NO872687L

NO872687L - SUPER CRITICAL FLUID EXTRACTION OF ANIMAL DERIVATIVE SUBSTANCES.

Info

Publication number: NO872687L
Application number: NO872687A
Authority: NO
Inventors: Ahmad Reza Kamarei; Norman D Hanson
Original assignee: Angio Medical Corp
Priority date: 1985-10-31
Filing date: 1987-06-26
Publication date: 1987-06-26
Also published as: DK333887A; DK333887D0; NO872687D0

Description

Foreliggende oppfinnelse angår ekstrahering av ønskede stoffer fra komplekse komponenter der det ønskede stoff er en del. Spesielt angår oppfinnelsen ekstrahering via superkritiske eller subkritiske fluider. Disse fluider benyttes for å ekstrahere ønskede stoffer fra animalvev, -celler og The present invention relates to the extraction of desired substances from complex components of which the desired substance is a part. In particular, the invention relates to extraction via supercritical or subcritical fluids. These fluids are used to extract desired substances from animal tissue, cells and

-organer.-organs.

Superkritiske fluider, SCF, har hatt og fortsetter å ha stor oppmerksomhet. Kort sagt er et superkritisk fluid en gass underkastet temperatur og trykk over grenser kjent som kritisk trykk, Pc, og kritisk temperatur, Tc. Over disse punkter har disse SCF andre egenskaper enn de har i gass-formig tilstand. Fig. 1 viser et diagram over de superkritiske betingelser for en gass, CO2. Forskjellige betingelser er nødvendige for å gi andre superkritiske fluider slik man f.eks. kan se i tabell 7 nedenfor. Supercritical fluids, SCF, have had and continue to have a lot of attention. In short, a supercritical fluid is a gas subjected to temperature and pressure above limits known as critical pressure, Pc, and critical temperature, Tc. Above these points, these SCFs have different properties than they have in the gaseous state. Fig. 1 shows a diagram of the supercritical conditions for a gas, CO2. Different conditions are necessary to produce other supercritical fluids such as e.g. can be seen in table 7 below.

En egenskap som er karakteristisk for SCF er øket oppløs-ningsevne. Det er kjent at SCF ved temperaturer og trykk over Pc og Tc,-ekstraherer stoffer som ellers kun kan fjernes ved giftige, prohibitivt kostbare eller ineffektive midler. F.eks. har forskjellige kjemiske oppløsningsmidler slik som kloroform og metanol vært brukt. Mens bruken av disse produkter gir gode resultater er og har faren vært at rester av oppløsningsmidlet enten er toksisk per se eller er promotere for sykdommer (f.eks. kan de være karsinogene). A property that is characteristic of SCF is increased resolving power. It is known that SCF at temperatures and pressures above Pc and Tc extracts substances that can otherwise only be removed by toxic, prohibitively expensive or ineffective means. E.g. different chemical solvents such as chloroform and methanol have been used. While the use of these products gives good results, the danger is and has been that residues of the solvent are either toxic per se or promoters of diseases (eg they can be carcinogenic).

SCF-ekstrahering har vært spesielt brukbar for å oppnå aromatiske og lipidkomponenter fra plantevevet. F.eks. tyr oljeindustrien i stor grad til prosesser der vegetabilske oljer slik som soyabønner-, bomullsfrø- og maisoljer fjernes fra de vegitative komponenter. Kaffeindustrien bruker superkritiske prosesser for å fjerne kaffein fra kaffe, og smaksekstrahering ved bruk av SCF har vært anvendt f.eks. på humle, vegetabilier, frukt (citroner) og krydder. Slik man skal kunne forvente fra bruken av SCF som smaksekstraktor har de vært benyttet for å ekstrahere duftemner også. SCF extraction has been particularly useful for obtaining aromatic and lipid components from the plant tissue. E.g. the oil industry largely resorts to processes where vegetable oils such as soybean, cottonseed and corn oils are removed from the vegetative components. The coffee industry uses supercritical processes to remove caffeine from coffee, and flavor extraction using SCF has been used e.g. on hops, vegetables, fruit (lemons) and spices. As can be expected from the use of SCF as a flavor extractor, they have also been used to extract fragrances.

Det er viktig å merke seg at mens SCF har vært og i utstrakt grad benyttes for ekstrahering av vegetabilier og frø, har ingen benyttet prosessen for ekstrahering av komponenter fra animalske stoffer slik som organer, vev eller celler. It is important to note that while SCF has been and is widely used for extracting vegetables and seeds, no one has used the process for extracting components from animal substances such as organs, tissues or cells.

Det er ønskelig å ha en effektiv metode for ekstrahering av animalsk vev på grunn av brukbarheten til det oppnådde materiale. Spesielt når det gjelder lipidholdige molekyler inneholder animalvevangiogene faktorer, hormoner, regulator-iske molekyler osv. Lipidrike organer slik som lever, hjerne, nyrer og epitelvev er rike kilder for nødvendige og ønskelige biologiske produkter. En metode for ekstrahering av disse via superkritiske prosesser har til nå manglet. It is desirable to have an efficient method of extracting animal tissue because of the usability of the material obtained. Especially when it comes to lipid-containing molecules, animal tissue contains angiogenic factors, hormones, regulatory molecules, etc. Lipid-rich organs such as the liver, brain, kidneys and epithelial tissue are rich sources of necessary and desirable biological products. A method for extracting these via supercritical processes has been lacking until now.

Således er det en gjenstand for foreliggende oppfinnelse å tilveiebringe en metode for å oppnå ønskede komponenter av animalvev, celler og organer ved bruk av superkritiske fluider. Det er videre en gjenstand for oppfinnelsen å oppnå komplekse og enkle animalavledede lipidholdige stoffer ved superkritiske prosesser. Thus, it is an object of the present invention to provide a method for obtaining desired components of animal tissue, cells and organs using supercritical fluids. It is also an object of the invention to obtain complex and simple animal-derived lipid-containing substances by supercritical processes.

Ytterligere en gjenstand for oppfinnelsen er å oppnå amino-syre-, nukleotid- og sakkeridholdige stoffer såvel som andre molekyler ved bruk av superkritiske prosesser. A further object of the invention is to obtain amino acid, nucleotide and saccharide-containing substances as well as other molecules using supercritical processes.

Hvordan disse og andre gjenstander ifølge oppfinnelsen oppnås vil fremgå av den følgende beskrivelse. How these and other items according to the invention are achieved will be apparent from the following description.

Superkritisk gassekstrahering har vært kjent i teknikken i lang tid. Anvendelsen har imidlertid vært begrenset til plantevev. Stahl , et al, Agricultural and Food Chemistry 28:1153-1157 (1980), beskriver ekstrahering av frøolje (f.eks. soyabønneolje) ved bruk av superkritisk COg. Trykket som beskrives ligger i området 350-700 bar, og temperaturen fra 20-40°C. Kun soyabønner, solsikkefrø og rapsfrø ekstraheres ved prosessen og under de betingelser som er gitt ovenfor, i Chemistry and Industry (June 19, 1982), s. 399- 402, beskriver Calame et al isolering av luft- og smaks-ekstrakter ved bruk av superkritisk C02. Ekstraher ingen er begrenset til syrin (34°C, 90 bar); citronskall (40°C, 300 bar); svart pepper (60°CC, variert trykk); og mandler (40°C, Supercritical gas extraction has been known in the art for a long time. However, the application has been limited to plant tissue. Stahl, et al, Agricultural and Food Chemistry 28:1153-1157 (1980), describes the extraction of seed oil (eg, soybean oil) using supercritical CO 2 . The pressure described is in the range 350-700 bar, and the temperature from 20-40°C. Only soybeans, sunflower seeds and rapeseed are extracted by the process and under the conditions given above, in Chemistry and Industry (June 19, 1982), pp. 399-402, Calame et al describe the isolation of air and flavor extracts using supercritical C02. Extract none is limited to lilac (34°C, 90 bar); lemon peel (40°C, 300 bar); black pepper (60°CC, varied pressure); and almonds (40°C,

600 bar). Det bemerkes at Calmane et al angir at teknikken har plass for utvikling, dette på side 401. 600 bar). It is noted that Calmane et al indicate that the technique has room for development, this on page 401.

Federal Register, 48:57269 (nr. 251, des. 29, 1983), antyderFederal Register, 48:57269 (No. 251, Dec. 29, 1983), suggests

at "Food and Drug Administration", FDA, har angitt C02, N2, He, C3H3, C4H10»iso-C^io og N20 som sikre humannæringsmid-delbestanndeler. Denne rapport tolkes dit at den antyder at FDA vil betrakte reguleringer i forbindelse med superkritiske C02, på et senere tidspunkt. that the Food and Drug Administration, FDA, has designated C02, N2, He, C3H3, C4H10, iso-C20 and N2O as safe human food constituents. This report is interpreted to indicate that the FDA will consider regulations related to supercritical C02 at a later date.

Ytterligere -anvendelser av superkritisk COg er angitt hos Vollbrecht, Chemistry and Industry 19:397-399 (June, 1982) Additional uses of supercritical COg are set forth in Vollbrecht, Chemistry and Industry 19:397-399 (June, 1982)

(ekstrahering av humle, ingen trykk- eller temperaturdata er gitt). Friedrich, et al, Jaocs 59(7) :288-292 (1982) (soyabøn-ner, heksan eller C02ble benyttet); Fillippi, Chemistry and Industry t9:390-394 (juni, 1982) (C02oppløsningsmiddelegen-skaper; ingen lære om ekstrahering); Johnson, Food Engineer-ing News (s. 1, 8-10, 15) (Nov. 1983), (bomul1sfrøolje-ekstrahering) ; Friedrich, et alJaocs:61(2) 223-228 (soya-bønneekstrahering); Bulley, et al Jaocs 61:8:1362-1365 (august, 1984) (kanolafrøekstrahering); Christianson, et al., (extraction of hops, no pressure or temperature data provided). Friedrich, et al, Jaocs 59(7):288-292 (1982) (soybeans, hexane or CO 2 were used); Fillippi, Chemistry and Industry t9:390-394 (June, 1982) (CO 2 solvent properties; no extraction teaching); Johnson, Food Engineer-ing News (pp. 1, 8-10, 15) (Nov. 1983), (cottonseed oil extraction); Friedrich, et alJaocs:61(2) 223-228 (soybean extraction); Bulley, et al Jaocs 61:8:1362-1365 (August, 1984) (canola seed extraction); Christianson, et al.,

J. Food Sei 49:229-232 (1984 ) (maisspireekstrahering); Snyder et Jaocs 6lfl2) (desember, 1984) (soyabønneekstraher-ing); Friedrich, et al, J. Agr. & Food Chem (januar-februar 1982) s. 192-193 (soyabønneekstrahering). Det er bevilget patenter, spesielt på superkritisk ekstrahering av kaffebøn-ner, i denne forbindelse skal det henvises til US-PS 4.255.461 (60-100°C, 200 atm); Roseleius, et al, US-PS 4.25 5.458 (C02, N20, SF6, Xe, CF4, CH4, C2H6, C2H4, cyklo-C3H8, 50-300 bar trykk); Zasel, U.S. 4.247.570 (dekaffeiner-ing, 32°C-140°C, 75-350 atm). J. Food Sei 49:229-232 (1984 ) (maize sprout extraction); Snyder et Jaocs 6lfl2) (December, 1984) (soybean extraction); Friedrich, et al., J. Agr. & Food Chem (January-February 1982) pp. 192-193 (soybean extraction). Patents have been granted, especially on supercritical extraction of coffee beans, in this connection reference should be made to US-PS 4,255,461 (60-100°C, 200 atm); Roseleius, et al, US-PS 4.25 5.458 (CO 2 , N 2 O, SF 6 , Xe, CF 4 , CH 4 , C 2 H 6 , C 2 H 4 , cyclo-C 3 H 8 , 50-300 bar pressure); Zasel, U.S. 4,247,570 (decaffeination, 32°C-140°C, 75-350 atm).

To US-patenter er bevilget med henblikk på bruk av superkritisk gass som anvendt på dyrevev. US-PS 4.280.961 beskriver en fremgangsmåte der dyrefett separeres fra kjøtt ved produktet slik som involler, skrapfett osv. Metoden angår f.eks. ekstrahering av renset nyrefett eller spekklignende materiale, men beskriver ikke rensede lipider eller lipidholdige stoffer. Som kjent i teknikken er, selv om fett inneholder lipider, de to typer ikke-ekvivalente. Schneider nevner i det sistnevnte patent ingen parametre for ekstraheringen. Two US patents have been granted for the use of supercritical gas as applied to animal tissue. US-PS 4,280,961 describes a method in which animal fat is separated from meat by the product such as viscera, scrap fat, etc. The method concerns e.g. extraction of purified kidney fat or lard-like material, but does not describe purified lipids or lipid-containing substances. As known in the art, although fats contain lipids, the two types are non-equivalent. In the latter patent, Schneider does not mention any parameters for the extraction.

US-PS 4.466.923 beskriver anvendelse av temperaturer ut over 60° C og trykk ut over 550 bar for å oppnå lipider fra lipidholdige ^stoffer. De eneste eksempler som gis er imidlertid vegetabilske frø. Ved de beskrevne trykk- og temperatur-områder, ville fagmannen forvente at mere delikat animalsk materiale disintegrerer. US-PS 4,466,923 describes the use of temperatures above 60° C and pressure above 550 bar to obtain lipids from lipid-containing substances. However, the only examples given are vegetable seeds. At the pressure and temperature ranges described, the person skilled in the art would expect more delicate animal material to disintegrate.

Således er det i teknikken ikke funnet noen lære eller noen antydninger at superkritisk gassekstrahering kan benyttes for å oppnå ønskede lipidholdige stoffer fra dyrevev, -celler og Thus, no teachings or any hints have been found in the art that supercritical gas extraction can be used to obtain desired lipid-containing substances from animal tissues, cells and

-organer. Som de følgende eksempler viser er dette mulig. Fig. 1 gjengir grafisk faseendringene hos en gass, C02, med en beskrivelse av de betingelser der gassen blir et superkritisk fluid, SCF.; Fig. 2 viser en eksemplifisering av en ekstrahering med en apparatur for bruk ved ekstrahering av ønskede stoffer fra animalavledede produkter. -organs. As the following examples show, this is possible. Fig. 1 graphically reproduces the phase changes of a gas, C02, with a description of the conditions under which the gas becomes a supercritical fluid, SCF.; Fig. 2 shows an example of an extraction with an apparatus for use in extracting desired substances from animal-derived products.

Seks prøver ble valgt for analyse: homogenater av svineadi-poseyev, svineomentum og bovinomentum,, og CMFr-ekstrakter av hver av disse. Six samples were selected for analysis: homogenates of porcine adi-poseyev, porcine neomentum and bovine mentum, and CMFr extracts of each of these.

Homogenatprøvene ble fremstilt ved tilsetning av destillert vann i to ganger vedvolumet. Homogeniseringen ble gjennomført ved sentrifugering ved 22.ooo omdr. pr. minutt i 90 sek. The homogenate samples were prepared by adding distilled water to twice the wood volume. The homogenization was carried out by centrifugation at 22,000 rpm. minute for 90 sec.

fulgt av f rysetørking overnatt. Klorof orm/metanolf raks jon CCMFr)-prøvene ble preparert ved å tilsette 4 ganger volumet followed by freeze-drying overnight. The chloroform/methanol fraction (CCMFr) samples were prepared by adding 4 times the volume

av fosfatbuffret saltoppløsning, PBS, med homogenisering og sentrifugering som angitt ovenfor. Dette ga en lipidkake som så gjenvinnes og ekstraheres med 10 ganger volumet av klorof orm/metanoloppløsningsmiddel, 2:1 v/v. Sentrifugering og fordamping av oppløsningsmidlet fulgte deretter med derpå følgende gjenvinning av filtrert, viskøs supernatant. of phosphate buffered saline, PBS, with homogenization and centrifugation as indicated above. This gave a lipid cake which was then recovered and extracted with 10 times the volume of chloroform/methanol solvent, 2:1 v/v. Centrifugation and evaporation of the solvent then followed with subsequent recovery of filtered viscous supernatant.

En prosessutviklingsenhet, PDU, som vist i fig. 2, benyttes. Kort sagt består denne av en ekstraktor og tre separatorer som befinner seg i en ovn med på forhånd bestemt temperatur. A process development unit, PDU, as shown in fig. 2, is used. In short, this consists of an extractor and three separators located in an oven with a predetermined temperature.

Et superkritisk oppløsningsmiddel, i dette tilfelle COg, pumpes inn i ekstraktoren 101 og strømmer deretter sekven-sielt gjennom separatorer 102, 103 og 104 og til utgangsbe-holderen 105. Trykket holdes ved tilbaketrykksregulatorer 106-109. Gass, f.eks. COg, ved omtrent vanlig atmosfærisk trykk, går ut gjennom ventilen 110 når ekstraheringen er ferdig. A supercritical solvent, in this case COg, is pumped into the extractor 101 and then flows sequentially through separators 102, 103 and 104 and to the output container 105. The pressure is maintained by back pressure regulators 106-109. Gas, e.g. COg, at approximately normal atmospheric pressure, exits through valve 110 when extraction is complete.

EKSEMPEL 1EXAMPLE 1

Prøver ble smeltet i en separat, nitrogenspylt pvn ved ca. 40°C, og så overført til ekstraktoren 101. Beholderne 102-104 ble spylt med lavtrykks COg og ble så bragt til de temperaturer og trykk som er angitt i tabellene 1-6. COg ble deretter pumpet inn i en mengde av ca. 0,3 lb/min. gjennom systemet inn til en<*>vekt på ca. 200 ganger prøven var pumpet. Trykket ble avlastet og prøvene i hver av beholderne og ekstraktoren fjernet, veiet og analysert. I disse forsøk ble det observert at nær og over det kritiske punkt til den benyttede COg øket oppløsningsevnen med en økning i temperaturen, ved konstant densitet og med øket densitet ved konstant temperatur. F.eks. felte en andel av prøven, oppløst i ekstraktoren ved 3500 psig, ut ved 1500 psig i den første separatorbeholder. Ytterligere reduksjoner i trykk/densitet forårsaket at ytteligere fraksjoner presipiterte inntil, ved omgi vel sestrykk, den superkritiske gass ikke inneholdt oppløst materiale. Samples were melted in a separate, nitrogen-purged pvn at approx. 40°C, and then transferred to the extractor 101. The containers 102-104 were flushed with low-pressure COg and were then brought to the temperatures and pressures indicated in tables 1-6. COg was then pumped into an amount of approx. 0.3 lb/min. through the system into a<*>weight of approx. 200 times the sample was pumped. The pressure was relieved and the samples in each of the containers and the extractor were removed, weighed and analyzed. In these experiments, it was observed that close to and above the critical point of the COg used, the dissolving ability increased with an increase in temperature, at constant density and with increased density at constant temperature. E.g. precipitated a portion of the sample, dissolved in the extractor at 3500 psig, out at 1500 psig into the first separator vessel. Further reductions in pressure/density caused further fractions to precipitate until, at ambient pressure, the supercritical gas contained no solute.

Restene som oppnås fra ekstraktoren, ble funnet å være uoppløselige i COg. Polare andeler av materialer slik som gangl i os i der, var forventet å forbli i fraksjonen mens ekstraktfraksjonene var forventet å være rike på nøytrale, ikke-polare komponenter. Dette har vist seg å være tilfelle da den uoppløselige rest finnes å inneholde polare stoffer slik som gangliosider mens ikke-polare stoffer slik som triglyserider finnes i ekstraktene. Mens en enkelt ekstraktor, tre separatorer og samlebeholdere og en utfellingsbehol-der benyttes i denne utførelsesform, vil fagmannen selvfølge-lig erkjenne lat antallet og kombinasjoner av hver av disse er et konstruksj-onsvalg. The residues obtained from the extractor were found to be insoluble in COg. Polar proportions of materials such as gangl i os i der were expected to remain in the fraction while the extract fractions were expected to be rich in neutral, non-polar components. This has been shown to be the case as the insoluble residue is found to contain polar substances such as gangliosides while non-polar substances such as triglycerides are found in the extracts. While a single extractor, three separators and collection containers and a precipitation container are used in this embodiment, the person skilled in the art will of course recognize that the number and combinations of each of these is a design choice.

POLAR IKKE- POLARPOLAR NON- POLAR

Tabell 1Table 1

Materiale: Porsinhomogenisert adiposevevMaterial: Porcine homogenized adipose tissue

BETINGELSERCONDITIONS

Separator Separator Separator Separator Separator Separator

EKSEMPLENE 2- 18 EXAMPLES 2-18

Prosedyren i eksempel 1 benyttes for å ekstrahere de ønskede komponenter som finnes i andre dyrevev, -organer og -celler. F. eks. benyttes sentralnervesystemvev og -organer (hjerne, ryggmark , spinalfluid osv.); per if ernervesystemvev og-organer Ckranienerver, spinalnerver etc); myokardialt og vaskulært vev og -organer (hjerte, arterier og vener); sirkulatorisk vev og organer (blod, erytrocyter, leukocyter, blodplater og -plasma); lymfesystemvev og -organer (lymfe-kjertel, milt, tymus); åndedrettssystemvev og -organer (øvre luftkanal, lunger); fordøyelsessystemvev og -organer (inkludert munn, tenner, tunge, spyttkjertler, pharynz, esofagus, peritonium, mave, -tykk- og tynntarm, lever, galleblære og pankreas); skjelettvev og -organer (aksial- og appendikulær-skjelett, benmarg); muskler (glatte og skjelett); endotelisk og epitelisk vev; membraner, omentum og kartiligenvev (sener, ligamenter, ledd); oppfattelsesorganer (øyne, øre, nese, tunge); endokrint eller annet glandulært vev (tyroidkjerte-len, paratyroidkjertelen, pituitærkjertelen, adrenalkjerte-len); urinvev og organer (nyrer, urinblære, urinrør osv.); reproduktive organer og vev (testikler, ovarier osv.) og adiposevev slik det inneholdes i subkutane og i indre organer såvel som biologiske eksudater slik som f.eks. urin, svette, sæd, melk osv. I hvert tilfelle blir et superkritisk fluid valgt som, under superkritiske betingelser, fjerner den ønskede komponent eller komponenter (f.eks. lipidholdige molekylæe proteiner osv.) med minimal skade på den resulter-ende ekstrakt. The procedure in example 1 is used to extract the desired components found in other animal tissues, organs and cells. For example central nervous system tissues and organs are used (brain, spinal cord, spinal fluid, etc.); per if nervous system tissues and organs Ccranial nerves, spinal nerves etc); myocardial and vascular tissues and organs (heart, arteries and veins); circulatory tissues and organs (blood, erythrocytes, leukocytes, platelets and plasma); lymphatic system tissues and organs (lymph gland, spleen, thymus); respiratory system tissues and organs (upper airway, lungs); digestive system tissues and organs (including mouth, teeth, tongue, salivary glands, pharynx, esophagus, peritoneum, stomach, large and small intestines, liver, gall bladder and pancreas); skeletal tissues and organs (axial and appendicular skeleton, bone marrow); muscles (smooth and skeletal); endothelial and epithelial tissues; membranes, omentum and cartilaginous tissue (tendons, ligaments, joints); organs of perception (eyes, ears, nose, tongue); endocrine or other glandular tissue (thyroid gland, parathyroid gland, pituitary gland, adrenal gland); urinary tissues and organs (kidneys, bladder, urethra, etc.); reproductive organs and tissues (testicles, ovaries, etc.) and adipose tissue as contained in subcutaneous and internal organs as well as biological exudates such as e.g. urine, sweat, semen, milk, etc. In each case, a supercritical fluid is selected which, under supercritical conditions, removes the desired component or components (e.g. lipid-containing molecular proteins, etc.) with minimal damage to the resulting extract.

Eksempelene 19- 66Examples 19-66

De følgende gasser i tabell 7 benyttes i superkritiske ekstraheringsprosesser på stoffene som beskrevet i eksemplene 1-18. Ikke alle disse gasser er ønskelige for hvert vev og hver ønsket ekstrakt. Hvis f. eks. den kritiske temperatur ligger over den temperatur ved hvilken en ønsket ekstrakt er funksjonell blir denne gass ikke benyttet. For fagmannen er det imidlertid en lett oppgave å bestemme hvilken gass som kan anvendes eller som er ønskelig, basert på de kjente egenskaper for vev og ønskelige komponenter såvel som gass-spesifikasjoner inkludert de superkritiske temperaturer og trykk. De stoffer som kan ekstraheres ved bruk av de her beskrevne fremgangsmåter inkluderer men er ikke begrenset til komplekse lipider slik som acylglyseroler, fosfoglyserider, sfingoli-pider og vokser; enkle lipider, slik som terpener, pigmenter, steroider og deres alkoholer (steroler), prostaglandiner osv. Glykolipider, lipoproteiner, membransupramolekylære komplekser og deres metabolske mellomprodukter, være disse katabolske eller anabolske, og metabolske produkter av disse molekyler, såvel som molekyler oppvarmer seg på en måte tilsvarende lipider, kan oppnås på en måte tilsvarende det som sies i eksempel 1. The following gases in table 7 are used in supercritical extraction processes for the substances as described in examples 1-18. Not all of these gases are desirable for every tissue and every desired extract. If e.g. the critical temperature is above the temperature at which a desired extract is functional, this gas is not used. For the person skilled in the art, however, it is an easy task to determine which gas can be used or which is desirable, based on the known properties of tissue and desirable components as well as gas specifications including the supercritical temperatures and pressures. The substances that can be extracted using the methods described here include but are not limited to complex lipids such as acylglycerols, phosphoglycerides, sphingolipids and waxes; simple lipids, such as terpenes, pigments, steroids and their alcohols (sterols), prostaglandins, etc. Glycolipids, lipoproteins, membrane supramolecular complexes and their metabolic intermediates, be these catabolic or anabolic, and metabolic products of these molecules, as well as molecules heating up on in a way similar to lipids, can be obtained in a way similar to what is said in example 1.

Ytterligere molekyler kan også oppnås ved fremgangsmåten ifølge oppfinnelsen. For eksempel kan "proteinholdige" stoffer slik som aminosyreholdige stoffer (inkludert ikke-prot einaminosyrer ) , ol igopeptider , peptider, polypeptider, hormoner, proteiner, enzymer, antistoffer, fraksjoner og komponenter av disse såvel som metabolske mellomprodukter og produkter oppnås. Mens valget av SCF og reaksjonsparametre vil variere avhengig av stoffet som skal ekstraheres, vil fagmannen være istand til å bestemme hvilke reaksjoner og betingelser som må brukes. Additional molecules can also be obtained by the method according to the invention. For example, "proteinaceous" substances such as amino acid-containing substances (including non-protein amino acids), oligopeptides, peptides, polypeptides, hormones, proteins, enzymes, antibodies, fractions and components thereof as well as metabolic intermediates and products can be obtained. While the choice of SCF and reaction parameters will vary depending on the substance to be extracted, one skilled in the art will be able to determine which reactions and conditions must be used.

Sakkar ider inkludert mono-, di-, oligo- og polysakkarider såvel som glykoproteiner kan ekstraheres på denne måte også. Igjen kan metabolske mellomprodukter og andre produkter oppnås på samme måte. Saccharides including mono-, di-, oligo- and polysaccharides as well as glycoproteins can be extracted in this way as well. Again, metabolic intermediates and other products can be obtained in the same way.

Nukleotidfamilien av molekyler inkludert pyriner og pyrimi-diner og alle molekyler inneholdende nukleinsyrebaser, nukleoslder (ribonukleosider og deoksyribonukleosider) , nukleinsyrer, supramolekylære komplekser av nukleinsyrer og proteiner, viruser osv. såvel som disses mellomprodukter og produkter, metabolske produkter kan også oppnås. The nucleotide family of molecules including pyrins and pyrimidines and all molecules containing nucleic acid bases, nucleosides (ribonucleosides and deoxyribonucleosides), nucleic acids, supramolecular complexes of nucleic acids and proteins, viruses, etc. as well as their intermediates and products, metabolic products can also be obtained.

I tillegg kan stoffer som ikke er grupper i noen av de ovenfor angitte "familer" oppnås. Disse inkluderer alle fetter og/eller vannoppløselige vitaminer, smaksstoffer, smakspotensiatorer, disses mellomprodukter, både katabolske og anabolske, og propdukter også. In addition, substances that are not groups in any of the above stated "families" can be obtained. These include all fat and/or water-soluble vitamins, flavorings, flavor enhancers, their intermediates, both catabolic and anabolic, and prop-products as well.

Det skal ikke antas at metoden kan benyttes kun for å oppnå ønskede produktr. Uønskede stoffer slik som toksiner, alergener osv. kan fjernes fra en prøve ved å følge oppfinnelsen. Såvel vil fagmannen merke seg at metoden har anvendelse for biologiske renseprosesser der det er nødvendig å fjerne uønskede stoffer. - It should not be assumed that the method can only be used to achieve desired products. Unwanted substances such as toxins, allergens etc. can be removed from a sample by following the invention. The person skilled in the art will also note that the method has application for biological cleaning processes where it is necessary to remove unwanted substances. -

Forskjellige metoder kan benyttes for å fremstille stoffer som benyttes i ekstraheringsprosessen inkludert, men ikke begrenset ti*l, maling, knusing, oppkutting, høy- og lav-trykkspressi-ng, kryooppmaling, flakdannelse, sonikering, frysing, frys-tin-behandling, frysetørking, emulgering, homogenisering, filtrering, høyhastighetsblanding, sentrifugering, celleseparering, mekanisk separering, termisk behandling og andre fysikalske behandlinger; kjemisk behandling slik-som behandling med uorganiske og organiske syrer, baser, oppløsningsmidler, overflateaktive midler, farvestof-fer, ioniseringsstrålingsbehandling; enzymatisk behandling slik som endogen og/eller eksogenenzymatisk behandling og en hvilken som helst kombinasjon av mer enn én av de ovenfor angitte metoder for behandling av prøven. Various methods can be used to produce substances used in the extraction process including, but not limited to, grinding, crushing, cutting, high and low pressure pressing, cryo-grinding, flake formation, sonication, freezing, freeze-thaw treatment, freeze-drying, emulsification, homogenization, filtration, high-speed mixing, centrifugation, cell separation, mechanical separation, thermal treatment and other physical treatments; chemical treatment such as treatment with inorganic and organic acids, bases, solvents, surfactants, dyes, ionizing radiation treatment; enzymatic treatment such as endogenous and/or exogenous enzymatic treatment and any combination of more than one of the above methods for treating the sample.

Prøven behøver ikke å behandles før SCF-ekstrahering men kan f.eks. behandles etter ekstrahering når stoffene allerede er fjernet fra prøven. Videre vil fagmannen se at forskjellige kombinasjoner av SCF kan benyttes ved forskjellige anvendelser for generelle formål. SCF kan kombineres eller kan benyttes en etter en i sekvenser eller trinn. The sample does not need to be processed before SCF extraction, but can e.g. processed after extraction when the substances have already been removed from the sample. Furthermore, the person skilled in the art will see that different combinations of SCF can be used in different applications for general purposes. SCF can be combined or can be used one by one in sequences or steps.

I praksis benyttes ved ekstrahering ved bruk av SCF forskjellige modifiserere eller hjelpestoffer for å optimalisere prosessen. Disse kan øke oppløseligheten og forbedre selekti-viteten og utbyttene ved ekstraheringene. Eksempler på slike stoffer er vann, alkoholer slik som etanol og n-propanol, ketoner, slik som aceton og andre. In practice, when extracting using SCF, different modifiers or auxiliaries are used to optimize the process. These can increase the solubility and improve the selectivity and yields of the extractions. Examples of such substances are water, alcohols such as ethanol and n-propanol, ketones such as acetone and others.

Slik fagmannen vil erkjenne, kan denne metode benyttes ved fremgangsmåter andre enn animalvevekstrahering. Den har anvendelsesområder på et hvilket som helst område der separering av forskjellige komponenter er ønskelig eller nødvendig. I eksperimentelle prosesser der separering av en blanding av polare og ikke-polare stoffer er vanskelig kan f.eks. ekstrahering med SCF muliggjøre denne rensing av biologiske stoffer slik som medikamenter og andre farmasøy-tiske produkter, kosmetika, næringsmidler, vitaminprodukter osv. ved hjelp av fremgangsmåten. I industriell skala kan en hvilken som* helst kjemisk proses som krever molekylær separering gjennomføres ved bruk av den her antydede fremgangsmåte . As the person skilled in the art will recognize, this method can be used for methods other than animal tissue extraction. It has applications in any area where separation of different components is desirable or necessary. In experimental processes where separation of a mixture of polar and non-polar substances is difficult, e.g. extraction with SCF enable this purification of biological substances such as drugs and other pharmaceutical products, cosmetics, foodstuffs, vitamin products, etc. using the method. On an industrial scale, any chemical process requiring molecular separation can be carried out using the method suggested here.

Oppfinnelsen skal ikke være begrenset av de gitte eksempler, fagmannen vil erkjenne at variasjoner kan gjennomføres og oppfinnelsens grense er de ledsagnde krav. The invention shall not be limited by the examples given, the person skilled in the art will recognize that variations can be implemented and the limit of the invention is the accompanying claims.

Claims

1. Method for obtaining components of animal material, characterized in that it comprises bringing material into contact with at least one supercritical fluid under conditions that favor extraction of the components, and collecting the components.

2. Method according to claim 1, characterized in that a number of supercritical fluids are used.

3. Method according to claim 1, characterized in that the components comprise lipid-containing molecules.

4- Method according to claim 1, characterized in that the at-components comprise amino acid-containing molecules.

5. Method according to claim 1, characterized in that the components comprise molecules containing saccharide residues.

6. Method according to claim 1, characterized in that the components comprise nucleotide-containing molecules.

7. Method according to claim 3, characterized in that the components comprise complex lipids including acylglycerols, phosphorus orglycerides, sphingolipids, waxes and anabolic and catabolic intermediates and products of complex lipid molecules.

8. Method according to claim 3, characterized in that the components comprise molecules comprising simple lipids including terpenes, pigments, steroids, sterols, prostaglandins and catabolic and anabolic intermediates and products of the simple lipids.

9. Method according to claim 3, characterized in that said lipid-containing molecules comprise glycolides, lipoproteins, cell membrane supramolecular complexes and catabolic and anabolic intermediates and products of said lipid-containing molecules.

10. Method according to claim 4, characterized in that the amino acid-containing molecules comprise peptides, oligopeptides, polypeptides, hormones, proteins, enzymes, antibodies, amino acid-containing fraction of molecules and catabolic and anabolic intermediates and products of the amino acid-containing molecules.

11. Method according to claim 5, characterized in that said molecules containing saccharide residues comprise mono-, di-, oligo- and polysaccharides, glycoproteins and catabolic and anabolic intermediates and products of the molecules.

12. Method according to claim 6, characterized in that the nucleotide-containing molecules comprise nucleosides, nucleic acids, protein-nucleic acid complexes, viruses, nucleotide-containing organisms and catabolic and anabolic intermediates and products of said molecules.

13. Method according to claim 1, characterized in that the components include vitamins and catabolic and anabolic intermediates and products of the components.

14. Process according to claim 1, characterized in that the components include fragrances, fragrance potentiators and catabolic and anabolic intermediates and products of the components.

15. Method according to claim 1, characterized in that it comprises removing unwanted components from a sample.

16. Method according to claim 15, characterized in that said components comprise toxins.

17. Method according to claim 1, characterized in that the components comprise molecules soluble in organic solvents.

18. Method according to claim 1, characterized in that the components comprise molecules soluble in inorganic solvents.

19. Method according to claim 1, characterized in that the animal material comprises animal tissue.

20. Method according to claim 1, characterized in that the animal material comprises cells.

21. Method according to claim 1, characterized in that the animal material comprises exudates.

22. Method according to claim 1, characterized in that the animal material comprises animal organs.

23. Method according to claim 1, characterized in that the animal material comprises animal internal organs.

24. Method according to claim 1, characterized in that the animal material comprises animal nerve tissue.

25. Method according to claim 1, characterized in that the animal material comprises animal muscle tissue.

26. Method according to claim 1, characterized in that the animal material comprises animal adipose tissue.

27. Method according to claim 1, characterized in that the animal material comprises animal cartilaginous tissue.

28. Method according to claim 1, characterized by. that the animal material comprises animal glandular tissue.

29. Method according to claim 1, characterized in that the animal material comprises animal epithelial tissue.

30. Method according to claim 1, characterized in that the animal material comprises animal endothelial tissue.

31. Method according to claim 1, characterized in that the animal material comprises animal myocardial tissue.

32. Method according to claim 1, characterized in that the animal material comprises animal vascular tissue.

33. Method according to claim 1, characterized in that the animal material comprises animal circulatory tissue.

34. Method according to claim 1, characterized in that the animal material comprises animal lymphatic tissue.

Method according to claim 1, characterized in that the animal material comprises animal respiratory tissue.

36. Method according to claim 1, characterized in that the animal material comprises animal digestive organ tissue.

37. Method according to claim 1, characterized in that the animal material comprises animal skeletal tissue.

38. Method according to claim 1, characterized in that the animal material comprises animal sense organ tissue.

39. Method according to claim 1, characterized in that the animal material comprises animal urinary tract tissue.

40. Method according to claim 1, characterized in that the animal material comprises animal reproductive tissue.

41. Method according to claim 1, characterized in that the supercritical fluid is a gas.

42. Method according to claim 41, characterized in that the gas is an elementary gas.

43. Method according to claim 41, characterized in that the elementary gas is an inert gas.

44. Method according to claim 41, characterized in that the gas contains carbon.

45. Method according to claim 41, characterized in that the gas is a noble gas.

46. Method according to claim 41, characterized in that the gas is carbon dioxide.

47. Method according to claim 41, characterized in that the gas is an alkane gas.

48. Method according to claim 41, characterized in that the gas is an alkene gas.

49. Method according to claim 41, characterized in that the gas is an alkyne gas.

50. Method according to claim 41, characterized in that the gas is a nitrogen-containing gas.

51. Method according to claim 50, characterized in that the gas is selected from among ammonia, nitrogen oxide, nitrogen dioxide and nitrous oxide.

52. Method according to claim 41, characterized in that the gas contains silicon.

53. Method according to claim 52, characterized in that the gas is selected from silane, chlorotrifluorosilane and silicon tetrafluoride.

54. Method according to claim 41, characterized in that the gas contains sulphur.

55. Method according to claim 54, characterized in that the gas is selected from the group of sulfur dioxide and sulfur hexafluoride.

56. Method according to claim 41, characterized in that the gas contains hydrogen.

57. Method according to claim 41, characterized in that the gas contains halogen.

58. Method according to claim 41, characterized in that the gas is water vapour.

59. Method according to claim 1, characterized in that the method further comprises the addition of a modifying or accompanying aid to the supercritical fluid.

60. Method according to claim 59, characterized in that the agent is water.

61. Method according to claim 59, characterized in that the agent is alcohol.

62. Method according to claim 61, characterized by. that the alcohol is ethanol.

63. Method according to claim 41, characterized in that the alcohol is n-propanol.

64. Method according to claim 59, characterized in that the agent is a ketone.

65. Method according to claim 64, characterized in that the ketone is acetone.