EXTRACTION METHOD OF ACTIVE MOLECULAR STRUCTURES FROM NATURAL RESINS AND/OR ESSENTIAL OILS
Kind Code:
A1
An extraction method is described of terpenes and terpenoids from natural resins, such as myrrh, incense, dacryodes, dammar, propolis and/or essential oils, by means of extraction with polar and/or semi-polar solvents in the presence of a rotating magnetic field.
Inventors:
Fochesato, Antonio (Via R. Rompato 19, Schio, I-36015, IT)
Application Number:
PCT/EP2004/007295
Publication Date:
01/20/2005
Filing Date:
07/02/2004
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Export Citation:
Assignee:
CALEGARO SILVER SARDA S.R.L. (Prolungamento Via Natta Casic, Cagliari, I-09132, IT)
Fochesato, Antonio (Via R. Rompato 19, Schio, I-36015, IT)
Fochesato, Antonio (Via R. Rompato 19, Schio, I-36015, IT)
International Classes:
B01D11/02; B01D11/04; C11B9/02
Attorney, Agent or Firm:
Susanetto, Carlo (Cantaluppi & Partners S.r.l, Via Matteotti 26, Padova, I-35137, IT)
Claims:
CLAIMS
1. A method for the extraction of terpenes and/or ter penoids from natural resins or essential oils by means of extraction with polar and/or semipolar solvents in the presence of a rotating magnetic field.
2. The method according to claim 1, characterized in that the natural resins are incense, myrrh, dacryo des, dammar and/or propolis.
3. The method according to claim 1, characterized in that the polar and/or semipolar solvent is selected from ethanol and mixtures of ethanol/ethyl etha noate.
4. The method according to claim 3, characterized in that the polar and/or semipolar solvent is pharma ceutical ethanol.
5. The method according to claim 1, characterized in that the solvent is present in a quantity varying from 10 to 90% by weight.
6. The method according to claim 1, characterized in that the rotating magnetic field has an intensity ranging from 500 to 3000 Gauss, preferably from 1500 to 3000 Gauss.
7. The method according to claim 1, characterized in that the extraction is carried out at a temperature ranging from 30°C to 75°C, preferably from 35°C to 60°C.
8. The method according to claim 1, characterized in that the extraction is carried out for a time rang ing from 15 to 120 minutes, preferably from 30 to 60 minutes.
9. The method according to claim 1, characterized in that the extraction is carried out at a temperature ranging from 35°C to 60°C, for a time ranging from 30 to 60 minutes, with a rotating magnetic field which has an intensity varying from 1500 to 3000 Gauss.
10. Alcohol and/or hydroalcohol solutions which can be obtained with the method according to any of the previous claims, characterized in that they contain free molecular structures of sesquiterpenes, terpe nes, triterpenes.
11. The solutions according to claim 10, characterized in that they are dispersible in air within a tem erature range of 40°C to 90oC.
12. The solutions according to claim 11, characterized in that they are dispersible in air within a tem perature range of 80°C to 90°C.
13. The solutions according to claim 11 or 12, charac terized in that they are dispersible in air with the use of thermoemanators or electroemanators.
14. The solutions according to claim 10, characterized in that they are used in a mixture with each other, in a mixture with all types of essential oil, in any proportion, and/or in a mixture with water, up to a maximum of 25% of distilled water, whatever the pro portion of the solutions between each other may be.
15. The solutions according to any of the claims from 10 to 14, characterized in that they are solutions in ethanol and/or ethyl ethanoate, in any proportion.
16. The solutions according to any of the claims from 10 to 15, characterized in that they contain aero dispersible compounds, terpenes and/or terpenoids, with a molecular weight which varies within the range of MW 136 (monoterpenes) to MM 532 (pentacy clic triterpenes).
17. The solutions according to claim 10, characterized in that they contain fractions of terpenes and/or terpenoids extracted from incense in a percentage of between 15 and 65% by weight, fractions of terpenes and/or terpenoids extracted from myrrh in a percent age of between 15 and 65% by weight.
18. The solutions according to claim 17, characterized in that they contain Hyssopus officinalis decumbens or Hyssopus officinalis aristatu. green tangerine, fractions of terpenes and/or terpenoids extracted from myrrh and fractions of terpenes and/or terpe noids extracted from incense.
19. The solutions according to claim 18, characterized in that they contain Hyssopus officinalis decumbens or Hyssopus officinalis aristatus, green tangerine, fractions of terpenes and/or terpenoids extracted from myrrh and fractions of terpenes and/or terpe noids extracted from incense in proportions equal to 1: 0.3 : 5: 5.
20. The solutions according to claim 10, characterized in that they contain fractions of terpenes and/or terpenoids extracted from incense, myrrh and propo lis in a mixture with each other.
21. The solutions according to claim 10, characterized in that they are topically applied.
1. A method for the extraction of terpenes and/or ter penoids from natural resins or essential oils by means of extraction with polar and/or semipolar solvents in the presence of a rotating magnetic field.
2. The method according to claim 1, characterized in that the natural resins are incense, myrrh, dacryo des, dammar and/or propolis.
3. The method according to claim 1, characterized in that the polar and/or semipolar solvent is selected from ethanol and mixtures of ethanol/ethyl etha noate.
4. The method according to claim 3, characterized in that the polar and/or semipolar solvent is pharma ceutical ethanol.
5. The method according to claim 1, characterized in that the solvent is present in a quantity varying from 10 to 90% by weight.
6. The method according to claim 1, characterized in that the rotating magnetic field has an intensity ranging from 500 to 3000 Gauss, preferably from 1500 to 3000 Gauss.
7. The method according to claim 1, characterized in that the extraction is carried out at a temperature ranging from 30°C to 75°C, preferably from 35°C to 60°C.
8. The method according to claim 1, characterized in that the extraction is carried out for a time rang ing from 15 to 120 minutes, preferably from 30 to 60 minutes.
9. The method according to claim 1, characterized in that the extraction is carried out at a temperature ranging from 35°C to 60°C, for a time ranging from 30 to 60 minutes, with a rotating magnetic field which has an intensity varying from 1500 to 3000 Gauss.
10. Alcohol and/or hydroalcohol solutions which can be obtained with the method according to any of the previous claims, characterized in that they contain free molecular structures of sesquiterpenes, terpe nes, triterpenes.
11. The solutions according to claim 10, characterized in that they are dispersible in air within a tem erature range of 40°C to 90oC.
12. The solutions according to claim 11, characterized in that they are dispersible in air within a tem perature range of 80°C to 90°C.
13. The solutions according to claim 11 or 12, charac terized in that they are dispersible in air with the use of thermoemanators or electroemanators.
14. The solutions according to claim 10, characterized in that they are used in a mixture with each other, in a mixture with all types of essential oil, in any proportion, and/or in a mixture with water, up to a maximum of 25% of distilled water, whatever the pro portion of the solutions between each other may be.
15. The solutions according to any of the claims from 10 to 14, characterized in that they are solutions in ethanol and/or ethyl ethanoate, in any proportion.
16. The solutions according to any of the claims from 10 to 15, characterized in that they contain aero dispersible compounds, terpenes and/or terpenoids, with a molecular weight which varies within the range of MW 136 (monoterpenes) to MM 532 (pentacy clic triterpenes).
17. The solutions according to claim 10, characterized in that they contain fractions of terpenes and/or terpenoids extracted from incense in a percentage of between 15 and 65% by weight, fractions of terpenes and/or terpenoids extracted from myrrh in a percent age of between 15 and 65% by weight.
18. The solutions according to claim 17, characterized in that they contain Hyssopus officinalis decumbens or Hyssopus officinalis aristatu. green tangerine, fractions of terpenes and/or terpenoids extracted from myrrh and fractions of terpenes and/or terpe noids extracted from incense.
19. The solutions according to claim 18, characterized in that they contain Hyssopus officinalis decumbens or Hyssopus officinalis aristatus, green tangerine, fractions of terpenes and/or terpenoids extracted from myrrh and fractions of terpenes and/or terpe noids extracted from incense in proportions equal to 1: 0.3 : 5: 5.
20. The solutions according to claim 10, characterized in that they contain fractions of terpenes and/or terpenoids extracted from incense, myrrh and propo lis in a mixture with each other.
21. The solutions according to claim 10, characterized in that they are topically applied.
Description:
EXTRACTION METHOD OF ACTIVE MOLECULAR STRUCTURES FROM NATURAL RESINS AND/OR ESSENTIAL OILS The present invention relates to an extraction method of active molecular structures from natural resins and/or essential oils.
In particular, it relates to a method for the ex- traction of terpenes and/or terpenoids from natural res- ins and/or essential oils.
Natural resins, such as myrrh, incense, dacryodes, propolis, are substances which contain a high quantity of terpenes and terpenoids.
The types of terpenes contained in these natural resins are mainly : sesquiterpenes (C15 structure) diter- penes (C20 structure) and triterpenes (C30 structure).
In the above resins, together with these active sub- stances, there are also sugars starches and Rosinic res- ins (rosins) and the active substances are imprisoned in- side these polymeric components.
In particular, DSC tests effected with a Perkin-
Elmer Pyris apparatus on the above natural resins, in- cense, myrrh, dacryodes and propolis, have shown that these resins have a polymeric-type structure. The poly- meric structure is characterized by a Tg equal to 80°C for incense, a Tg equal to 60°C for myrrh and a Tg equal to 85°C for dacryodes.
The DSC plots show that incense has a structure of the amorphous type, with a melting point of 130°C, myrrh has a semi-crystalline structure with a melting point of 140°C, dacryodes has a semi-crystalline structure with a melting point of 250°C.
Among the three resins examined, only dacryodes has the capacity of crystallizing at a temperature of 180°C.
The dacryodes resin consequently behaves like a polymer with phase transition, i. e. an amorphous polymer which crystallizes when subjected to heat.
The traditional extraction method of the active sub- stances contained in these resins is extraction in a va- pour stream. The eatraction system with supercritical CO2 has also been recently introduced and used for these sub- stances. The extraction method with supercritical COs al- lows the extraction of a higher number of active sub- stances, with a greater yield, but creates problems of reaction between the substances themselves. For this rea- son, the most widely used method is still vapour stream
extraction.
Through vapour stream extraction, it is possible to extract the following mixtures of compounds, also called essential oils, contained in incense, myrrh, dacryodes and in propolis: A) INCENSE: the essential oils extracted in a vapour stream from incense prove to consist of: -ALPHA PINENE monoterpene ; - ALPHA THUJENE monoterpene ; - LIMONENE monoterpene; - P-CIMENE monoterpene; <BR> <BR> <BR> <BR> <BR> - BETA-MYRCENE monoterpene ; <BR> <BR> <BR> <BR> <BR> <BR> - GURJUNEN sesquiterpene ; - OLIBANOL ketonic alcohol with small quantities of other substances. The average extraction yield is equal to 1. 70%-2% by weight.
B) MYRRH : the essential oils of myrrh extracted in a va- pour stream prove to consist of: - FURANEUDESMA-ALPHA-1,3-DIENE sesquiterpene; <BR> <BR> <BR> <BR> <BR> - CURZERENE sesquiterpene ; <BR> <BR> <BR> <BR> <BR> <BR> <BR> - LINDESTRENE sesquiterpene ; - ELEMENE sesquiterpene; - COPAENE terpenoid; - 1-CADINOL sesquiterpenoid; - GERMACRENE sesquiterpene
with small quantities of other substances. The average extraction yield is equal to 2. 80%-3% by weight.
C) DACRYODES: extraction in a vapour stream did not allow an essential oil of the commercial type to be obtained.
D) PROPOLIS : - 2-methyl-3-butenol - 3-penten-2-ol -3-methyl-buten-1-ol - toluene - 2-methyl-buten-l-ol - ethyl-benzene <BR> <BR> <BR> <BR> <BR> - 1, 3o5o7-cyclooctatetraene <BR> <BR> <BR> <BR> <BR> <BR> -2-buten-1-ol-3-methyl-acetate<BR> <BR> <BR> <BR> <BR> <BR> - benzaldehyde <BR> <BR> <BR> <BR> <BR> <BR> - beta-myrcene - octanal Beta-myrcene is the only terpene extracted, with a total extraction of 10 compounds and an average extrac- tion yield of 6%.
Scientific literature (1998) indicates the following compositions for incense and myrrh : - Incense : 50-70% BOSWELLIC ACIDS ; 4-8% PINENE-PHELLANDRENE-TERPENE ALCOHOLS 20% RUBBER SUBSTANCES; POLYSACCHARIDES.
The active substances therefore form 73-75% by weight, whereas the inert substances form 28-30% by weight with respect to the total weight of the resin.
From what is specified above, it can be deduced that the major components of incense are Bosellic acids which are triterpenes.
- Myrrh : 30% DELTA ELEMENE ; 10% ALPHA COPAENE; 12% FURANEUDESMA-1, 3-DIENE ; 4% LINDESTRENE ; 12% ISOFURANGERMACRENE AND CURZERENONE ; 20% TRITERPENES ; 12% STARCHES AND VEGETABLE PROTEINS.
The active substances therefore form 40% by weight, whereas the inert substances form 60% by weight with re- spect to the total weight of the resin.
From what is specified above, it can be deduced that triterpenes represent a high quantity in myrrh, equal to 20% by weight of the resin.
From a simple comparison with the data indicated above relating to products extracted in a vapour stream from incense and myrrh, it can be observed that the triterpenes have not been extracted in incense, whereas the triterpenes and part of the sesquiterpenes have not been extracted in myrrh.
The objective of the present invention is therefore to find a new extraction method which allows the disad- vantages of the known art indicated above to be overcome.
The object of the present invention consequently re- lates to an extraction method of terpenes and/or terpe- noids from natural resins, such as myrrh, incense, dac- ryodes, dammar, propolis and/or essential oils, by means of extraction with polar and/or semi-polar solvents in the presence of a rotating magnetic field.
The polar and/or semi-polar solvent is preferably selected from ethanol and ethanol/ethyl ethanoate mix- tures.
Even more preferably, the polar and/or semi-polar solvent is ethanol, in particular pharmaceutical ethanol.
Pharmaceutical ethanol refers to extremely pure neu- tral ethanol 96% vol. with a maximum content of contami- nants equal to 0.058 mg/1.
The solvent is present in a quantity which varies from 10 to 90%.
The rotating magnetic field has an intensity varying from 600 to 3000 Gauss, preferably from 1500 to 3000 Gauss.
The extraction is effected at a temperature which ranges from 30°C to 75°C, preferably from 35°C to 60°C and is carried out for a time ranging from 15 to 120 min-
utes, preferably from 30 to 60 minutes.
An extraction method by means of extraction with ethanol at a temperature ranging from 35°C to 60°C, for a time varying from 30 to 60 minutes, with a rotating mag- netic field having an intensity ranging from 1500 to 3000 Gauss, is particularly preferred.
Very surprisingly in fact, the method according to the present invention allows all the active substances present in incense, myrrh, dacryodes and propolis, to be extracted, with the removal of the rubbery part, starches, sugars and vegetable proteins.
It allows the extraction of compounds having molecu- lar weights higher than 600 dalton.
Similarly, the method according to the present in- vention allows essential oils to be obtained, in which terpenes and/or terpenoids, flavonoids and steroids are present as free molecules. The essential oils, in fact, obtained by extraction in a vapour stream or with super- critical CO2 are not aerodispersible or at least only to a minimum degree.
A fundamental advantage of the extraction method ac- cording to the present invention is that it allows solu- tions to be obtained at different concentrations. By varying the ratio between time/temperature/magnetic field intensity it is in fact possible to cover the whole
range of extractions of all types of resins and/or essen- tial oils partially soluble in ethanol and ethyl etha- noate and mixtures of these two solvents in any propor- tion.
A further object of the present invention also re- lates to the alcohol and/or hydro-alcohol solutions ob- tained with the extraction method according to the pres- ent invention, containing free molecular structures of sesquiterpenes, terpenes, triterpenes, terpenoids, flavo- noids and/or steroids.
In particular, these solutions are dispersible in air within the temperature range of 40°C to 90°C, pref- erably with the use of thermo-emanators or electro- emanators. They are preferably perfectly dispersible in air within the temperature range of 80 to 90°C.
An object of the present invention also relates to the solutions obtained with the extraction method accord- ing to the present invention mixed with each other, mixed with all types of essential oil, in any proportion, and/or mixed with water, up to a maximum of 25% of dis- tilled water, whatever the proportion of the solutions with each other may be.
Solutions are preferred, obtained with the extrac- tion method according to the present invention, in a mix, comprising fractions of terpenes and/or terpenoids ex-
tracted from incense in a percentage ranging from 15 to 65% by weight, fractions of terpenes and/or terpenoids extracted from myrrh, in a percentage of between 15 and 60% by weight.
Solutions are also preferred obtained by means of the extraction method according to the present invention, which includes fractions of terpenes and/or terpenoids extracted from incense, myrrh and propolis in a mix.
In particular, solutions are also preferred contain- ing Hyssopus officinalis decumbens or Hyssopus officina- lis aristatus, green tangerine, fractions of terpenes and/or terpenoids extracted from myrrh and fractions of terpenes and/or terpenoids extracted from incense, in particular proportions equal to about 1: 0.3 : 5: 5.
The characteristics and advantages of the extraction method according to the present invention will be more evident from the following detailed and illustrative de- scription, referring to the following examples.
EXAMPLE 1.
A general description follows of the various phases.
Grinding of the resin: a) grinding of the resins to a particle size of 20 mi- crons using a mill with cross blades. The grinding chamber of the mill is maintained at a temperature
of about 15°C to prevent the formation of large- dimensioned agglomerations during the grinding proc- ess ; b) the ground product is stored in air-tight contain- ers; c) the grinding is effected under a suction hood to prevent the depositing of fine powder which, in the case of incense, could cause flammability phenomena in air (flashing).
Preparation of the solution to be subjected to the extraction process: The resins thus ground are mixed with pharmaceutical ethanol in a ratio of 10: 1 (ten parts of ethanol per one part of resin). The resins are then left to soak in the ethanol for 15 minutes in sealed glass container.
Extraction : The ethanol-resin solution is placed in a Pyrex glass flask with a flat bottom and positioned on a heat- able magnetic stirrer.
The flask is closed with a sealing glass plug and parafilm.
A magnetic anchor coated with Teflon is placed in the solution. The temperature is regulated at 50°C and the stirring rate is regulated at the maximum value (1, 500 rpm).
These operating conditions are maintained for 60 minutes.
The temperature is then brought to 23°C and the stirring rate is decreased (500 rpm).
These conditions are maintained for 15 minutes.
The solutions are left to rest for 30 minutes at room temperature.
The magnetic field is then induced by the rotation of a permanent magnet having a field intensity equal to 3000 Gauss.
Filtration: The solutions obtained are filtered using filter pa- per for qualitative analysis with a filtering capacity of 5 microns, in order to prevent the passage of any parti- cle remaining in solid form.
The solutions are left to rest for 15 minutes and subsequently filtered again using the same type of fil- ter.
Results : Limpid solutions with an amber-yellow colour are ob- tainede absolutely free of impurities and particulate.
Analysis of the solutions obtained Analysis of the alcohol solutions thus obtained was effected by means of Mass gas-chromatography (GC/MS and GC/FID) using a Hewlett-Packard HP 6890 instrument with
Supelco SBP-5 and Supelco-wax 10 columns.
Solution prepared starting from incense.
The extraction method according to the present in- vention starting from incense as natural resin produced a solution in which the following compounds were identi- fied: alpha pinene, alpha thujene, (+) limonene, p-cymene, beta-myrcene, 2-propyl-5-methylnaphthoquinone, furandiene, cembrene A (cembra-3,7, 11, 15-tetraone), l-isopropenyl-3-propenylcyclopentane, germacrene A, 2-N-butyl-8-N-hexyl-1,2,3,4-tetrahydronaphthalene, verticellol (Verticilia (20) 7,11-triene), aciphyllylic alcohol, pentacyclododecane, globulol lH-cyclopropen (e) azulenef 17-alpha-methyl-5-alpha-androstane, 1, 5-dimethoxy-2, 6-bis (prop-2'-enyl) anthraquinone, E-Ocimenone 2, 6-dimethyloctane, Trunculin-F methyl estere
4, 7-methanoisobenzofuran-1-ol 1,2, 3°4, 7, 7° hexahydro Eicosane, 4,8-decadienoic acid 2-acetyl 2,5, 9 trimethyl Longicyclene, 7-hydroxyoctanoic acid 2TMS, Aristolone 2H-cyclopropene-naphthalen-2-one, beta-boswellic acid, acetyl beta-boswellic acid, 11-keto-beta-boswellic acid, acetyl 11-keto-boswellic acid, trans-octahydro-5, 5-dimethyl-8a-tosyloxymethyl 2 11-H indene (1.2-B) quinoxalin-11-11H-indenol 3A-hydroxyolean-12-en-24 boswellic acid, Total active products present: 32.
Extraction yield: 56% out of a total of 72-75% of active substances.
Solution prepared starting from myrrh.
The extraction method according to the present in- vention starting from myrrh as natural resin produced a solution in which the following compounds were identi- fied: alpha-terpinene para-mentha 1,3-diene, beta-bourbonene, delta-elemene, trans-caryophyllene,
6-ethenyl-4,5, 6,7-tetrahydro 5-benzofuranacetic acid, 1-cyclohexene-1-carbonaldehyde, gamma-cardinene, germacrene B 2-ter-butyl-1, 4-naphthoquinone, (+) hepi-bicyclosesquiphellandrene, furaneudesma-1, 3-alpha diene, Ledene alcohol, alpha-longipinene, alpha-copaene, 1-ethyl-1, 2, 3,4-tetrahydro 2-naphthalenemethanol, benzylthio-6-methylluracin 2 (1H) pyrimidinone, 1, 3-dimethyl-5-azulenecarbaldehyde, dibenzofuranol-1, 4-dicarbaldehyde 1,3-naphthalenate, germacrene A, 2-isopropyl-4,7-dimethyl-1-naphthol, <BR> <BR> <BR> <BR> <BR> bicyclogermacrene,<BR> <BR> <BR> <BR> <BR> <BR> lindestreney<BR> <BR> <BR> <BR> <BR> <BR> 4-(phenylmethyl) phenol, amino-3 (1) imidazo (4,5H) quinoline, methyl 5-hydroxy-3-oxo-5-phenylpentanoate, 2-cyclohexyl-1-(lH-imidazol-2-yl) ethanone beta-amirine, (Urs-12-ene, 3-methoxy, 3 beta) beta-commiphoric acid.
Total active products present : 28.
Extraction yield: 35% out of a total of 40% of active substances.
Solution prepared starting from dacryodes.
The extraction method according to the present in- vention starting from dacryodes as natural resin produced a solution in which the following compounds were identi- fied: delta-elemene, alpha-copaene, beta-bourbonene, beta-elemene, trans-caryophillene, alpha-amorphene, beta-silenene, germacrene B, caryophyllene oxide, epicurzerenone, 1,4-dimethyl-7- (1-methylethyl)-azulen-2-ol, <BR> <BR> <BR> <BR> <BR> alpha-cadinolg<BR> <BR> <BR> <BR> <BR> <BR> furandiene,<BR> <BR> <BR> <BR> <BR> <BR> 2-isopropylquinoline-beta-D6, vulgarol A, phenoxazone, seychelene, 1-naphthalenemethanol alpha, alpha-dimethyl-5,
2,3 dihydroindole-4-ol-2-one, 2-pentyl-lH-quinolin-4-one, 1, 1, 4, 6, 7-pentamethyl-2, 3-dihydroindene, dibenzothiophene, germacrene A, 2-isopropyl-4, 7-dimethyl-l-naphthol, 4-quinolinol, 2 pentyl-N-oxide, methyl-3-methyl-2-thiophenecarboxylate, 2 (5H) furanne, 3 butyryl-5-hexyl-4-hydroxy-5-methyl du- vantriendiol, 1, 2-epoxynonane, (+) alpha-ciperone, 6, 6-dimethylundecane ltlldyl bis (2-hydroxy-5- (2-carot)- 1, 4-dienaldehyde.
Total active products present: 32 Extraction yield: 13%.
It is not possible to calculate a ratio between the extracted quantities and the total quantity of active substances as no composition for this resin has been cited in literature.
The triterpene compounds were identified not only with the GC/MS system but also by means of gas chromato- graph in liquid phase (HPLC) in UV light in a wave length of 210 nm.
EXAMPLE 2
A general description of the different phases is provided.
Grinding of the resin a) Propolis is in the physical form of very consistent waxy cakes. Grinding is effected in order to obtain an average diameter of the particles ranging from 80 to 100 microns, making use of a crossed blade mill.
The grinding chamber of the mill is maintained at a temperature of 15-18°C to prevent the formation of large dimensional clumps during the grinding. b) The ground product is stored in a hermetically sealed container. c) Grinding is effected under a fume hood in order to prevent deposition of fine powders which can give rise to flashing phenomena.
Preparation of the solution to be subjected to the ex- traction process : The propolis thus ground is mixed with pharmaceuti- cal ethanol in the ratio 7: 1 (seven parts of ethanol per one part of resin). The resin is left to soak in ethanol for 30 minutes in a sealed glass container.
Extraction: The ethanol-propolis solution is poured into a Pyrex glass flask with a flat bottom and placed on a heated magnetic stirrer. The flask is closed by means of a
sealing glass plug and parafilm. A magnetic, Teflon- lined anchor is placed in the solution. The tempera- ture is regulated at 40°C and the stirring rate at the maximum value (1, 500 rpm).
These operating conditions are maintained for 45 min- utes. The temperature is then lowered to 23°C and the stirring rate to 500 rpm. These conditions are maintained for 30 minutes at room temperature. The magnetic field is then induced by the rotation of a permanent magnet with an intensity of 3,000 Gauss.
Filtration The solution obtained is filtered by means of a paper filter for qualitative analysis having a filtering capac- ity of 5 microns, in order to prevent the passage of any small particles in solid form.
The solution is left to rest for 15 minutes and then filtered again.
The resulting filtrate is left to rest for a further 15 minutes and is then filtered again, still using the same type of filter.
Results : A limpid brown-coloured solution is obtained, abso- lutely free of impurities and particulate.
Solution prepared starting from propolis.
The extraction method according to the present inven-
tion, starting from propolis as natural resin, gave a so- lution in which the following compounds were identified by means of GC/MS and GC/FID analysis: guaiole; 10-epi-gamma-eudesmole; beta-eudesmole ; 2-propenoic acid; isopropyl tetradecanoate; alpha-eudesmole ; 8-beta-H-cedran-8-ole; ethyl ester of heptanoic acid ; n-pentacosane ; butyl ester of hexadecanoic acid ; n-tetracosane ; butyl stearate ; androstan-4-en-3-one; hexacosane; <BR> <BR> <BR> <BR> <BR> benzo [c] naphthol [2a 1-p] crysene ; <BR> <BR> <BR> <BR> <BR> <BR> dihydroxymethylcalcone ; colesteryl myristate ; Total active products present : 17.
Extraction yield: 80% out of a total of 85-90% of ac- tive substances.
A propolis solution is then obtained, by means of the method of the present invention, in which terpenes, ter-
penoids, flavonoids and steroids are present as free molecules.
In particular, the propolis solution analyzed above, refers to a particular type of propolis produced in Northern Italy, more precisely in the Asiago/Lavorano- Folgaria plateau. It is known that propolis has a compo- sition which varies according to the type of flora of the area where the beehives are positioned.
Miscibility of the solutions.
The solutions extracted prove to be miscible with each other in all proportions.
Miscibility with essential oils.
The solutions extracted prove to be miscible with all types of essential oils in any proportion with each other.
Miscibility with water.
The solutions extracted prove to be miscible with water up to a maximum of 25% of distilled water whatever the proportion of the solutions with each other.
Evaporation and aero-dispersion The solutions extracted are completely aero- dispersible within a temperature range of 40°C to 90°C with the use of thermo-emanators or electro-emanators as described in Italian patent Nr. 1287235.
Concentration
The extraction method according to the present in- vention also allows solutions to be obtained at different concentrations.
By varying, in fact, the ratio between time/temperature/magnetic field intensity, it is possible to cover the whole range of extractions of all types of resins and/or essential oils partially soluble in: - Ethano 1 (CH3CH20H) - Ethyl ethanoate (CH3CO2CH2) and mixtures of these two solvents in any proportion with each other.
This characteristic makes the extraction method ac- cording to the present invention extremely versatile and elastic and allows the extraction and preparation of al- cohol and/or hydro-alcohol solutions containing the ac- tive principles in different concentrations according to the end-use of the product.
Specific characteristics of the alcohol and/or hy- dro-alcohol solutions obtained with the extraction method according to the present invention are the following.
Contrary to normal essential oils extracted in a va- pour stream, or with supercritical C02 and normal alcohol and/or hydro-alcohol solutions obtained from natural res- ins, the solutions extracted from natural resins and/or essential oils, produced with the method according to the
present invention, allow not only monoterpenes and some sesquiterpenes, but also all (C15) sesquiterpenes, (C20) diterpenes and (C30) triterpenes, flavonoids and ster- oids, to be dispersible in air.
This is possible because the single solute molecular structures are free molecules. The magnetic field, in fact, releases the molecular structures of the terpenes and/or terpenoids from the polymeric part of the resins made up of starches, rubber based on isoprene and rosinic resin (rosin).
Analogously, in the case of essential oils, the pro- cess according to the present invention allows solutions of essential oils to be obtained, in which the single solute molecular structures, terpenes and/or terpenoids, are free molecules.
This was not possible so far with the known extrac- tion systems including systems using N2 and HzO in super- critical form.
The essential oils and mixtures of compounds ex- tracted from resins in a vapour stream or with super- critical gases are not made up of free molecules and when they are heated to temperatures exceeding 50°C, they in- teract forming new molecular structures, in particular compounds of the rubber type (isoprene, rosin) which cross-link forming a solid polymeric structure which is
therefore absolutely non-aero-dispersible.
The method according to the present invention forms an absolute novelty in the field of terpene and/or terpe- noid extraction from natural resins such as incense, myrrh, dacryodes, dammar, propolis.
The method according to the present invention very surprisingly also allows triterpenes such as boswellic acids and commiphoric acids, having the formula C30H4803, C3oH4805, C30H5003, to be aero-dispersed in a controlled quantity and time.
Aero-dispersion of the solutions.
The method according to the present invention very surprisingly allows solutions to be obtained, containing aero-dispersible compounds with a vast rang of molecular weights from MW 136 (monoterpenes) to MW 532 (pentacyclic triterpenes), and with a molecular weight higher than 600 dalton.
It is therefore possible to contemporaneously dis- perse in air terpenes and/or terpenoids with a different molecular weight, which varies within the range of HN 136 to MN 532.
This means that all terpenes and/or terpenoids con- tained in natural resins or in essential oils can be dis- persed in the air in a different time depending on the temperature.
In practice, the dispersion time at 50°C proves to be 240 minutes, whereas at 90°C it is 60 minutes.
In particular the following compositions are de- scribed, obtained by mixing the fractions of terpenes and/or terpenoids extracted by means of the method ac- cording to the present invention.
Composition nr. 1 A composition is described, obtained by mixing the following components in the amounts indicated with re- spect to 1 ml of composition. Incense and myrrh are the terpene and/or terpenoid fractions extracted from incense and myrrh according to the method of the present inven- tion, as described in example 1, whereas Hyssopus offici- nalis decumbens consists of the fractions extracted ac- cording to the method of the present invention from the essential oil Hyssopus officinalis decumbens.
Hyssopus officinalis decumbens 0. 0271 ml ; green tangerine 0.0075 ml ; myrrh 0.125 ml ; incense 0.125 mlg ethanol 0.5854 ml ; distilled water 0.130 ml.
Composition nr. 2 A composition is described, obtained by mixing the fol- lowing components in the amounts indicated with respect
to 1 ml of composition. Incense and myrrh are the terpene and/or terpenoid fractions extracted from incense and myrrh according to the method of the present invention, as described in example 1, whereas Hyssopus officinalis aristatus consists of the fractions extracted according to the method of the present invention from the essential oil Hyssopus officinalis aristatus.
Hyssopus officinalis aristatus 0.0271 ml; green tangerine 0.0075 ml; myrrh 0.125 ml; incense 0.125 ml; ethanol 0.5954 ml; distilled water 0.120 ml.
Composition nr. 3 A composition is described, obtained by mixing composi- tion nr. 1 and the fractions of terpenes and/or terpe- noids extracted from propolis as described in example 2.
The values are volume percentages.
Composition nr. 1 80% ; propolis 20%.
Composition nr. 4 A composition is described, obtained by mixing com- position nr. 2 with the fractions of terpenes and terpe- noids extracted from propolis as described in example 2.
The values are volume percentages.
Composition nr. 2 80% ; propolis 20%.
In particular, the compositions according to the present invention can be used as aero-dispersible solu- tions and as solutions for topic use.