SK280689B6 - Process for extracting tetraploid camomile enriched with bisabolol - Google Patents

Abstract

Process for extracting tetraploid camomile of cultural plant species (Chamonilla recutia L., source plant with Matricaria chaminilla L., Asteraceae) enriched with bisabolol, of which the flowers dried at 40 Degree Celsius contain at least 100 mg per cent of chamazulene, at least 200 mg per cent (-)-alpha- -bisabolol and less than 50 mg per cent of other bisaboloids excepted cultivator of Manzana camomile wherein percentage of (-)-alpha-bisabolol in essential oil is determined in 1.000 to 10.000 individual plants of diploid or tetraploid population of camomile and the plants of camomile are selected in which (-)-alpha-bisabolol is the main component of the essential oil, and in which flowers dried at 40 Degree Celsius contain at least 100 mg per cent of chamazulene, at least 50 to 100 mg per cent of (-)-alpha-bisabolol and less than 50 mg per cent of other bisaboloids, these plants are separated from the initial population, multiplied and are subjected to various selection and propagation steps and seedcorn can be acquired for propagation and/or for camomile drug from this sort of process of extracting camomile.

Description

Technical field

The present invention relates to a process for the preparation of a novel bisabolol-rich tetraploid camomile of the plant species Chamonilla recutita L., synonymous with Matricaria chamonilla L., Asteraceae, whose flowers, dried at 40 ° C, contain at least 100 mg% chamazulene , at least 200 mg% of (-) - α-bisabolol and less than 50 mg% of other bisaboloids, with the exception of the variety of camomile Manzan.

BACKGROUND OF THE INVENTION

Chamomile blooms (Chamonilla recutita (L.) Rauschert, synonymous with Matricaria Chamonilla, L.) have a wide therapeutic use due to their antiphlogistic and spasmolytic effect and form an important part of the herbal medicinal raw material range. Particular therapeutic importance is ascribed to the active ingredients of (-) - α-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 substances of chamazulene and (-) - α-bisabolol is available, or the active substances are missing or present in very small quantities, in which cases the main constituent of essential oil consists of less active bisaboloids (bisabolonoxide A or B, bisabololoxide A).

DEGUMILL (a variety of camomile, protected and listed in West German Patent 24 02 802; a protected variety registered in the former GDR under the name Degumill; tal. Patent No. 1,035,096) is known as a variety of camomile, which contains at the same time a high (-) - α-bisabolol and chamazulene content. The chamazulene content of this camomile variety DEGUMILL is stable only if accidental crosses or foreign pollination by other varieties of camomile are avoided, the content of bisabolol and chamazulene being substantially lower, and 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 tetraploid forms of camomile are known (for example, BODEGOLD, GDR; POHOHELICKÝ, CR and SR; ZLOTY LAN, Poland; BK-2, Hungary). These tetraploid forms have a satisfactory chamazulene content, but the important active ingredient (-) - α-bisabolol is only very small in these tetraploid species, and the other bisaboloids (bisabol A and B as well as bisabolonoxide), which account for more than half essential oils in known tetraploid camomile species.

SUMMARY OF THE INVENTION

It has now been found that quite generally it is possible to obtain from the naturally occurring populations of camomile, whether diploid or tetraploid cultivated camomile, by a certain method according to the invention biscuitol-rich tetraploid camomile plants which on the one hand are no longer susceptible to foreign pollination. they also have a high content of chamazulene and (-) - α-bisabolol, while the content of (-) - α-bisabol significantly exceeds the content of chamazulene and at the same time other bisaboloids are virtually absent or only to a small extent. It is also important that the high content of the main active ingredients (-) - α-bi-sabolol 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 of the plant camomile (Chamonilla recutita L., synonymous with Matricaria chamonilla L., Asteraceae), the dried flowers of which at 40 ° C contain at least 100 mg chamazulene, at least 200 mg% of (-) - α-bisabolol and less than 50 mg% of other bisaboloids, with the exception of the Manzana variety, which is based on the determination of 1000 to 10 000 individual plants of the diploid or tetraploid camomile population. (-) - .alpha.-bisabolol content and select those camomile plants in which (-) - .alpha.-bisabolol is a major constituent of essential oil and wherein the flowers, dried at 40 DEG C., contain at least 100 mg% chamazulene, at least 50- 100 mg% of (-) - 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, optionally, from the plants of the resulting camomile, seed for propagation and / or the camomile drug is obtained, using a diploid camomile from which the Degumill variety is excluded as the starting camomile, and tetraploidization takes place after all selection steps have been completed. by chemicals at temperatures ranging from 0 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 lowered temperatures ranging from 0 to 35 ° C 5 ° C, using the decapitation-callus method or using anthracite cultures.

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

(a) selecting at least 10 individuals of tetraploid plants according to the active substance content; j. 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, current flowering time, uniformity of base branch, as well as flower size from 20 to 40 mm, cloning the plants thus selected and the seed obtained from the plants obtained by cloning,

(b) growing progeny from seed obtained under paragraph

(a), the subsequent selection referred to in paragraph (a) and the harvesting of seed from the plants so selected,

(c) repeating the measures referred to in paragraph three to five times

b),

(d) growing offspring from seed obtained under paragraph

(c) selecting these offspring according to (a), 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, camomile according to the invention is distinguished by a surprisingly high content of the active ingredient (-) - α-bisabolol (at least 200 mg%), while the content of other bisaboloids (e.g. bisaboloxides) is very low and less than 50 mg. %. Known and described in the literature the camomile all have a very high content of bisaboloids and partially bisabolonoxides, which generally make up 50% of the essential oil of the drug (i.e., about 1% essential oil contains about 500 mg per 100 g of drug).

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 (-) - α-bisabolol being constant (i.e. homozygous).

In contrast, the evaluation of individual plants of known tetraploid varieties of camomile shows that individual individuals belong to different types in terms of substance content, which means that a single random collection of the drug produced from them has a qualitatively and quantitatively different inhomogeneous result.

In addition, the camomile of the invention surprisingly differs from known tetraploid camomile varieties, for example, by one or more of the following properties: improved seed germination, less unnecessary green matter (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. These advantages are particularly pronounced when, in addition to the selection for active ingredients, one or more of the above-mentioned features are selected, seed is obtained from the selected plants, a new selection is made from the offspring thus obtained and these measures are carried out three to five times.

Examples of starting diploid camomiles include, for example, "Camomile of Argentine origin" (see L. Z Padula, RVD Rondina and JD Cousio, Quantitative evaluation of essential essential oil, total azulenes and chamazulene in German camomile grown in Argentina; Planta med. 30, 273 - 273). 280, 1976), as well as all camomiles having measurable concentrations of (-) - α-bisabolol (generally above 5% in essential oil) in essential oil. Possible diploid camomiles are described in the following literature: H. Schilcher, "More recent knowledge in qualitative determination of camomile oil or camomile flowers", Planta med. 23, 132-

144 (1973); O. Motl, M. Felklová, V. Lukeš and M. Jasicová “On gas-chromatographic analysis and chemical types of camomile oil”. Arch. Pharm. 310, 210-215 (1977); Ch. Franz, J. Ilôlzl and A. Vômel "Preliminary morphological and chemical characterization of several populations and variations of Matricaria chamonilla L.", Acta Horí., 73, 109 -

114 (1978).

Camomile essential oil usually consists of the main constituents of famesen, spatulenol, chamazulene, one of the four bisaboloids of (-) - α-bisabolol, bisabololoxide A, bisabololoxide B or bisabolonoxide with respect to the diploid individual plants; in a mixed sample of the population, as well as in tetraploid individuals, the different bisabolides may be side-by-side) as well as the spiroether. Said substances together constitute 70 to 80% of the essential oil. Thus, starting diploid chamomiles are particularly preferred, wherein (-) - α-bisabolol is the predominant component (more than half the amount of said substances).

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

Examples of tetraploid basil leaves are: Bodegold (GDR) varieties,

Pohorelicky (CSSR), Zloty Lan (Poland), BK-2 (Hungary). These species are described in the following literature: M. Chladek and V. Kosova, Pharmazie 13, 712-713 (1958), W. Czabajska, Diss. Poznan (1963); W. Poethka and P. Bulin Pharm. Z. Halle, 108, 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 (-) - α-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 (-) - .alpha.-bisabolol (relative to the dry weight of the flowers dried at 40 DEG C.). that seed of such a camomile is seeded in the case of a diploid starting camomile and the seed is trapraploidized (irrespective of and / or knowledge of the bisabolol content of the parent plants). In the case of the starting tetraploid camomile, the seed described above is then immediately selected and propagated.

Suitable diploid and tetraploid starting chamomiles are selected by screening individual plants. In general, it is necessary to screen 1000 to 10,000 individuals from the initial population and to find several individuals having a high (-) -? -Bisabolol content according to the above criteria and which are advantageous as the initial chamomile. Seeds are obtained in the usual manner from such selected individuals and this is tetraploidized. After this process, the tetraploid plants are selected from the remaining diploid plants. From the tetraploid plants thus obtained after possible previous propagation, all plants having at least 100 mg% chamazulene, at least 200 mg% (-) - α-bisabolol and less than 50 mg% other bisaboloids in flowers dried at 40 ° C are now selected. . These flowers are harvested at the stage of development when 30 to 70% of all tubular flowers are open. This may optionally be followed by other selection and propagation steps that affect, for example, the following characteristics or properties: the term of simultaneous flowering, uniform base branching and narrow flowering zone (i.e. 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 shall be made for those plants whose flowers dried at 40 ° C contain at least 100 mg% chamazulene, at least 200 mg% (-) - α-bisabolol and less than 50 mg% of other bisaboloids, these flowers being harvested at an evolutionary 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 already mentioned).

A particularly preferred camomile is obtained when selecting (according to the invention) plants (known or prepared according to the invention) having a chamazulene content of at least 200 mg%, preferably 250 mg%, and having a content selected from chamazulene and (-) - α-bisabolol. The (-) - α-bisabolol is at least 300 mg%, preferably 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, claw buds or tissues of diploid starting plants of camomile) by chemicals, X-rays, gamma rays or UF-rays. It can also be carried out by the method of decapitating callus, by an anther culture or after

By applying high or low temperatures to the camomile plants, or to parts of plants or to the mesh of camomile plants. Here, for example, reference may be made to Wem Gottschalk's book "Die Bedeutung der Polyploidie für die Evolution der Pflanzen", Gustav Fischer Verlag, Stuttgart, 1976, in particular pages 13 to 22.

Tetraploidization using chemicals

Suitable tetraploidization chemicals include, for example: colchicine, acenaphthene, alkaloids such as atropine, veratrine, nicotine, sanguinarine, benzene derivatives, biphenyl and phenanthrene, naphthalene and naphthalene derivatives, diphenylamine, tribromaniline, p-dichloronphenobenzofuran 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 convalamurine (heteroisanuxurine), heteroisuxine, heteroisuxine ethylphosphate, ethyl chloride, phenylhydroxide and phenyldinaphthylmethane disulfonate of mercury), chloroform, gas of gas (N ₂ O) as well as mixtures thereof. Pellets, compost and cow manure can also be considered.

For example, the action of the compounds described herein takes place at temperatures between 0 ° C and 35 ° C, preferably between 12 ° C and 30 ° C, especially at temperatures between 15 ° C and 25 ° C.

The application is carried out, for example, by treating the seeds, the tips of the shoots, the roots (especially the roots or roots of the germinating seedlings), the foreheads, the cut areas on the leaves or stems, the cell suspensions of meristemic tissues, callus cultures. field of armpit buds. Chemicals are usually used in the form of solutions in water, weakly alcoholic solutions (alcohol content is 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 per se, optionally under pressure (e.g. 1 to 100 kPa). The duration of action is, for example, 1 to 36, preferably 2 to 12, in particular 4 to 6 hours.

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 0.01-0.2%, in particular 0.02-0.1%, preferably in a 0.0% colchicine solution, or germinate five- to five-fold. The 7-day well-developed germinating seedling of the diploid chamomile (with the cotyledons down) immerses up to 0.01-0.2%, in particular 0.02-0.1%, preferably 0.0% colchicine solution. In the latter process, the ambient atmosphere should have a relative humidity of 100%. For example, the duration of action of colchicine is 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. For example, plants with treated roots or other portions thereof or treated germinating seedlings are pictographed in the plant hazel.

From the treated seeds or germinating seedlings, the plants are grown (for example in a greenhouse: at a temperature between 18-25 ° C by day and 10-16 ° C at night), plants whose pollen is about 1.5 times larger are selected. as pollen of the starting material, possibly having a chromosome number of 36 in somatic cells.

If other plant parts (above ground or ground) are treated, only shoots, roots or flowers (seeds that have come from such treated parts) are later examined for the tetraploidization carried out. If, for example, the treatment of the shoot or claws is carried out, then only a sprout formed from this cobweb or from this bud and the flowers formed thereon, or the seeds per number of chromosomes.

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 UF-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 further than 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.

Suitable temperatures are: 0 to 5 ° C, preferably 0.5 to 4 ° C, in particular 2 ° C. These temperatures are, for example, exposed to: swollen seeds, germinating seedlings, shoots and meristem meshes. Treatment time: for example 1 to 100, preferably 20 to 40 days. Plants thus treated, or parts thereof, are further treated in the same way as after treatment with chemicals.

Decapitation-callus method (decapitation = cutting off the head or tip (tips) or back cut). The decapitation is carried out, for example, in young plants on 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.

Millenium culture (preparation of plants with a single number of pollen vesicles chromosomes followed by spontaneous or artificial tetraploidization). Closed tubular flowers are harvested from flowering plants whose pollen (pollen cams) are in the first stage of pollen mitosis. The rods are removed using a micromanipulator and transferred to Petri dishes, which are filled, for example, with Nitsch and Nitsch broth (Table 1). The petri dishes are then stored in the culture chamber for 16 hours at 28 ° C in the daytime, at 20 ° C at night. After about 4 weeks, the mildew begins to burst and the plants grow. These are haploid in the diploid parental share, and dihaploid in the tetraploid parental share: they are transplanted, for example, into the garden soil

EN 280689 Β6 and the greenhouse are allowed to bloom. These (di) haploid plants are sterile, but can be converted to polyploid forms by treatment of the ends of the shoots, roots or stems with chemicals such as colchicine, producing homozygous plants which can then be further propagated by seeds. For further treatment see chemicals treatment, for example with colchicine.

Table 1

Nitsch and Nitsch Environment (Science, 1969)

Mg / 1

KNO ₃ 950

NH4NO3720

MgSO ₄ .7H ₂ O185

CaCl ₂ 166

KH ₂ PO ₄ 68

MnSO ₄ .4H ₂ O25

H3BO310

ZnSO ₄ .7H ₂ O10

Na ₂ MoO. 2H ₂ O 0.25

CuSO ₄ .5H ₂ 0,025 ml solution from 7,45 g ethylenediaminetetraacetic acid disodium salt and 5,57 g FeSO ₄ .7H ₂ O per 1000 ml myo-inosities100 glycine2 nicotinic acid5 pyridoxine-HCl0,5 thiamine-HCl0,5 acid folic 0.5 biotin 0.05 sucrose 20 g broth (DIFCO-Bacto-Agar) 8 g indolylacetic acid 0.1 pH is adjusted to 5.5.

By measuring the size of the pollen 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, while the other bisaboloids (especially bisabololoxides). it should be below 50 mg% (with respect to 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 day length for a minimum of 14 hours after removal of the already blooming inflorescences. they are in bloom or before disintegration. After drying at an air temperature between 20 and 35 ° C, seed is obtained for propagation of the camomile according to the invention.

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

(a) approximately the current flowering time;

(b) uniform base branches and a narrow zone of flowers, about 10 cm, in particular 5 cm,

(c) large inflorescences with an outside diameter of about 30 mm (20 to 40 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 steps of selection and propagation.

In order to obtain a camomile which, in addition to containing at least 100 mg% of chamazulene and at least 200 mg% of (-) - α-bisabolol (relative to dried inflorescences), also provides high flower yield and is more suitable for machine harvesting, the following procedure is recommended: (as described above) are propagated by vegetative divers and monitored for 3 to 5 generations while still being selected according to the above criteria and, if applicable, additional criteria (a) to (c). The plants grown according to these criteria are cloned, seed is obtained from the cloned plants, from which the obtained plants are again selected according to the stated minimum chamazulene and bisabolol content, as well as other criteria (a) to (c), and the last seed is recovered.

Sequence - selection according to the stated minimum chamazulene content of 100 mg% and (-) - α-bisabolol of at least 200 mg% (other bisaboloids below 50 mg%) as well as according to other criteria a) to c) up to five times.

This is followed by another sequence of sowing - selection, as described previously (or 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 (clonal 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 of from 10 to 15 ° C, preferably 12 ° C. For cloning or multiplication, divers from shoot leaves and especially short shoots (side shoots) are suitable. They are rooted at temperatures of 12 to 18 ° C, preferably 15 ° C, and at 12 to 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, clayey 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 territories (for example, Northern Hemisphere, temperate to subtropical regions).

The novel camomile according to the invention belongs to the species of plant camomile, botanical designation Matricaria chamonilla L. (synonym Chamonilla reeutita (L.), Rauschert, and is defined by the active substance data in point 1 of the definition of the subject matter of the invention. (-) - α-bisabolol and bisaboloid oxides always refer to the developmental stage of flowers where 30 to 70%, in particular 40 to 60% of all tubular inflorescence flowers are open, that is, the flowers used to determine the active substance content were harvested to this term and then were 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, this means that, for example, 100% or up to 100% of all tubular inflorescences are open (for example, in full bloom) and / or harvested flowers are dried at higher temperatures as 40 ° C, the content of the active ingredients (-) - α-bisabolol can be

And chamazulene lower, since at higher temperatures and / or at a later harvest, more significant decomposition of azulene and bisabolol may occur.

The term "other bisaboloids" in point 1 of the definition of the subject matter of the invention includes in particular: (-) - α-bisabololoxide A and B; (-) - α-Bisabolone Oxide A; the other constituents of the essential oil of the camomile variety of the invention are En-In-dicycloether, farnesene, spatulol and, in moderate concentrations, various volatile terpene hydrocarbons.

For example, camomile flowers of the invention dried in a (cabinet) oven at 40 ° C (harvested as indicated) between 100 to 200 mg% chamazulene, between 200 to 450 mg% (-) - α-bisabolol and little, that is to say 5 to 50 mg% of other bisaboloids based on 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, for example, at a temperature of 35 to 50 ° C (drugs) 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, Pohofelic large-flowered); In particular, they differ from them in that in the camomile according to the invention (-) - .alpha.-bisabolol is the main constituent of the flowers of the flowers and, moreover, the content of other bisaboloids (bisabolooxide A and B as well as bisabolonoxide) is clearly lower. Other components of camomile essential oil are famesen, 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 soil microorganisms); no agrotechnical and cultivation measures are necessary; only a long day with more than 13 hours of maximum day length 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 (-) - α-bisabolol 200 mg%, while the content of other bisaboloids (especially bisabololoxides) is below 50 mg% (all values with respect to dried inflorescences), only those plants that are selected shall be selected

- 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 generations, the selection being always made 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 again selected according to the above criteria and the seeding sequence

- selection - seed harvesting is repeated three to five times.

The drug prepared from the camomile obtained according to the invention has a maximum active ingredient content of chamazulene and (-) - α-bisabolol when the collection of the camomile inflorescences takes place in the vegetation stage, for example 30 to 60% tubular flowers, preferably 40 to 60%, In general, 50% of the tubular inflorescence flowers are open and drying is carried out at an air temperature of at most 50 ° C, for example 35 to 50 ° C, in particular 40 ° C.

Drying can be carried out either by supply air or 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. Drying may be performed spontaneously or artificially (for example, artificially heated warm air). The yield of the active substances is the smallest 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 (-) - α-bisabolol, depends on the vegetation stage at which the flowers are present 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 wherever possible. The vegetation stage of the flowers is characterized by how many% of tubular flowers of the inflorescence 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 with 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, in most plants 40 to 60% of all tubular plants are open at the same time. flowers. This makes it possible to obtain 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 (-) - α-bisabolol content to the flowering stage at the time of harvest and the drying temperature of the drug is shown, for example, in the following table (where the content of other bisaboloids is in all cases less than 50%).

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table

Drying temperature / air temperature / Vegetation stage of the flowers in the des harvest 30 to 70 9 in all tubular inflorescences open 70 to 100 9 in all tubular flowers of inflorescence flowers open Content of active substances in the judged material Content of active substances in the material to be weighed up Deň: 32 ° C Chamazulene: at least 100 g% (-) - alpha-bisabolol: at least 200 cng% Chamazulene: at least 40 mg% of (-) - alpha-bisabolol; at least 120 ngt Content of active substances in alcoholic extract: Content of active substances in alcoholic extract: up Deň: 32 ° C Chamazulene: at least 5.0 mg% (-) - alpha-bisabolol: at least 10.0 mgft Chamazulene: OGT of 2.0 (J-SLFE-bieabolol: less than 6 ₍₀ ogi Obeah of active substances in aušenon material: Content of active substances in the material to be weighed: between 50 and 70 ° C Chamazulene: at least 50 mg Chamazulene: at least 30 mgX temperature Vegetation stage of flowers at harvest time euienia / air temperature / 30 to 70 16 all tubular inflorescences open 70 l and 100% in all tubular flowers of your open flower <-) - alpha-Bisabolol: at least 150 ag9 C -) - alpha-bisabolol: at least 100 mg between 50 to Content of active substances in alcoholic extract; Content of active substances in alcoholic extract; 70 ° C Chamazulene: at least 2.5 C -) - alpha-bisabolol: at least 7,5 og% chamazulene; at least 1.5 mgjU (r) -alpha-bieabolol: at least 10.0

Consequently, from the camomile used to produce the compositions of the invention, for example, the harvesting takes place in the vegetation 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 is obtained having a chamazulene content in each case of at least 30 mg% and a content of (-) -? -bisabolol of at least 100 mg%, 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% (-) - α-bisabolol and less than 10% other bisaboloids.

The determination of (-) - .alpha.-bisabolol as well as other constituents for camomile is carried out in the usual gas chromatographic manner.

The exact method of analytical determination is given in Annex A (after the text of Example 4).

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

Matricin has pharmacological activity close to chamazulene. From this precursor, matrix, for example, is instantly chamazulene when heated (for example by steam distillation to prepare tea infusion). 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 (-) - α-bisabolol, chamazulene and other camomile ingredients. Thus, for example, the extract obtained from the camomile according to the invention, for example by extraction with an alcohol or a water-alcohol mixture, or by extraction with supercritical gases, can be used to obtain camomile extracts or extracts. Furthermore, camomile essential oil, (-) - α-bisabolol, chamazulene and other camomile ingredients can be obtained 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% (-) - α-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 the extraction 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, and mixtures 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 per part of dried material are used, in particular 3 to 10 parts of solvent per part of dried material.

Fresh camomile flowers or frozen camomile flowers (in case fresh flowers have been frozen) may also be used for the production of extracts, in particular alcoholic extracts.

The oil prepared from the drug contains at least 3.5% chamazulene, preferably at least 5% chamazulene and at least 10% (-) - α-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 35-60 ° C), pentane, xylene, decalin, the organic phase is dried (e.g. over sodium sulfate), and the organic solvent is removed gently by a method (for example by distillation on a rotary evaporator or by 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.

SK 280689-6

When the camomile flowers are harvested at the vegetation stage when 30 to 100% of tubular inflorescences are 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% (-) -a-bisabolol and less than 50 mg% of other bisaboloids and silica obtained by steam distillation from this drug contained at least 3.5% chamazulene, at least 10% (-) - α-bisabolool and less than 10 mg% other bisaboloids and alcoholic yield prepared by extraction with lower alkanols contains at least

1.5% chamazulene, at least 5.0 mg% (-) - α-bisabolol and less than 10 mg% other bisaboloids.

When camomile flowers are harvested at the vegetation stage, when 30 to 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% (-) - α-bisabolol and less than 50 mg% of the other bisaboloids and the silica prepared therefrom contains at least

3.5% of chamazulene, at least 10 mg% of other bisaboloids, and alcoholic extract of camomile made with lower alcohols contains 1.5 mg% of chamazulene, 5.0 mg% of (-) - α-bisabolol and 10 mg% of other bisaboloids.

If camomile flowers are harvested in 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% (- ) α-bisabolol and less than 50 mg% of other bisaboloids, and the resulting silica contains not less than 3,5% chamazulene, not less than 10% of (-) - α-bisabolol and less than 10% of other bisaboloids and of which the alcoholic extract obtained contains 2.5 mg% chamazulene, at least

7.5 mg% (-) - α-bisabolol and less than 10.0 mg% 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% (- (a) -a-bisabolol and less than 50 mg% of other bisaboloids, possibly prepared therefrom, containing at least 3,5% chamazulene, at least 10% of (-) - α-bisabolol and less than 10% other bisaboloids and an alcoholic extract prepared therefrom The drug contains at least 1.5% chamazulene, at least 10 mg% (-) - α-bisabolol and less than 10.0 mg% other bisaboloids.

DETAILED DESCRIPTION OF THE INVENTION

Example 1 (The starting romance is tetraploid)

The individual plant variability test of 10,000 plants of the tetraploid camomile variety described above

I. Sarkany (Herba Hungar. 4 (1), 125-169 (1965)), it was found that about 1000 individuals had one plant having a high chamazulene content of 20% by weight and a high content ( -) - α-bisabolol, while at the same time the content of other bisaboloids, in particular (-) - α-bisabolonoxide, is very low (less than 5% by weight). Subjects were selected and subjected to the following steps:

Stage 1

From the progeny (as described) of the selected tetraploid camomile plants were selected those individuals that

A) bloom at approximately the same time,

B) have a uniform base branch and a narrow flower zone, about 10 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 a minimum content of 150 mg% of chamazulene and 300 mg of bisabolol and their content of other bisaboids (especially bisabololoxides) is clearly below 50 mg. (All figures 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 trimmed 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.

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 plant parts, preferably the ends of the shoots or the buds of the claws, are suitable for establishing a camomile culture.

After rinsing the plants with H ₂ O ₂ , under aseptic conditions in the "laminar flow" (high-performance filter with low turbulence displacement filter) the shoot ends or armpit leaf buds are removed and transferred to nutrient reagent containers such as Murashig and Skoog (Physiol 15, 473-497 (1962). The reagent containers are placed in an air-conditioned space at 12-18, preferably 16 hours of day length (attained by fluorescent lamps) at a light intensity of 500 to 1000 lux, preferably 1000 to 3000 lux and a temperature of 15 to 30 ° C, preferably 22 to 27 ° C. It is hardly obvious that the explants grow well, they are transferred to the culture medium, but with a higher concentration of cytokinin * (e.g. 30 mg / 16 6-isopentenyladenine) and with little or no auxin (0 to 0.3 mg / 1 indolylacetic acid) ). This is followed by the growth of stems (axes) and advent organs as well as the proliferation of the armpit buds. These can be harvested and grown as described above.

For the multiplication of plants in larger numbers, the explanatory notes (third passage, ie the 3rd generation) are transferred to a culture medium containing 10 mg / l of indolylacetic acid or 3-indolylbutyric acid or 0.1 after growing and forming the leaves in the first mentioned medium. up to 0.3 mg / l of α-naphthylacetic acid. At the same time, the plants are rooted, after about 4 weeks they can be grown in sterilized garden soil (steamed for 12 hours at 120 ° C) in pots and in a greenhouse (under usual conditions for seedlings).

Murashig and Skoog environment:

mg / 1

NH ₄ NO ₃ 400 Ca (NO ₃ ) ₂ .4H ₂ O 144 kno ₃ 80 kn ₂ after ₄ 12.5 MgSO ₄ .7H ₂ O 72 KC1 65 NaFe EDTA 25 H ₃ BO ₃ 1.6 MnSO ₄ .4H ₂ O 6.5 ZnSO ₄ .7H ₂ O 2.7

SK 280689-6

,70,75 indolylacetic acid2,0 furfuryladenine0,1 thiamin0,1 nicotinic acid0,5 pyridoxine0,5 glycine2,0 myo-inosit100 acid hydrolyzate1000 sucrose 2% purified agar powder 1%

Note to previous explanation: ^x Cytokines are phytohormones that promote cell division.

Stage 2

The plants obtained in step 1 bloom together under isolated conditions in a greenhouse. The plants stood in 11 cm pots filled with garden soil at 18-24 ° C by day and 12-14 ° C at night. The length of day was at least 14 hours and was achieved by additional lighting of 200 Watl / m ² in the winter half of the year. Water was supplied as needed.

Of the total number of selected individuals, inflorescences that bloomed 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 with a sieve machine.

Stage 3

A random sample was taken from the seed obtained in Stage 2 and about 2000 offspring were grown (ecological conditions as input 2) and selected according to the same criteria A) to D) of 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.

Position I

450mNN, 48.5 ° N / l, 5µ

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

Abbreviations explained:

NN = North altitude ° N = North latitude (n-degree) ° O = East longitude (n-degree).

Habitat II

200 NN, 42 ° N / 1 ° C,

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 tested and evaluated for homogeneous growth, flower size and harvest date, in addition random flower samples were examined for active substance content. From the field experiment, individuals corresponding to the above-mentioned parameters from step 1 were selected again. Seeds corresponding to step 2 were obtained from these.

Stage 5

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

Stage 6

About 1,500 individuals were grown from seed obtained under Stage 5 (ecological conditions as input 2) and selected according to the same principle as in Stage 1. From the selected plants, 34 individuals were selected and cloned according to step 1. Ten 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 propagation of camomile according to the invention.

Plant flowers from this seed for multiplication (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 immediately dried at 40 ° C in a drying cabinet for 72 hours , containing, for example, with respect to the weight of the dried flowers (dry matter), for example, at least: 150 mg% of chamazulene, 300 mg% of (-) - α-bisabolol and a maximum of 50 mg% of other bisaboloids.

Further exemplary data for plants obtained according to the invention:

1. Growth

Stem: straight, little branched

2. Olistenie

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 (inflorescences): outside diameter about 30 mm, inside diameter about 15 mm; weight of single inflorescence (dried): about 45 mg; flower head, length (mm): about 700, but depending on the location (cultivation) (climate), sowing time, soil, fertilization, plant treatment, weather;

beginning of flowering (counted from 1 January): about 160 days (sowing September, location Freising, Federal Republic of Germany, otherwise dependent on these factors).

Flowers: mid-June; inflorescence without stem, essential oil content (% dry matter): about 1.0%; azulene content in essential oil: at least 15%.

Disintegration of dried inflorescences, flower drug: slight if harvested before opening last tubular flowers;

pollen diameter: about 30 µm; seed length: about 1.25 mm; number of chromosomes in somatic cells: 4n = 36.

4. Fetus weight 1000 seeds (TKM): 0.06-0.13 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 growth (equilibrium) with a narrow flowering zone, thus particularly suitable for mechanized harvesting, large flower baskets (inflorescences), medium yield; basically branched form (three to five times).

Obtaining a drug

The camomile obtained according to this example is sown in the usual field. As soon as the plants develop and the inflorescence 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 distances of the harvesting machine being set to collect only inflorescences whose tubular flowers are 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 and from the crops and dried to 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.

After about 2-3 days, drying is complete. To remove portions of the flower and waste, the camomile drug is sieved and pressed into bales.

Analysis silica 960 mg% chamazulene 162 mg% (-) - α-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 the flower and flower waste, the drug is screened and then compressed into bales. Analysis values for such a drug are as follows: silica 870 mg% (-) - α-bisabolol 197% chamazulene 94 mg%.

Example 2 (The starting romance is tetraploid)

An individual plant variability test carried out on 10,000 plants of the I. sarkany tetraploid vermilion variety (Herba Hungar. 4, 1, 125-169, 1965) found that about 1,000 individuals had one plant having a high chamazulene content in essential oil. The content of other bisaboloids, in particular (-) - .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 at least 100 mg% and (-) - α-bisabolol of 200 mg%, or having a content of other bisaboloids (especially bisabololoxides) below 50 mg%, were selected from the progeny of the selected 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 in step 1, then they were in a greenhouse and bloomed together under isolated conditions. 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 ² . Water was supplied as needed.

From the total of the 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 cleaned with a shaker.

Step 2 provides seed which is a 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 a time when 30 to 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% (-) - α-bisabolol and at most 50 mg% 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. As soon as the plants develop and the inflorescences are in the vegetation stage, when about 50% of the tubular flowers are open, the inflorescences are harvested using a camomile harvesting machine, the ridge distances of the harvesting machine being adjusted to tear off only the flower heads (inflorescences). whose tubular flowers are 50% open. 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 2-3 days, drying is complete. In order to remove the stem parts and flower waste, the camomile drug is sieved on sieves and pressed into bales at the foot.

Analysis of silica chamazulene (-) - α-bisabolol

940 mg%

132 mg%

243 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, 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 at the foot.

Values determined by analysis of such a drug are, for example, as follows: silica chamazulene (-) - α-bisabolol

775 mg% mg% 163mg%.

SK 280689-6

Example 3 (The starting romance is diploid)

Using the variability assays of individual diploid camomile plants containing a small amount of bisabolol but not without bisabolol as described by Ch. Franz a

J. Hozl and A. Vomel and Acta Horticulturae 73, 109-114 (1978) found that up to 20% of all plants had an essential oil content of about 20% by weight and / or a high content of 50% by weight of (-) - and - while the content of other bisaboloids (especially (-) - α-bisabolonoxide) 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, relative humidity: about 60%, daylight duration: 14 hours.

Germinating plants were observed until flowering, the success of polyploidization was evaluated by comparative pollen and seed size measurements as well as by the number of plant chromosomes (F _r progeny = first progeny grown from seed, which was confirmed as tetraploid colchicine plants).

Tetraploidization can also be carried out 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 germinate leaves in 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% relative atmospheric humidity. After treatment, the germinating seedlings were washed several times with water and plated in hazelnuts. This was followed by further treatments as with seeds.

Pollen size was measured using a Leitz binocular microscope with a micrometric eyepiece and micrometric object carrier.

Chromosome counting is performed at the tip of the roots: fresh green root tips are taken from young greenhouse-grown plants or divers and dipped in 0.002 molar hydroxycholine solution for 5 hours and then placed in IN IIC1 for 15 minutes. For examination, about 1 mm of the root apex is stained with 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 pollen diameter is about 50% greater than that of the diploid starting material (about 30 instead of 20 pm) and those where the chromosome number was doubled to 36 (the diploid starting material is equal to 18) are tetraploid. These are selected. About 0.1 to 0.5% of the seeds or germinating seedlings were tetraploidized and flowering intact plants were developed as described above. Now steps 1 and 2 correspond to example 2.

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

910 mg% 117 mg% 252 mg%.

Obtaining a drug

The camomile obtained according to this example is sown in the usual field. When the plants develop and the inflorescences are in the vegetation stage when about 50% of the 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 50% open. 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 of silica chamazulene (-) -? -Bisabolol

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 ch. Franz, J. HOzl and A. Voel in Acta Horticulturae 73, 109-11 (1978) found that a maximum of 20% of all individuals had a chamazulene content of about 20% by weight and / or a high content of (-) - a- bisabolol 50% by weight, while at the same time the content of other bisaboloids, (especially (-) - α-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.

Steps 1 to 6, corresponding to Example 1, are further followed.

For example, the flowers of the resulting camomile contain at least 150 mg% of chamazulene, 300 mg% of (-) - α-bisabolol and at most 50 mg% of other bisaboloids) harvested while 30 to 70% of tubular flowers are open and then 72 hours of drying in oven (40 ° C).

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 the usual field. As soon as 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 50% open. 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 in a well ventilated place in a layer with a thickness of about 10 cm.

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

SK 280689-6

analysis silica 1020 mg% chamazulene 173 mg% (-) - a-bisabolol 418 mg% Annex A

The starting material is the camomile drug obtained according to the invention. Only those inflorescences whose 30 to 70%, in particular 40 to 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, 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, possibly adhered to the apparatus, is rinsed in an additional amount of pentane. To remove any residual water, add a small amount (to the tip of a spoon) of anhydrous sodium sulphate to the solution and then filter through a sintered glass filter having a porosity of D ₃ or D ₄ . After evaporating pentane at 40 ° C in a water bath and drying in a desiccator, the amount of essential oil is determined gravimetrically.

The chamazulene and bisabolol are then determined in the oil thus obtained (about 20 mg).

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 by filter (eg Eppendorf).

Wavelength: 578 nm

Cuvette: 1 cm Specific chamazulene extinction (1 g / 100 ml; 1 cm /: 20.8 Compensation liquid: n-hexane or cyclohexane Chamazulene content found in mg / 100 g: 120 E ₅₇₈ .

If a photometer with a filter is not available for measurement, this measurement may also be performed using a spectral photometer as follows:

Measuring device: spectral photometer (eg PM QII or PM QIII "Zeiss")

Wavelength: 605 nm Cell: 1 cm

Specific iamination of chamazulene (1 g / 100 ml; 1 cm): 24.5 Compensation fluid: n-hexane or cyclohexane Found chamazulene content in mg / 100 g: 102 E ₆₀₅ .

Bisabolol is then determined in the measured solution. Gas-chormatogratic 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; steel 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 ° C

Column: 25-220 ° C

Temperature programming: 4 ° / minute Sample solution: chamazulene measurement solution used. Reference solution: dissolve about 15 mg of standard bisabolol in cyclohexane to a volume of 25 ml.

Injected amount: 5 μΐ of sample solution and reference solution

Evaluation: Evaluation is performed by comparing the peaks (peak areas).

Bisabolol content in mg per 100 g of drug:

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:

Equipment: Packard, model 7721, 800 series, possibly Erba Fractovap, 2350 series

Columns: 3 m / 2 mm glass columns in diameter; 2 m / 2 mm in diameter

Filling: 3% methylphenylsilicone rubber "OV 1" on silanized "Gaschrom Q" 125 to 150 pm as carrier

Carrier gas: 30 ml / minute N ₂ Temperature program: 80 to 180 ° C, 2.5 (3 / ° C) minute Injector / detector temperature: 200 ° C Detector: flame ionization detector Injection amount: about 2 µl approx. 1: 50 diluted oil

The evaluation is partly without, partly with an internal standard. As the internal standard, methyl lauric acid or hexadecane is suitable for chamazulene-poor essential oils, optionally without chamazulene. For oils with a content of more than 5%, this should be preferred as an internal standard, and the determination is 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 the camomile drug thus obtained containing 105 mg% of chamazulene and 212 mg% of (-) - α-bisabolol are extracted with 2100 g of an aqueous solution of ethanol (40% by weight of ethanol) 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% (-) - α-bisabolol 10.2 mg%

Example 6

Preparation of fresh camomile extract

758 g of fresh camomile flowers harvested according to Example 1 at the time 30 to 70% of tubular flowers were opened (74% water content) 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: 11.7 mg% (-) - α-bisabolol 16.3 mg%

Claims (2)

PATENT CLAIMS A process for the preparation of a novel bisabolol-rich tetraploid camomile of the plant camomile (Chamonilla recutita L., synonymous with Matricaria chamonilla L., Asteraceae), the dried flowers of which at 40 ° C contain at least 100 mg% chamazulene , at least 200 mg% of (-) - α-bisabolol and less than 50 mg% of other bisaboloids, with the exception of manzana, characterized in that for 1000 to 10 000 individual plants of the diploid or tetraploid camomile population, it is determined in essential oil (-) - α-bisabolol content and select those camomile plants in which (-) - α-bisabolol is the major constituent of essential oil and in which the flowers, dried at 40 ° C, contain at least 100 mg% chamazulene, at least 50 to 100 mg% (-) - α-bi-sabolol and less than 50 mg% of other bisaboloids, these plants are separated from the initial population, multiplied and subjected to further selection and multiplication steps and optionally, from the plants of the resulting camomile, seed for propagation and / or the camomile drug is obtained, wherein in the case of a diploid camomile from which the Degumill variety is excluded as the starting camomile, tetraploidisation by means of chemicals is performed after all selection steps. temperatures ranging from 0 to 35 ° C, by gamma-rays, X-rays or UV rays at temperatures ranging 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 anthrax cultures. 2. The method of claim 1, wherein the selection or propagation comprises the steps of: (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 and, depending on the size of the flowers, from 20 to 40 mm, cloning the plants so selected and obtaining seed from the cloning plants, (b) growing progeny from seed obtained under paragraph (a), the subsequent selection referred to in paragraph (a) and the harvesting of seed from the plants so selected, (c) repeating the measures referred to in paragraph three to five times b), (d) growing offspring from seed obtained under paragraph c), selecting these offspring according to paragraph a), cloning the plants thus selected and obtaining seed from the plants obtained by cloning.