DE10308298A1 - Production of a plant preparation useful for treating cancer comprises extracting glucosinolates from plant material and enzymatically hydrolyzing the glucosinolates to isothiocyanates - Google Patents

Abstract

Production of a plant preparation (P1) comprises preparing material from a glucosinolate-containing plant species, inactivating endogenous thioglucosidases in the material, extracting the glucosinolates, enriching the glucosinolates by freeze concentration, enzymatically hydrolyzing the glucosinolates to isothiocyanates or stable storage forms thereof, and optionally enriching the hydrolysis products. Independent claims are also included for: (1) plant preparation P1 in which the hydrolysis products comprise more than 50 mole% isothiocyanates or stable storage forms thereof; (2) production of a plant preparation (P2) by preparing material from a glucosinolate-containing plant species, inactivating endogenous thioglucosidases in the material, extracting the glucosinolates and freeze drying the extract; (3) production of a kit by producing P1 or P2 as above; producing a preparation comprising a thioglucosidase/glucohydrolase, a buffer, optionally an enzyme inactivator and optionally an acid; and packaging the two preparations for mixing immediately before, during and/or after use; (4) kit produced as above; (5) production of a plant preparation (P3) by preparing material from a glucosinolate-containing plant species, hydrolyzing glucosinolates in the material with endogenous glucosidases after comminuting the material, optionally adding water, extracting the hydrolysis products and enriching the hydrolysis products by freeze concentration at constant pH; (6) plant preparation P3. ACTIVITY : Cytostatic. No biological data given. MECHANISM OF ACTION : None given.

Description

The present invention relates to a herbal preparation from Brassica ingredients and a process for their production. In addition, the present concerns Invention a pharmaceutical containing the herbal preparation Prevention composition or / and treatment of cancer.

In Germany, 300 people fall ill each year 000 people from cancer and about every fifth German dies from one Tumor disease, a number that will undoubtedly be in the next few years will increase. In addition to a genetic disposition for development There are a number of tumors in scientific literature of environmental influences, such as ionizing radiation and carcinogenic agents, or factors lifestyle, such as tobacco and alcohol consumption, as essential determinants for the development from cancer. Studies from the past decade point to this indicates that up to two thirds of the tumor disorders also influence factors related to food.

With food, a variety potentially mutagenic and procarcinogenic substances added. At the same time, however, there are also substances with anticarcinogenic effects fed. For a variety the risk of disease is related to human tumor diseases inversely proportional to the consumption of fresh vegetables and fruit. This has been particular for carcinomas of the mouth, the esophagus and the gastrointestinal tract, breast, lungs or other organs demonstrated.

Depending on the consumption of vegetables from the Brassicaceae plant family, especially white cabbage, Brussels sprouts and especially broccoli, has been a decrease in humans Statistical incidence of a number of different tumor diseases detected (Verhoeven et al., Cancer Epidemol. Biomarkers Prev. 5 (1996), 733-748). This anti-carcinogenic effect of vegetables from the Brassicaceae plant family is responsible for the presence of Bound glucosinolates.

If the Brassica plant cell or when eating Brassica vegetables the glucosinolates by the plant's own enzyme myrosinase or by the bacteria formed in the intestine of the consumer Enzyme myrosinase hydrolyzed to the corresponding isothiocyanates. newer Experimental studies showed that the enzymatic Hydrolysis from Brassica glucosinolates emerging isothiocyanates for the anticarcinogenic effect of Brassicaceae are immediately causal. Especially for Methylsulfinylalkyl isothiocyanates and indolyl isothiocyanates could both a so-called chemopreventive effect after exposure to carcinogens (Mehta et al., Carcinogenesis (1995) 399-404; Stoner and Morse, Cancer Lett. 114: 113-119 (1997); Talalay and Zhang, Biochem. Soc. Trans. 24: 806-810 (1996); Zhang et al., PNAS USA 91: 3147-3150 (1994); London et al., Lancet 356 (2000), 724-729) as well as a direct anticarcinogenic effect (Musk and Johnson, Carcinogenesis 14 (1993), 2079-2083).

Depending on the reaction conditions, such as for example the pH value, the reaction temperature and the presence of metal ions, such as Iron ions, or the presence of nitrate or nitrite, can contribute to the enzymatic hydrolysis of the glucosinolates in addition to the corresponding ones However, isothiocyanates also undesirable reaction products, such as for example thiocyanates, nitriles, epithonitriles or oxazolidinthione arise (Lange et al., nutritional survey 39: 292-296 (1992); Uda et al., Agric. Biol. Chem. 50: 2735-2740 (1986); Griffith et al., J. Horticult. Sci. Biotech. 73: 1-18 (1998); Benn, Pure appl. Chem. 49 (1977), 197-210). These connections can too undesirable Side effects and relevant health disorders, such as morphological changes the liver (Fenwick et al., Toxicants of plant origin. Vol. II. Glycosidases. CRC Press, Boca Raton, Florida USA (1989), 1-41), hypothyroidism by inhibiting iodine uptake or iodine accumulation in the epithelial cells the thyroid (Fenwick et al., (1989), supra) or by inhibiting the formation of the thyroid hormone Thyroxine (Ahlin et al., Acta Veter. Scand. 35 (1994), 37-53; Bachelard and Trikojus, Nature 185 (1960), 80-82).

The main reason for the anticarcinogenic effects of certain Brassica ingredients seems to be an induction of the xenobiotic metabolism. The xenobiotic metabolism detoxifies foreign substances with potentially mutagenic and carcinogenic properties, the so-called xenobiotics, such as pharmaceuticals or environmental toxins (Posner et al., J. Med. Chem. 37 (1994), 170-176; Zhang et al. , (1994), supra). The xenobiotics are chemically converted by two functionally connected enzyme systems of the metabolism in such a way that an accelerated excretion of the same results from the body. Oxidation or reduction is initially carried out by phase 1 enzymes, ie by enzymes from the cytochrome P450 group (Murray, Int. J. Mol. Med. 3 (1999), 227-238; Williams, J. Florida MA 78: 517-519 (1991); Sligar, 1999 Biochemistry 34: 71-83; Omiecinski et al., Toxicol. Sci. 48: 151-156 (1999). Subsequently, phase II enzymes, for example quinone reductase or gluthation-S-transferase, conjugate with endogenous ligands, for example with gluthation, which leads to an accelerated excretion from the body (Wattenberg, Cancer Res. 45 (1985), 1-6; Fahey et al., PNAS USA 94: 10367-10372 (1997).

Recent studies have shown that Methylsulfinylalkyl-isothiocyanates selectively stimulate the phase II enzymes of the xenobiotic metabolism. For example, mammalian cells show which have been treated with methylsulfinylalkyl isothiocyanates, decreased sensitivity to the mutagenic and carcinogenic Properties of various xenobiotics (Talalay et al., Toxicol. Lett. 82-83 (1995), 173-179; Zhang et al., 1992 PNAS USA 89: 2399-2403; Zhang et al., (1994), supra).

But also for the indolyl isothiocanates and their decay products are known to have anticarcinogenic effects. In particular, indole-3-carbinol (glucobrassicin) and two others resulting reaction products, ascorbigen and diindolylmethane (Chen et al., Carcinogenesis 19 (1998) 1631-1639; Preobrazhenskaya et al .; Pharmac. Ther 60 (1993), 301-313) have an anti-carcinogenic effect in cell cultures as well as in animal experiments Effect especially on breast tumors on (Cover et al., J. Biol. Chem. 1998, 273: 3838-3847; Grubbs et al., Anticancer Res. 15 (1995), 709-716).

In the presence of L-ascorbic acid by enzymatic hydrolysis from the glucosinolate glucobrassicin resulting indolyl isothiocyanate indole-3-carbinol (brassicin) the more oxidation-stable fat-soluble Ascorbigen implemented. In the gastrointestinal tract of the consumer, Ascorbigen is spread over several Intermediates converted to diindolylmethane and 5,11-H-indolo- [3,4-b -] - carbazole. This both compounds represent ligands of the so-called aromatic hydrocarbon Responsiveness Receptors and lead through interaction with this Receptor immediately activates the enzymes of the xenobiotic Metabolism (Gillner et al., MoI. Pharmacol. 44 (1993), 336-345; Bjeldanes et al., PNAS USA 88: 9543-9547 (1991); Chen et al., (1998), supra).

In addition to inducing xenobiotic metabolism, especially the phase I enzymes, are said compounds also show direct anticarcinogenic effects, which appear primarily Interaction with tissue estrogen receptors (Michnovicz and Bradlow, J. Natl. Cancer Inst. 82: 947-949 (1990); Liu et al., J. Natl. Cancer Inst. 86 (1994) 1758-1765).

Also for the glucosinolate sinigrin, for example in white cabbage contain, anticarcinogenic effects were determined experimentally, the exact mechanism of which has not yet been clarified (Tanaka et al., Carcinogenesis 8 (1990) 1403-1406).

Although so far various Brassica ingredients or has identified their hydrolysis or degradation products, the for the anticarcinogenic effects of Brassicaceae seem to be responsible is still unclear whether the anticarcinogenic effects of Brassicacea on the mechanism of action of individual ingredients or their hydrolysis or Degradation products or on the synergistic effect of various of these compounds with different mechanisms of action based. To optimally from the health-protective effect To benefit from the Brassicaceae, the Brassica ingredients or their degradation or / and hydrolysis products according to their natural Occurrence in these plants will be included. Given the Low levels of the compounds mentioned in Brassica vegetables must regularly be substantial Amounts of these plants are ingested to be sufficient with food to benefit from the anticarcinogenic effects of Brassicaceae. On such regular, possibly more daily Vegetable consumption from the Brassicaceae plant family, for example cabbage plants, contradicts the current, statistically verifiable eating habits in the western industrialized countries (Lange et al., Nutritional Review 39: 252-257 (1992).

Alternatively, the health-protecting effect of the Brassicaceae ingredients can also be brought about by regular use of preparations with concentrated natural Brassica ingredients. Various products with Brassica ingredients have therefore been developed in recent years, e.g. sprouts from Brassica vegetables and extracts thereof (US Patents 5,725,895, 5,968,505 or 5,968,567), Brassica powder and pressed tablets, isothiocyanate dry extracts ( EP-A-0 772 976 ), Powder products from freeze-dried broccoli sprouts or synthetic products. However, due to unsatisfactory production or isolation processes, these products have only a low content of the relevant health-protecting Brassica ingredients and / or additionally contain a number of undesirable, potentially health-impairing ingredients. Furthermore, the previously known manufacturing or insulation processes are very complex in terms of process technology, ineffective and associated with considerable costs.

One object of the present invention was therefore to provide a method for producing herbal preparations with Brassica ingredients, in which the disadvantages of the prior art are at least partially avoided. Herbal preparations of Brassica ingredients with improved quality should also be provided, which can be used for external or internal use in humans. In particular, the new preparations should contain highly concentrated methylsulfinylalkyl isothiocyanates and / or indolyl isothiocyanates or substances which use the latter heat-labile Brassica ingredients as stable stores have shapes.

• (a) Bereitstellen von Material aus mindestens einer Glucosinolat-haltigen Pflanzenspezies,
• (b) Inaktivieren von endogenen Thioglucosidase(n) in dem Material,
• (c) Extrahieren der in dem Material enthaltenen Glucosinolate,
• (d) Anreichern der Glucosinolate mittels Gefrierkonzentration,
• (e) enzymatisches Hydrolysieren der Glucosinolate zu den entsprechenden Isothiocyanaten und gegebenenfalls stabilen Speicherformen davon und
• (f) gegebenenfalls Anreichern der Hydrolyseprodukte.

This object is achieved by a method for producing a herbal preparation, comprising

• (a) providing material from at least one plant species containing glucosinolate,
• (b) inactivating endogenous thioglucosidase (s) in the material,
• (c) extracting the glucosinolates contained in the material,
• (d) enriching the glucosinolates by means of freezing concentration,
• (e) enzymatic hydrolysis of the glucosinolates to the corresponding isothiocyanates and, if appropriate, stable storage forms thereof and
• (f) optionally enriching the hydrolysis products.

The method according to the invention allows production of products from Brassica vegetables with concentrated, health-protecting natural isocyanates or with stable storage forms of it, such as ascorbigen. The through the inventive method available preparations can both in liquid Form as well as in the form of dry products. The vegetable according to the invention preparations point towards Preparations from fresh ripe Brassica vegetables and conventional products with Brassica ingredients and their manufacturing processes have significant advantages.

That for step (a) of the method according to the invention Plant material used is preferably made from Brassica vegetables or Brassica plant parts with a high content of methylsulfinylalkyl glucosinates and / or Indolyl glucosinolates compiled. Furthermore, the use preferred by plant material that has a low content of undesirable Ingredients, e.g. certain here undesirable glucosinolates, such as 3-butenyl-glucosinolate (gluconapine), 4-pentenyl-gucosinolate (progoitrin), further on thiocyanates, nitriles, epithonitriles, oxazoline dithiones, in particular L-5-vinyloxazolidin-2-thione, contains or of these substances preferably free is. About that it is also convenient to use a material with a low content of nitrate and nitrite, in particular with a maximum nitrate content of 100 mg / kg, in order to To prevent the development of procarcinogenic nitrogen compounds (Lüthy et al., Mitt. Geb. Lebensm. Hyg. 75: 101-109 (1984); Wakabayashi et al., Mutagenesis 1 (1986), 423-426).

The material can, for example from plants of the order Capparales, of which selected from the Capparaceae, Brassicaceae or Cruciferae, Koeberliniaceae, Moringaceae, Resedaceae and / or Tovariaceae. It is preferred to use Broccoli, white cabbage, Red cabbage, cauliflower, kohlrabi, savoy and / or Chinese cabbage. Especially Broccoli and Brussels sprouts are preferred because they are not only health-related desirable Contain glucosinolates in sufficient concentration, but at the same time are essentially free of unwanted glucosinolates, in particular by Progoitrin.

The one for selection in the method according to the invention Parts of plants used should contain the desired glucosinolates in high concentration and at the same time undesirable ingredients, in particular Nitriles, in a comparatively low concentration. So In broccoli plants the florets are used in particular, while the stems are preferred be discarded as they have a higher proportion than the florets contain nitriles.

As material for the method according to the invention plant species containing glucosinolate are suitable in all Stages of development, from seeds to mature plants. Because ripe Plants completely as a basic material are sufficient, their use allows a cheaper one Generation of end products compared to such processes, in which only plants an early Stage of development can be used. Because the composition of the various glucosinolates in Brassica vegetables while the growing season and varies from species to species can by Selection of an appropriate vegetable base material or mixtures the creation of product variants using several vegetable base materials with different compositions of Brassica ingredients.

The method according to the invention allows use of wild forms of vegetables or crossing products with the commonly grown ones Low-glucosinolate varieties to a higher content of glucosinolates to ensure already in the vegetable base material. By mixing different partial batches of vegetable raw materials can the content of Brassica ingredients can be adjusted to a at best little variance between different production batches to ensure. This allows a compared to conventional manufacturing processes improved constancy of the content of ingredients and thus a Generation of higher quality Products.

The vegetable plants used for the process according to the invention can be grown in soil or other suitable nutrient media which preferably have no or only a low content of nitrate and / or nitrite. In this way, starting material can be provided which has a maximum nitrate content of 100 mg / kg. When the plants are grown, nitrogen fertilization is preferably not carried out, but sulfur fertilization in order to increase the concentration of sulfur-containing glucosinolates in the Brassica plants. A low content of nitrates and nitrite in the cultivation media or in what was used for cultivation This, as well as the avoidance of nitrogen fertilization, reduces the nitrite content of the plants. This prevents the formation of undesirable procarcinogenic nitrogen compounds in the plant material (Wakabayashi et al., Proc. Jap. Acad. 61 (1985) 190-192; Wakabayashi et al., (1986), supra).

The used for the method according to the invention Can plant additionally during the Cultivation exposed to special conditions to reduce the glusosinolate content increase in vegetable raw material. This effect can be caused by an increase the ambient temperature during of the entire cultivation to 25 to 30 ° C, a termination of irrigation about 100 hours before harvest, an artificial exposure Light in the visible spectrum and / or by light with an additional UV component at wavelengths between 100 and 280 nm, about at least 8 hours a day in the last 4 days before harvest or by a combination of the above-mentioned measures.

Step (b) of the method according to the invention includes inactivating endogenous thioglucosidases, in particular the endogenous myrosinase of the plant starting material suitable processes, in particular by heating, for example at a temperature of 95 ° C up to 100 ° C for one Time enough to get the endogenous thioglucosidases as possible Completely irreversibly inactivating, but at the same time small enough to avoid a loss of glucosinolates, particularly indolyl glucosinolates to keep low. The duration of the heating is preferably from 1 s to 420 s. By the way, the content of the in some Cabbage types, undesirable Glucosinolates Progoitrin reduced, whose hydrolysis products the thyroid function in humans can.

Inactivation of endogenous thioglucosidase The plants are therefore made to prevent them from growing out the glucosinolates during their subsequent extraction from the plant material (see step (c)) undesirable compounds by hydrolysis under uncontrolled conditions form such as thiocyanates, nitriles and epithonitriles. Besides, will prevented by inactivating the enogenous thioglucosidases, that hydrolysis products of glucosinolates with other phytonutrients uncontrolled chemical before removing them from the production batch Make connections.

This is compared to conventional Manufacturing process reduces the content of undesirable ingredients and thus higher quality Products.

The heating is preferably carried out by immersion blanching in hot water. An irreversible inactivation the endogenous enzymes can, in principle, also be used by others Methods, e.g. Steam blanching or heating by microwave radiation, respectively. A cool or freezing the intact plants only leads to one reversible inactivation of the enzymes, as well as immersion in ethanol. These methods are therefore unsuitable. Another advantage of heating is that the vegetable raw material in a single Additional production step is cleaned and made low in germs. This is particularly preferred immersion blanching in such a way that the material over a Period between 1 min and a maximum of 15 min, preferably between 3 min and 5 min at a temperature of 95-100 ° C is heated.

By means of a control procedure can, in the sense of optimization, the shortest necessary heating duration to complete. Enzyme inactivation while minimizing the loss of thermally labile indolyl glucosinolates be determined. For this purpose, samples from the production batch heated for different lengths of time and then thoroughly Crushing the material conditions for hydrolysis by endogenous Plant enzymes produced. Since equimolar in the hydrolysis of glucosinolates glucose is also released to other hydrolysis products, can be determined by determining the sugar content in the samples and checked after heating, whether further endogenous thioglucosidases are effective. The so determined shortest duration necessary Heating time to complete Enzyme inactivation, at the same time loss of heat labile Indolyl glucosinolates are then minimized the entire production batch applied.

Step (c) of the method according to the invention involves extracting the glucosinolates obtained in the plant material. The material is first crushed. The plant material is preferably comminuted with the addition of water or an aqueous medium, which is free of metal ions, especially iron ions, so that an aqueous Plant porridge is obtained. The temperature during the grinding is preferably between 0 ° C up to 4 ° C, again about the loss of heat labile indolyl glucosinolates, in particular of glucobrassicin.

The duration of the shredding process is preferably 6 minutes or less. The aqueous medium can optionally an enzyme activator, such as L-ascorbic acid, and a buffer for Setting a desired one pH, e.g. a citrate buffer and / or a carbonate buffer, such as a sodium hydrogen carbonate buffer, and / or a physiological one compatible Acid, e.g. Citric acid.

Then the material is preferably subjected to a vortex extraction, the water-soluble glucosinolates of the plant starting material being dissolved out of the plant cells and into a suitable solvent with the highest possible efficiency due to mechanical action, in particular shear forces for example water or a buffered aqueous medium. However, even under optimized experimental conditions, only 30% of the glucosinolates from mature Brassica plants can be converted into an aqueous solution (Fahey et al., (1997), supra). Despite this comparatively poor solubility for glucosinolates and their cleavage products, water is used as the extraction agent in the process according to the invention, because other non-aqueous extraction agents are either not approved under food law or hinder the subsequent enzymatic hydrolysis.

The extraction is preferred at a temperature between 0 ° C and 8 ° C, particularly preferably between 0 ° C and 4 ° C, carried out, in turn, the loss of heat unstable Minimize indolyl glucosinolates, especially on glucobrassicin. The vortex extraction is preferably carried out in such a way that the plant material using centrifugal and shear forces with more than 5000 rpm, preferably at 10,000 to 12,000 rpm above, at least Crushed and stirred for 3 min is going to make the water soluble Ingredients, especially the glucosinolates, if used of water as a solvent possible, from the plant cells ..

Other extraction methods, such as percolation and conventional extraction, can be used however less suitable.

The taking into account the properties relative completeness of the extractant used the glucosinolate extraction from the plant material can be based on of samples to be examined in which under uniform conditions in terms of reaction time, temperature, pH value and amount added of exogenous enzyme, L-ascorbic acid and / or glutamic acid and buffers before or after the addition of exogenous enzyme to the glucose content is determined and as long as thereafter a new extraction process the production batch takes place until the glucose concentration in the Samples no longer rise after adding exogenous enzyme. Also this control method is based on the equimolar one already explained above Release of glucose during the hydrolysis of glucosinolates. This Investigation guaranteed as far as possible leaching the glucosinolates from the plant material in the solvent used Water in the sense of optimizing the extraction process.

After extraction and before enzymatic Hydrolysis and are preferably coarse and fine plant components separated so that an essentially solids-free solution is obtained. By separating the insoluble Vegetable solids are primarily prevented from doing so in the subsequent hydrolysis of the glucosinolates react chemically with the resulting hydrolysis products. The separation the solids can be filtered and conventional, for example or mechanical separation or decanting, where appropriate the finely dispersed, chlorophyll-containing plant components after removal from the production batch, initially using suitable processes at an elevated Dried temperature of preferably up to a maximum of 80 ° C to a solid mass and to a later one Time to be added to the end product of the process can. The separation of solid plant components particularly preferably comprises first filtration to remove coarse constituents, followed by a mechanical separation of fine plant parts. The rough ingredients are discarded because they contain vegetable frameworks that intestinal motility in humans affect or bloating and promote flatulence can, especially insoluble ones Non-starch polysaccharides such as cellulose, hemicellulose and lignins.

Step (d) of the method according to the invention includes an enrichment of the glucosinolates by freezing concentration to a pre-selected degree of concentration. Enrichment preferably takes place due to the freezing concentration the glucosinolates, especially the indolyl glucosinolates, e.g. of 3-indolyl glucosinolate (Glucobrassicin) and / or the methylsulfinylalkyl glucosinolates, e.g. of glucoraphanin, at least three times, particularly preferred at least five times the initial concentration in aqueous Extract.

The freeze concentration enables one effective, inexpensive and gentle enrichment of the water-soluble glucosinolates. Beyond that in particular, process-related losses of heat-labile indolyl glucosinolates were reduced. The freezing concentration is preferably carried out in such a way that the water of the glucosinolate-rich solution in a temperature range between 0 ° C and the eutectic solidification temperature of the solution frozen out to ice crystals will do this afterwards using separating presses, centrifuges and / or washing columns from the remaining, now concentrated solution are separated. in the Contrary to that some enrichment by heat drying and / or negative pressure significant losses in glucosinolates during freeze drying and reverse osmosis from economic or technical establish are unsuitable.

Even with different glucosinolate concentrations in different batches of the vegetable raw material an enrichment of the glucosinolates by means of freezing concentration a predetermined value. This makes it constant the concentration of Brassica ingredients ensured in the end products.

A significant qualitative advantage of the products available through freeze concentration compared to preparations from fresh ripe Brassi Ca vegetables or conventional products with Brassica ingredients is that the products according to the invention have health-protecting methylsulfinylalkyl isothiocyanates and / or indolyl isothiocyanates or their cleavage products in aqueous solution, that is to say in the form which is directly absorbed from the gastrointestinal tract Tract allows. In contrast, the glucosinolates in fresh, ripe Brassica vegetables or in conventional products with Brassica ingredients only have to pass into an aqueous solution after being absorbed into the gastrointestinal tract before they can be absorbed.

In one embodiment of the method can by heating the glucosinolate-containing liquid before and / or after a reduction in the concentration of indolyl glucosinolates, especially of glucobrassicin, to a predetermined value and thus a change the concentration ratio of the various glucosinolates in favor of the methylsulfinylalkyl glucosinolates be reached. For this purpose, the preparation is preferably at 70 ° C to 100 ° C, especially preferably to at least 80 ° C, heated.

To determine the required Heating time can Several samples were taken in advance and each over a defined period be heated between 3 min and 30 min. Then the Content of indolyl glucosinolates, in particular on glucobrassicin, determined by suitable methods and the heating time of that sample at which the predetermined one Reduction in the content of indolyl glucosinolates or glucobrassicin has occurred, on the processing batch of the further manufacturing process applied.

The enzymatic hydrolysis according to step (e) of the method according to the invention is preferably carried out by at least one exogenous thiol glucosidase / glucohydrolase (Myrosinase E.C.3.2.3.1), preferably of vegetable origin. The for the Enzyme used in enzymatic hydrolysis is particularly preferred a myrosinase made from parts or whole plants of Raphanus sativus (radish) or Sinapis alba (white mustard) in any stage of development is available from seeds to mature plants. The hydrolytic activity this enzyme can by adding activators, such as ascorbic acid and / or glutamic acid, be increased in the production approach. The use of enzyme from plants suitable for consumption has considerable advantages in food technology and nutritional Respect.

The hydrolysis process takes place under Conditions where a possible high degree of implementation of the glucosinolates contained in the production batch to the preferred hydrolysis products. The Degree of conversion of the methylsulfinylalkyl and / or indolyl glucosinates to the corresponding isothiocyanates or to others desired Fission products such as ascorbigen is preferably at least 50%. The completeness hydrolysis of the glucosinolates after one or each addition of Myrosinase can be checked using a control procedure in which the glucose content of the reaction batch before and a few Minutes after adding the enzyme and from this the so-called Net glucose concentration, defined as the difference in glucose concentration between the two samples, is calculated, and only then Myrosinase is added again when the net glucose concentration is> 0. The principle the procedure has already been explained.

The indolyl isothiocyanate glucobrassicin is in watery solution relatively unstable, but can be converted to ascorbigen with L-ascorbic acid in a subsequent step continue to react, as a stable storage form of this indolyl compound can be viewed. The L-ascorbic acid can in the process according to the invention the process solution before and / or during be added to the enzymatic hydrolysis. The addition of L-ascorbic acid to Incidentally, production approach reduced the temperature optimum of the myrosinase from a range around 60 ° C to around 35 ° C, whereby again heat loss minimized by indolyl glucosinolates, especially glucobrassicin become.

Hydrolysis of glucobrassicin takes place under the natural unaffected Conditions of autolysis of shredded plant material, for example in the production of conventional products, in food preparation or after consumption in the gastrointestinal tract of the consumer, indol-3-ylmethyl is formed in addition to the ascorbigen Oligomers, their nutritional Importance and their potential pharmacological effects currently are unknown. It is therefore from a nutritional point of view reasonable the proportion of these oligomers in herbal preparations, the for intended for human use, by suitable hydrolysis conditions in favor of the ascorbigenes. The method according to the invention Accordingly, it is preferably carried out under such conditions at which generate undesired, because potentially health-impairing hydrolysis products, for example, nitriles and thiocyanates are prevented. Accordingly, the end products of the process according to the invention are distinguished across from conventional products and preparations from fresh ripe Brassica plants due to a lower content of unwanted Ingredients from what an additional qualitative advantage represents.

In the process according to the invention, the enzymatic hydrolysis of the glucosinolates takes place with the addition of a suitable buffer, the pH being adjusted to a desired value. Furthermore, the temperature is set to a desired value. Doing so will be preferred Wisely selected values in which the desired products are produced in high yield as free as possible from undesired by-products.

In one embodiment, the pH is below Addition of a buffer preferably in a range from 6.8 to 7.5, particularly preferably in a range from 6.9 to 7.1, for example about 7.0. Subsequently is preferably repeated once or continuously with continuous stirring added the enzyme. contains the watery solution in relevant concentrations of L-ascorbic acid, either from the outside or originating from the plant material used itself, should in the solution a temperature between 32 ° C and 60 ° C, preferred of around 35 ° C, be maintained until essentially complete hydrolysis the one in the solution contained methylsulfinylalkyl glucosinolates has taken place, especially for the corresponding isothiocyanates, z. B. from glucoraphanin to sulforaphane. If the solution does not contain L-ascorbic acid, then should have a temperature of 60 ° C be maintained.

In another embodiment will first L-ascorbic acid added, preferably in at least an equimolar amount to that in the solution contained glucobrassicin (3-indolylmethyl glucosinolate), and below the pH of the solution with the addition of a buffer, optionally a physiologically acceptable one Acid, for example citric acid, set to a range from 4.0 to 5.5, for example to 5.0. Then will preferably with continuous stirring one or more times Enzyme added. A temperature between 32 ° C to 60 ° C, preferred of about 35 ° C, upright obtained until essentially complete hydrolysis of the in the solution contained glucobrassicin, preferably to indole-3-carbinol (brassicin, 3-indolylmethyl isothiocyanate), and its subsequent conversion to Ascorbigen.

In another embodiment of the method according to the invention can reduce the content by heating the hydrolyzate on indolyl isothiocyanates, in particular on indole-3-carbinol (brassicin, 3-indolylmethyl isothiocyanate) to a predetermined value and thus a change in the concentration ratio of the various isothiocyanates in favor of the methylsulfinylalkyl isothiocyanates can be achieved. The heating is preferably carried out at a temperature of at least 70 ° C over a defined period.

The one for setting a desired one concentration ratio the isothiocyanates in the solution the required heating time can be determined, for example that several samples are taken in advance and each one defined period, for example between 3 and 30 minutes, heated be, then the content of indolyl isothiocyanates, especially indole-3-carbinol (Brassicin, 3-indolylmethyl isothiocyanate), by means of suitable methods, for example spectrophotometric, is determined. The heating time of the sample at which the predetermined reduction in the content of indolyl isothiocyanates has occurred, is then applied to the processing batch.

The through the enzymatic hydrolysis emerging products can according to step (f) the method according to the invention if necessary be enriched. This enrichment can, for example by a further freeze concentration, for example performed as previously described can be. The pH value must be taken using suitable measures to be kept constant to prevent acid-induced decay to avoid previously hydrolytically generated ascorbigens. alternative or additionally The enrichment of the hydrolysis products can be converted into a non-aqueous solvent include. This can be done, for example, by liquid: liquid solvent extraction, being the non-aqueous solvent from olive oil, coconut oil, soybean oil, palm oil, sunflower oil, peanut oil, sesame oil, others vegetable or animal oils as well as from others for suitable lipids or mixtures suitable for human consumption. The resulting oily Enable product forms a faster and more complete Absorption of the isothiocyanates or the fat-soluble ascorbigen in the gastrointestinal tract of the consumer. Alternatively, you can also watery Compositions or dry extracts are made. The production dry extracts can be freeze-dried.

Optional after hydrolysis Freezing concentration that occurs may be physiological in advance compatible polar solvent, preferably ethanol, are added. This will enrichment Hydrophobic isothiocyanates simplified and bacterial contamination of the preparations prevented. this leads to for an improved shelf life of the end products.

The method according to the invention allows production of a product in different forms, e.g. than concentrated aqueous Solution, as watery-alcoholic Solution, as a dry extract or as an oily one solution or suspension. The product can still other substances, especially pharmaceutically acceptable ones Carrier- and / or auxiliary substances added become. The product can also be wrapped in suitable e.g. Soft gelatin capsules. Furthermore, substances which the product quality additionally improve, e.g. previously separated and dried finely dispersed Plant fractions are added.

Other substances which can optionally be added to the end products of the process according to the invention are substances which can reduce the activity of enzymes of the cytochrome P-450 group, for example wise plant components, such as lemon oil, orange oil, components of the grapefruit or rosemary plant or such plant components that contain zinc aspartate. In preparations of this type, a predominant stimulation of phase II enzymes of the health-protecting and detoxifying xenobiotic metabolism results from the methylsulfinyfalkyl isothiocyanates in the herbal preparation according to the invention.

The present invention relates to also a vegetable one obtainable by the method described above preparation with methylsulfinylalkyl and / or Indolyl isothiocyanates and, where appropriate, their stable storage forms, like ascorbigen. The methylsulfinylalkyl isothiocyanates thus produced are present or the ascorbigen preferably in a molar concentration of more than 50% of all glucosinolate hydrolysis products in front. Also preferably contains the herbal preparation a maximum nitrite content of 0.1 mg per kg or l. Furthermore identifies potentially harmful substances, selected Thiocyanates, nitriles, epithonitriles and oxazolidine ions, furthermore on indol-3-ylmethyl oligomers from the hydrolysis of glucobrassicin, in a lower molar concentration, preferably in at least a 50% lower molar concentration than this after an autolysis of crushed material at least one plant species containing glucosinolate under natural, unaffected Conditions arise, such as those associated with autolysis of the plant material during the shredding for food preparation or when biting through Animals rule. The herbal preparation according to the invention is preferably at least substantially free of glucosinolates.

• (a) Bereitstellen von Material aus mindestens einer Glucosinolat-haltigen Pflanzenspezies,
• (b) Inaktivieren der endogenen Thioglucosidase(n) in dem Material,
• (c) Extrahieren der in dem Material enthaltenen Glucosinolate und
• (d) Herstellen eines Trockenextraktes von Glucosinolat-haltigem Pflanzenmaterial durch Gefriertrocknung.

Another aspect of the present invention is a process for the preparation of a herbal preparation which contains a high proportion of plant material containing glucosinolate, and preferably can be used for the preparation of a plant preparation containing isothiocyanate and optionally ascorbigen, as described above. This method preferably comprises the steps:

• (a) providing material from at least one plant species containing glucosinolate,
• (b) inactivating the endogenous thioglucosidase (s) in the material,
• (c) extracting the glucosinolates and
• (d) Preparation of a dry extract of glucosinolate-containing plant material by freeze-drying.

On preferred conditions to carry out the individual process steps is based on the above statements directed.

Another aspect of the invention relates to various methods of producing products that spatial separate from each other containing a preparation Brassica glucosinolate and a preparation containing a thioglucosidase / glucohydrolase (Myrosinase), preferably of vegetable origin, as described above, contain. Both preparations which may be in the form of a kit, are preferably immediate matched before use. This leads to a hydrolytic de novo production of isothiocyanates and / or ascorbigens from the Brassica glucosinolates.

For example, the kit can be an in represent two compartments of subdivided containers, their contents mix with each other when emptying. Another one embodiment for oral administration, the two components are mixed with subsequent hydrolysis of the glucosinolates only in the gastrointestinal tract of the consumer. This can be done, for example, with soft gelatin capsules that are soluble in digestive juice can be realized with two compartments, the glucosinolates or the enzyme spatially included separately. By appropriate selection of the wrapping material can the anatomical location of the merging of the two components and thus the physical conditions are determined under where the hydrolysis of the glucosinolates takes place. For example can be prevented by using gastric juice resistant capsules that the hydrolysis of the glucosinolates in the acidic environmental conditions of the stomach takes place where due to the low pH from 1.0 to 1.5 potentially unwanted Hydrolysis products of glucisinolates can be formed.

Both partial preparations of the kit can if necessary also frozen separately and in a frozen form in a common Hüllung be packed so that they can be defrosted before, during and / or after intended use mix and thereby hydrolytically isothiocyanates and / or ascorbigen arise.

• (a) Bereitstellen von Material aus mindestens einer Glucosinolat-haltigen Pflanzenspezies,
• (b) Hydrolysieren der in dem Material enthaltenen Glucosinolate durch die endogene(n)Thioglucosidase(n) nach Zerkleinern des Materials,
• (c) Extrahieren der Hydrolyseprodukte nach fakultativer Wasserzugabe und
• (d) Anreichern der Hydrolyseprodukte im Extrakt durch Gefrierkonzentration unter Konstanthalten des pH-Wertes.

Yet another aspect of the invention is a method comprising:

• (a) providing material from at least one plant species containing glucosinolate,
• (b) hydrolyzing the glucosinolates contained in the material by the endogenous thioglucosidase (s) after crushing the material,
• (c) extracting the hydrolysis products after optional water addition and
• (d) Enrichment of the hydrolysis products in the extract by freezing concentration while keeping the pH constant.

In this process, the glucosinolates are first hydrolysed to isothiocyanates and / or ascorbigens and then the hydrolysis products are enriched by freeze concentration. On the preferred implementation of the individual process steps is referred to the above statements. The time saving associated with this modification of the production process reduces process-related losses in heat-labile indolyl glucosinolates, in particular in glucobrassicin, the precursor of ascorbigen. This process variant results in a higher content of health-protecting ascorbigen in the end product.

Furthermore, the present concerns Invention of a vegetable plant obtainable by this process variant Preparation, which preferably has the features as described above.

Yet another subject of present invention is a pharmaceutical composition comprising a herbal preparation or a kit of herbal preparations with ascorbigen and / or isothiocyanates, obtainable by a process as previously described, and pharmaceutically acceptable carriers and adjuvants. The composition is for an outer or suitable for internal use in medicine or veterinary medicine. In particular the composition becomes a prevention and / or treatment of cancer.

The composition can optionally be in Form of liquids, Capsules, tablets, coated tablets, etc. for internal use, furthermore formulated as an aerosol for oral or pernasal inhalation. Furthermore, the composition in the form of lotions, ointments, Creams, drops, wet compresses and sprays for external Application, especially on skin or mucous membranes. Of course other types of formulation as well as the addition to food or nutritional supplements conceivable.

Furthermore, the invention is intended to the examples below explained become.

Examples

Example 1: Production an aqueous concentrated ascorbigen solution

• (1) Zunächst wurde eine konzentrierte wässrige Glucosinolat-Lösung aus Broccoli-Pflanzen hergestellt: Insgesamt 26 kg Broccoli (Brassica oleracea botrytis cymosa var. Italica; Herkunftsland: Spanien) wurden in kochendem Wasser über 4 min blanchiert, anschließend die Stängel entfernt. Die verbliebenen 13 kg Broccoli-Röschen wurden nach initialer Zugabe von insgesamt 1 l Wasser bei einer Temperatur von 4 °C über 10 min in einem Cutter/Mixer (Rotor GT 95; Schlaeffli & Mauerer; Interlaken, Schweiz) mit 12 000 U/min zu einem feinen Pflanzenbrei dispergiert. Anschließend wurden mittels Scheidepresse und nachfolgender Filtration zunächst die grobdispersen Feststoffe, danach mittels Separation (Westfalia LWA 205) die feindispersen Schwebstoffe abgetrennt. Den hierdurch erzeugten 6,07 l einer wässrigen feststofffreien Lösung wurde im Weiteren durch Gefrierkonzentration mittels Zentrifuge schrittweise Wasser entzogen, bis das Volumen zuletzt noch 1,0 l betrug. Proben der wässrigen Lösung wurden unmittelbar bei minus 20°C eingegefroren. In diesen Proben wurde mittels HPLC (Methode nach ISO 9167-1) der Gehalt an Glucobrassicin und 4-Hydroxyglucobrassicin bestimmt.
• (2) Die verbliebenen 1,0 l der konzentrierten wässrigen Glucosinolat-Lösung wurden unmittelbar anschließend auf 35 °C erwärmt und nach Zugabe von L-Ascorbinsäure sowie Natriumhydrogencarbonat ein pH-Wert von 5,0 eingestellt (RQflex 2; Merck, Darmstadt Deutschland). Danach wurden 750 g gereinigte Raphanusprossen mittels Cutter/Mixer (Rotor GT 95) zerkleinert, die Flüssigkeit abgepresst und anschließend mittels Filtration die Feststoffe entfernt. Der Nachweis von Myrosinase in den hierdurch erzeugten 350 ml einer feststofffreien, wässrigen Flüssigkeit erfolgte mittels Glucose-release-Methode (mod. nach VanEtten et al., J. Agr. Food. Chem. 22 (1974), 483-487). Diese Myrosinase-haltige Flüssigkeit wurde der wässrigen konzentrierten Glucosinolat-Lösung sukzessive vollständig zugegeben und die entstandene Mischung über zwei Stunden unter den oben angegebenen Bedingungen langsam gerührt. Proben dieser wässrigen Lösung wurden unmittelbar bei minus 20°C eingefroren. Die zu einem späteren Zeitpunkt mittels Spektrophotometrie durch Doppelbestimmung ermittelte Konzentration an Ascorbigenen in diesen Proben betrug 24,3 ± 0,02 mg pro Liter.

The solution according to Example 1 was prepared in two steps.

• (1) First, a concentrated aqueous glucosinolate solution was made from broccoli plants: A total of 26 kg of broccoli (Brassica oleracea botrytis cymosa var. Italica; country of origin: Spain) were blanched in boiling water for 4 min, then the stems were removed. The remaining 13 kg broccoli florets were, after the initial addition of a total of 1 liter of water at a temperature of 4 ° C. for 10 minutes, in a cutter / mixer (Rotor GT 95; Schlaeffli &Mauerer; Interlaken, Switzerland) at 12,000 rpm dispersed into a fine vegetable pulp. Subsequently, the coarsely dispersed solids were separated using a separating press and subsequent filtration, then the finely dispersed suspended solids were separated using separation (Westfalia LWA 205). Water was gradually removed from the 6.07 l of an aqueous solids-free solution produced in this way by freezing concentration by means of a centrifuge until the volume finally reached 1.0 l. Samples of the aqueous solution were immediately frozen at minus 20 ° C. The content of glucobrassicin and 4-hydroxyglucobrassicin was determined in these samples by means of HPLC (method according to ISO 9167-1).
• (2) The remaining 1.0 l of the concentrated aqueous glucosinolate solution were immediately heated to 35 ° C. and, after addition of L-ascorbic acid and sodium bicarbonate, a pH of 5.0 was set (RQflex 2; Merck, Darmstadt Germany) , Then 750 g of cleaned Raphanus sprouts were crushed using a cutter / mixer (Rotor GT 95), the liquid was pressed off and the solids were then removed by filtration. The detection of myrosinase in the 350 ml of a solids-free, aqueous liquid produced in this way was carried out by means of the glucose release method (mod. According to VanEtten et al., J. Agr. Food. Chem. 22 (1974), 483-487). This myrosinase-containing liquid was successively completely added to the aqueous concentrated glucosinolate solution and the resulting mixture was slowly stirred for two hours under the conditions specified above. Samples of this aqueous solution were frozen immediately at minus 20 ° C. The concentration of ascorbigens in these samples determined at a later point in time by means of spectrophotometry by double determination was 24.3 ± 0.02 mg per liter.

Example 2: Production an oily one preparation with enrichment of the ascorbigen formed by hydrolysis

The in 500 ml of the aqueous solution Brassica ingredients contained in Example 1 were obtained by solvent extraction transferred into 130 ml of olive oil, accordingly a water: oil ratio of around 4: 1.

The ascorbigen concentration in the aqueous Phase after solvent extraction, again using spectrophotometry by double determination, was found to be 16.0 ± 0.0 mg per liter. The content of ascorbigen in the oily product example 2 is calculated derive from the difference in concentration in the aqueous Phase before and after solvent extraction to 31.0 ± 0.02 mg per liter of oil.

rating of the examples

The concentration information for individual Glucosinolates and their hydrolysis products in mature Brassica plants differ considerably in various references. These differences are largely determined by a different method selection, in particular by the use of different preparative extracting agents on the plant material: While the extraction rate of the glucosinolates contained in mature Brassica plants or their hydrolysis products when using the solvent mixture dimethyl sulfoxide / dimethylformamide / acetonitrile is 100%, it is reduced to about 80% when using boiling methanol and even to about 30% when using water as an extractant (Fahey et al., (1997), supra). However, all non-aqueous extractants of Brassica ingredients are either not approved under food law or they hinder the enzymatic hydrolysis of the glucosinolates. So if preparations for human use are to be made, only water remains as the only suitable extraction agent for Brassica ingredients.

Because even under the optimal in vitro conditions a laboratory extraction only around 30% of the ingredients mature Brassica plants are transferred into an aqueous solution and thus extracted can, it can be concluded that after eating Brassica plants also no more than a third of that in the plant material analytically detectable glucosinolates or their hydrolysis products in the gastrointestinal tract in an aqueous and thus resorbable solution passes.

Will that in the examples according to the invention with 24.3 mg per liter determined concentration of ascorbigens the content in the literature with 7 to 18 mg per kilogram of Ascorbigen in the autolysate of mature broccoli plants (Hrncirik et al. Eur. Food Res. Technol. 212 (2001), 576-581), see above the following results: The concentration of ascorbigens is in the aqueous Extracts of Example 1 formally 1.3 times to 3.4 times higher than in the information in the literature on the ascorbigen content in the autolysate ripe broccoli plants. Taking into account the above-mentioned relationships, according to which only around a third of those contained in Brassica plants Glucosinolates or their hydrolysis products in an aqueous and thus directly absorbable solution passes from the gastrointestinal tract, is calculated for the example product 1 even a 4 to 9 times higher ascorbigen concentration compared to the information in the literature on the ascorbigen content in the Autolysate of mature broccoli plants.

In the oily preparation according to the invention In Example 2, the asorbigen content is 31 mg per liter 1.7 times to 4.4 times higher than in the literature mentioned above on the total content of ascorbigens in the autolysate of ripe broccoli plants.

Ascorbigens are lipid soluble and can in an oil solved can also be absorbed directly from the gastrointestinal tract. Therefore can also for the oily preparation according to the invention with ascorbigen a comparison with the proportion dissolved in water be carried out on ascorbigen in autolysate of mature plants: is calculated for the sample product 2 in the oily preparation even a 5-fold to 11-fold higher ascorbigen concentration compared to the literature on the ascorbigen content in the Autolysate of mature Brassica plants.

Claims (56)

Process for the preparation of a herbal preparation, full (a) Providing material from at least one containing glucosinolate Plant species, (b) inactivating endogenous thioglucosidase (s) in the material (c) extracting those contained in the material glucosinolates, (d) enriching the glucosinolates by means of freezing concentration, (E) enzymatic hydrolysis of the glucosinolates to the corresponding ones Isothiocyanates and possibly stable storage forms thereof and (f) optionally enriching the hydrolysis products. A method according to claim 1, characterized in that the material contains methylsulfinylalkyl glucosinolates and / or indolyl glucosinolates. Method according to one of claims 1 or 2, characterized in that that the material is free from 3-butenyl glucosinolate (gluconapine) and / or 4-pentenyl glucosinolate (Progoitrin). Method according to one of claims 1 to 3, characterized in that that the material has a maximum nitrate content of 100 mg / kg. Method according to one of claims 1 to 4, characterized in that that the at least one plant species containing glucosinolate is selected from Plants of the order Capparales, of which selected from the Capparaceae, Brassicaceae or Cruciferae, Koeberliniaceae, Moringaceae, Resedaceae and / or Tovariaceae. A method according to claim 5, characterized in that the at least one plant species containing glucosinolate is selected from broccoli, white cabbage, Red cabbage, cauliflower, Brussels sprouts, broccoli, kohlrabi, savoy and / or Chinese cabbage. Method according to one of claims 1 to 6, characterized in that that the material comes from mature plants. Method according to one of claims 1 to 7, characterized in that that irreversible inactivation of the endogenous thioglucosidase (s) done by heating. A method according to claim 8, characterized in that heating at a temperature of 95 ° C to 100 ° C for a period of 1 second up to 420 seconds. A method according to claim 8 or 9, characterized in that heating by immersion blanching, steam blanching or / and Microwave heating takes place. Method according to one of claims 1 to 10, characterized in that that after inactivation, a shredding of the plant material he follows. A method according to claim 11, characterized in that the crushing of the plant material with the addition of water or an aqueous one Medium. A method according to claim 12, characterized in that the water or the aqueous medium is free of metal ions. Method according to one of claims 1 to 13, characterized in that that the extraction is carried out at a temperature of 0 ° C to 8 ° C, preferably from 0 ° C to 4 ° C. Method according to one of claims 1 to 14, characterized in that that the extraction involves a vortex extraction. Method according to one of claims 1 to 15, characterized in that that after extraction, a separation of the shredded plant material he follows. A method according to claim 16, characterized in that coarsely dispersed plant material is separated by filtration becomes. A method according to claim 17, characterized in that finely dispersed plant material is separated by decanting becomes. Method according to one of claims 1 to 18, characterized in that that the freezing concentration enriches the glucosinolates, in particular of 3-indolylmethyl glucosinolate or of methylsulfinylalkyl glucosinolates at least five times the initial concentration in aqueous Extract is done. Method according to one of claims 1 to 19, characterized in that that the content of indolyl glucosinolates before or / and after Enrichment of the glucosinolates by exposure to temperature, in particular by heating to at least 70 ° C, is reduced. Method according to one of claims 1 to 20, characterized in that that the enzymatic hydrolysis of the glucosinolates by at least an exogenous myrosinase, preferably by a plant myrosinase, he follows. Method according to one of claims 1 to 21, characterized in that that the enzymatic hydrolysis after adding an enzyme activator, especially of L-ascorbic acid and / or glutamic acid, he follows. Method according to one of claims 1 to 22, characterized in that that the enzymatic hydrolysis at a temperature of 30 ° C to 37 ° C, preferably from 35 ° C, he follows. Method according to one of claims 1 to 23, characterized in that that the enzymatic hydrolysis after adding a buffer selected from Citrate buffer and / or sodium hydrogen carbonate. Method according to one of claims 1 to 24, characterized in that that the enzymatic hydrolysis takes place under conditions in which resulting 3- (hydroxymethyl) indole isothiocyanate (3-indolylcarbinol, 3-indolylmethanol, indole-3-carbinol, I3C) with L-ascorbic acid at least partially Ascorbigen is implemented. Method according to one of claims 1 to 25, characterized in that that the enzymatic hydrolysis after adding a physiologically acceptable Acid, especially citric acid, he follows. Method according to one of claims 1 to 26, characterized in that that the enzymatic hydrolysis at a pH of 4.0 to 5.5 he follows. Method according to one of claims 1 to 26, characterized in that that the enzymatic hydrolysis at a pH of 6.8 to 7.5 he follows. Method according to one of claims 1 to 28, characterized in that the enzymatic hydrolysis for a conversion of more than 50% of the total molar concentration of indolyl and / or methylsulfinylalkyl glucosinolates to the corresponding methylsulfinylalkyl isothiocyanates or leads to corresponding stable forms of storage, such as ascorbigenes. A method according to claim 29, characterized in that the enzymatic hydrolysis to an essentially complete implementation which leads to glucosinolates. Method according to one of claims 1 to 30, characterized in that that after the enzymatic hydrolysis an enrichment of the hydrolysis products he follows. Method according to one of claims 1 to 31, characterized in that that the enrichment of the hydrolysis products includes a freezing concentration. Method according to one of claims 1 to 32, characterized in that that the enrichment of the hydrolysis products turns into a non-aqueous solvent includes. A method according to claim 33, characterized in that the enrichment of the hydrolysis products by liquid: liquid solvent extraction he follows. A method according to claim 34, characterized in that the non-watery solvent selected is made from olive oil, coconut oil, Soybean oil, palm oil, sunflower oil, peanut oil, sesame oil, others for the suitable human lipids or mixtures thereof. Method according to one of claims 1 to 35, characterized in that that the herbal preparation is processed into a dry extract. A method according to claim 36, characterized in that the dry extract is made by freeze drying. A method according to claim 1 or 37, characterized in that of the preparation Substances are added that improve product quality. A method according to claim 38, characterized in that of the preparation Material from finely dispersed plant fractions is added. A method according to claim 38 or 39, characterized in that that of the preparation added at least one substance which is the activity of enzymes of the cytochrome P-450 group. A method according to claim 40, characterized in that this substance is selected is made from citrus oil, Orange oil, Ingredients of the grapefruit, the rosemary plant and / or zinc aspartate. Herbal preparation, available by the method according to one of claims 1 to 41, characterized in that that they are isothiocyanates or their stable storage forms, such as ascorbigens, in a molar concentration of more than 50% of the molar Has total concentration of the glucosinolate hydrolysis products. Herbal preparation according to claim 42, characterized in that it has a nitrite content of has a maximum of 0.1 mg per kilogram or liter. Herbal preparation according to claim 42 or 43, characterized in that they contain substances, selected from thiocyanates, nitriles, epithonitriles and oxazolidine ions, in a lower molar concentration than after an autolysis of material from at least one containing glucosinolate Plant species among natural uncontrolled conditions after shredding the plant material. Process for the preparation of a herbal preparation, full (a) Providing material from at least one containing glucosinolate Plant species, (b) inactivating the endogenous thioglucosidase (s) in the material (c) extracting those contained in the material Glucosinolates and (d) preparing a dry extract of Plant material containing glucosinolate by freeze-drying. A method of making a kit comprising two herbal preparations with indolyl and / or Methylsulfinylalkyl glucosinolates and a thioglucosidase / glucohydrolase, full (a) Prepare a preparation with highly concentrated Indolyl and / or Methylsulfinylalkyl glucosinolates by a method according to claim 1 (a) to (d) or 45, (b) preparing a preparation, containing a thioglucosidase / glucohydrolase, a buffer substance, preferably selected from citrate buffer and / or carbonate buffer, optionally an enzyme activator and optionally a physiologically acceptable acid and (c) packing both preparations to a common composition, however, such that these preparations immediately before, during and / or mix according to the intended use. A method according to claim 46, characterized ge indicates that preparation (b) contains L-ascorbic acid and the pH is 4.0 to 5.5, so that when mixed with preparation (a), ascorbigens are formed hydrolytically. A method according to claim 46, characterized in that the pH of the preparation (b) is 6.8 to 7.5, so when mixed with preparation (a) Hydrolysed methylsulfinylalkyl isothiocyanates. Method according to one of claims 46 to 48, characterized in that that step (c) is a spatial separate packaging of both preparations. Method according to one of claims 46 to 48, characterized in that that step (c) separate freezing of the preparations after (a) and (b) and packaging of both frozen preparations (a) and (b) in a common enclosure. Kit comprising two herbal preparations, available by a method according to any one of claims 46 to 50. Process for the preparation of a herbal preparation, full (a) Providing material from at least one containing glucosinolate Plant species, (b) hydrolyzing the glucosinolates contained in the material by the endogenous glucosidase (s) after crushing the material, (C) Extract the hydrolysis products after optional water addition and (d) Enrichment of the hydrolysis products in the extract Freezing concentration while keeping the pH constant. Herbal preparation, available by a method according to claim 52. Herbal preparations or kits of herbal preparations with ascorbigen or isothiocyanates, obtainable by a process according to one of the claims 1 to 41, 46 to 50 or 52, characterized in that they are free of flatulence-producing, insoluble, non-starch polysaccharides, such as cellulose, hemicellulose and lignins. A pharmaceutical composition comprising a herbal preparation or a kit of herbal preparations with ascorbigen or isothiocyanates, available by a method according to any one of claims 1 to 41, 46 to 50, or 52 or according to one of the claims 42 to 44, 51, 53 or 54, and pharmaceutically acceptable carriers and Aids. A pharmaceutical composition according to claim 55 for prevention and / or treatment of cancer.