FI81007C - Procedure for Obtaining a Substance with Anti-Inflammatory Effect in Matricaria chamomilla - Google Patents

Description

1 81007

A method for obtaining an agent having an anti-inflammatory effect from Matricaria chamomilla

The invention relates to a process for obtaining an agent having an anti-inflammatory effect, Matricaria chamomilla.

Medicinal and cosmetic products made from 10 mycorrhizae of Chamomilla recutita (L.) Rauschert, synonymous with Matricaria chamomilla L. have a wide range of uses due to their anti-inflammatory and antispasmodic effect. Of particular importance are the active ingredients (-) - α-bisabolol and camazulene. A substance made from a chamomile sauna should therefore contain as much of these two substances as possible.

A method has now been invented for preparing a new substance using a tetraploid chamomile sauna, which substance has advantageous properties.

It is an object of the present invention to provide ai-20 having an anti-inflammatory effect, in particular due to the high content of (-) - α-bisabolol and camatsulene, the content of camatsulene being at least 30 mg%, the content of (-) - α-bisabolol being at least 100 mg% and other bisaboloids less than 25 mg%. The term "other bisaboloids" means in particular (-) - α-bisabolol oxide A and B and (-) - α-bisabolone oxide A.

The method according to the invention is characterized in that the content of essential oil (-) - α-bisabolol is determined in each case from 1 000 to 10 000 individual plants of the diploid or tetraploid chamomile population and the chamomile plants in which the (-) - α-bis-abolol represents the major component of the essential oil, in the case of tetraploid starting chamomile, the flowers dried at 40 ° C 35 contain at least 100 mg% of camatsulene, at least 200 mg% of (-) - α-bisabolol and less than 2 81007% than 50 mg% other bisaboloids, and in the case of diploid starting coma, flowers dried at 40 ° C contain at least 100 mg% of camazulene, at least 50-100 mg% of (-) - α-bisabolol and less than 50 mg% of other 5 bisaboloids , and provided that the diploid parent chamomile does not have the chamomile variety Degumill, the chamomile isolated from the initial population is further grown and at least 10 individuals are selected from these plants. with a simultaneous flowering period, with a mycelium size of 20 to 40 10 mm and an active substance content of at least 100 mg% of camazulene, at least 200 mg% of bisabolol and less than 50 mg% of other bisaboloids (all calculated on the dry matter) , possibly these selected diploid or tetraploid plants are finally cloned, 15 and (a) seeds are obtained from the plants thus obtained, (b) progeny of the seeds obtained in (a) are grown, finally the active substance concentration and the simultaneous flowering period, regular branching from 20 roots and 20 to 40 mm, as described above, the plants thus selected may be cloned and seeded, (c) the procedure in (b) shall be repeated 3 to 5 times, (d) the progeny of the seed obtained in (c) shall be grown, the progeny being selected in accordance with point (a), any plants so selected may be cloned and, in the case of diploid chamomile, these are finally tetraploidized by chemicals in the temperature range from 0 to 35 ° C, by gamma rays, X-rays or UV radiation at 0 to 35 ° C, using higher temperatures of 30 to 50 ° C, lower temperatures of 0 to 5 ° C or a decapitation tilting method or a ponsivil-35 scheme, which is finally tetraploidised and the plants thus obtained are harvested flowers which are in a growth stage of 3 81 007 h, where 30 - 100% of the mycelium of the mycera has been opened and dried at a temperature of not more than 70 ° C to obtain a dry material containing not less than 30 mg% of camazulene, not less than 100 mg of (-) - α-bisabolo-5 and less than 50 mg-% of other bisabolites; and possibly from the dry material thus obtained by distilling in the presence of water an essential oil containing not less than 3,5% of camazulene, not less than 10% of (-) - α-bisabolol and less than 10% of other bisabolols, or by extraction with lower alcohols an alcoholic chamomile sauna extract containing at least 1.5 mg% of camazulene, at least 5.0 mg% of (-) - α-bisabolol and less than 10 mg% of other bisabolites.

The production method of the present invention uses a tetraploid chamomile sauna in which the main component of the essential oil is (-) - α-bisabolol.

From these chamomile saunas, plants are selected whose flowers, dried at 40 ° C, contain at least 100 mg of camazulene, at least 200 mg of (-) - α-bisabolol and at most 50 mg of other bisaboloids (at the time of flower harvest, where 30-70% of the hornbeam flowers of Mycorrhea bloom), and this is accompanied by other selection and growth stages (strain growth for vegetatively propagated cross-25 fertilizer).

Examples of suitable tetraploid chamomile sauna plants are the chamomile sauna varieties Bodagols (German Democratic Republic), Pohorelicky (Czechoslovakia), Zloty Lan (Poland) and BK-2 (Hungary).

30 These varieties have been described in the literature in the following publications: M. Chlädek and V. Kosova, Pharmazie 13, (1958), p. 712 -713; W. Czabajska, Diss. Poznan (1963); 35 W. Poethke and P. Bulin, Pharm. ZHalle 108 (1969), p. 813-823; and I. Särkäny, Herb. Hungar. 4 (1) (1965), ss. 125 - 169.

4 81007

Otherwise, tetraploid chamomile sauce parent plants are considered to be equivalent in terms of (-) - α-bisabolol content to diploid chamomile sauna parent plants (see below).

The starting plant of a tetraploid chamomile sauna can also be obtained from diploid chamomile saunas, in which (-) - α-bisabolol represents the main component of the essential oil. Such diploid chamomile saunas are tetraploidized (gene mutation), the tetraploid plants thus obtained are selected, and from these selected tetraploid plants the plants with a minimum content of 100 mg of catamazulene in flowers dried at 40 ° C are again selected, (-) - a-b is 200 mg-% and the amount of other bisaboloids is at most 50 mg-% (flowers 15 are harvested at the time 30-70% of the mycorrhizal horn flowers are opened), and possibly other selection and growth steps are associated with the examples.

The starting plants for diploid chamomile saunas are obtained, for example, from the following chamomile saunas: 20 "Chamomile Argentinischer Provenienz" [see L. Z. Padula, R. V. D. Rondina and J. D. Coussio; Quantitative Determination of Essential Oil, Total Azulenes and Chamazulene in German Chamomille, Matricatia chamomilla, Cultivated in Argentina; Planta med. 30 (1976), ss. 273-280]; 25 as well as all chamomile saunas with clearly high concentrations of (-) - α-bis-abolol in their essential oils (usually more than 5% of the essential oil). Examples include diploid chamomile saunas described in the following references: H. Schilder; "Neuere Erkenntnisse bei der Qualitätsber-teilung von Kamilleöl bezieungsweise KamilleblUten", Planta med. 23 (1973), ss. 132 - 144; 0. Motl, M. Felklovä, V. Lukes and M. Jasicovä; "For 35 gas chromatographic analysis and chemical nitrogen of chamomile oil", Arch. Pharm. 310 (1977), ss. 210-215; and li 5 81007

Ch. Franz, J. Hölzl and Vömel; "Preliminary Morphological and Chemical Characterization of Some Populations and Varieties of Matricaria chamomilla L.", Acta Hort. 73 (1978), p. 109 - 114.

5 The use of the chamomile variety DEGUMILL as a starting plant also gives, for example, a tetraploid chamomile glass union with a camatsulene content of at least 150 mg% and a bisabolol content of at least 200 mg%, in case the chamomile flowers are corrected at a growth stage of only 30-70% m The Tor vices are opened and the drying is carried out at an air temperature of at most 50 ° C (DE patent 2,402,802).

In accordance with this publication, the 15 selection and growth steps subsequently added can be used in this context to further increase the mentioned bisabolol and azulene values and ·

The essential oil of chamomile sauna usually consists of the main components, farnesene, spatulenol, 20 camazulene and one bisaboloid among four bisaboloids [(-) - α-bisabolol, bisabolol oxide A, bisabolol oxide B or bisabolone oxide, in the case of a single isis; mixed samples as well as tetraploid individuals may contain more than 25 bisabolites in parallel] as well as spiroethers. Said substances make up 70-80% of the essential oil at best. Therefore, diploid chamomile saunas are particularly suitable, in which (-) - α-bisabolol forms the main component (more than half of the sum of the above-mentioned substances). Suitable starting plants for the chamomile sauna are, in particular, diploid chamomile stands, in which, for example, (-) - α-bisabolol constitutes at least 40% of the essential oil or at least 90% of the bis-abolooids.

6 81007

Particularly suitable are diploid or tetraploid chamomile populations, which usually contain at least 100 mg% of camazulene and 50-100 mg-% of (-) - α-bisabolol 5 in the solids samples, based on the dry matter of flowers dried at 40 ° C. ); this means that from such populations of chamomile sauna in the case of the starting plant of diploid chamomile sauna, seeds are obtained and these seeds are tetraploidized (disregarding and / or not knowing the bisabolol content of the host plants). In the case of the starting plant of a tetraploid chamomile sauna, the seeds described above are used immediately for the subsequent selection and growth stages.

Suitable diploid and tetraploid starting plants for chamomile-15 saunas are selected through a study of individual plants. In general, it is necessary to study, for example, 1,000 to 10,000 individuals from a single parent population in order to find a few individuals with a high (-) - α-20 bisabolol content according to the above criteria and suitable as starting plants for chamomile saunas. Seeds are then obtained from such selected individuals in a conventional manner and these are tetraploidized. After completion of the tetraploidization, the tetraploid plants are selected from other diploid plants. From the tetraploid plants thus obtained, possibly after prior addition, all plants are now selected whose flowers, dried at 40 ° C, contain at least 100 mg% of camazulene, at least 200 mg% of (-) - α-bisabolol and less than 50 mg of % 30 other bisabolites. Harvesting of these flowers is carried out at the developmental stage, when 30 to 70% of all mycorrhizal horn flowers have opened. Possibly, this can be accompanied by other selection and growth stages that improve, for example, the following characteristics and characteristics: concurrent flowering season; even, branching from the root, and a narrow flowering zone (this means better li 7 81007 suitable for mechanical harvesting), large mycorrhizae, better preservation of the herb, especially the fragrant aroma.

In the case of starting from 5 tetraploid chamomile saunas from the beginning, only plants with at least 100 mg% of camazulene, at least 200 mg% of (-) - α-bisabolol and less than 50 mg-% of other bisabolites, in which case 10 of these flowers are also harvested at the stage of flower development, in which 30-70% of the mycorrhizal horn flowers have opened. This may be accompanied by other selection and growth phases that result in other improvements (as mentioned above).

It is particularly preferred that such plants (known or prepared according to the method) are selected on the basis of the content of camatsulene and (-) - α-bisabolol. with a content of at least 200 mg of camazulene, predominantly 250 mg, and with a content of (-) - α-bisabolol-20 of not less than 300 mg, predominantly 400 mg (with a content of other bisabolols of 50 mg below) .

Tetraploids are obtained by treating plant parts (seeds, roots, shoot shoots, embryos and abrasive buds) or the starting plants of diploid chamomile saunas with chemicals, X-rays, gamma rays or UV rays. This can also be done by preparing pony cultures and subsequent tetraploids, or by using high or low temperatures on chamomile sauna plants or plant parts or chamomile sauna tissues.

8 81007

Manufacture of tetraploids by chemicals.

Suitable chemicals for the preparation of tetraploids are, for example, colchicine, acenaphthene, alkaloids, such as atropine, veratrine, nicotine, sanguinarine, benzene, diphenyl and phenanthrene derivatives, naphthalene and naphthalene derivatives, diphenylamine, diphenylamine, tribromenamine, , methylnaphthoquinone, methylnaphthohydroquinone, salicylic acid and related compounds, hexachlorhexane, methamphetamine (hydrochloride), alkyl alkali carbamate such as isopropyl sodium carbamate, phenylurethane, cononeine, cacodyl acid salts, e.g. sodium salt, e.g. heterouxin, germisane (phenylmercuric brentscatechin), organic mercury compounds such as ethylmercuric phosphate, ethylmercuric chloride, phenylmercuric hydroxide, phenylmercuric dinaphthylmethane disoforms, and Oilcakes, compost and cow dung are still in question.

The treatment with the substances mentioned here takes place, for example, at temperatures of 0 to 35 ° C, in particular at 12 to 30 ° C, in particular at 15 to 25 ° C.

The application is carried out, for example, by treating seeds, shoots, roots (especially root tips or roots of cotyledons), fetuses, leaf or stem cross-sections, cell suspensions of wound cell cultures or wound cork cultures, or by spraying on the base of the stem or in the area of abrasive buds. Chemicals are generally used in the form of 30 aqueous solutions, low-alcohol (usually less than 5% alcohol) or weakly acidic solutions.

It 9 81007 ten format. The pH of weakly acidic solutions is, for example, 5.5-6.5, in which case the acidification takes place by means of lower, organic, aliphatic acids, such as acetic acid. In the case of alcoholic solutions, they can also be weakly acidic. The concentrations of the chemicals in these solutions can be, for example, 0.01-0.5%, in particular 0.02-0.2%, in particular 0.05-0.1%. Substances in gaseous form are used as such, possibly under pressure (e.g. 100-1000 kPa). The processing time is, for example, 1-36, mainly 2-12, in particular 4-6 hours.

The most effective concentrations, as well as the exposure time, would be expedient to be examined in each case by means of preliminary experiments.

Particularly preferred is, for example, treatment with colchicine at 0-35 ° C, in particular at 12-30 ° C, in particular at 15-25 ° C. This can be done, for example, by allowing the seeds of the starting plants of the diploid chamomile saunas to swell in 0.01 to 0.2%, in particular 0.02 to 1%, mainly 0.05% colchicine. - in solution or that the germinated, well-developed embryos of the parent plant of the diploid chamomile sauna (5 to 7 days old) (embryo-leaves downwards) are immersed in 0,01 to 0,2%, in particular 0,02 to 0,1%, primarily to a 0.5% solution of colchicine-25. In the latter method, the relative humidity of the ambient air should be 100%. The length of the colchicine treatment is, for example, 3 to 36 hours, especially 4 to 10 hours. When embryos are used, an exposure time of up to 10 hours is usually sufficient. When seeds are used, the spawning time may possibly be extended to 36 hours.

After chemical treatment, swollen seeds, embryos or other parts of the plant are rinsed several times with water. For example, swollen seeds are sown. Plants with treated roots or other plant parts, for example, are replanted in plant boxes. The plants are grown from seeds or cotyledons treated in this way (e.g. in a greenhouse; temperature 18-25 ° C during the day and 10-16 ° C at night), and plants with pollen about 1 1/2 times higher than the pollen of the starting material or somatic cells are selected. the chromosome number is 36. In the case of chemical treatment of other parts of the plant (above-ground or underground), tumors, roots or flowers / seeds originating exclusively from the treated parts are examined for 10 subsequently formed tetraploids. When the shoot or leaf blight treatment is carried out, only the new plant resulting from this leaf blight or this shoot and the flowers or seeds formed in it shall be examined for chromosome number.

Measurement of pollen size and counting of chromosomes can be performed, for example, as described in Example 3.

Manufacture of tetraploids by irradiation.

The effect is applied, for example, to the seeds or the root tips at a temperature of 0-35 ° C, in particular 10-30 ° C, in particular 15-25 ° C. Amount of radiation: 5-50 Krad. These are mainly gamma and X-ray rays. Examples of UV rays are rays with a wavelength of 400-30 nm, mainly 350 nm.

25 Plants or parts of plants so irradiated are then treated in the same way as in the case of chemical treatment.

Use of high and low temperatures.

Suitable high temperatures are, for example, temperatures of 33-50 ° C, in particular 42-45 ° C. For example, swollen seeds, cotyledons, tumor canopies, and growth cell tissue are placed at these temperatures. The duration of the treatment is, for example, 1-48, preferably 12-24 hours.

35 Examples of suitable low temperatures are 0-5 ° C, in particular 0.5-4 ° C, in particular 2 ° C. these

II

11 81007 temperatures include swollen seeds, cotyledons, cotyledons and growth cell tissue. The duration of the treatment is, for example, 1-100, mainly 20-40 days. The plants or parts of plants thus treated are further treated in the same way as in the case of chemical treatment.

Hetene pony culture (inbreeding of plants with a simple chromosome sequence by hetene pony followed by the formation of tetraploids by chemicals 10 and irradiation).

Closed horn flowers are collected from the flowering plants, the stamens of which are in the stage before the first pollen mitosis. Stool strains are taken from the flower buds using a micro-manipulator and transferred to Petri-15 dishes filled with Nitsch and Nitsch medium (Table 1). Petri dishes are then stored in a culture room with 16-hour days at 28 ° C during the day and 20 ° C at night. After about 4 weeks, the petioles begin to open and the plants grow. In the case of di-20 ploid plants, these are haploid and in the case of tetraploid plants these are dihaploid; after the roots have formed, they are transferred to the garden, for example, and made to bloom in a greenhouse. These (di) haploid plants are sterile, however, polyploid plants can be formed by treating the shoots, roots and stems with chemicals such as colchicine to form homozygous plants which can then be propagated by seed. See further treatment for chemical treatment (eg colchicine treatment).

12 81 007

table 1

Nitsch and Nitsch medium (Science, 1969) mg / l KN03 950 NH-NO-3 720

MgSO 4 x 7 H 2 O 185

CaCl2 166 KH2PO4 68

MnSO 4 x 4 H 2 O 25 h 3bo3 10

ZnSO 4 x 7 H 2 O 10

Na2Mo04 x 2 Η2 <3 0.25

CuSO 4 x 5 H 2 O 0.025 5 ml of a solution of 7.45 g of the disodium salt of ethylenediaminetetraacetic acid and 5.57 g of FeSO 4 x 7 H 2 O filled to 1000 ml with myo-inositol 100

Glycine 2

Nicotinic acid 5

Pyridoxine HCl 0.5

Thiamine HCl 0.5

Folic acid 0.5

Biotin 0.05

Sucrose 20 g

Finished medium (DIFCO-Bacto-agar) 8 g

Indoleacetic acid 0.1.

The pH of the medium was adjusted to 5.5 by 13 81 007

From the chamomile sauna plants selected by calculating the pollen size and / or chromosome number, which can be obtained according to the possibilities described above, those plants are then selected with a minimum content of 100 mg camatsulene 5 and a minimum bisabolol content of 200 mg, while other bisabolols (especially bisabolol oxides) should be below 50 mg% (for dried flowers, see Example 1).

10 These plants, thus selected, after harvesting all the mycelium which has already erupted in the flower, are allowed to stop flowering in a greenhouse at a daytime temperature of 18 to 24 ° C and a night temperature of 12 to 14 ° C for at least 14 hours, during which time four during a week 15 all mycelium that has withered or is just about to crumble is collected for seed intake. After drying at an air temperature of 20-35 ° C, for example, a seed crop of such a chamomile sauna is obtained.

20 The following may still be used as selection criteria: (a) approximately simultaneous flowering; b) a flat, branching branch and a narrow flowering zone, about 10, in particular 5 cm; (C) large mycelium with an outside diameter of about 30 mm (20 to 40 mm), in particular 25 to 35 mm.

The use of additional criteria a), b) and / or c) will lead to, for example, higher flower and herb yields and better suitability for mechanical harvesting.

In the case of already known tetraploid chamomile saunas as starting plants, the selection described above, as well as possibly other selection and growth steps, takes place in the same way.

In order to obtain a chamomile sauna which, except that it contains at least 100 mg% of camatsulene and at least 14 81 007 200 mg% (-) - Oi-bisabolol (for dried mycelium ^ still gives a high flower yield and is better suited for mechanical harvesting, the following procedure is recommended : selected plants (as described above) are vegetatively propagated from cuttings and selected over 3-5 generations, the selection always being made according to the above criterion or possibly additional criteria a) -c). The plants selected on the basis of these criteria are cloned, the cloned plants yield 10 seeds, the plants obtained from these are again selected on the basis of the minimum concentrations of camatsulene and bisabolol given above and possibly additional criteria a) to c), and the latter are again harvested.

Period, sowing choice, in accordance with the above minimum content of 100 mg% of camatsulene 15 and 200 mg% of (-) - Qi-bisbolol (less than 50 mg of other bisabolites) as well as any additional criteria (a) to (c), - seed yield intake, can be repeated 3-5 times.

This is once again accompanied by a cycle, sowing selection, 20 as well as above (possibly cloning) - seed yield.

The last seed yield obtained is then the seed used to increase the chamomile growth.

Propagation by cuttings is carried out as follows: For propagation by cuttings, the parent plants (clonal parent plants) must form dwarf shoots without flower buds under short-day conditions.

This takes place in a greenhouse in the winter half of the year, without additional lighting or in an air-conditioned chamber, with a day-30 length of 6-10, mainly 8 hours and a temperature of 10-15 ° C, mainly 12 ° C. Leaf, tumor and special dwarf cuttings (side-shoots) suitable for cloning or propagation. They take root at 12-18 ° C, mainly at 15 ° C and with a day length of 12-16, at the front-35 end for 14 hours in a dense atmosphere (about 100%

II

15 81 007 humidity). For example, a 1: 1 peat-sand mixture can be used as a base; but pure quartz sand, rock wool plug substrate, peat plug substrate, etc. are also suitable as such.

5 For sowing, the following substrates are suitable for sowing: garden soil, humus-rich, medium-heavy clay soil, clayey or humus-rich sandy soil.

Sowing can be done in a greenhouse or also in the open. The germination and growth of the plants take place, for example, at a temperature of 12 to 24 ° C, in particular 18 to 20 ° C. In the case of sowing in the open, sowing is mainly in autumn (September / October) or spring (March-month / April). These deadlines refer to all cultivation areas in question (e.g., northern hemisphere-15 koa, temperate to subtropical climates).

It is particularly advantageous if the following procedural conditions are available:

Tetraploid chamomile sauna plants with a camatsulene content of at least 100 mg and a minimum content of (-) - Oi-bi-20 sabolol of 200 mg, while other bisaboloids (especially bisabolol oxides) are less than 50 mg (all values for dried mycorrhizae) only those which: - bloom approximately simultaneously, 25 - are evenly branched, from the root and have a narrow flowering zone, about 10 cm in diameter, and - large mycorrhiza, about 30 mm in diameter.

Selected plants are propagated vegetatively by cuttings and selected over 3-5 generations, with 30 selections always made according to the above criteria. Selected plants are propagated vegetatively (cloned), allowed to wither together, and seeds are taken from them. The plants obtained from the seeds are again selected according to the above criteria and the period, 35 sowing-selection-seed intake-sowing is repeated 3-5 times.

16 81 007

The chamomile sauna used in the method according to the invention is a crop quality of an authentic chamomile sauna, the botanical name of which is Matricaria chamomilla L. (synonymous with Chamomilla 5 recutita (L.), Rauschert). The material, dried at a temperature not exceeding 70 ° C, obtained from the flowers of this chamomile sauna, harvested at the stage of growth in which 30 to 100% of the mycelium horns have opened, contains at least 30 mg% of camatsulene, at least 10 100 mg% ( -) - Qi-bisabolol and less than 50 mg-% of other bisaboloids.

In the event that the chamomile sauna flowers are harvested at a time when the flower development phase is still continuing, this means that, for example, 100%, or up to 100%, of all mycorrhizal horn flowers have opened (e.g. full flowering stage) and / or that the harvested flowers are dried at 40 ° C, the concentration of the active ingredients (-) - Ct-bisabolol and camatsulene may be lower, because at higher temperatures and / or later during harvesting, azulene and bisabolols may decompose more strongly.

For example, chamomile sauna flowers (assembled as described above) dried in an oven at 40 ° C contain 100-200 mg% camazulene, 200-450 mg% (-) - CC bisabolol and only a small amount, i.e. -50 mg-%, other bisabolites in terms of dry matter (calculated on the absolute dry weight of the flowers). This absolute dry weight is determined by drying a separate sample of kamo-30 millasaunion flowers in an oven at 105 ° C to constant weight (72-96 hours). For example, the active ingredient content of flowers dried at 35-50 ° C is then calculated at 105 ° C relative to the declared dry matter.

A particularly active substance is obtained (having the highest possible content of the active substances camatsulene and (-) - Ct-17 81 007 bisabolol) when the mycelium of the chamomile sauna is collected in a growth phase in which, for example, 30-70%, mainly 40-60% of mycorrhizal horn flowers, that a total of 50% is opened and the drying is carried out at an air temperature of at most 50 ° C, for example 35-50 ° C, in particular 50 ° C. For example, flowers dried in an oven at 40 ° C (collected when 40-60% of the mycorrhizal horn flowers have opened) contain at least 100 mg%, for example 100-150 mg% of camazulene, at least 300 mg%, for example 200-300 mg -% (-) - O, -bisabolol and only a small amount (i.e. 5-50 mg-% of bisaboloids in the dry matter (calculated on the absolute dry weight of the flowers) This absolute dry weight is determined by drying a separate sample of chamomile sauna flowers in an oven at 105 ° C to a constant weight of 25 (72-96 hours) For example, the active ingredient content of flowers dried at 35-50 ° C is then calculated relative to the dry matter of the flowers at 105 ° C.

The drying of the chamomile sauna flowers can take place either by means of a mechanical air supply or also by drying in a var-20, possibly also in the sun, in which case it must be noted, however, that the heat supply does not exceed the amount required for complete drying. It is also preferred that the control weighing ensures that the constant weight is reached. The drying can be carried out independently or artificially (for example by means of artificially heated air). The yield of active ingredient is highest in the absence of sunlight, mainly at 40-60 ° C, especially at 40-50 ° C. The drying phase should take place as soon as possible or as soon as possible after harvest. Drying 30 should be performed in thin layers, for example 5-20 cm, mainly 10 cm thick. Drying is also possible in well-ventilated halls with an air temperature of 20-30 ° C. In general, the air temperature for drying should not be higher than 6 ° C. Preferred 35 is an air temperature of 35-50 ° C.

18 81 007

The concentration of the active ingredient of the substance according to the invention in relation to the main active ingredients camatsulene and (-) - OC bisabolol depends on the growth stage in which the flowers of the chamomile sauna are harvested and the drying of the harvested flowers. The higher the drying temperature, the more strongly the active substance decomposes; it means that the lower the active ingredient content of the dried flowers. It is also based on the fact that direct sunlight has a detrimental effect on drying and should be avoided as far as possible.

The growth stage of a flower is characterized by the percentage of mycorrhizal horn flowers that have opened at a given time (in this context, at the time of harvest).

15 It is also possible to harvest flowers in which, for example, 30-50%, 30-70%, 40-60%, 60-100% or 90-100% of the mycorrhizal horn flowers have opened; the concentration of the active substance depends on the stage of the flowers in each case, in the chamomile saunas used according to the present invention it is at a maximum when 40-60% of all horn flowers have opened and is lower in both less and more opened mycorrhizal horn flowers.

The great advantage of the described chamomile is that it withers the individual plants to wither at the same time, i.e. that the majority of the plants formed from a given sowing have the same flowering stage; it means that in incomparably most plants, 40 to 60% of all mycorrhizal horn flowers are opened. Complete processing of mycorrhizae at the optimal time is thus possible. Kamo-30 millasaunio, which is used for the preparation of the substance according to the invention, is therefore particularly suitable for mechanical harvesting.

The relationship between the concentrations of camatsulene and (-) - Oi bisabolol, on the one hand, at the time of harvest, on the other hand, and at the drying temperature used to obtain the substance, is shown in the following table (other bisaboloids are less than 50% in all cases).

Il 19 O 1 Π O 7 u I uu /

Drying- Flower growth stage at the time of repair | ämp ^ ila 30-70% of mycera 70-100% of all mycorrhiza, „....,> Of all hornbills hornbills have opened- 1α" Ρ * ιΐ3) has opened nut

Concentration of active substance - Concentration of active substance in the dried matter: oral content:

Not higher Carazulene: Camatsulene: than 50¾ Not less than 100 mg-% Not less than 40 mg-% (-) - Qf-bisabolol: (-) - Φ-bisabolol: Not less than 200 mg-% Not less than 120 mg-%

Active substance Concentration of active substance in alcoholic extract: extract:

Not higher Camatsulene: Camatsulene: than 50¾ Not less than 5.0 mg-% Not less than 2.0 mg-% (-) - ^ - bisabolol: (-) - Qt-bisabolol: Not less than 10.0 mg-% Not less than 6.0 mg -%

Concentration of active substance in the dried substance: content in the dried substance: Between Camazulene: Camatsulene: 50-70 ° C Not less than 50 mg-% Not less than 30 mg-% (-) - ^ - bisabolol: (-) - Ct -bisabolol: Not less than 150 mg% Not less than 100 mg

Concentration of active substance in the alcoholic extract: concentration in the alcoholic extract: Between Camazulene: Camazulene: 50-7QOC Not less than 2,5 mg-% Not less than 1,5 mg-% of (-) - β-bisabolol: (-) - Cl-bisabolol: Not less than 7,5 mg% Not less than 10 mg% It follows that a canine sauna used for the production of a substance, for example in a collection which takes place during the growth phase, in which 30 to 100% of the horn flowers have opened and where drying can take place at air temperature up to 70 ° C, a dried substance is obtained, the camatsulene content of which in any case rises to at least 30 mg% and the (-) - Oo-bisabolol content is at least 35 100 mg%, while the other bisaboloids are always less than 50 mg%.

20 8 1 007 The content of other bisaboloids in these alcoholic extracts is always less than 10 mg%.

Essential oils prepared from dried material in accordance with the condition-5 ratios given in the table above always have a content of at least 3.5% of camatsulene, at least 10% of (-) - O-bisabolol and less than 10% of other bisabo -loideja.

The chamomile sauna used in the method of the present invention has a moderately late harvest time, a uniform growth height with narrow flower zones and a large mycelium, and is therefore particularly suitable for mechanical harvesting. In addition to this, the plants of the new tetraploid chamomile sauce sown at the same time generally practically wither together at the same time, so that harvesting also becomes very simple and easier. In addition, these new chamomile saunas give a great harvest. The bisabolol and camazulene-containing substances prepared from this chamomile sauna, according to the present invention, which have an anti-inflammatory effect, are used, for example, in products according to diet theory, in cosmetics and in the form of tea preparations. Thus, it can be in question; from the dried material 25 obtained according to the invention, for example, by chamomile sauna baths or chamomile sauna extracts with extraction or extraction with supercritical gases with alcohols or aqueous alcohol mixtures. Fresh or frozen (in the case of fresh flowers frozen) flowers of a chamomile sauna can also be used to prepare extracts, especially alcoholic extracts.

In addition, chamomile oil, (-) - Λ-bisabolol, camazulene and other chamomile-containing substances can be obtained from the dried material.

Thus, the alcoholic extracts of the dried material mentioned in claim 1 contain at least 1.5 mg% of predominantly 5.0 mg% of camazulene and at least 5.0 mg% of predominantly at least 10.0 mg% of (-) - ^ - bisabolol in an alcohol-5-containing herbal extract and other bisaboloids less than 8 mg-%.

Such extracts are prepared in a manner conventional for the preparation of extracts.

For extraction, for example, mixing devices can be used, for example so-called pool mixing devices, percolators and other suitable extraction devices. During the extraction, the temperature rises to, for example, 10-50 ° C. Cooling is not necessary.

Suitable solvents are, in particular, straight-chain or branched, monohydric or polyhydric alcohols having 1 to 6 carbon atoms, such as, for example, methanol, ethanol, propanol (2), butanol, glycerin, isopropylidene-Gly-serol and the like, such as mixtures of these solvents with water.

Mixtures of these solvents can also be used.

The minimum amount of solvent is 2 parts of solvent per part of dried substance. Generally, 2 to 20 parts of solvent per part of dried substance are used, especially 3 to 10 parts of solvent per part of dried substance.

The essential oil obtained from the dried material prepared according to the invention contains at least 3.5%, in particular at least 5% of camazulene and at least 10%, in particular at least 15% of (-) - Cfc bisabolol and less than 10% of other bisabolites.

The essential oil is generally prepared by heating the dried substance to boiling point with water, for example in the presence of ascorbic acid (e.g. as a salt, especially the sodium salt), at a pH of 4-6, predominantly 22 81 007 5-5.5. The pH is adjusted with an acid, for example hydrochloric acid.

For example, 10 to 50 parts by weight of water and possibly 0.1 to 5 parts by weight of ascorbic acid are used per part by weight of dried substance.

Usually heated for 2-8 hours. The resulting aqueous distillate is shaken several times with a lower aliphatic liquid hydrocarbon (e.g. petroleum ether (e.g. b.p. 35-60 ° C), pentane, xylene, 1Q decalin), the organic fraction is dried (e.g. with sodium sulfate) and the organic fraction is carefully removed. (for example by distillation on a rotary evaporator or by distillation at 40-70 ° C, preferably at 50-60 ° C.) In the case of a high-boiling solvent, the distillation is carried out in vacuo.

The determination of camatsulene is performed spectrophotometrically, in a manner conventional for chamomile extracts.

The determination of (_) - Λ-bisabolol and other substances contained in chamomile oil is carried out by conventional gas chromatographic methods.

Annex A contains a detailed description of the analytical methods.

The active substance camatsulene is not present in the flowers of the chamomile sauna as such, but in the form of a matrix-25 of sesquiterpene lactone. Matrix has a pharmacological effect similar to that of camatsulene. For example, this matrix precursor produces camazulene immediately upon heating (e.g. steam distillation, tea extraction). Therefore, it has become customary to report the concentration of camatsulene formed from the matrix and not the concentration of the matrix in the chamomile sauna.

If the flowers of the described tetraploid chamomile sauna are collected in a growth phase in which 30-100% of the mycorrhizal horn flowers have opened and the flowers are dried at air temperature up to 70 ° C, the substance obtained contains, for example, 23 23 007 mg-% (_) _ Oi-bisabolol and less than 50 mg-% of other bisa-boloids, and the essential oil prepared therefrom by steam distillation contains at least 3.5% camazulene, 5 at least 10% (-) - £ Z / -bisabolol, and less than 10% of other bisaboloids, and the alcoholic extract obtained by extraction with lower alcohols contains at least 1.5 mg% of camazulene, at least 5.0 mg% of (-) - <3> bisabolol and less than 10 mg% of other bisaboloids.

10 If the flowers of the described tetraploid chamomile sauna are assembled at a growth stage in which 30-70% of the mycorrhizal horn flowers have opened and the flowers are not dried at an air temperature higher than 50 ° C, then the substance thus obtained contains, for example, at least 100 mg-% -% (-) - Ot-bisabolol and less than 50 mg-% of other bisaboloids and, possibly, the essential oil prepared from it contains at least 3,5% of camazulene, at least 10% of (-) - Qi-bisabolol and less than 10% of other bisaboloides and, and the alcoholic extract contains at least 5.0 mg% of kamatsu-20 len, at least 10.0 mg% of (-) - Cfc bisabolol and less than 10.0 mg% of other bisaboloids.

If the flowers of the described tetraploid chamomile sauna are collected in a growth phase in which 30-70% of the mycorrhizal horn flowers have opened and the flowers are dried at an air temperature of 50-70 ° C, the substance thus obtained contains, for example, at least 50 mg% of mg-% of (_) _CX »-bisabolol and less than 50 mg-% of other bisabolites and, optionally, the essential oil prepared therefrom contains at least 3.5% of camazulene, at least 30 10% of (-) - 05-bisabolol and less than 10% other bisa bolooids, and, and the alcoholic extract contains at least 2.5 mg% of camazulene, at least 7.5 mg% of (-) - β-bisabolol and less than 10.0 mg% of other bisabolites.

If the flowers are assembled at a growth stage in which 70-100% of the 35 mycorrhizal horns have been opened and the flowers are dried in air at a temperature not exceeding 50 ° C, then the substance thus obtained contains, for example, at least 40 mg% of camatsuene, at least 120 mg-% of (-) - Qt-bisabolol and less than 50 mg-% of other bisaboloids, the essential oil possibly prepared therefrom containing at least 3,5% of camazolene, at least 10% of (-) - G-bisabolol and less than 10% of other bisabolites and the alcoholic extract contains at least 2.0 mg% of camazulene, at least 6.0 mg% of (-) - Q1 · bisabolol and less than 10.0 mg% of other bisabolites.

1Q If the flowers of the described tetraploid chamomile sauna are collected in a growth phase in which 70-100% of the mycorrhizal horn flowers have opened and the flowers are dried at an air temperature between 50-70 ° C, then the substance thus obtained contains, for example, at least 30 mg mg-% 15 (“) -Ot-bisabolol and less than 50 mg-% of other bisaboloids, the essential oil possibly made from this containing at least 3.5% camazulene, at least 10% (-) - Ot -bisabolol and less than 10% % of other bisabolites and the alcoholic extract contains at least 1.5 mg% of kamatsu-20, at least 10 mg% of (-) - Ot bisabolol and less than 10.0 mg% of other bisabolides.

Example 1

Intake of an anti-inflammatory agent from the parent plant of a tetraploid chamomile sauna: The chamomile sauna obtained as described below is sown in a conventional manner in agriculture. As soon as the plants have developed and the mycorrhizae are in a growth stage in which about 50% of the hornflowers have opened, these mycorrhizae are corrected with a chamomile sauna picking machine, 3q where the distance of the pickling machine combs is set The crop is transported as quickly as possible to a shady place and spread there in a thin layer for further processing. The mycelium 35 is then removed from the stems in a sieve and dried to a constant weight of 25 81 007. Weight loss rises to about 80%. Drying is carried out in the shade, in a well-ventilated place, on top of the trusses, with a layer thickness of about 10 cm.

After about 2-3 days, drying ends. To separate the stem-5 parts and the crushed flower, the dried material is sieved and then compressed into balls.

Analysis:

Essential oil 960 mg-%

Bamatsulene 162 "10 (-) - ^ -bisabolol 330"

In the case where the drying is performed in a fixed conveyor dryer with artificially heated air at a temperature of 50 to 70 ° C, the drying is completed after 4-5 hours. To separate the stem parts and the flower-15 crush, the dried material is screened and then compressed into balls.

Examples of analytical values for such a substance are: Essential oil 870 mg-%

Camazylene 94 "2Q (-) -? - Bisabolol 197" The starting plants of the tetraploid chamomile sauna obtained are obtained as follows: by means of a selection test on individual plants carried out on 10,000 plants which were 25 tetraploid chamomile sauna varieties. Särkäny photographed / Herba Hungar. 4 (1), 125-169 (1965), it was found that per about 1000 units a plant with a high content of camatsulene in the essential oil of about 20% by weight and a high content of (-) - CC-bisabolol of 50% by weight was obtained. the content of bisaboloids (especially (-) - o & --bisabolite oxide) is very low (less than 5% by weight) These individuals were selected and treated in the following steps: 26 81 007

Phase 1:

From the progeny of the selected tetraploid chamomile sauna plants (as described above), individuals were selected that 5 A) flowered approximately simultaneously, B) branched evenly, from the root, and had a narrow flower zone, about 10 cm, primarily 5 cm, C) with large mycorrhizae , with an outer diameter of about 3Q mm, mainly 25-35 mm, 10 D) with a minimum concentration of 150 mg% for camatsulene and 300 mg% for bisabolol or more and with a clear concentration for other bisaboloids (especially bisabolol oxides) Below 50 mg%. (All values are for mycera-15, which were dried at 40 ° C and collected at the stage when 30-70% of all mycorrhizal horn flowers were opened).

These plants were cloned. In addition, the plants (parent plants used for cloning) were cut so that the shoot 20 became about 15 cm long, brought to a daily length of 8-10 hours and 12-14 ° C to form new vesicles (to form short side tumors). The dwarf shoots were cut and placed in a peat-sand mixture. At approximately 100% relative humidity, at an air temperature of 15 ° C for 25 and 14 hours of daytime, cuttings took 7-14 days to take root.

Instead of the described conventional cloning (plug propagation), in vitro propagation of plant-divisible tissue parts (so-called growth cell-30 propagation) can also be used. Different parts of the plant are suitable for setting up a chamomile sauna plantation, mainly head tumors or difficult to grow.

After the plants were rinsed with HjOj under aseptic conditions in a "laminar flow" (high power 35 fluid filter with low turbulence exhaust thrusts), the sprout tips of the leaf swabs are removed and transferred to test tubes filled with Phisol. 51, 473-497, 1962). The test tubes are placed aerated. In a room with a day length of 12-18, mainly 16 hours (achieved by means of fluorescent light tubes), the light intensity is 500-10,000 lux, mainly 1000-3000 lux and the temperature is 15-30 ° C, mainly 22 -27 ° C. 5 As soon as the grafts grow well, they are transferred to the above-mentioned medium, which, however, has a higher concentration of cytokinins (cytokinins are plant hormones that promote cell division) (e.g.

30 ml / l 6-isopentenyladenine) and little or no 10 auxins (10-0.3 mg / l indoleacetic acid). In the future, the spines will stretch and form late organs as well as more awkward buds. These can be removed and grown as mentioned above.

In order to propagate the plants to a greater extent, certain grafts (stage 3 = 3rd generation) are transferred after rooting and cultivation with the first medium to a medium containing 10 mg / l indoleacetic acid or 3-indolebutyric acid or 0.1-0 , 3 mg / l -naphthylacetic acid. In this case, the plants 20 take root and after about 4 weeks can be planted in pots filled with sterilized garden soil (steamed for 12 hours at about 120 ° C) and further grown in a greenhouse (under conditions suitable for conventional propagation by cuttings).

25 Medium according to Murashigen & Skoog: - _mg / 1_mg / 1 NH4NO3 400 Indoleacetic acid 2.0

Ca (NO2) 2'4H2 ° 144 Furfuryladenine 0.1 KN03 80 Thiamine 0.1 30 Κί ^ ΡΟ ^ 12.5 Nicotinic acid 0.5

MgSO 4 ^ f ^ O 72 Pyridoxine 0.5 KCl 65 Glycine 2.0

NaFe-EDTA 25 myo-inositol 100 Z '; H ~ BCU 1.6 Casein hydroly- 1000. 35 sent. **; MnSO 4 • 4H 2 O 6.5 Sucrose 2%: ZnSO 4 • 7H 2 O 2.7 Purified agar powder 1%] KJ 0.75 28 81 007

Step 2:

The plants obtained in step 1 withered under isolated conditions, in the greenhouse at the same time. The plants were then in 11 cm pots filled with garden soil at a daytime temperature of 18-24 ° C and a night temperature of 12-14 ° C. The length of the day was at least 14 hours, and in the winter half of the year it was achieved with additional lighting 2 (200 watts / m). Irrigation was performed as needed.

10 For a period of 4 weeks, the mycelium that had flowered and was about to fall was collected from the selected individuals for seed intake. Drying was performed in a well-ventilated hall at an air temperature of 20-30 ° C; then the seeds were separated with a sieve (5 x 0.4 mm) and subsequently cleaned with a whisk.

Step 3:

A seed test was taken from the seed crop from step 2, and nojn 2000 progeny were grown from this (ecological conditions as in step 2) and selected according to the similar criteria of step 1 A) -D). For individuals so selected, the procedure was as in step 2.

Step 4: 25 Part of the seed yield according to Step 3 was sown in the autumn in two different growing areas.

Habitat I

450 m NN, 48.5 ° north latitude / ll, 5 ° east longitude, 750 mm annual rainfall, humid-temperate 30 climate, January -10 to 0 ° C, July +10 to + 20 ° C.

li 29 81 007

Habitat II

200 m NN, 42 ° north latitude / 10 east longitude, 400 min annual rainfall, Mediterranean climate, January 0 to + 10 ° C, July +20 to + 30 ° C.

5 Sowing was carried out in both growing areas at the end of September / beginning of October.

The field test conditions thus obtained were retested and evaluated for homogeneous growth, flower size, and date of harvest, in addition, flower test-10 samples were examined for active ingredient concentration. Individuals that met the parameters mentioned in step 1 were re-selected from the field growth. From these, seeds are obtained according to the last sentence of step 2.

Step 5: 15 Using the seeds of Step 4, Steps 3 and 4 were repeated in the above order and as shown. NN = altitude above sea level.

Step 6:

From the seeds obtained from step 5, about 20,500 individuals were grown (ecological conditions as in step 2) and selected according to the same principle as shown in step 1. From the plants thus selected, 34 individuals were selected and cloned according to step 1. Every tenth plant of each clone was planted by random distribution of them at a distance of 40 x 30 cm to open ground (site I, see step 4) in an isolated site. The soil was loess, pH 7.0. Planting was carried out in early June, the first seed harvest was obtained in mid-July, after which the plants were cut, they bloomed again 30 and produced a second seed harvest from mid-August to the end of August.

Seed harvesting of this material was performed according to step 2.

The seeds according to step 6 are the seeds used for sowing the chamomile sauna according to the invention in question.

The flowers of these seeds (sowing in September / October, harvested at the beginning of June-5 of the following year) are picked when 30-70% of the horn flowers are open and then immediately dried in an oven at 40 ° C for 72 hours. contain, based on the weight of the dried flowers (dry matter), for example at least 150 mg% of camazulene, 300 mg% of (-) - O-10 bisabolol and up to 50 mg% of other bisaboloids.

Example 2

Intake of an anti-inflammatory agent from the tetraploid stem of a chamomile sauna:

The chamomile sauna, obtained as described below, is sown in a conventional manner, according to the farming method. As soon as the plants have developed and the mycorrhizae are in the growth stage in which about 50% of the hornbills have opened, these mycorrhizae are corrected by a chamomile sauna picking machine. The harvested crop is transferred to a shady place as soon as possible and spread there for further processing into a thin layer. The mycelium is then separated from the stems in a sieve and dried to constant weight. The weight loss is about 80%. Drying is carried out in the shade, in a well-ventilated place with drying racks, with a layer thickness of about 10 cm.

Drying ends after 2-3 days. To separate the stem parts and the crushed flower, the dried material is sieved and then compressed into balls.

Analysis:

Essential oil 940 mg-%

Camazulene 132 "(_) _ Ot-bisabolol 243" 35 If drying is carried out, for example, in a solid solution

II

3i 81007 in a belt dryer, using artificially heated air, at a temperature between 50 ° C and 70 ° C, then drying is completed after about 4-5 hours. To remove the stem parts and the crushed flower, the dried material 5 is sieved and then compressed into balls.

Examples of analytical values for such a substance are: Essential oil 775 mg%

Camazulene 65 "(_) _ Ot-bisabolol 163" 10 The tetraploid chamomile sauna stock used is obtained as follows:

By means of a single plant variation experiment performed on 10,000 plants of tetraploid chamomal quality described by I. Särkäny / kerba Hungar. 4 (1), 125-169 15 (1965)], it was found that about 1000 individuals form a plant with a high content of camatsulene in essential oils of 20% by weight and a high content of (-) - <X-bisa bolol. is 50% by weight, whereby at the same time the amount of other bisaboloids (especially (-) - Ct-bisabolone oxide) is very low (less than 5% by weight). These individuals were selected and subjected to the following procedures:

Measure 1:

From the progeny of the selected tetraploid chamois-25 millasauna plants described above, those individuals were selected that achieve a minimum concentration of 100 mg% for camatsulene and 200 mg% or more for (-) - Cfc-bisabolol and less than 50 mg of other bisaboloids. %. All values refer to flower mycorrhizae, which were dried at about 40 ° C and harvested at a stage when about 30-70% of all mycelium horns had opened.

Measure 2:

All open flower buds were removed from the plants obtained according to step 1, after which the plants were transferred to a greenhouse and stopped flowering under isolated conditions. The plants were then located in 11 cm pots filled with garden soil at a daytime temperature of 18-24 ° C and a night temperature of 12-14 ° C. The length of the day was at least 14 hours and was achieved in the winter half of the year with additional lighting (200 o watts / m). Irrigation was performed as needed.

Of the total number of individuals selected, the 10 mycorrhizae that had withered and were just about to decompose were continuously collected for 4 weeks to obtain seeds. Drying was performed in a well-ventilated hall, at an air temperature of 20-30 ° C; the seed was then removed by sieving in a filling sieve (5 x 0.4 mm) and then purified on a whisk 15 (Steigsichter).

The seeds obtained after step 2 are the seeds of the chamomile sauna used in the method according to the invention. Flowers of plants obtained from this seed (sowing in September / October, harvested at the beginning of June of the following year), 20 picked at the time when 30-70% of the horn flowers have opened and immediately dried in an oven at 40 ° C for 72 hours , containing, by weight of dried flowers, not less than 100 mg of camazulene, not less than 200 mg of (-) - C * bisabolol and not more than 50 mg of other bisabolites.

Example 3

Intake of an anti-inflammatory agent from a diploid chamomile sauna stem:

The chamomile sauna obtained by the method described above is sown in accordance with conventional agricultural practice. As soon as the plants have developed and the flower buds are in a growth phase in which about 50% of the horn flowers have opened, these flower buds are harvested with a chamomile sauna picking machine, the distances of the combs of the picking machine being set so that only those flower buds

II

33 81 007 of which 50% of the horn flowers have been opened are pulled out. The crop is transported as quickly as possible to a shady place and spread there in a thin layer for further processing. The mycelium is then removed from the mycelium in a sieve apparatus and the mycelium is dried to constant weight. The weight loss is about 80%. Drying is carried out in the shade, in a well-ventilated place, on a drying grate, in a layer about 1 cm thick.

Drying ends after 2-3 days. To remove the stem parts and crushed flowers, the dried material is sieved and then compressed into balls.

Analysis:

Essential oil 910 mg-%

Camazulene 117 "15 (-) - (X -bisabolol 252" The diploid chamomile sauna stock used is obtained as follows:

Using a single-plant variation test performed on diploid, low-bisabolol, but not bisabolol-free chamomile saunas

Franz, Ch., J. Hölzl and A. Vömel describe in Acta Horticulurae 73, 109-114 (1978) that up to 20% of all individuals in essential oils had a camatsulene content of about 20% by weight and (-) - Cfci -bisabolol content was high, 50% by weight, whereby at the same time the content of other bisabolols (especially (-) - O 2 -bisabolone oxide) is very low. These individuals were selected and formed into tetraploids as follows:

Seeds from selected chamomile saunas were placed on filter paper impregnated with 0.05% aqueous colchicine and left at room temperature (20 ° C) for 6 hours. They were then removed from the filter paper, rinsed several times with water, seeded in a seed box (greenhouse): soil: peat-sand mixture 1: 1, temperature: 35 18-20 ° C, relative humidity: about 60%, daytime cooking time. 007 in notable lighting: 14 hours.

Germinating plants were observed until flowering, the result of polyploidization was determined by a comparative measurement of pollen and seed size and by counting the chromium-5 somes of the plants (F ^ progeny = progeny grown from seeds of colchicine-treated plants proven to be tetraploid).

Tetraploid formation can also take place as follows: 1Q Well-developed chamomile seedlings, 5-7 days old, germinated with filter paper immersed in water, were placed at room temperature (20 ° C) for 4-6 hours, cotyledons down, 0.05%: colchicine solution. Sensitive locust seedlings were then treated with caution; to avoid drought-15 damage in them, the relative humidity of the surrounding atmosphere must be close to 100%. After treatment, the embryos were rinsed several times, with water, and replanted on plant trays. Further processing was performed as for seeds.

Pollen size measurements are performed with a Leitz binocular examination microscope with a micrometer measuring eyepiece and using micrometer object glasses.

Chromosomes are counted from root tips: From seedlings or cuttings grown in a greenhouse, kootacin 1-2 cm 25-length, fresh root tips are placed in 0.002 molar hydroxyquinoline solution for 5 hours and then in 1N HCl for 15 minutes. For examination, approximately 1 mm of root tips are stained with 2% orsein acetic acid and examined under a microscope using oil immersion.

30 Cells in mitosis can thus be used to determine the 4-fold chromosome sequence of somatic cells (4n = 36).

Plants with a pollen diameter of about 50 '& larger than the pollen diameter with the diploid starting material (about 30 instead of about 20) and in which the chromosome sequence of 35 somatic cells was doubled to 36

II

35 81007 (with a corresponding number of diploid starting material of 18) were tetraploid. These plants were selected. About 0.5% of the seeds or cotyledons formed tetraploids by the above method and formed flowering, undamaged plants. This is now related to Example 2 corresponding to steps 1 and 2, for example.

The flowers of the chamomile saunas thus obtained contain, for example, not less than 100 mg% of camazulene, 200 mg% of 10 (-) - β-bisabolol and not more than 50 mg% of other bisaboloids (harvested at 30-70% of the horn flowers used for 72 hours in an oven at 40 ° C).

Example 4 15 Intake of an anti-inflammatory agent from a diploid chamomile sauna stem:

The chamomile sauna, obtained as described below, is sown according to conventional farming methods. As soon as the plants have developed and the flower-20 mycelium is in a growth phase in which about 50% of the horn flowers have opened, these flower mycorrhizae are harvested with a chamomile-picking machine, the comb distances of the pick-up machine being set so that only 50% . The crop is transported as quickly as possible to a shady place and spread there in a thin layer for further processing. The strainers are then removed from the stems in a strainer and dried to constant weight. The weight loss is about 80%.

Drying is carried out in the shade, in a well-ventilated place, on top of a drying grate, with a layer thickness of about 10 cm.

Drying ends after about 2-3 days. To remove the stem parts and crushed flower, the dried material is sieved and then compressed into balls.

36 81 007

Analysis:

Essential oil 1020 mg-%

Camazulene 173 "(-) - Ot bisabolol 418" 5 The used, diploid chamomile sauna starting plants are obtained as follows:

Using a single-plant variation test performed on diploid, low-bisabolol but non-bisabolol-free chamomile saunas by Franz, Ch. # J. Hölzl and A.

Vömel, in Acta Horticulturae 73, 109-114 (1978), found that up to 20% of all individuals had an essential oil content of camatsulene of about 20% by weight and / or a content of (-) - bisabolol as high as 50% by weight. -%, at the same time the concentration of other bisaboloids, in particular (-) - oo-bisabolone oxide) is very low. These individuals were selected and formed into tetraploids exactly as in Example 3, and tetraploid individuals were selected. 20 About 0.1-0.5% of seeds or cotyledons formed tetraploids and developed flowering, undamaged plants.

This is now related to, for example, steps 1-6, corresponding to Example 1.

The flowers of the chamomile sauna thus obtained contain, for example, at least 150 mg% of camatsulene, 300 mg% of (-) -? - bisabolol and at most 50 mg% of other bisabolols (harvested at a time when 30-70% of the horn flowers have opened, and the crop has been dried for 72 hours in an oven at 40 ° C).

The other properties correspond to those of the chamomile sauna obtained in Example 1.

Example 5

Preparation of the extract Chamomile-herbal chamomile 35 Chamomile flowers for the herb to be used are harvested according to Example 1 at a time when 30-70%

II

37 81 007 horn flowers have opened, and are dried in air at a temperature below 50 ° C.

400 g of the chamomile herb thus obtained, containing 105 mg% of camatsulene and 212 mg% of (-) -? - bisabolol, are extracted with 2100 g of aqueous ethanol (40% by weight of ethanol) in a pool mixer with a speed of 30 rpm / min 3 hours. The herbal product is then squeezed and the extract is filtered.

The content of active substances in the extract is determined in 10 conventional ways: camatsulene: 3.7 mg-% (-) - G'-bisabolol: 10.2 mg-%

Example 6

Preparation of essential chamomile oil 200 g of the dried chamomile flowers obtained in Example 1 (drying at 50 ° C protected from sunlight) are reacted in a 5 l round-bottomed flask with 3.6 l of water and 2 g of sodium ascorbate; the pH of the mixture is adjusted to 5.0 IN with Heli. Boiling hops are added to the mixture and the mixture is heated to boiling point. About 1.2 l of distillate is obtained in about 3 hours.

After completion of the distillation, the distillate is shaken 3 times with 100 ml of petroleum ether and dried over anhydrous sodium sulfate. The dried solution is filtered, after which the solvent is removed by distillation on a rotary evaporator.

The content of camazulene and (-) - Qi-bisa bolol in the resulting essential oil is then determined.

Yield: 1.44 g of essential oil 30 Mathematene: 3.6% of oil (-) - Oi-bisabolol: 10.2% of oil

Appendix A

Intake of essential oil The starting material is a dried material obtained from a chamomile sauna, which according to the present invention is suitable for the production of an anti-inflammatory agent. only mycorrhizae with 30-70%, in particular 40-60%, of the horn flowers are used in the production of the oil. Drying is performed in an oven at 40 ° C for 72 hours.

The essential oil is obtained from the dried material by the following two-hour steam distillation process: 2.0 g of undiluted, dried flowers are placed in a one-liter round-bottomed flask with 250 ml of desalinated water and refluxed for two hours on a Clevenger ( steam distillation recovery apparatus for the quantitative determination of small quantities of essential oil). The collection vessel contains 1 ml of pentane per analysis. The recovery rate is 40 + 4 drops / -15 minutes. At the end of the distillation, the oil dissolved in the pentane is drained as anhydrous as possible into the test tubes, and any remaining essential oil adhering to the apparatus is rinsed with pentane. To remove any residual water, a spatula tip is added to the solution with dried Ν32εο®, and the solution is then filtered under suction through a glass sintered suction filter with a pore size of D 3 or D 4 in bottles with a helical mouthpiece. After removal of the pentane by evaporation at 40 [deg.] C. in a water bath and drying in a desiccator, the amount of oil is determined gravimetrically.

The oil thus obtained (about 20 mg) is then assayed for camatsulene and bisabolol.

Spectrophotometric determination of camatsulene Solution to be measured: 30 Dissolve the whole of an essential oil (about 20 mg) obtained from 2 g of dried material (obtained as described above) in 25 ml of n-hexane or cyclohexane.

Filter photometer (eg Eppendorf).

Il 39 81 007

Wavelength: 578 nm Cuvette: 1 cm

Specific extinction of camatsulene (1 g / 100 ml: 1 cm): 20.8 Compensation fluid: n-hexane or cyclohexane 5 Observed concentration of camatsulen mg / 100 g: 120 E ^ g

In the absence of a filter photometer, the determination may also be performed with a spectrophotometer:

Measuring device: Spectrophotometer (eg PM Q II / or PM Q 10 HI "ZEISS")

Wavelength: 605 nm Cuvette: 1 cm

Specific extinction of camatsulene (1 g / 100 ml; 1 cm): 24.5 Compensation fluid: n-hexane or cyclohexane Observed concentration of camatsulen mg / 100 g: 102 Egg ^

The bisabolol is then determined from the solution to be measured.

Gas chromatographic determination of bisabolol and other bisaboloids: 20 Gas chromatography: Hewlett Packard model 5750, Erba Fracto-vap 2350 or similar device Detector: Flame ionization detector Carrier gas: Helium

Column: 0.32 cm (4/8 "); 200 cm2 of steel 25 Column packing: 3% nitrile silicone rubber" XE 60 "with diatomaceous earth silanized" Cromosorb WAW HP ": 11a 125-150 pm, as support Temperature:

Detector: 320 ° C 30 Injection housing: 220 ° C Column: 85-220 ° C Temperature programming: 4 ° / min Sample solution: Use a standard solution close to kamatsuylene broth 35 Reference solution: Dissolve about 15 mg of standard bisabolol in 81 007 40 cyclohexane 25 ml

Amount to inject: Sample solution and reference solution, 5 μΐ each

Measurement: The measurement is made by comparison of the peak areas 5

Bisabolol content: mg / 100 g dried flowers: 10 x weighed batch (reference) / mg7 x surface area (sample)

Area (comparison)

The determination of other bisaboloids can likewise be carried out by gas chromatography, for example under the following measurement conditions:

Device: Packard, model 7721, series 800 or Erba Fractovap series 2350

Columns: Glass columns, 3 m / 2 mm in diameter; 2 m / 2 mm in diameter Filling: 3% methylphenylsilicone rubber "OV 1" with diatomaceous earth silanized "Gaschrom Q", 125-150 μm, as carrier Carrier gas: 30 ml / minute N2

Temperature programming: 80-180 ° C, 2.5 (3) ° C / min 20 Injector / detector temperature: 200 ° C

Detectors: Flame ionization detector

Injected amount: 1:50 diluted essential oil about 2 μl

The measurement is performed partly without an internal standard-25 slide, partly with an internal standard. Suitable lauric acid methyl ester or hexadecane as internal standard for oils with or without camatsulene, oils with a camatsulene content of more than 5%, is considered to be the better internal standard, the concentration being determined photometrically (at 578 nm).

Claims (5)

1. A method for obtaining a substance with anti-inflammatory effect from Matricaria chamomilla, characterized in that it is decided separately from 1,000 to 10,000 individual plants of a diploid or tetraploid chamomile population ( -) -a-bisabolol content of the ethereal oil and selects the chamomile plants of which (-) -a-bisabolol constitutes the main component of the ethereal oil, in which case the starting chamomile is tetraploid, the flowers dried at 40 ° C at least 100 mg% camazulene, at least 200 mg% (-) - α-bisabolol and less than 50 mg% other bisaboloids, and if the starting chamomile is diploid, the flowers dried at 40 ° C contain at least 15 100 mg% camazulene, at least 50-100 mg% (-) - a-bisabolol and less than 50 mg% other bisaboloids, and provided that the diploid starting chamomile is not the camomile variety Degumill, they further propagate from the starting chamomile separated chamomiles and from these plants well-2 Emit at least 10 individuals who have common flowering time, whose flower heads are 20 - 40 mm in size and whose active ingredient is at least 100 mg% camazule, at least 200 mg% bisabolol, and less than 50 mg% of other bisaboloids (all calculated on the basis of dry matter), possibly cloning these selected diploid or tetraploid plants, and a) taking seeds from the well-received plants, b) growing offspring of the seeds obtained in (a); finally, on the basis of the content of active substance, and coinciding flowering time, regular branching from the root and 20 - 40 mm size on the flower heads select as indicated above, possibly cloning the selected plants and taking seeds from them, c) repeats the measures in paragraph b) 3 - 5 times, d) grows offspring to the seeds obtained in paragraph c), selects according to paragraph a), possibly clones the 46 46 00 007 selected plants and takes the seeds of the selected seeds the planes, and in the case of diploid chamomiles, the final tetraploidize them by gamma rays, X-rays or UV radiation at 0 - 35 ° C, using higher temperatures of 33 - 50 ° C, at lower temperatures ρέ 0-5 ° C or by means of the decapitation callus method or uterine bud cultivation, the final gene is tetraploidized and takes care of the flowers of the well-received plants, which are in a plant stage where 30 - 100% of the flower heads' tube flowers are opened, and dry them at a temp. perature not exceeding 70 ° C, to obtain dry material containing at least 30 mg% of camazule, at least 100 mg% (-) - a-bis-abolol and less than 50 mg% of other bisaboloids; and optionally from the well-obtained dry material, prepare ethereal and yes by distilling in the presence of water, which oil contains at least 3.5% camazule, at least 10% (-) - α-bisabolol, and less than 10 other bisaboloids, or prepare an alcoholic chamomile extract by extracting with lower alcohols, which extract contains at least 1.5 mg% camazule, at least 5.0 mg% (-) - α-bisabolol and less than 10 mg% other bisaboloids . 2. A method according to claim 1, characterized in that the flowers are utilized in a plant stage, where 30 - 70% of the flower heads' tube flowers are opened, the flowers are dried at an air temperature not exceeding 50 ° C and the thus obtained the dried material contains at least 100 mg% of camazule, at least 200 mg% of (-) - α-bisabolol and less than 50 mg% of other bisaboloids, and the ethereal oil, possibly prepared from it, contains at least 3.5% of camazule , at least 10% (-) - α-bisabolol and less than 10 other bisaboloids, and the alcoholic extract prepared therefrom contains at least 5.0 mg% camazule, at least 10.0 mg% (-) -a-bisabolol and less than 10.0 mg-% of other bisaboloids. li * 7 81007 Method according to one or more of the above claims, characterized in that the flowers are used in a plant stage, where 30 - 70% of the flower head tube balloons are opened, the flowers are dried at an air temperature of 50 - 70 ° C, and so on. the thus obtained dried material contains at least 50 mg% camazule, at least 150 mg% (-) - α-bisabolol and less than 50 mg% other bisaboloids, and the ethereal oil, possibly prepared from it, contains at least 3 , 5% camazulene, at least 10% (-) - α-bisabolol and less than 10 other bisaboloids, and the alcoholic extract prepared therefrom contain at least 2.5 mg% camazulene, at least 7.5 mg% (-) -a-bisabolol and less than 10.0 mg-% of other bisaboloids. Method according to one or more of the preceding claims, characterized in that the flowers are utilized in a plant stage, where 70 - 100% of the flower heads' tube flowers are opened, the flowers are dried at an air temperature not exceeding 50 ° C, and the sealant-dried dried material contains at least 40 mg% of camazule, at least 120 mg% (-) - α-bisabolol and less than 50 mg% of other bisaboloids, and the ethereal oil, possibly prepared from it, contains at least 3.5% camazule, at least 10% (-) - α-bisabolol and less than 10 other bisaboloids, and the alcoholic extract prepared therefrom contain at least 2.0 mg% camazule, at least 6.0 mg% (-) -a-bisabolol and less than 10.0 mg% of other bisaboloids. Method according to one or more of the above claims, characterized in that the flowers are used in a plant stage, where 70 - 100% of the flower heads' tube flowers are opened, the flowers are dried at an air temperature of 50 - 70 ° C, and the thus obtained dried material contains at least 30 mg% of camazule, at least 100 mg% (-) - α-bisabolol and less than 50 mg% of other bisaboloids, and the ethereal oil, possibly prepared from it, contains at least 3.5% camazule, at least 48 81 007 10% (-) - α-bisabolol and less than 10 other bisaboloids, and the alcoholic extract prepared therefrom contain at least 1.5 mg% camazule, at least 10.0 mg% (-) - α-bisabolol and less than 10.0 mg% of other bisaboloids.