SK944585A3 - Process for extracting tetraploid camomile enriched with bisabolol - Google Patents

Abstract

A process for extracting camomile together with bisabol uses as the source plant a specially selected strain of Matrica Chamomille of which the flowers dried at 40 DEG C contain at least 100 mg% of chamazulene and at least 200 mg% of (-)-???-bisabolol, the content of the other bisaboloids being below 50 mg%, wherein an initially tetraploidized diploid camomile or a tetraploid camomile, in which (-)-???-bisabolol is the main component of the essential oil, is subjected to various selection and propagation steps.

Description

Process for preparing a new tetraploid camomile rich in bisabolol

Technical field

BACKGROUND OF THE INVENTION The present invention relates to a process for the preparation of a novel bisabolol-rich tetraploid marigold of the plant species Chamomilla recutiťa L., synonymous with Matricaria chamcmilla L., Asteraceae, whose flowers dried at 40 ° C contain at least 100 mg% chamazulene. , at least 200 mg% of (-) - from β-bisabolol and less than 50 mg% of other bisaboloids, with the exception of manzana.

I

BACKGROUND OF THE INVENTION

The inflorescence preparations of Chamomilla recutitea (L.) Rauscnert, synonymous with Matricaria Chamomilla, L., have a wide therapeutic use due to their anti-inflammatory and spasmolytic effect and form an important part of the plant medicinal raw material assortment. Particular therapeutic importance is ascribed to the active ingredients of (-) alpha-bisabolol and chamazulene. Consequently, a satisfactory camomile drug should have a high content of both substances.

The naturally occurring diploid camomiles are very heterogeneous with respect to the active ingredient content. Only one of the important active ingredients of chamazulene and (-) alpha-bisabolol is available, or these are missing or present in very small amounts, in which case the main constituent of the essential oil consists of less active bisaboloids / bisabolonoxide A or B, bisabololoxide A /.

Under the designation DEGUMILL / this is a variety of camomile, protected and listed in West German Patent 24 02 802; o a protected variety registered in the former GDR by the name Degumill; tal. U.S. Pat. No. 1,035,096 / is known a variety of camomile, which contains at the same time a high content of (-) - alphabisabolol and chamazulene. The chamazulene content of this rumarpecker variety DEGUMILL is stable only if accidental crosses or foreign pollination with other varieties of camomile are avoided, the content of bisabolol and chamazulene being substantially lower, and of the wild active camomile poor in general. The risk of cross-pollination is almost everywhere, as wild camomile occurs practically in general. In addition, various tetrsploid forms of camomile are known / for example BODEGOLD, NDR; POHOÉELICKÝ, CR and SR; ZLOTY LIN, Poland; Bl <-2, Hungary /. Although these tetrapioid forms have a satisfactory chamazulene content, the important active ingredient (-) - alpha-bisabolol is only very small in these tetraploid species, and the other bisaboloids (bisabolol A and B as well as bisabolonoxide) predominate which they form more than half the essential oils of the known tetraploid camomile species.

SUMMARY OF THE INVENTION

It has now been found that quite generally it is possible to use naturally. According to the invention, a marbled populations of camomile, whether diploid or tetraploid cultivated camomile, in some way according to the invention, obtain bisabolol-rich tetraploid camomile plants which, on the one hand, are susceptible to foreign pollination of naturally occurring camomiles. high content of chamazulene and (-) - alpha-bisabolol, while the content of C -) - alpha-bisabolol significantly exceeds the content of chamazulene and at the same time other bisaboloids are practically absent or only to a small extent.

It is also important that the high content of the main active compounds C-.beta.-bisabolol and chamazulene in the camomile according to the invention is permanent, that is to say, it is maintained at every propagation.

SUMMARY OF THE INVENTION The present invention provides a process for the preparation of a novel bisabolol-rich tetraploid camomile (Chamonilla recutita L., synonymous with chamonilla L., Asteraceae), of which the dried flowers contain at least 100 mg% chamazulene relative to dry matter. , at least 200 mg% of C - 1 - .beta.-bisabolol and less than 50 mg% of other bisaboloids, with the exception of varieties of creeping rhizome, which consists in the determination of 1 000 to 10 000 essential oils containing C -) - .beta.-bisabolol and selected those camomile plants in which C -) - .beta.-bisabolol is a major constituent of essential oil and whose flowers, dried at 40 DEG C., contain at least 100 mg% chamazulene, at least 50 to 100. mgX (-α-bisabolol and less than 50 mg% of other bisaboloids), these plants are separated from the initial population, multiplied and subjected to further selection and, if appropriate, multiplication step and, where appropriate: from the plants of the resulting camomile, seed for propagation and / or camomile drug is obtained, using a diploid camomile from which the Degumill variety is excluded as the starting camomile, is completed after all selection steps have been completed tetraploidization by chemicals at temperatures ranging from D to 35 ° C, by gamma-rays, X-rays or UV rays from 0 to 35 ° C, by elevated temperatures ranging from 33 to 50 ° C, by reduced temperatures ranging from 0 to 5 ° C, by the decapitation-callus method or by anthropogenic cultures.

Selection and multiplication in the method of the invention consists of the following steps:

- 4 a / selection of at least 10 individuals of tetraploid plants as active substance content, i. at least 100 mg% of chamazulene, at least 200 mg% of bisabolol and less than 50 mg% of other bisaboloids, all values are based on dry matter, at the present time of flowering, at the uniformity of basic branching, and by flower size from <0 to 40 b) growing offspring from seed obtained in paragraph a), and then selecting the seed stage and / and obtaining seed from such selected c) three to five times the repetition of the steps of the paragraph b / jl

d) growing offspring from the seed obtained in paragraph c), selecting these offspring from the stage and /, cloning the plants thus selected and obtaining seed from the plants obtained by cloning.

Surprisingly, the novel tetraploid camomile according to the invention has a number of novel advantageous properties in comparison with the known camomile or camomile varieties. In particular, it is no longer susceptible to foreign pollination by naturally occurring populations of camomile (wild camomile) as the known camomile variety DEGUMILL. Compared to diploid and tetraploid varieties of camomile, the scent of the invention is surprisingly high in (-) - alpha-bisabolol (at least 200 mg%), while the content of other bisaboloids (eg bisabololoxides) is very low and is less than 50 mg% . Known and described in the literature the camomile all have a very high content of bisaboloids and partially bisabolone oxides, which generally make up 50% of the essential oil of the drug

- 5 (i.e., at a content of 1% essential oil, a content of about 500 mg per 100 g of the drug).

I

Furthermore, the camomile according to the invention is, in contrast to the previously known tetraploid camomile varieties, homogeneous with respect to the substances contained therein, the properties of chamazulene and (-) - alpha-bisabolol being constant (i.e. homozygous).

On the other hand, the evaluation of individual plants of known tetraploid varieties of camomile shows that the individual ^- individual ^- belongs to different types in terms of substance content, which means that a single random collection of the drug produced from them shows a qualitatively and quantitatively different inhomogeneous result.

In addition, the camomile of the invention surprisingly differs from the known tetraploid camomile varieties, for example, by one or more of the following properties: improved seed germination, less unnecessary green matter content, i.e.. higher flower yield /; lower water content of the flowers (higher yield of dried flowers and shorter drying times); more suitable for mechanized harvesting because the flowers are significantly localized in one plane and most of them bloom at the same time (thus allowing a single harvesting term); better drug shelf life (lower propensity for disintegration and waste generation); extraordinary aromatic typical camomile fragrance. The aforementioned advantages will in particular arise when, in addition to the selection for active ingredients, one or more of the above-mentioned features are selected, the selected plants are seeded, the offspring thus obtained are re-selected and these measures are carried out three to five times.

Possible default diploid camomiles include, for example, Camomile of Argentine origin ”

- 6 / see L. Z. Padula, R. V.-D. Ro'ndiňa and J. D. Coussió / 'quantitative -'. essential oil / total azulene and '-; chamazulan-y by German' -currant 'grown in-Argentina; -Plan-. The honey. 30, pages 273-230> 1976), as well as all the camomiles (which have a measurable concentration in the essential oil); silica /. ' Optionally, diploid vermicelli as described below are contemplated; H. - Schhilchare, - (Newer knowledge of the quality of essential oil), possibly of the flower plant Planta medí - '23j-32 - Ί44-71973 /; Kot 0. Kotlý; M.Artilkló '+. y. Lukeš and M.Jasičová K '' gas-chromatographic analysis of color, color;

PWlzl and A ' H. Schhilcher, thy determination, rumaneal chak, va, V.

and to the chemical types of camomile "essential oil."

310, 210-215 (1977); Ch. Franz, J common morphological and chemical characterization of several pulsations and variations of Matricaria chamomile L., Acta. , '- 73, 109-114 / 1978 //

Camomile essential oil typically consists of the main constituents of farnesene, spatulenol, charaazulene, one of the four bisaboloids / (-) - alpha-bisabolol, bisabololoxide A, bisabololoxide B or bisabolonoxide, as far as diploid individual plants are concerned; in the population sample, as well as in. tetraploid individuals may find various bisabolides side by side as well as spiroether. Said substances together constitute 70 to 80% of the essential oil. Thus, the starting diploid chamomiles are particularly preferred, wherein C - .alpha.-bisabolol is the predominant component (more than half the amount of the above).

Particularly suitable are diploid camomiles as starting materials in which, for example, (-) - alpha-bisabolol constitutes at least 40% of the essential oil or at least 50% of the bisaboloids.

Examples of tetraploid basil leaves are: Bodegold (GDR), Pohorelický (Czechoslovakia), Zloty Lan (Poland), BK-2 (Hungary). These species are described in the following literature: M. Cnládek and V. Košová, Pnarmazie 13, 712-713 (1958), W. Czabajska, Diss. Poznan (1963); W. Poethka and P. Bulin Pharm. Z. Halle, 103, 813-823 (1969); I. Sarkany, Herb. Hungar, 4 (1), 125-169 (1965). Essentially, the same applies to the tetraploid starting chamomiles with respect to the content of (-) - alpha-bisabolol as to the diploid starting chamomiles.

Particularly suitable are diploid and tetraploid camomile populations which, as a rule, contain at least 100 mg% of chamazulene and 50 to 100 mg% of (-) - alphabisabolol / with respect to the dry weight of the flowers dried at temperature. 40 ° C (i.e. seed from such populations of camomile is obtained in the case of diploid starting camomile and seed is tetraploidized (irrespective of and / or knowledge of the parent plant bisabolol content). In the case of a starting tetraploid camomile, the seed described above is then subjected to selection and propagation immediately.

Suitable diploid and tetraploid starting chamomiles are selected by screening individual plants. In general, it is necessary to investigate between 1,000 and 10,000 individuals from the initial population and to find several individuals having a high (-) - alpha-bisabolol content according to the above criteria and which are advantageous as the starting romance. Seeds are obtained in the usual manner from such selected individuals and this is tetraploidized. At the end of this process, the tetraploid plants are selected from the remaining diploid plants. All plants having at least 100 mg% chamazulene, at least 200 mg% (-) - alpha-bisabolol and less than 50 mg% other bisaboloids in the flowers dried at 40 ° C are selected from the tetraploid plants thus obtained after possible previous propagation. . These flowers are harvested at the stage of development when 30 to 70% of all tubular flowers are open. This may be followed by further selection and propagation steps which affect, for example, the following characteristics or, if appropriate, the following characteristics: tertiary of simultaneous flowering, uniform base branching and narrow flowering zone (ie better suitability for mechanized harvesting), large inflorescences, better drug shelf life, especially aromatic scent.

When starting in advance from tetraploid camomile, only the aforementioned selection of those plants whose flowers dried at 40 ° C contain at least 100 mg% of chamzulene, at least 200 mg% of (-) - alpha - bisabolol and.

- 9 less than 50 mg% of other bisaboloids, the collection of these flowers being carried out at the developmental stage where 30 to 70% of all tubular flowers are open. Also in this case, further selection and propagation steps may follow, possibly leading to further improvements (as mentioned above).

A particularly preferred camomile is obtained when selecting (according to the invention) those plants (known or prepared according to the invention) whose chamazulene content is at least 200 mg%, preferably 250 mg%, and selected according to the chamazulene and (-) - alpha-bisabolol content. the content of (.-) - alpha-bisabolol is at least 300 mg%, with the advantage of 400 mg% (the content of other bisaboloids is still below 50 mg%).

Tetraploidization can be accomplished in a manner known per se by affecting plant parts / seeds, roots, shoot ends, germs, armpit pucks or tissues of diploid romanian / chemical starting plants, X-rays, gamma rays or UF-rays. It can be carried out further by a callus decapitation method, an anthropogenic culture, or by applying high or low temperatures to the camomile plants, possibly to plant parts or to the camomile plant tissues. Here, reference may be made, for example, to V / erner Gottschalk's book "Die Eedeutung der Polyploidie fttr die Evolution der Pflanzen", Gustav Fischer Verlag, Stuttgart, 1976, in particular pages 13 to 22.

Tetraploidization using chemicals:

Examples of suitable tetraploidization chemicals are: colchicine, acenaphthene, alkaloids such as atropine, veratrine, nicotine, sanguinarine, benzene derivatives, biphenyl and phenanthrene, naphthalene and derivatives of naphthalene, diphenylamine, tribromaniline methyl, p-dichloro-benzoquinone, p-dichloro-benzoquinone, salicylic and related substances, hexachlorohexane, methamphetamine hydrochloride, alkali metal alkylcarbamates such as sodium isopropylcarbamate, phenylurethane, cacodylic acid salts (e.g. sodium salt), lily of the valley lysides such as convalarine, convalatoxin and convalaminuromine (heteroisanuxurine) heteroisuxine ethylphosphate, ethyl chloride, phenylhydroxide and phenyldinaphthylmethane disulfonate mercury (chloroform, gas of naphtha (N _o 0)). than '

Also mixtures of these substances. Cakes, compost and cow manure can also be considered.

The action of the substances mentioned herein takes place, for example, at temperatures between 0 and 35 ° C, preferably between 12 and 30 ° C, in particular at temperatures of 15 to 25 ° C.

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The application is carried out, for example, by treating the seeds, the tips of the shoots, the roots (in particular the root endings or roots of the germinating seedlings), the sponges, cut areas on leaves or stems, cell suspensions of meristemic tissues, callus cultures. into the area of the armpit buds. Chemicals are usually used in the form of solutions in water, weakly alcoholic solutions (alcohol content generally below 5%), or in the form of weakly acidic solutions. The pH of weakly acidic solutions is, for example, between 5.5 and 6.5, the acidification being carried out with lower aliphatic acids than acetic acid. If alcohol solutions are used, they may also be weakly acidic. The concentration of chemicals in these solutions may be, for example, 0.01 to 0.5%, preferably 0.02 to 0.2%, in particular 0.05 to 0.1%. The gaseous substances are used as such, optionally under pressure (e.g. 1 to 100 kPa). The duration of action is, for example, 1 to 36, preferably 2 to 12, especially 4 to 6 hours. '

1

The most effective concentration as well as the duration of action should be suitably tested in preliminary experiments.

For example, the action of colchicine at temperatures between 0 and 35 ° C, preferably at 12 to 30 ° C, in particular at 15 to 25 ° C, is particularly favorable. This can be done, for example, by allowing the seeds of the diploid starting chamomile to swell in 0.01 to 0.2%, in particular 0.02 to 0.1%, preferably in a 0.0% colchicine solution, or germinate five- The well-sprouted seedlings of the diploid chamomile (with the key leaves down) dipped to 0.01 to 0.2%, in particular 0.02 to 0.1%, preferably 0.0, of co-chicin solution. in the latter method, the ambient atmosphere should have a relative humidity of 100%. The duration of action of colchicine is, for example, 4 to 10 hours. When germinating seedlings are used, an exposure time of up to 10 hours is generally sufficient. If seed is used, the treatment time can optionally be extended up to 36 hours.

After exposure to chemicals, the swollen seeds, germinating seedlings or other plant parts are washed several times with water. The swollen seeds are then sown, for example. Plants with treated roots or other parts thereof or treated germinating seedlings, for example, are picked up in plant hazel. From the treated seeds or germinating seedlings the plants are grown (for example in a greenhouse: at a temperature between 18 and 25 ° C by day and 10 to 16 ° C at night), plants whose feathers are about 1.5 times larger are selected. as the starting material, possibly having a chromosome number of 36 in somatic cells.

If other plant parts (above ground or ground) are treated, only the shoots, roots or flowers / seeds that have arisen from the chemical treated parts are subjected to a later examination for the tetraploidization carried out. For example, if the shoot or claws are treated, then only the shoot formed from the claw or the bud and the flowers formed thereon or the seeds for the number of chixmosomes are examined.

Pollen size measurement and chromosome counting can be performed, for example, as described in Example 3.

Tetraploidization by radiation:

The treatment is carried out, for example, on seeds or root ends at temperatures between 0 and 35 ° C, preferably at 10 to 30 ° C, in particular at 15 to 25 ° C. Amount of irradiation: 5 to 50 Krad. Gamma rays and X-rays are preferred.

Suitable UV-rays are those having, for example, a wavelength of 400 to 30 nm, preferably 350 nm.

The irradiated plants or plant parts are then treated as far as after treatment with chemicals.

Use of high and low temperatures:

Suitable temperatures are, for example, temperatures between 33 and 45 ° C. These temperatures are, for example, exposed to: swollen seeds, germinating seedlings, sprouts and meristem meshes. treatment time: for example 1 to 48, preferably 12 to 24 hours.

As a low temperature are: 0-5 C, preferably 0.5 ° to 4 ° C, in particular the ^'Q _2; C. These temperatures are exhibited, for example: swollen seeds, germinating seedlings, shoots and

- 13 meristem meshes. Treatment time: for example 1 to 100, preferably 20 to 40 days. Plants thus treated or parts thereof are treated in the same way as after treatment with chemicals.

Decapitating callus method (decspitating = cutting off the head or tip / tip) or back-cutting /. The decapitation is carried out, for example, in young plants at the stem, preferably at the end of the vegetation cone, after the formation of 4 to 6 leaves or even on leaf stems or side shoots. The pucks formed on the wounded tissue (callus tissue), or shoots, are separated, brought to roots, grown in pots, and the plants converted to the tetraploid form are selected in the same way as under the action of chemicals.

Anther culture (preparation of plants with a single number of pollen vesicle chromosomes followed by spontaneous or artificial tetraploidization). Closed tubular flowers are harvested from flowering plants whose anthers (pollen sacs) are in the first stage of pollen mitosis. The anthers are removed using a micromanipulator and transferred to Petri dishes, which are filled, for example, with Nitsch and Nitsch broth (Table 1). Petri dishes are then stored in the culture chamber for 16 hours at 28 ° C in the daytime. ° C, at night at 20 ° C. After about 4 weeks, the anthers begin to burst and the plants grow. These are haploid in the diploid parental fraction, dihaploid in the tetraploid parental fraction: after the root system has been established, they are transplanted, for example, into the garden soil and allowed to bloom. These (di) haploid plants are sterile, but can be converted to polyploid forms by treating the ends of the shoots, roots or stems with chemicals such as colchicine, producing homozygous plants which can then be further propagated by the seeds. in the treatment of chemicals, for example with colchicine.

Table 1

Nitsch and Nitsch / Science, 1969 / mg / l

KN0 ₃ 950 4.ΝΟ _ί 4 3 720 MgSO ₄ .7H ₂ O 185 CaCl ₂ 166 kh ₂ Mon ₄ I 68 MnSO ₄ .4H ₂ O 25 h ₃ or ₃ 1 0 ZnS0 ₄ .7H _? 0 1 0 Na ₂ Mo0 ₄ .2H ₂ 0 0.25 CuSO ₄ .5H ₂ O 0,025

ml of a solution of 7.45 g of ethylenediaminetetraacetic acid disodium salt and 5.57 g of FeSO4 · H2O per 1000 ml of myo-inosities 100 glycine2 nicotinic acid5 pyridoxine-HCl0.5 thiamine-HCl.

Table 1 - continued mg / l folic acid biotin sucrose broth / DIFCO-Bacto-Agar / indolylacetic acid

0.5

0.05 g

The pH of the medium is adjusted to 5.5.

By measuring the size of the pee and / or by counting the chromosomes of the selected tetraploid camomile plants that can be obtained by the above options, then plants having a minimum chamazulene content of 100 mg% and a minimum bisabolol content of 200 mg% are selected, in particular other bisaboloids. bisabololoxides (should be below 50 mg%) relative to the dried flowers (see Example 1).

The plants thus selected are allowed to bloom in a greenhouse at 18 to 24 ° C by day and at 12 to 14 ° C at night and daytime for a minimum of 14 hours after removal of the already blooming inflorescences, all seeds being harvested for 4 weeks. inflorescences that are in bloom or before disintegration. After drying at an air temperature of between 20 and 35 ° C, seed is obtained for propagation of camomile according to the invention.

In addition, the following criteria may be considered as further criteria for this selection:

a) approximately the present flowering time, b) a uniform base branch and a narrow zone of flowers, about 10 cm, especially 5 cm, c) large inflorescences with an outside diameter of about 30 mm (20 to mm), in particular 25 to 35 mm.

The use of these additional criteria a /, b / and / or c / leads, for example, to high flower and drug yields and is more suitable for machine harvesting.

If starting from the known tetraploid camomile, the selection described above is carried out, as well as, optionally, further selection and propagation steps.

In order to obtain a camomile which, in addition to containing at least 10 mg of chamazulene and at least 200 mg% of (-) - alpha-bisabolol (with regard to dried inflorescences), also provides high flower yield and is more suitable for dissemination, the following approach is recommended: , plants (as described above) and. multiply by vegetative divers and are monitored during 3. to 5 & energy, while still making a selection according to the above criteria, possibly additional criteria &. The plants grown according to these criteria are cloned, from the cloned plants seed is obtained, from which the obtained plants are again selected according to the stated minimum chamazoulene and bisabolol content, as well as other criteria a / to c /, and from the latter the seed is recovered.

Sequence - selection according to the above-mentioned minimum chamazulene content of 100 mg% and C -) - alpha-bisabolol of at least 200 mg% (other bisaboloids below 50 mg%), as well as according to 3 other criteria and / to c / repeat three to five times.

This is followed by one more sowing sequence, as described previously (possibly cloning), - seed harvesting.

Then, the seed obtained is seed for multiplying camomile according to the invention.

Diving propagation is carried out as follows: For diving propagation, the starting plants (clone mother plants) must produce short shoots without buds under short day conditions. This occurs in the winter of the year in a greenhouse without additional illumination or in air-conditioning chambers at a day length of 6 to 10, preferably 8 hours, and temperatures from 1 ° C to 15 ° C, preferably 12 ° C. For cloning or for multiplication, divers from the shoots of shoots and in particular short shoots (side shoots) are suitable. They are rooted at temperatures of 12-18 ° C, preferably 15 ° C, and a bottom length of 12-16, preferably 14 hours, in a saturated atmosphere (about 100% relative humidity). For example, a 1: 1 mixture of peat and sand; Also suitable is pure quartz sand, asbestos diving cubes, peat diving cubes, etc.

For example, the following soils are suitable for sowing: garden soil, humous medium-heavy clay soil, clay or humous sandy soil. Sowing can be carried out in greenhouses or in the field. The temperatures for germination and plant growth are between 12 and 24 ° C, in particular 18 to 20 ° C. If sowing in the open field takes place, it is preferably sown in autumn / September / October / or early spring / March / April /. These terms apply to all eligible areas (eg Northern Hemisphere, temperate to subtropical areas).

The novel camomile according to the invention belongs to the species of the plant camomile, botanical designation Matricaria chamomilla L. (synonym Chamomilla recutita / L..), Rauschert, and is defined by the active substance data in point 1 of the definition of the subject invention. The indicated levels of active substances of chamazulene, (-) - alpha-bisabolol and bisaboloidoxides always refer to the developmental stage of the flowers, with 30 to 70 in particular 40 to 60% of all tubular inflorescences open, ie the flowers used to determine the content The active ingredients were collected by this date and then dried in a box dryer at 40 ° C for 72 hours.

If the camomile flowers are harvested at a time when the development stage of the flowers has progressed further, this means that, for example, 100% or up to 100% of all tubular inflorescences are open (eg in full bloom) and / or harvested flowers are dried at higher temperatures above 40 ° C, the content of the active substances C -? - bisabolol and chamazulene may be lower, since at higher temperatures and / or at a later harvest, the decomposition of azulene and bisabolol may be more pronounced.

The term ostatné 'other bisaboloids in point 1 of the definition of the invention is understood in particular to mean: (-) - alpha-bisabololoxide A and B; (-) - alpha-bisabolone oxide A; The other constituents of the essential oil of the camomile variety of the invention are the N, N-dicycloether, farnesene, spatulenol and, in moderate concentrations, various volatile terpenic hydrocarbons.

For example, camomile flowers dried in an oven (box) at 40 ° C / harvested as above / between 100 to 200 mg% chamazulene, between 200 to 450 mg% (-) - alpha-bisabolol and little, that is, 5 to 50 mg% of other bisaboloids relative to the dry matter, which means relative to the absolute dry weight of the flowers. This absolute dry weight is determined by drying with a separate sample of camomile flowers in a drying cabinet at 105 ° C until constant weight (72-96 hours). The active ingredient content of the flowers dried at, for example, 35 to 50 ° C (drug) is converted to the dry weight of the flowers determined at 105 ° C.

In its phenotype, the camomile obtained according to the invention is similar to the previously known tetraploid camomile varieties (e.g. Bodegold, Zloty Lan, BK-2, Pohoøelický velkvětý); They differ in particular from the fact that (-) - alpha-bisabolol is the main constituent of the essential oils of the flowers of the invention according to the invention and, moreover, the content of other bisaboloids (bisabololoxide A and B as well as bisabolonoxide) is clearly lower. Other components of camomile essential oil are farnesene, spatulenol and En-In-dicycloether.

The camomile obtained according to the invention can be grown on all soils except those containing more than 20% organic matter (humus with soil microorganisms); no agrotechnical and cultivation measures are necessary; only a long day with more than 13 hours of maximum length of day is necessary for cultivation, that is to say, in particular, temperate zones and subtropical areas are suitable for cultivation.

The camomile obtained according to the invention usually has further advantages: higher yield, medium late harvest, uniform height, narrow flower zones and large inflorescences; thus, an exceptional suitability for mechanized harvesting. In addition, plants sown at the same time usually bloom practically together at the same time, which greatly simplifies and facilitates harvesting.

Finally, the above advantages are apparent when the following process conditions are used: from tetraploid camomile plants having a chamazulene content of at least 100 mg # and (-) - alpha-bisabolol 200 mg%, while the content of other bisaboloids (especially bisabololoxides) is below 50 mg % (all values with respect to the dried inflorescences), only those plants that are selected are selected

- they bloom at about the same time

have a uniform base branch and a narrow zone of flowers - about 10 cm, as well as

- large inflorescences with an outside diameter of about 30 mm.

The selected plants are propagated by vegetative divers and monitored for 3 to 5 genarations, selected according to the above criteria. The selected plants are propagated vegetatively (cloned), allowed to bloom together and seed is harvested from them. The seeds obtained from the seeds are selected again according to the above criteria and the seeding-selection sequence is repeated three to five times.

The drug prepared from the camomile obtained according to the invention has a maximum content of active ingredients of chamazulene and (-) - alpha-bisabolol when <3 the camomile inflorescence is harvested at a vegetation stage where, for example, 30-70% of the yellow flower tubes, preferably 40- 60%, i.e. generally 50% of tubular inflorescence flowers are open and drying is carried out at an air temperature of not more than 50 ° C, for example 35 to 50 ° C, in particular 40 ° C.

Drying can be carried out either by the supplied air, but also by shading, but it must be ensured that the heat supplied does not exceed the amount required for complete drying. It is also advantageous to ensure that a constant weight is obtained by means of checkweighing. The drying may be carried out spontaneously or artificially (for example, artificially heated warm air). The yield of the active substances is greatest in spontaneous drying under the exclusion of sunlight, preferably at 40 to 60 ° C, in particular at 40 to 50 ° C. The drying process should be carried out as soon as possible after harvesting. The drying should take place in thin layers, for example 5 to 20 cm, preferably at a thickness of 10 cm. Drying can also be carried out in well-ventilated halls at temperatures between 20 and 30 ° C. The air temperature for drying should not exceed 60 ° C. For example, a temperature between 35 and 50 ° C is favorable.

The content of the active ingredients in the drug prepared from the camomile obtained according to the invention, in particular the main active ingredients, chamazulene and (-) - alpha-bisabolol, depends on the vegetation stage at which the flowers are at the time of harvest and before drying the harvested flowers. The higher the drying temperature, the more pronounced the decomposition of the active ingredients, i.e. the lower the active ingredient content in the dried flowers. For the same reason, direct sunlight has an adverse effect and should be avoided as far as possible. The vegetation stage of the flowers is characterized by how many% of tubular flowers are open at a certain time (here, the harvest time is meant). Flowers can also be harvested if, for example, 30 to 50%, 30 to 70%, 40 to 60%, 60 to 100% or 90 to 100% tubular inflorescence are open. The content of the active substances depends on the stage at which the flowers are always present and is highest when 40 to 60% of all tubular flowers are open, and is lower for less or more open tubular flowers.

Thus, a great advantage of the camomile obtained according to the invention is that it is evenly flowering, i.e. the predominant number of plants after sowing has the same flowering stage, which means that, for example, most plants have 40 to 60% of all tubular flowers open at the same time. . This makes it possible to achieve a perfect collection of inflorescences at the optimum time with the camomile obtained according to the invention and this camomile is suitable for mechanized harvesting.

The relationship of the chamazulene and (-) - alpha-bisabolol content to the flowering stage at the time of harvest and to the drying temperature of the drug is shown, for example, in the following table (the content of other bisaboloids in all cases being less than 50%).

table

Drying temperature (air temperature)

Vegetation stage of flowers at harvest time up to 70% of all 70 to 100% of all tubular flowers tubular flowers

Table continuation

Content of active substances in the dried material

Content of active substances in the dried material

up Deň: 32 ° C Chamazulene: at least 100 mg% (-) -alpha-bisabolol: at least 200 mg% Chamazulene: at least 40 mg% (-) - a-bisabolol: at least 120 mg% Content of active substances in alcohol extract: Content of active substances in alcohol extract: up Deň: 32 ° C Chamazulene: at least 5.0 mg% (-) - a-bisabolol: at least 10.0 mg% Chamazulene: at least 2.0 mg% (.-) - alpha-Bisabolol: at least 6.0 mg% Active ingredient content in the dried material; Content of active substances in the dried material: between 50 and 70 ° C Chamazulene: at least 50 mg% Chamazulene: at least 30 mg%

temperature Vegetation stage of flowers at harvest time drying / temperature 30 to 70% of all 70 to 100% of all air / tubular flowers blooming tubular flowers-blooms- • openness openness (-) -alpha-bisabolol: <-? - a-bisabolol: at least 150 mg% at least 100 mg%

between 50 to Content of active substances in alcohol extract: Content of active substances in alcohol extract: 70 ° C chamazulene: chamazulene: at least 2.5 mg # at least 1.5 mg # (-) - a-bisabolol: (-) - a-bisabolol: at least 7.5 mg # at least 10.0 mg #

Consequently, from the vermilion used to manufacture the compositions of the invention, for example, harvesting takes place at the returning stage when 30 to 100% of tubular inflorescences are open and drying can be carried out at air temperatures up to 70 ° C. , a dry material having at least 30 mg # of a chamazulene is obtained and whose (-) - alpha-bisabolol content is at least 100 mg of a #, while the content of other bisaboloids is less than 50 mg%.

In alcoholic extracts, the content of other bisaboloids is still less than 10 mg #.

The content of essential oil produced from dry material under the conditions in the above table is still at least

3,5% chamazulene, at least 10% (-) - alpha-bisabolol and not more than 10% other bisaboloids.

The determination of (-) - alpha-bisabolol, as well as other ingredients for camomile, is carried out by the usual gas-chromatographic method.

The exact method of analytical determination is given in Annex A / after [the text of example no. 4 /.

I

The active substance chamazulene is not found in camomile flowers per se but in the form of the sesquiterpenic lactone of matricin.

Matricin exhibits pharmacological activity close to chamazulene. This precursor forms matrix, for example, upon heating (e.g., steam distillation to prepare tea infusion), immediately chamazulene. It is therefore customary for camomile not to report the content of matricin, but the content of the chamazulene that forms.

Possible uses of the camomile obtained according to the invention are as follows: Obtaining the drug camomile, camomile oil, camomile extracts, as well as (, -) - alpha-bisabolol, chamazulene and other camomile ingredients. For example, it is possible to obtain the camomile extracts or extracts from the drug obtained from clove of the invention, for example, by extraction with an alcohol or an aqueous-alcohol mixture, or by supercritical gas extraction. It is also possible to obtain camomile essential oil, β -) - alpha-bisabolol, chamzulene and other camomile-containing substances from the drug.

Alcohol extracts of the drug contain, for example, at least 1.5 mg%, preferably 5.0 mg% chamazulene and at least 5.0 mg%, preferably at least 10 mg% -alpha-bisabolol in the alcohol extract of the drug and less than 8 mg% of the other bisaboloids . The production of such extracts is carried out in a conventional manner.

For example, instrumentation, for example so-called trough mixers, percolators and other suitable extraction devices, may be used for extraction. The temperature during extraction ¹ is, for example, 10 to 50 ° C. Cooling is not required.

Suitable solvents are, in particular, straight-chain or branched aliphatic mono- or polyhydric alcohols having 1 to 6 carbon atoms, such as methanol, ethanol, 2-propanol, butanol, glycerol, isopropylidene glycerol and others, as well as a mixture of these solvents with water.

Mixtures of these solvents may also be used. The minimum amount of solvent is 2 parts solvent per 1 part of dried material. Usually 2 to 20 parts of solvent are used per part of dried material, in particular 3 to 10 parts of solvent per part of dried material.

For the production of extracts, in particular alcohol extracts, it is also possible to use fresh camomile flowers or frozen camomile flowers (if the fresh flowers have been frozen).

The essential oil prepared from the drug contains at least 3.5% chamazulene, preferably at least 5% chamazulene and at least 10% (-) - alpha-bisabolol and less than 10% other bisaboloids.

The production of such essential oil is usually carried out by heating the drug with water, optionally in the presence of ascorbic acid (e.g. in the form of a salt, in particular sodium ascorbic acid) to boiling at a pH between 4 and 6, preferably 5 to 5.5. The pH is adjusted with an acid such as hydrochloric acid. For example, 10 to 50 parts by weight of water and optionally 0.1 to 1 part by weight of ascorbic acid are used per part by weight of the dried material.

Heating is carried out for 2 to 8 hours.

The aqueous distillate obtained is repeatedly shaken with a lower aliphatic liquid hydrocarbon such as petroleum ether (e.g. boiling point 35 to 60 °), pentane, xylene, decalin, the organic phase is dried (e.g. over sodium sulfate), and the organic solvent is removed. in a gentle manner (e.g. by distillation on a rotary evaporator or distillation at 40 to 70 ° C, preferably at 50 to 60 ° C). In the case of higher boiling solvents, this distillation is carried out under vacuum;

The determination of the constituents is carried out as previously described.

When the camomile flowers are harvested at the vegetation stage when 30-100% of tubular inflorescence is open, and the flowers are dried at an air temperature of up to 70 ° C, then the drug obtained contains at least 30 mg% chamazulene, at least 100 mg%. -alpha-bisabolol and less than 50 other bisaboloids and silica obtained by steam distillation from this drug contained at least 3.5% chamazulene, at least 10% (-3alpha-bisabolol and less than 10 mg% other bisaboloids and alcoholic extract prepared by extraction with lower contains at least 1.5% chamazulene, at least 5.0 mg% of C - alpha - bisabolol and less than 10 mg% of other bisaboloids.

When the camomile flowers are harvested at the vegetation stage, when 30-70% of tubular flowers are opened and the flowers are dried at a maximum of 50 ° C, then such drug contains, for example, 100 mg% chamazulene, at least 200 mg% (-) - alpha-bisabolol and less than 50 mg% of other bisaboloids and the silica obtained therefrom contains at least 3,5% chamazulene, at least 10 mg% of other bisaboloids, and the alcoholic extract of camomile produced with lower alcohols contains 1,5 mg% chamazulene, 5,0 mg% (- -alpha-bisabolol and 10 mg% of other bisaboloids.

If the flowers; they are harvested at the vegetation stage when 30-70% of tubular inflorescence is open and the flowers are dried at an air temperature between 50-70 ° C, then the drug obtained contains, for example, at least 50 mg% chamazulene, at least 150 mg% (-) - alpha -bisabolol and less than 50 mg% of other bisaboloids, and the silica obtained therefrom contains not less than 3,5% chamazulene, not less than 10% 6 -> - alpha-bisabolol and less than 10% other bisaboloids and of which the alcoholic extract obtained contains not less than 2,5 mg of chamazulene, at least 7.5 mg% of (-) - alpha-bisabolol and less than 10.0 mg% of other bisaboloids.

If camomile flowers are harvested in the vegetation stage, when 70-100% of tubular flowers are open and the flowers are dried at an air temperature between 50-70 ° C, then the drug thus obtained contains at least 30 mg% chamazulene, at least 100 mg% (- ) -alpha-bisabolol and less than 50 tngfr of other bisaboloids, possibly prepared therefrom, containing at least 3,5% chamazulene, at least 10% q -) - alpha-bissbolol and less than 10% other bisaboloids and alcoholic extract prepared from this drug it contains not less than 1,5% chaciazulene, at least 10 mg% (-) - alpha-bisabolol and less than 10,0 mg% other bisaboloids.

29 Examples

Example - 1 / The starting romance is tetraploid /

Testing the variability of individual plants from the number of 10,000 plants of the tetraploid camomile variety described

I. Sarkány / Herba Hungar. 4 (1), 125-169 (1965), it has been found that for about 1,000 individuals there is one plant having a high chamazulene content of 20% by weight and a high content of (-) - alpha-bisabolol and at the same time the content of other bisaboloids, in particular β-7-alphabisabolone oxide, is very low (less than 5% by weight). Subjects were selected and subjected to the following steps:

Stage 1:

From the offspring (as described above) the selected tetraploid camomile plants were selected those individuals that

A / they bloom at about the same time,

B) have a uniform base branch and a narrow flower zone, about cm, preferably 5 cm,

C / have large inflorescences with an outer diameter of about 30 mm, preferably 25 to 35 mm,

D / reach or exceed the minimum chamazulene content

150 mg% and bisabolol 300 mg% and their content of other bisabolo-30 loids (especially bisabolol oxides) are clearly below 50 mg. / All values refer to inflorescences dried at 40 ° C and harvested at a stage where 30 to 70 ·% of all tubular inflorescences are open. /

These plants were cloned. To this end, the plants (cloned mother plants) were cut to about 15 cm of shoot length, at 8 to 10 hours of day length and at temperatures of 12 to 14 ° C, to produce new shoots (short side shoots). These short shoots were trimmed and planted in a mixture of peat and sand. Rooting of seedlings / divers / at 100% relative humidity and a 14 day duration lasted 7 to 14 days.

| T

Instead of the above cloning (propagation by seedlings), in vitro propagation of the tissue sections of the plant capable of dividing (the so-called meristem division) was also used. Various parts of the plants, preferably the ends of the shoots or the buds of the claws, are suitable for establishing a camomile culture.

After rinsing the plants with aseptic conditions in a laminar flow / low turbulence displacement high-performance filter /, the shoot ends or armpit leaf buds are removed and transferred to nutrient reagent containers, for example according to Murashig and Skoog / Physiol. Plent. 15, 473-497, 1962]. The reagent vials are placed in an air-conditioned space at 12-18, preferably 16 hours of day length (attained by fluorescent lamps) at an illumination intensity of 100 to 1000 lux, preferably 1,000 to 3,000 lux and a temperature of 15 to 30 ° C, preferably 22 ° C. to 27 ° C. Once it is clear that the explanations grow well, they are transferred to the above-mentioned nutrient medium, but with a higher concentration of cytokinin * (e.g. 30 mg (16-isopentenyladenine) and with little or no auxin activity) 0 to 0.3 mg / l indolylacetic acid). This is followed by the growth of the stems and the reversing organs as well as the proliferation of the armpit buds are formed. These can be harvested and grown as described above. '

For the multiplication of plants to be multiplied, explanations / third passages, i. 3 · of the reproductive generation /, after growth and formation of leaves on the first mentioned medium, are transferred to a culture medium containing 10 mg / l of indolylacetic acid or 3-indolylbutyric acid or 0,1 to 0,3 mg / l of α-naphthyl acetic acid . The plants are rooted after about 4 weeks and can be grown in sterilized garden soil (steamed for 12 hours at 120 [deg.] C. (in pots and greenhouse) under conditions usual for seedlings).

- 32 Murashig and Skoog environment:

'tng / 1

NH.NO. 4 3 400 Ca / NO _3/2 .4H ₂ O 144 kno ₃ 80 kh ₂ Mon ₄ 12.5 MgSO ₄ .7H ₂ 0 72 KC1 65 NaFe EDTA 25 h ₃ or ₃ 1, 6 MnSO ₄ .4H ₂ 0 6.5 ZnSO ₄ .7H ₂ 0 2.7 KI 0.75 indolylacetic acid 2.0 furfuryladenín 0.1 thiamine 0.1 nicotinic acid 0.5 pyridoxine 0.5 glycine 2.0 myo-inositol 1 00 acid hydrolyzate 1 000 sucrose 2% Purified agar powder 1%

Note to previous interpretation above:

* Cytokinins are phytohornones that promote cell division.

Stage 2:

The plants obtained by the Stage 1 stage bloom together under isolated greenhouse conditions. The plants stood in 1 1 cm pots filled with garden soil at 18-24 ° C by day and 12-14 ° C at night. The length of the day was at least 14 hours and was achieved by additional lighting of 200 Watt / m ^ in the winter half of the year. Water was supplied as needed.

Of the total number of selected individuals, inflorescences that blossomed shortly before disintegration were continuously harvested over a period of 4 weeks to obtain seed. Drying took place in a well ventilated hall at an air temperature between 20 and 30 ° C; the seed was then sieved through a sieve (5 x 0.4 mm) and screened using a sieving machine.

Stage 3:

A random sample was taken from the seed obtained in Stage 2 and about 2,000 offspring were grown (ecological conditions as in Stage 2) and selected according to the same criteria A / D / Stage 1. The individuals so selected were followed by the procedure of step 2.

Stage 4:

The seed of step 3 was sown in autumn at two different sites.

- 34 Position 1 /

450 m NN, 48.5 ° N / 11.5 ° 0

750 mm total-year rainfall, humid temperate climate, January -10 to 0 ° C, July +10 to 20 ° C;

Abbreviations explained:

NN = North Altitude ° N = North latitude / n-degree / ° 0 = East longitude / n-degree /.

Position II /

200m NN, 42 ° N / 1 ° N,

400 mm of total annual rainfall, Mediterranean climate, January 0 to +10 ° C, July +20 to +30 ° C.

Sowing took place at both sites at the end of September / early October.

The results of the field trial thus obtained were examined and evaluated for homogeneous growth, flower size and harvest time, in addition random flower samples were examined for drug content. From the field experiment, individuals corresponding to the above-mentioned parameters from step 1 were again selected.

Second

Stage 5:

With seed of step 4, steps 3 and 4 were repeated in the sequence and procedure indicated.

Stage 6:

From the seed obtained in step 5, about

50C individuals (ecological conditions as in step 2) and according to the same principle as in step 1 were selected. From the selected plants, 34 individuals were selected and cloned according to step 1. 10 plants of each clone were grown in random distribution at a distance of 40 x 30 cm freely in the field (site I, see step 4) in an isolated location. A soil permeable clay having a pH of 7.0; cultivation was carried out in early June, the first harvest in the middle of July, then the plants were trimmed, bloomed again and provided a second seed harvest from Wednesday to the end of August.

The seed obtained from this material was carried out as in step 2.

The seed obtained according to step 6 is seed for multiplying the camomile of the invention.

Plant flowers from this seed for propagation (sowing September / October, harvested at the beginning of June of the following year), which are harvested at a time when 30 to 70% of tubular flowers are open and then lambs are dried at 40 ° C in a drying cabinet 72 hours, based on the weight of the dried flowers (dry matter), for example, at least: 150 mg% of chama zulen, 300 mg% of (-) - alpha-bisabolol and a maximum of 50 mg% of the other bisaboloids.

Further exemplary data for plants obtained according to the invention:

1. Growth

Stem: straight, little branched

2. Listing:

Leaf: 2 and 3 folded

Strength: medium feathered leaf, color medium green feathered leaf (center of stem); feathered composition; Medium to strongly folded.

3. Inflorescence

Flower heads: outer diameter about 30 mm, inner diameter about 15 am;

Weight of single inflorescence (dried):

about 45 mg;

Flowering stem, length (mm): about 700, but dependent on the site (climate), sowing time, soil, fertilization, plant treatment, weather;

Beginning of flowering / calculated from 1 January /:

about 160 days / sowing September, Freising,

Germany (Federal Republic of Germany, otherwise dependent on the above factors).

- 37 Flowers: middle of June (sentences above);

Flowering without stem, essential oil content (% dry matter): about 1.0%;

azulene content in essential oil: minimum 15

Disintegration of dried inflorescences, flower drug: slight, if harvested before opening last tubular flowers Diameter peíu: about 30 p®;

Seed length: about 1.25 mtn;

Number of chromosomes in somatic cells: 4n = 36.

4. Fetus weight 1,000 seeds (TKM): 0.06 to 0.13 g;

5. Germination (CF) about 75

6. Purity up to 95%.

7. Other features

Drying ratio of fresh: dry flowers = 5.5 to 6: 1, characteristic aroma of the drug, slightly aromatic, typical taste of tea infusion.

In addition, the following properties may exist:

Uniform height-balance / with narrow flower zone, especially suitable for mechanized harvesting, large flower baskets (inflorescences), medium yield; base branched form (three to five times).

Obtaining a drug:

The camomile obtained according to this example is sown in the usual field. Once the plants have developed and the quota is in the vegetation stage, when about 50% of the tubular flowers are open, the camomile is harvested with a camomile harvesting machine, with the ridge spacing of the machine set to pick only inflorescences whose tubular flowers are open. 50%. The crop is transported as quickly as possible to a shaded place and spread there in a thin layer until further processing. Thereafter, the inflorescences are removed by means of a sieve device. and dried to constant weight. The weight loss is about 80%. The drying takes place in the shade in a well ventilated place in a layer with a thickness of about 10 cm.

After about 2-3 days, drying is complete. To remove the parts of the herbs and the flowering plant, the rum leaf drug is sieved and pressed into bales.

Analysis of silica:. > 960 mg% chamazulene; 162 mg% (-) - alpha-bisabolol 330 mg%.

If the drying is carried out, for example, in a stationary belt dryer using artificially heated air at a temperature of between 50 and 70 ° C, then drying is completed after about 4 to 5 hours. To remove parts of herbs and flower waste, the drug is sieved and then pressed into bales. The analysis values for such a drug are as follows:

- 39

(-) - alpha-bisabolol: chamazulene:

870 mg%

197 mg% 94 mg%.

Example 2 (The starting romance is tetrapioid)

Individual plant variability test carried out on 10,000 plants of the tetraploid vermilion variety according to I. Sarkány / Herb Hungar. 4, 1, 125-169, 1965], it has been found that about 1,000 individuals have one plant having a high chamazulene content of 20% by weight and a high content of (-) - alpha-bisabolol: 50% and at the same time the content of other bisaboloids, in particular. The (-) - alpha-bisabolonoxide is very low (less than 5% by weight).

These individuals were selected and subjected to the following steps:

Stage 1:

Individuals having a minimum chamazulene content of 100 mg% and (.-) - .alpha.-bisabolol of 200 mg%, or having a content of other bisaboloids (especially bisabololoxides) of less than 50 mg, were selected from the progeny of the above-described tetraploid camomile plants. %. All values refer to inflorescences that have been dried at about 40 ° C and harvested at a stage where about 30 to 70% of all tubular inflorescences have been opened.

Stage 2:

All the flowering flower heads (inflorescences) were removed from the plants obtained from the Stage 1 stage, then they were in a greenhouse and flowering under isolated conditions annually. The plants stood in 11-cm pots filled with garden soil at a daily temperature of 18 to 24 ° C and a night temperature of 12 to 14 ° C. The length of the day was at least 14 hours and was achieved in the winter half with additional lighting of 200 W / m. Water was supplied as needed.

From the total of selected individuals, inflorescences that were blooming shortly before disintegration were continuously harvested over a period of 4 weeks to obtain seed. Drying was performed in a well ventilated hall at an air temperature between 20 and 30 ° C; then the seed was sieved using a sieve screen (5 x 0.4 mm) and further renumbered on a shaking screen.

Stage 2 provides seed which is seed for multiplying camomile for the processes of the invention.

Flowers from plants of this propagating seed (sowing September-October, harvested at the beginning of June of the following year), which are harvested at the time when 30 ε, ζ 70% of tubular flowers are open and which are then dried in a drying cabinet at 40 ° C 72 hours, containing at least 100 mg% chamazulene, at least 200 mg% <-) - alpha-bisabolol and at most 50 mg # of other bisaboloids, based on the weight of the dried flowers.

Obtaining a drug:

The camomile obtained according to this example is sown in the usual field. Once the plants have developed and flowering - the vegetation stage is about 50% of the tubular flowers open, these inflorescences are harvested using a camomile harvesting machine, with the ridge distances of the harvesting machine being set to tear off only the flowering heads / inflorescences whose tubular flowers are open to 50%. The harvested material is conveyed as quickly as possible to a shaded place and is then smeared in a thin layer until further processing. In a subsequent operation, the flower heads (inflorescences) in the sifter are removed from the stalks and dried until a steady weight is reached. The weight loss is about 80%. Drying takes place in the shade in a well ventilated place on platforms with a layer thickness of about 10 cm.

After about 2 to 3 days, drying is complete. To remove

sieves on the sieves and flower waste with a camomile and in the wrist is pressed into bales. analysis: silica: 940 □ g% chamzulén: 132 mg% (-) - a-bisabolol: 243 mg%.

If the drying is carried out, for example, in a stationary belt dryer using artificially heated air at temperatures between 50 and 70 ° C, the drying is completed after about four to five hours. To remove part of the stems and flower dust, the drug is sieved on sieves and then pressed into bales.

For example, the values obtained by analyzing such a drug are:

silica:

chamazulene (-) - alpha-bisabolol

775 tng

163 mg%.

Example 3 (The starting romance is diploid)

By means of variability assays of individual diploid camomile plants containing a small amount of bisabolol but not without bisabolol as described by Ch. Franz and J. Hazl and A. Vmelem and Acta Eorticulturae 73, 109-114 (1978) found that up to 20% of all growth was found. The liner has a content of about 20% by weight and / or a high content of 50% by weight ( -) - alpha-bisabolol, while at the same time the content of other bisaboloids (especially α-β-alpha-bisabolooxide) is very low. These individuals were selected and tetraploidized as follows:

Seeds of the selected camomile plants were seeded on filter paper soaked with a 0.05% aqueous colchicine solution and allowed to swell for 6 hours. They were then removed from the filter paper, washed several times with water and sieved in a greenhouse. Soil: 1: 1 peat / sand mixture, temperature: 18 to 20 ° C, real humidity: about 63%, daylight duration: 14 hours.

Germinating plants were observed until flowering, success, polyploidization was evaluated by comparative measurement of the size of the pea and seed, as well as the number of plant chromosomes (F ^ = the first progeny grown from 'seed' confirmed as tetraplid colchicine plants).

Tetraploidization can also be performed as follows:

Five- to seven-day well-developed camomile plants germinated on water-moistened filter paper were plated at room temperature (20 ° C) for 4 to 6 hours with the seed sheets down to a 0.05% colchicine solution. The sensitive germ roots were treated in order to prevent damage thereto, when drying, the ambient atmosphere must contain approximately 100% real atmospheric humidity. After treatment, the germinating seedlings were washed several times with water and plated in hazelnuts. This was followed by more treatments than seeds.

The size of the pei was measured using a Leitz binocular microscope with a micrometric measuring eyepiece and a micrometric object carrier.

Chromosome counting is done at the tip of the roots: Fresh green root tips 1 to 2 cm long were taken from young plants grown in the greenhouse or divers and immersed in a 0.002 molar solution of hydroxycholine for 5 hours and then placed in 1N HCl for 15 minutes. For screening, about 1 mm of the root apex was stained with a 2% orcein acetic acid solution and examined by oil microscopy. Thus, four times the number of chromosomes / somatic cells (4n = 36) can be determined in cells found in mitosis.

Those plants in which the diameter of the pee is about 50% greater than _{that of the} diploid starting material (about 30 instead of 20 pm) and those where the chromosome number has been doubled to 3.6 (for diploid starting material is44 leading number 18) are tetraploid . These are selected. Approximately 0.1 to 0.5 # of seeds or germinating seedlings were tetraploidized as described above and flowering intact plants were developed. Now steps 1 and 2 correspond to Example 2.

The flowers of the resulting camomile contain, for example, at least 100 mg of # chamazulene, 200 mg of # (-) - alpha-bisabolol, and at most 50 mg of # other bisaboloids / harvest at a time when 30 to 70 # of all tubular flowers are open and dried for 72 hours in an oven at 40 ° C /.

I

Obtaining a drug:

The camomile obtained according to this example is sown in the usual field. When the plants unfold and the inflorescences are in the vegetation stage, when about 50 # tubular flowers are open, these inflorescences are harvested mechanized, with the ridge distances of the harvesting machine selected to pick only those inflorescences whose tubular flowers are open at 50 # . Preferably, the harvested material is transferred to a shaded area and blurs in a thin layer there until further processing. Thereafter, the inflorescences are de-sieved by sieving the remnants of the stems and dried until they reach a constant weight. The weight loss is about 80%. The drying takes place in the shade in a well-ventilated place in a layer with a thickness of about 10 cm.

Drying is completed after about 2 to 3 days.

The drug is screened and then compressed into bales to remove stalk residue and flower waste.

Analysis:

silica chamazulene (-) - alpha-bisabolol

910 mg%

117 mg%

252 mg%

Example 4 (The starting romance is diploid)

Using the variability test of single diploid camomiles containing only a small amount of bisabolol but not without bisabolol, described by Ch. Franz, J. Hazl and A. Vämel in Acta Horticulturae 73, 109-11 (1978) found that a maximum of 20% of all individuals in the essential oil had a chamazulene content of about 20% by weight and / or a high content of? -J-alphabisabolol of 50% and the content of other bisaboloids, in particular (-) - alpha-bisabolonoxide, is very low. These individuals are selected and, exactly as shown in Example 3, are tetraploidized and the tetraploid individuals are selected. About 0.1 to 0.5% of the seeds or seedlings were tetraploidized and developed viable intact plants.

For example, grades 1 to 6, corresponding to Example 1, follow.

For example, the flowers of the camomile contained at least 150 mg% of chamazulene, 300 mg% of C - .alpha.-bisabolol and at most 50 mg% of other bisaboloids / harvest while 30 to 70% of tubular flowers are open and then 72 hours of drying in a drying cabinet at a temperature of 40 C /. .

- 46 The other characteristics correspond to the camomile obtained according to Example 1.

Obtaining a drug:

The camomile obtained according to this example is sown in a conventional manner in the field. Once the plants develop and the inflorescences are in the vegetation stage, when about 50% of the tubular flowers are open, these inflorescences are harvested by a harvesting machine, with the ridge distances adjusted to: tear only those inflorescences whose tubular flowers are open to 50%. The crop is transported as quickly as possible to a shaded place and there is blurred in a thin layer until further processing. The inflorescences are then de-sieved and sieved to constant weight. The weight loss is about 80%. The drying is carried out in the shade at a well ventilated place in a layer of about

0 cm.

After about 2 to 3 days, drying is complete, the camomile drug is sieved to remove stem residue and flower waste and then compressed into bales.

analysis:

silica 1020 mg% chamlulene 173 mg% (-) - alpha-bisabolol 418 mg%

Annex A

The starting material is the camomile drug obtained according to the invention. Only those flowers whose 30-70%, in particular 40-60%, of tubular flowers are open are used in drug production. Drying is carried out in a box dryer at 40 ° C for 72 hours. The essential oil of camomile flowers is obtained, as described below, by two-hour steam distillation of the drug:

2.0 g of the uncrumbed drug are mixed in a 1 liter round-bottomed flask with 250 ml of desalinated water and subjected to distillation on a Clevenger reflux apparatus (water steam distillation apparatus) for quantitative evaluation of small amounts of essential oils. 1 ml of pentane of analytical purity serves as a template. The reflux temperature (reflux rate) is 40 + 4 drops per minute. Upon completion of the distillation, the pentane dissolved silica, preferably anhydrous, is discharged into test tubes and the silica, optionally adhered to the apparatus, is rinsed to a further amount, pentane. To remove any residual water, add a small amount (per tip of the spoon) of anhydrous sodium sulfate to the solution and

by evaporating pentane at 40 ° C in a water bath and drying in a desiccator, the amount of essential oil is determined gravimetrically.

The oil thus obtained (about 20 mg) is then determined for chamazulene and bisabolol.

Spectral photometric determination of chamazulene

Specific solution: The entire amount of essential oil obtained from 2 g of the drug (about 20 mg) obtained as described above is dissolved in 25 ml of n-hexane or cyclohexane.

Measuring device: Photometer with filter / eg Eppen- '.

Dorf /.

Wavelength:

578 nm

cell:

I

Specific extinction cm chamazulene / 1 g / 100 ml; 1 cm / 20.8

Compensation liquid: n-hexane or cyclohexane

Found content:

in ^{1 20 E} 578 chamazulene mg / 100 g:

a photometer with a filter is available for measurement, this measurement can also be performed using a spectral photometer as follows:

unless

Measuring device:

Spectral photometer / for example

PM QII or PM QIII Zeiss’7

Wavelength:

605 nm

cell:

cm

Specific extinction of chamzulene / 1g / 100 ml; 1 cm /:

24.5

Compensation fluid:

n-hexane or cyclohexane

Found chamazulene content in mg / 100 g:

^{'02 E} 6C5

Bisabolol is then determined in the measured solution.

- 49 Gas-chromatographic determination of bisabolol and other bisaboloids

Gas chromatograph: Hewlett Packard model 5750, Erba Fractovap 2350 or similar device detector: flame ionization detector Carrier gas: helium column: 3.175 mm (= 1/8 inch); 200 cm; Ssteel Column fill: 3% nitrilsilicone rubber XE 60 on silanized silica gel Chromosorb WAW HP 125 to 150 pm as carrier Detector temperature; 320 ° C injection block: 220 DEG column: Mp 85-220 ° C Temperature programming: 4 ° / minute Sample solution: a solution is used to measure chamazulene Reference solution: about 15 mg of Standard Bisabolol is dissolved in cyclohexane to a volume of 25 ml Injected amount: 5 µl of sample solution and reference solution

evaluation: Evaluation is performed by comparing the peaks / peak areas /. I Bisabolol content, in mg per 100 g of drug: 10 x weight / comparison / / mg / x area / sample / area / comparison /

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

Appliances: Packard, model 7721, 800 series, optionally Erba Fractovap, series 2350 Columns: glass columns 3o / 2 mm in diameter; 2 m / 2 mm in diameter Filling: 3% methylphenylsilicone rubber. ´ OV 1 on silanized Gaschrom Q ”125 to 150 μαι as carrier Carrier gas: 30 rpm N g Temperature program: 80 to 180 ° C, 2.5 / 3 / ° C / minute Injector / detector temperature: 200 ° C detector: flame ionization detector

Injected amount: about 2 jl of about 1:50 diluted oil

The evaluation is partly without, partly with an internal standard. As the internal standard, methyl laurate or hexadecane is suitable for chamazulene-poor essential oils, optionally without chamazulene. For oils with a content of more than 5%, this is to be preferred as the internal standard, the determination being carried out photometrically (at 578 nm).

Example 5

Production of camomile drug extract

The camomile flowers for the drug used are harvested according to Example 1 at a time when 30-70% of the tubular flowers are open and are dried at a temperature not exceeding 50 ° C.

400 g of camomile drug containing

105 mg% of chamazulene and 212 mg% of (-) - alpha-bisabolol are extracted with 200 g of an aqueous ethanol solution (40% ethanol by weight) in a trough mixer at 30 rpm for 3 hours. The drug material is then compressed and the extract is filtered.

In the extract, the active substance content is determined in a known manner:

chamazulene: 3.7 mg% (-) - alpha-bisabolol 10.2 mg%.

Example 6

Preparation of fresh camomile extract

I

758 g of fresh camomile flowers harvested according to Example 1 at the time 30 to 70% of tubular flowers were opened (water content 74%) are extracted in a trough mixer at 65 rpm for 30 minutes with 510 g of ethanol () 84% by weight. The remainder of the drug is compressed and the extract filtered.

The extract is determined in a manner known per se by the content of active substances:

chamazulene:

(-) -. alpha.-Bisabolol

11,7 Og%

16.3 mg #.

Claims (2)

PATENT CLAIMS First Process for the preparation of a new bisabolol-rich tetraploid camomile of the plant camomile Chamonilla recutita L. synonymous with the chamonilla L., Asteraceae), which contains dried flowers at 40 ° C. based on dry matter at least 100 mg% chamazulene at least 200 mg% C-1-α-bisabolol and less than 50 mg% of other bisaboloids. except for the variety of camomile Planzana, in that u 1,000 to 10,000 Single plants of diploid or tetraploid camomile populations determine the content of C-) - bis-bisabolol in essential oil and select those camomile plants for which ζ -> - α-bisabolol is the major constituent of essential oil and for which flowers dried at 40 ° C, containing at least 100 mg% of chamazulene, at least 50 to 100 mg% of C - .alpha.-bisabolol and less than 50 mg% of other bisaboloids, these plants are separated, multiplied from the initial population and subjected to further selection and multiplication steps; and where appropriate, seed for propagation and / or camomile drug is obtained from the plants of the resulting camomile, using, where the starting camomile is used, Even the diploid camomile, which excludes the Degumill variety, will be tetraploidized with chemicals at temperatures between 0 and 35 ° C, by gamma-ray, X-ray or UV radiation at 0 to 35 ° C after all selection steps have been completed. , by means of elevated temperatures in the range of from 33 to 50 ° C, by means of lower temperatures in the range of from 0 to 5 ° C, by the decapitation-callus method or by anthropogenic cultures. Method according to claim 1, characterized in that the selection or multiplication consists of the following steps: (a) selecting at least 10 individuals of tetraploid plants according to the active substance content, that is to say at least 100 mg% chamazulene, at least 200 mg% bisabolol and less than 50 mg% other bisaboloids, all based on dry matter, current flowering time, as well as a flower size of 20 to 40 mm, cloning the plants so selected and obtaining seed from the cloning plants, (b) growing offspring from the seed obtained in accordance with paragraph (a), the subsequent selection referred to in paragraph (a) and obtaining from seed from the plants so selected, (c) three to five times the repetition of the measures referred to in paragraph (b); (d) growing offspring from the seed obtained under (c), selecting said offspring under (a), cloning the plants thus selected and obtaining seed from the plants obtained by cloning.