PREPARATION AND USE OF PLANTS OF IMPROVED BIOLOGICAL VALUE

Abstract: The invention relates to a process for preparing plants or parts of plants of improved biological value which comprises growing plant seed, tissue or cell in a culture medium completed with one or more trace elements in a concentration of 10<-9>-10<-1> M. The obtained plant, part or extract of plant of high trace-element-content in form of dietetic, pharmaceutical or cosmetical composition.

Description: PREPARATION AND USE OF PLANTS OF IMPROVED BIOLOGICAL VALUE The invention relates to a cultivating process for preparatiπg plants of improved biological value, first of all of plant germs, as well as plant organs, tissues and cells and the use of the thus-prepared active ingredient as cosmetical, dietetic and pharma- ceutical compositions. All living organisms consist of the elements of the periodic system. Eleven elements of this system can be found in the living creatures in great amount, these are: carbon, hydrogen, oxygen, nitrogen, sulphur, cal- cium, phosphorus, potassium, sodium, chlorine, magne¬ sium. Further 16 elements can be found in the organism of living creatures only in a small amount, these are the so-called trace elements, such as iron, zinc, copper, manganese, nickel, cobalt, molybdenum, selenium, chromium, iodine, fluorine, tin, silicon, vanadium, arsenic, lithium. In addition to these elements the presence of boron is absolutely necessary for growing of the higher plants. Most of the above elements are between the atoms No. 23 and 34. In this region the gallium and germanium can also be found, the biological role of which has been unknown and is being clarified lately. From the above it can be seen that the presence of element ' s, first of all of trace elements in a suitab¬ le amount is very important for life. In spite of the different artificial manures and fertilizers the trace elements-content of soil is more and more decreasing nowadays owing to the agricultural producing which has grown to huge dimensions. As a consequence troubles arise in the trace element supply of the soil - plant - human and animal food chain, which leads to an increased lack of trace elements. The most important point here is the trace element supply of the plant both for people and the animals for this is the first source where people and the animals can get trace elements from. Four main factors influence the trace element content of plants: a) genus, species b) ' type of soil, where the plant is growing c) climatic and seasonal circumstances during growing d) stage of development of the plant. In the specification according to the invention thos possibilities have been optimalized and used that certain plant species are able in themselves or by a gradual accustoming to enrich certain trace elements in their tissue in biological active form and high coπceπt- ration. Astragalus racemosus and Neptunia amplexicaulis are able for example to accumulate selenium. Nyssa sylvatica gathers cobalt from the same soil a hundred times better than other plants do. Similar differences can be observed between legu¬ minous plants, Gramineae and cereals growing on the same soil, to the advantage of leguminous plants in case of Ca, Mi, Fe, Cu and Zn. Gramineae and cereals can gather Mn, Mo and especially silicon much better. During development these elements can decrease and increase in the whole plant or may become stable. In the next step the trace element amount already decreases in the human and animal organism too, which may cause different symptoms. These symptoms can be (detailed literature can be found: Trace elements in Human and Animal Nutrition, Eric. J. Underwood; Academic Preww New York, San Francisco, London, 1977) and the same time the biolo¬ gical advantages of the trace elements used according to the process of the invention can be summarized as follows: The presence of iron (Fe ) in the human organism is of especially great importance, its whole amount is 4 to 5 g. which can be found mainly in the hae oglobine and myoglobine, flavopr oteins contain less than 1 % of same. Lack of iron may cause serious diseases in forma¬ tion of blood and this is the most obvious sign of lack of iron, but there are other lack symptoms such as palpitation, sore tongue, stomatitis, dysphagia. Copper is also important in blood formation, its whole amount runs to 80 mg. Several proteins of copper content are known such as erythrocuprine, cerebracuprine, hepatocupriπe , cytocuprine. These enzymes have a sig- nificant role in the oxydatiπg mechanisms and in the inhibition and formation of free radicals. It has effect on the hair growth, for the disulphide bounds will be formed only in its presence during the keratin synthesis, and as it is part of the polyphenyloxidase enzyme form- ing tyrosiπe into melanine it has a role in becoming gray. The biological role of nickel proved to be signi¬ ficant first of all in case of myocardial iπfarct and burning lesions. Lack of nickel also decreases the work- ing of liver hydrogeπase. Manganese can be found in the organism in a total amount of 12-20 mg. , first of all in the mitocondriums . In lack of manganese the growth of bones decreases as the synthesis of coπdroitine sulphate and hexcsamine decreases, ataxia arises, the reproduction functions sustain injuries, the lipid metabolism alters and the cholesterol synthesis decreases. The active part of se¬ veral enzymes as hydrolases, quiπases, decarboxylases, traπsferases etc. decrases. Molybdenum can be found in every tissue in a low concentration. The flavoprotein and xantinoxidase enzyme are metalloenzyme containing molybdenum, its lack causes caries- Selenium can be found in the liver and kidney in the greatest amount. Its importance in the oxidative mechanism was recognized lately. It is ingredient of the glutation peroxidase which protects the organism from the unbeπeficial peroxides: 2 GSH + H 2 0 2 -Sly..§ gn.pergxidase__ QSSG + 2 ^ GSSG + NADPH 2 GSH + NA[}p+ Therefore selenium protects the cells directly from oxi¬ dation, the exudative diathesis can be prevented by add¬ ing selenium or vitamine E. The effect of selenium is similar to that of vitamine E known as aπtioxidant, they both protect the biological membraπs from the oxidative attack and prevent the exudative diathesis. The cyto- chromes containing selenium play presumably a role in the oxidation-reduction mechanisms of tricarboxylic acid cycle. Its lack causes disorders in growth, sexual troubles, -pancreatic fibrosis hepatosis detetica, exuda- tive diathesis. The examinations of the previous years have shown the very important role of selenium in prevention of cancer (Mark F. Mc. Carty, Medical Hypothesis 1_4, 213- -225, 1984). According to the test results the addition of 600 meg/day of selenium protects people from several chemical inductors causing cancer and supports the work¬ ing of the DNA repair. During the examinations it became obvious that the effectivity of organically bound sele¬ nium is the same in spite of the fact that its toxicity significantly decreased. This examination was carried out by using sαdiumselenit and yeast fungus enriched with selenium (Schwarz K., 60-day toxicity trial with selenium yeast II. Dec. 21, 1975 unpublished manuscript). A significant amount of silicon can be found in the skin, this element is integral part of mucopoly- saccharides, being essential structural components of the skin. According to the examinations of Schwarz (Proc. Nat Acad. Sci. USA 7_0_ 1608, 1973) not less than 330-554 ppm of bound silicon can be found in hyaluronic acid. At the same time it was also proved that the c< chains of collagen contain at least 3-6 atom of silicon. Silicon is essential for bone formation and growth, it has effect directly for the amount of phosphorus. 70-80 % of iodine can be found in the thyroid gland, in lack of iodine the cellularic oxidation suffers injuries, respiration increases in the mitocoπdriums. It has an effect on the differentiating and growth of cells, inducing the RNA and protein synthesis by controll ing the gen expression. The fluorine-content of the organism is concent¬ rated in the bones and teeth and inhibits the caries and osteoporosis. Chromium can be found in the organism only in a very small amount its role is, however, significant. Typical symptoms of chromium-deficiency are: backwardness of growth, disorders of the metabolism of gluco, lipid and protein. The role of tin is less known but it is absolutely necessary for growth. Vanadium is present in a very low concentration. It has effect on the reproduction and the lipoid eta- bolism. It hinders the cholesterol synthesis and the phospholipoid and cholesterol level of plasm and reduces the cholesterol concentration of the aorta. It protects against caries. In a small amount (30 ppb) arsenic could have an adavaπtageous effect to growth, in higher concentration is as selenium antagonist disadvantageous. The physiological role of lithium in the living organism is unknown though it has been used for a long time for treating psychosis (manic-depression). Its acti- vity for the herpes viruses was recognized lately. The role of boron by mammalia is unknown but it is essential because of its role in the flavonoid synthesis with higher plants. Germanium was known until the late years as being necessary in the organisms of mammalia. According to the newest search results it plays a significant role in the oxidative mechanisms, its role is similar to that of se¬ lenium. It has a strong effect against tumours and is espe- cially good for the strong metastatising and hard influ- encable Lewis-Lung carcinoma in mice ( umaπo N., Ishikawa T., Koiπumaru S., et al. Antitimour effect of organoger- manium compound G-132 on the Lewis-Lung carcinoma /3LL/ in C57 PL/6/86/ mice Tohoku J. Exp. Med. 1985 146/1 94-104.). Here we would especially remark that first of all the representative of the organic germanium compounds have been examined and found to be effective. Germanium presumably has its required effect by modifying the immun response (Sato I., Yaπ B.D., Nishimura I., Tanaka N. Inhibition of tumor growth and metastasis in associa¬ tion with modification of immune response by novel or¬ ganic germanium compound 3. Biαl. Response Modif. 1985 4/2, 159-168). Magnesium has a various role in the organism.It takes part in the working of several enzymes. The most significant is to ensure the activity of these taking part in the πucleinacid,protein synthesis. Calcium is present in the organism in a. relative great amount, its role in the bone and tooth formation is ' significant. The aluminum needs of the organism is less than 1 ,ug per day. It is the promoter of succiπe dehydro- genase enzyme. The importance of cobalt in the blood formation is known, the cobalt and iron are transportated together. It is ingredient of vitamine B, 2 promoting the transfor¬ mation of methylmalonyl CoA into succinyl CoA. 5-Methyl- -tetrahydrofolcyl:homocysteine methyltransferase is also an important enzyme catalysing the formation of ethio- πiπe from ho ocysteine. Daily needs: 3 mg./kg. One fifth of zinc can be found in the skin. Its importance is great in the liver where it is present in the metallothionine. A relative great amount is in the eyes (139-436 ppm) , in the testis (120 ppm) , in the prostate (859 ppm), in the nail (93-292 ppm) and in the hair (92-255 ppm). In blood it is found bound to «λ macroglobuline and albumine. Stress decreases the Zn level. In lack of zinc the growth is backward as the activity of timidiπe chiπase is decreasing and also the DNA synthesis and cell division are decreasing, alopecia, dermatitis, deformations on the skin, chronic skin ulceration arises. - a - The hyaluroπic acid amount in the skin also decrases (Thomson, R.W. 3 . Nutr. 105, 154, 1975), the collagen synthesis is reduced (McClaiπ BBA 3_3__, 457, 1973 . Zinc is therefore of great importance for it plays a part in the collagen, DNΛ , RNA and protein syn¬ thesis, with other words the presence of zinc is necessa for the progresses of gen activation. Further enzymes containing zinc are the pankreas carboxypeptiase. lactas dehydrogeπase (LDH), alonic acid dehydrogeπase J«1DH) , alcohol dehydrogenase (ADH), alkalic phosphatase. 15-25 mg. of zinc is necessary a day for people. At last we mention the precious metals as gold, silver, platinum direct the trace element effects of which are unknown but they are used as medicines. In for of silver protein silver is used mainly for disinfection gold by rheumatic articular diseases and platiπa for treating cancer. From the above it is clear that the maint nance of the suitable level and the supplying of trace element in the living and first of all in the human organism, are of primary importance. The healing of several unknown diseases can be achieved many times by the addition of the suitable trac element into the organism. According to the invention trace elements are en¬ riched in the organism of the suitable plant arz used as cosmetics, dietetic compositions or pharmaceutical com¬ positions in a biologically valuable form, that is in a form being directly usable for the organism of -ammalia. According to what kind of trace elements it co ~ :ains the composition is suitable for the medical treatment and healing of the above biological disorders crtailed by the trace elements. It is known that administering the trace elements in the form of inorganic salt is not suitably effective for the human organism as it cannot use this form suitab¬ ly (e.g. iron compositions). A more advantageous form is when the metal ion in question is bound to a carrier mole- cule which after getting into the human organism is ab¬ sorbed and gets to the active centrum of the given enzyme; it activates the enzyme so that it can fulfil it function in the organism. In case to the contrary the microelement in question (metal ions) leaves the organism without being used. According to the process of our invention micro¬ element-carrier complexes are prepared by using different plants, medicinal plants, agricultural plants or feral plants or parts of plants which organic complexes comple- tely fulfil the above requirements. According to the invention seeds, tissues or cells of different plants are grown in a culture medium completed with trace elements in a concentration of 10 -9-10-1 M. From the plants, plant tissues or cells of high trace element-content dietetic compositions, pharmaceutical com¬ positions or cosmetics containing trace elements of high biological value are prepared by processing or after extraction. During growing (germinating) the seeds according to the invention and the cultivation of the plant tissues and cells according to the invention a solid or a liquid culture medium is used. The following culture media can preferably by used: White, Murashige-Skoog, Gambαrg (B5) and that of Nitsch- -type, ("Tissue Culture Methods and Applicaitons" , Edited by Perul F. Kruse, JR. and M.K. Patterson, J.R. Academic Press, New York and London, 1973; Murashige, T.: Nutrition of Plant Cells and Organs in vitro. 3_, 81-85, 1973; B.V. Conger et al: "Cloning Agricultural Plants via in vitro Techniques", CRC Press, 1981; P.H.Calcoot: "Continuous Cultures of Cells", CRC Press, 1981). Any elements which can be found in a little amount in living organism and which have biological activity can be used as trace element. Preferred trace elements are selected from group consisting iron, zinc, copper, manganese, nickel, cobalt, molybdenum, selenium, chromium, iodine, fluorine, tin, silicon, vanadium, arsenic, boron, lithium, gold, silver, platinum, bromine, gallium, ger- maπium, magnesium, calcium, and aluminum. This enumera¬ tion is not of restricting nature. The trace elements are added into the culture mediu in form of organic or inorganic compound. Ionic compounds can preferably be used such as the suitable halogenides in case of metallic trace elements such as tiπchloride, lithiumchloride or goldchloride, other mineral acidic salts, as πickelsulphate, chromosulphate or silver nit¬ rate, furthermore compounds of covalent bond, e.g. arsene trioxide and organic compounds such as selenocysteine. For administering the trace elements water of na¬ tural origin, as sweet water, mineral water, thermal wa¬ ter, medicinal water or the mixture thereof can complete¬ ly or preferably partly be used, which contain the requir ed trace elements. The given water can be used as the solvent or diluent of the culture medium used. Waters of natural origin such as the water of Parad, Heviz and Margitsziget (Margaret Island, Budapest), Hungary and Postyeπ, Czechoslovakia, as well as the mineral water named Mohai Agnes can preferably be used. The great advantage of the waters of natural ori¬ gin is that they show up an individual trace elemeπt- -composition, which has been formed in itself in the nature during a long period and in addition to the. traceable trace elements of known biological activity (see the enumeration above) they contain further, un¬ known trace elements which cannot be aπalized yet accord ing to 'the present state of prior art. By using the natural waters the advantageous trace element compositio and the activity of the trace elements which are present in the nature but have not been tested yet, are utilized. One or more elements of the natural trace element compo¬ sition can be enriched by an extra addition in the requir ed manner. During germination or cultivation the concentration range is registered, where the plants are growing well and their trace-elemeπt-coπtent is higher than the natu¬ ral value. The plants being able to develop also by a high trace-elemeπt-content are selected then cloned, subjected to further cultivation at the given high trace element concentration, than if required, they are further in form of a tissue- or a cell culture, preferably in a suspension culture. Continuous accustoming is used by certain plants storing hard the trace elements, that is the given trace element coπceπtraitoπ is not added at once into the cul¬ ture medium but divided it is gradually increased for 1-28 days until the required level is achieved. According 'to our experiments after 28 days the absorbtioπ of trace elements does not increase significantly. The natural level of trace elements is increased by this process ' generally by 4-5 orders. This means that starting from the concentration of 10 M of the natural level the concentration of 10 " M is πeared depending on the given trace element and plant. Germination is carried out in a culture medium completed with trace element pref-erably at a temperature of 5-35 °C for 1 to 3 weeks under aseptic or sterile con- ditions. The cultivation of the plant tissues and cells is carried out preferably at a temperature of 10-30 °C for 7 to 28 days under sterile conditions by using lighting periods of 16 hours, if necessary. In principle any plant can be used according to our invention. First of all plants containing high amount of trace elements belonging to the legumes (Legumiπαsae- sal) , papilionaceae (Fabaceae) and gramineae (Gramiπales) can preferably be used. The usable plants are for example as follows: bean (Phaseolus vulgaris), pea (Pisum sativum) , lens (Lens culiπaris), Astragalus racemosus, Neptunia amplexicaulis, white lupin (Lipiπus albus) , wheat (Triticum aestivum) , barley (Hαrdeum nodosum) , rye (Secale cereale) , oat (Ave- πa sativa), rice (Oryze sativa), mais (Zea mays), millet (Paπicu miliaceum ) rye grass (Lolium pereπne) as well as garlic (Allium sativum), onion (Allium cepa) , radish (Raphanus sativus), gipsy-wort (Lycopus exaltatus), borag (Borago officinalis) , evening primrose (Oeπothera bieππis syo bean (Glycine soja), cowslip (Primula veris), Lathyru sativus, sorghum (Sorghum vulgare) . The above enumeration is not of re-strictiπg nature. From the above plants very hardy entities and tissue or cell cultures can be prepared by cloning, which grow well by a trace-elemeπt-conteπt of high coπceπtraito they absorb the trace elements into their organism and build them in. After accumulation, drying and storing the substance of high biological value can be used as supplement, medicine and cosmetics. • The plant material of improved biological value wit high trace-elemeπt-content obtained according to the process of the invention is processed for use. If desired germ is decomposed to its parts, i.e. roots, stem and leaves are separated from each other then dried, pregroun and micronised. The thus-obtained powder can directly be used. In case of a tissue- or cell culture, the culture is dried, the excess substance such as sugar and other culture medium residue is removed by washing with water then dried and processed in analogues way as germ. Drying is preferably carried out by air current at a temperature up to 60 C. The pregrinding is carried out until a grain size below 0.1 mm, is achieved, while micronizing is carried out until 1 , grain size is achieved. The powder thus obtained is converted known per se into the usual compositions as tablets, capsules, soft gelatine capsules, dragees, pirules, granulates, aero- sole, syrup, emulsion and suspension for using it as dietetic or pharmaceutical composition by means of intoxic pharmaceutically acceptable carriers or solvents. The concentration of the plant substance of high trace-element-coπtent is about 0.5 to 90 mass% relating to the whole mixture, i.e. it is a value being necessary for achieving the desired level of dose. These compositions can be prepared for example by mixing a plant powder with a solvent and/or with a carrier optionally by using emulsifying and/or dispersing agents, whereas in case of using water as diluting agent, as auxiliary solvent optionally organic solvents can also be used. The following auxiliary agents can be used: e.g. water, intoxic organic solvents, such as paraffines (e.g. oil fraπtiαπs), plant oils (e.g. hazel-nut oil or sesame oil), alcohols (e.g. ethanol, glycerine), solid carriers, for example natural rock-dusts (e.g. caoliπe, clay, talc, chalk), artificial inorganic powders .(e.g. silicic acid of high dispersion degreee or silicates), sugars (e.g. saccharose, lactose, glucose), emulsifying agents (e.g. poly/oxy-ethylene/-fatty acid ester, poly/oxy-ethylen/-fatty alcohol ether, alkyl-sulphoπate, arylsulphoπate) , dispersing agents (e.g. lignine, sul- phite-sewage base, methyl-cellulose, starch and poly/vi- πyl-pyrrolidoπe/) and sliding materials (e.g. magnesium- -stearate, talc, stearic acid and sodium-lauryl-sulphate) The substances are administered in usually way, preferably orally. Of course, in case of oral admiπistra- tion the tablet may contain in addition to the above-men¬ tioned carriers other materials, such as sodium-citrate, calcium-carbonate and dicalcium-phosphate together with other auxiliary agents, such as starch, preferably potato starch, gelatine and other similar materials. For forming tablets sliding materials, such as magπesium-stearate, sodium-lauryl-sulphate and talc can also be used.. In case of an aqueous suspension for oral administration the acti ingredients can be mixed in addition to the above mention auxiliary agents with different flavouring and colouring substances. For cos etical purposes the plant powder of high trace-element-coπteπt is concerted into cosmetical compos tions in a way known per se. The following compositions are preferably be used: body lotions, face creams, hydrat ing creams, regenerating creams, creams for treating wriπckles of the eye, different gels and jellies, mascs as well as shampoos, bath foams and dentifrices. For preparing cαsm-etical compositions the liquid carriers, such as water, alcohol (e.g. ethanol and glyce- riπe) , paraffiπe oil, plant oils (such as sunflower oil), synthetic oils (e.g. silicon oil), glycerine-esters (such as glyceriπe-moπostearate) and fatty alcohols (e.g. cetylalcohol) , furthermore emulsifying agents, such as ionic or non-ionic emulsifying agents (e.g. poly/oxi-ethy lene/-fatty acid ester, poly/oxy-ethyleπe/-fatty alcohol ether, alkyl-sulphonate and aryl-sulphonate), gel forming agents (e.g. algiπates, polyviπylcarboxylic acid) further more other additives, as rancidity-inhibiting agents (as butylhydroxy-toluol) , conserving agents, essence of perfume, or flavouring agents and colouring ageπst can be used. In addition to the plant powder the compositions according to the invention can contain other known active ingredients. As especially preferable effect can be achie ed by using germ oils of physiological activity. Cold- -pressed plant oils, such as maisger oil, wheat germ oil puπpkiπ-seed oil, oil of evening primrose, soy bean oil as well as the mixture thereof can preferably be used. The invention is demonstrated more detailed by the following Examples. Example 1 Beans (Phaseolus vulgaris) sterilized by a 3 % hydrogenperoxide are subjected to germination on a liquid containing 0.5 mM of CaSO^, which liquid also contains cobalt chloride as trace element enrichment in a 10 -9-10-1 concentration line in increasing order. Ger¬ mination is carried out with 50 seeds per concentration for 10-14 days at a temperature of 25 °C. The highest cobalt concentration is thereafter stated which the bean can bear by good growing. Leaves and roots of the here grown germ plants are furthered, cloned under steile conditions. The root and leaf-parts are grown on a modifie solid White cultur medium completed with the highest cobalt concentration already stated. The composition of the White culture medium is the following: 0.36 g./l. of agπεsiumsulphate, 0.26 g./l. of calciumπitrate, 0.2 g./l. of sodiumsulphate, 0.08 g./l. of potassium- nitrate, 0.065 g./l. of potassiu chloride, 0.165 g./l. of sodium dihydrogeπphosphate, 3 mg/1. of manganese- sulphate, 0.5 mg./l. of ziπcsulphate, 0.5 mg./l. of boron acid, 0.025 mg./l. of copper sulphate, 1 mg./l. of iron citrate, 7.5 mg./l. of glycine, 1.15 mg./l. of πyacine, 0.25 mg./l. of thamiπe HC1, 0.25 mg./l. of Pyridoxiπe HC1, 0.25 ml. of calcium pantotheπate, 2 mg./l of indolacetic acid, 20 g./l. of saccharose, 8 g./l. of agar-agar. Growing is carried out at a temperature between 20-30 °C with redividing per 14-21 days, lighting was use periodically in every 16 hours. From the leaf and root parts a suspension culture was prepared on the liquid Gamborg (B5) culture medium without agar-agar and they are grown here in a continuous ly working cultivating reactor. After the suitable incubation time (14-21 days at 20-30 °C) plant parts containing great amount of cobalt are gathered by centrifuging or filtrating, decomposed, dried and prepared for use. Example 2 The process of Example 1 is followed with the difference that_ Gamborg (B5) solid culture medium is used - in every case. Example 3 The plant seeds fertilized by using a 3 % hydrogen peroxide are subjected to germination in a plastic tray of 25 x 40 x 10 cm, where an acidproof steel net is strained at 8 cm. The plant seeds are placed depending on their size (50 to 250 pieces) , and the trays are fille till line of the net with a sterile liquid culture medium The liquid culture medium contains 0.5 mM of CaSO, and in addition until a concentration of 10 -9-10-1 M one or more trace elements and/or up to 0-100 % mineral water, thermal water, medicinal water or the mixtures thereof. The end concentration of trace elements is achieved during 1-21 days by administration per 1 to 5 days. The seeds are lighted by neon lights during the whole period of germination, using lighting periods of 16 hours. Temperature: 5-35 °C. After the germination is finished the plants are dried and milled to 1 um in a ball mill. The plant cell are so decomposed. The grist thus obtained can directly be used for prepar¬ ing the compositions. Example 4 The process according to Example 3 is followed. The germed plant is bean (Phaseolus vulgaris). Starting concentration of zinc sulphate as trace element is 1.8 x 10 -5 M, the end concentration is 1.8 x 10-2 M. Example 5 The process according to Example 3 and 4 are follow with the difference that mineral water of Parad (Hungary) is added to the germinating solvent in an amount of 80 %. Example 6 The process according to Example 3-4 are followed with the difference that medicinal water of Heviz (Hungar are used as germinating solvent in an amount of 100 %. Example 7 The process according to Examples 3-4 are followed with the difference that thermal water of Margitsziget (Margaret Island, Budapest, Hungary) is used as germinat- ing solvent in an amount of 90 %. Example 8 The process according to Examples 3-4 are followed with the difference that an amount of 50 % mineral water of Ha duszoboszlό (Hungary) and 30 % mineral water named Mohai Agnes are used as germinating solvent. Number of Trace Starting End Examples Plant element concentration concentration Bean (Phaseolus vulgaris) iron citrate ,7x10 M copper sulphate xlO M zinc sulphate 8x10 M 10 Bean (Phaseolus vulgaris) nickel sulphate lO M 11 Mais (Zea mays) manganese chloride ,2x10 M 12 Avena sativa sodium molybdenate xlO M 13 Rice (Oryza sativa) sodium molybdenate xlO M manganese chloride xlO ' M C 14 Garlic (Alli'u sativum) sodium selenite xlO M I 15 Garlic (Allium sativum) sodium selenite xlO ' M germanium chloride xlO M 16 Astragalus racemosus germanium chloride xlO M 17 Neptunia amplexicaulis germanium chlorid xlO M 18 Onion (Allium cepa) germanium chloride xlO ' M 19 Wtiont (Triticum aesticuπi) sodium silicate xlO ' M 20 Rye grass (Lolium perenue) sodium fluoride 8 x 10 M 21 Gipsy wort (Lycopus exaltatus) germanium chloride M x l O ' Number of Trace Examples Plant element 22 White lupin (Lupinus albus) germanium chloride 23 Lathyrus sativus germanium chloride 24 Sorghum (Sorghum vulgare) sodium molybdenate 25 Barley (Hordeum nodosum) nickel sulphate 26 Rye (Secale cereale) chromosulphate 27 Radish (Raphanus sativus) platina chloride 28 Radish (Raphanus sativus) tin chloride 29 Barley (Hordeum nodosum) ammoniumvanada e 30 Borage (Borago officinalis) lithium chloride 31 Cowslip (Primula veris) lithium chloride 32 Evening primrose (Oenothera biennis) lithium chloride 33 Pea (Pisu sativum) gold chloride 34 Pea (Pisum sativum) silver nitrate 35 Lens (Lens culinaris) gallium sulphate 36 Pea (Pisum sativum) germanium chloride 37 Soy (Glycine soja) magnesium chloride calcium chloride Number of Trace Starting End Examples Plant element concentration concentration 38 Bean (Phaseolus vulgaris) cobalt chloride x H 2 0 4.2x10 ' M 39 Carrot (Daucus carota) sodium selenite xlO ,-2 M 40 Carrot (Daucus carota) germanium chloride xlO M Example 41 The plant parts, leaf and root pieces, prepared for cloning, are grown on sterile, modified solit White culture medium. The composition of the White culture medium is: 0.2 g./l. of sodium sulphate, 0.08 g./l. of potassium nitrate, 0.065 g./l. of potassium chloride, 0.165 g./l. of sodium dihydrogeπ sulphate, 3 mg./l. of manganese sulphate, 0.5 mg./l. of zinc sulphate, 0.5 mg. of boron acid, 0.025 mg./l. of copper sulphate, 1 mg./l. of iron citrate, 7.5 mg./l. of glycine, 1.25 mg./l. of πyacine, 0.25 mg./l. of thiamin HC1, 0.25 mg./l. of py- ridoxine HC1, 0.25 ml. of calcium pantothenate , 2 mg./l. of iπdol-acetic acid, 20 g./l. of saccharose, 8 g./l. of agar-agar. Growing is carried out at 20-30 °C with re- dividing per 14-21 days, the plants are lighted periodi¬ cally in every 16 hours. After the suitable amount of tissues is obtained a suspension culture is prepared from the leaf and root parts on the liquid culture medium Gamborg (B5) without agar-agar (Tissue culture Methods and Applications Edited by Perul F. Kruse, JR. and M.K. Patterson, 3 . R . Academic Press New York and London, 1973; Murashige, T: Nutrition of Plant Cells and Organs in vitro _9_, 81-85, 1973; Conger, B.V. et: Cloning Agri¬ cultural Plants via in vitro Techniques CRC Press 1981, Calcott, PH: Continuous Cultures of Cells CRC Press 1931) and they are grown here in a continuously working cul¬ tivating reactor. Incubation time is 7-28 days at 10-30 °C, the lighting periods are of 16 hours. The trace-element-coπtent of the liquid culture medium is increased between the concentration 10 -9-10-1 M by administration per 1-5 days until the end concentra¬ tion is achieved and/or the culture medium is completed from 0 to 100 % with mineral water, thermal water, mεdi- ciπal water or with the mixtures thereof. After the cultivation is finished the plant cells containing great amount of trace elements are gathered by centrifuging or filtration, they are decomposed, dried and prepared for 'use. Example 42 To 200 g. of lactose 20 g. of silicon acid of the trade-name Aerosil are added under stirring and 40 g. of plant active ingredient of Example 38 are added in form of flour with a 3 % humidity content, and it is homogenized in inert atmosphere. After moistening the mixture is stirred in a malaxator for 30 minutes until a paste-like mass is ob¬ tained; granule is prepared from the obtained mass by a known method. The granule is dried in fluid dryer in inert gas atmosphere. Drying is continued until the water content of -the sample decreases under the desired value measured by method of Karl-Fischer. After drying the granule is ground and filled into hard gelatine capsules of 250 mg. by known method. 1000 pieces of capsules are obtained soothing the symptoms of lack of cobalt. Example 43 40 g. of the plant active ingredient of Example 38 are homogenized with 220 g. of mais germ oil and in a cold pressed soft gelatine capsulating equipment it is filled into oval capsules of 250 mg. by known method. Example 44 40 g. of the plant active ingredient of Example 38 are homogenized with 140 g. of lactose, 20 g. of saccharose, 26 g. of potatoe starch, 4 g. of talc and 4 g. of magnesium stearate, then granulated and pressed into tablets of 250 mg. by known method. Example 45 The tablets prepared according to Example 44 are panned by a known method in a panning boiler and is coated with a yellow layer. Example 46 10 g. of the plant active ingredient prepared according to Examples 3-4 are suspended in 62.3 g. of water and heated to 70-75 °C and added to the fatty phase of 70 °C containing 7 g. cetyl alcohol, 10 g. of mais germ oil, 2 g. of vaseline, 5 g. of vaseline oil, 1 g. of white beeswax, 2.5 g. Tween 60. The emulsion is stirred until cooling. At a tempe¬ rature of 40 C the conserving agent (0.1 g.) and the flavour (0.1 g.) are added. A skin treating creame of high quality is obtained. Example 47 5 g. of cetyl alcohol, 10 g. of vaseline, 4 g. of vaseline oil and 2 g. of Tween 60 are melt together at 70-75 °C and the water phase being necessary to 100 mass%, prepared according to Examples 3-4 and contain ing 5.0 g. of plant active ingredient are added. The body lotion is stirred until cooling off, then preserv¬ ing agent and favouring agent are added to the composi¬ tion in 0.1 % and 0.15 %, resp. at 40 °C . The obtained composition is a body lotion with hydrating and skin regenerating activity of high quality. Example 48 5 g. of the plant active ingredient of Examples 3-4 are suspended in 100 g. of evening primrose oil which was preserved with 0.02 % of butyl hydroxytαluole . The composition is then filled into a 2 ml. tube-forming soft gelatine capsule. Ampouls are prepared by known method in a soft gelatine capsulating equipment. Example 49 5 g. of the active ingredient of Example 3-4 are suspended in water and added to 1 g. of Carbopol 940 which was swelled in 50 g. of water for 24 hours. Preserving agent content is 0.1 %. Thw two water-phases are mixed then stirred with 1.0 g. of triethanolamiπe until gel is formed. An excellent hydrating, unwrinckling gel is ob¬ tained. Example 50 10Q g. of polyvinylalcohol are swelled warm in 600 ml. of distilled water. The homogeneous solution is cooled to room temperature then 200 ml of 9 G - % alcohol and 10 g. of plant active ingredient prepared according to Examples 3-4 suspended in 190 g. of water are added. The obtained mask smoothens and hydrates the skin of face.- Example 51 To 1. g. of plant active ingredient prepared according to Example 3-4 93 g. of Texapon N 25 then 6 ml of a 10 mass% hypagine M are added. The obtained product is used as a hair tonic and shampoo.

Claims: CLAIMS 1. Process for preparing plants or parts of plants of improved biological value which comprises growing 5 plant seed, tissue or cell in a culture medium completed with one or more trace elements in a concentration of 10 "9 -10 _1 M. 2. A process according to claim 1 which comprises using one or more of the following elements as trace ele- -10 meπts: iron, zinc, copper, manganese, nickel, cobalt, molybdenum, selenium, chromium, iodine, fluorine, tin, silicon, vanadium, arsenic, boron, lithium, gold, silver, platinum, bromine, gallium, germanium, magnesium, calcium and aluminum. 15 3. A process as claimed in claims 1 to 2 which comprises inserting the trace elements into the culture medium in form of organic or inorganic compound. 4. A process as claimed in claims 1 to 2 which comprises inserting the trace elements partly or complete 20 ly into the culture medium in form of water of natural origin such as sweet water, mineral water, ' thermal water, medicinal water or the mixture thereof containing the desired trace elements. 5. A process as claimed in claims 1 to 4 which 5 comprises cultivating the plant seed, tissue or cell for 1 to 28 days in a culture medium completed with one or more trace elements the concentration of which gradually increases. 6. A process as claimed in claims 1 to 5 which 0 comprises using as plant bean (Phaseolus vulgaris), pea (Pisum sativum), lens (Lens culiπaris), Astragalus racemosus, Neptuπia amplexicaulis , white lupin (Lupiπus albus), wheat (Triticu aestivu ) , barley (Hordeum no¬ dosum), rye (Secale cereale), oat (Avena sativa), rice (Oryze sativa), mais (Zea mays), millet (Paπicum milia- ceum) , rye grass (Lolium perenne), garlic (Allium sativum onion (Allium cepa), radish (Raphanus sativus), gipsy-wor (Lycopus exaltatus), borge (Borago αfficinalis) , evening primrose (Qenotherea bienπis), soy bean (Glycine soja), cowslip (Primula veris), Lathyrus sativus, sorghum (Sor¬ ghum vulgare) . 7. A dietetic composition which comprises contain¬ ing a plant or part or extract of plant of high trace-ele meπt-coπteπt prepared according to claims 1 to 6. 8. Pharmaceutical composition which comprises con¬ taining a plant, part or extract of plant of high trace- -element-conteπt, prepared according to claims 1 to 6. 9. Cosmetical composition which comprises containin a plant, part or extract of plant of high tracε-elemeπt- -content prepared according to claims 1 to 6. 10. Use of the plant,and part or extract of plant of high trace-elemeπt-coπtent prepared according to claims 1 to 6 in form of dietetic, pharmaceutical or cosmetical composition.

Inventors: ABRAHAM ZSUZSA, BOGDANY LASZLO, BOGDANYNE FORGACS OLGA, BOGNAR JANOS, FABIAN MIKLOS, FABIANNE PEHAR KATALIN, KAPOVITS ISTVAN, KEVE TIBOR, KEVENE SZENTTORNYAI ANDREA, LONYAI PETER, ABRAHAM, ZSUZSA, BOGDANY, LASZLO, BOGDANYNE FORGACS, OLGA, BOGNAR, JANOS, FABIAN, MIKLOS, FABIANNE PEHAR, KATALIN, KAPOVITS, ISTVAN, KEVE, TIBOR, KEVENE SZENTTORNYAI, ANDREA, LONYAI, PETER
Applicants: BOGDANY LASZLO, BOGDANYNE FORGACS OLGA, BOGNAR JANOS, FABIAN MIKLOS, FABIANNE PEHAR KATALIN, KAPOVITS ISTVAN, KEVE TIBOR, KEVENE SZENTTORNYAI ANDREA, LONYAI PETER, ZSUZSA ABRAHAM, ABRAHAM, ZSUZSA, BOGDANY, LASZLO, BOGDANYNE FORGACS, OLGA, BOGNAR, JANOS, FABIAN, MIKLOS, FABIANNE PEHAR, KATALIN, KAPOVITS, ISTVAN, KEVE, TIBOR, KEVENE SZENTTORNYAI, ANDREA, LONYAI, PETER
Application year: 1988
Main classification: A01G31
IPC codes: A01G31, A01H4, A23L1, A61K8, A61K36, A61Q5, A61Q19, C12N5