_active Expired - Fee Related
- 1991-09-04 DK DK91916173.7T patent/DK0498870T3/en active
- 1991-09-04 AT AT91916049T patent/ATE124444T1/en not_active IP Right Cessation
- 1991-09-04 CA CA002073051A patent/CA2073051A1/en not_active Abandoned
- 1991-09-04 CA CA002073014A patent/CA2073014A1/en not_active Abandoned
Also Published As
| Publication number | Publication date |
|---|---|
| FR2666345A1 (en) | 1992-03-06 |
| US5412127A (en) | 1995-05-02 |
| DE69110856D1 (en) | 1995-08-03 |
| AU652182B2 (en) | 1994-08-18 |
| NO913453L (en) | 1992-03-05 |
| ATE124716T1 (en) | 1995-07-15 |
| DE69111048D1 (en) | 1995-08-10 |
| NO180494C (en) | 1997-04-30 |
| WO1992004430A1 (en) | 1992-03-19 |
| EP0498869B1 (en) | 1995-06-28 |
| ES2073770T3 (en) | 1995-08-16 |
| FR2666345B1 (en) | 1994-10-14 |
| EP0498870A1 (en) | 1992-08-19 |
| DK0498870T3 (en) | 1995-11-20 |
| DE69111048T2 (en) | 1996-01-04 |
| US5371255A (en) | 1994-12-06 |
| EP0498869A1 (en) | 1992-08-19 |
| FI914132A0 (en) | 1991-09-03 |
| DE69110856T2 (en) | 1995-12-07 |
| JPH05505849A (en) | 1993-08-26 |
| AU652180B2 (en) | 1994-08-18 |
| JPH05502910A (en) | 1993-05-20 |
| ES2073769T3 (en) | 1995-08-16 |
| DK0498869T3 (en) | 1995-10-30 |
| NO180494B (en) | 1997-01-20 |
| AU8511391A (en) | 1992-03-30 |
| WO1992004431A1 (en) | 1992-03-19 |
| CA2073014A1 (en) | 1992-03-05 |
| FI914132A (en) | 1992-03-05 |
| AU8517091A (en) | 1992-03-30 |
| NO913453D0 (en) | 1991-09-03 |
| EP0498870B1 (en) | 1995-07-05 |
| ATE124444T1 (en) | 1995-07-15 |
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| Date | Code | Title | Description |
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| FZDE | Discontinued |