Indigo-based dye with improved stability and improved dyeing properties

Abstract

The invention relates to a product, a kit and a method for dyeing keratin fibers, in particular human hair, using an indican-containing component that was pretreated at a temperature in the range of 60°C to 200°C, an enzyme mixture having cellulase activity, cysteine and isatin.

Description

[0001] Indigo-based dye with improved stability and dyeing properties

[0002] The invention relates to agents, kits and methods for coloring keratin-containing fibers, in particular human hair, using an indican-containing component which has been pretreated at a temperature in the range of 60 °C to 200 °C, an enzyme mixture with cellulase activity, cysteine ​​and isatin.

[0003] The desire to change one's hair color is a major need for many consumers. To satisfy this need, the cosmetics industry offers a diverse range of products. Hair dyes that achieve particularly long-lasting color with high coverage are usually oxidation dyes. These use oxidizing agents that can damage the hair structure. Certain cationic direct azo dyes are also capable of achieving hair color changes with excellent color fastness properties. However, these azo dyes are synthetic dyes.

[0004] A growing number of consumers are demanding hair dyes and hair coloring processes based on natural dyes, even though these products and processes are often inferior to the aforementioned products and processes in terms of authenticity, coverage and color variety.

[0005] In addition to dyeing with henna, which is obtained from the plant Lawsonia inermis, hair dyeing with plants from which indigo or indirubin can be produced has also been known for a long time.

[0006] State of the art

[0007] The production of storage-stable cosmetic products containing natural plant material is often difficult. The plant material contains enzymes that can react with other ingredients of the plant material or the cosmetic, even during storage. Therefore, there has been no lack of efforts in the prior art to increase the storage stability of cosmetics containing plant material. In dyeings with indigo or indirubin-producing plant materials, it is often observed that the coloration changes constantly over a period of up to 2 weeks after application of the dye, and the final color only persists after this period. From a concentration of 2 wt.-% indigo or indirubin producing plant powder in water, the color tone on the hair changes in the first few days after coloring from an initial turquoise-blue tone to a redder and darker tone, creating a purple color impression.

[0008] There has been no lack of efforts in the state of the art to improve the coloring of keratin fibers, especially human hair, with indigo.

[0009] US20040055096A1 discloses a dyeing agent for the production of which powdered leaves of an indigo-producing plant, such as Polygonum tinctorium, Strobilanthes cusia, Indigofera tinctoria or Isatis indigotica, whose natural enzyme content has been deactivated by heat treatment, are mixed shortly before use with a beta-glucosidase-containing component (e.g. fresh leaves of an indigo-producing plant which have not been heat-treated, or the pressed juice from these non-heat-treated leaves, optionally in freeze-dried form, or extracts of various mushrooms, such as enokitake (Flammulina velutipes) and shiitake (Lentinus Edodes), or extracts of corn leaves, apricot kernels or almonds). By separating the indigo precursor indican, which is present in the plants in the form of 3-indioxyl-ß-D-glucopyranoside, from the plant's own enzyme beta-glucosidase, an improved storage stability of the natural dye is achieved.In PCT application W02014104301A1, a similar approach to improved storage stability is disclosed, namely the combination of two different indigo powders, whereby the natural enzyme content in the first leaf powder component was deactivated by heat treatment, while the second leaf powder component was dried at temperatures below 50 °C and thus contained active enzyme.

[0010] WO2015082482A1 discloses a method for coloring hair using powdered plant parts from plants that produce indigo. After the indigo dyeing, which can be done in the presence of hydrogen peroxide, the hair is treated with a strongly alkaline composition. This is intended to accelerate the development of the final color tone and prevent color change after the actual dyeing process. However, the alkaline pH and the oxidizing agent damage the hair.

[0011] From DE4211450A1, non-oxidative hair dyes are known which contain isatin in combination with at least one amino acid, for example cysteine, or a water-soluble oligopeptide composed of 2 to 9 amino acids in an aqueous carrier.

[0012] Task

[0013] One object was to provide agents, kits, and methods for the non-oxidative coloring of keratin-containing fibers, in particular human hair, using plants that produce indigo or indirubin, with which a broader range of colorations can be achieved. In particular, colorations beyond the typical indigo blue, in particular colorations with a shift into warmer color ranges, should be provided. A further object of the present invention was to provide agents, kits, and methods for the non-oxidative coloring of keratin-containing fibers, in particular human hair, using plants that contain indican and produce indigo or indirubin, with which the final color result of the indigo coloration is obtained as soon as possible after completion of the actual coloring process.

[0014] A further object of the present invention was to provide agents, kits and methods for the non-oxidative coloring of keratin-containing fibers, in particular human hair, using plants containing indican and producing indigo or indirubin, with improved storage stability.

[0015] Surprisingly, it was discovered that with the dyeing agents, dyeing processes, and dyeing kits described in the patent claims, using parts or extracts of plants containing indican and producing indigo or indirubin, in combination with cysteine ​​and isatin, an intense magenta hair color could be achieved directly after the dyeing procedure. Furthermore, the final color result was achieved shortly after completion of the actual dyeing process, and this color result—unlike what is often typical for indigo dyeing—did not change in the days following completion of the dyeing process. Neither the initial turquoise-blue color characteristic of indigo nor the final purple tone formed on the hair fiber. The dyeing agents and dyeing kits according to the invention also exhibit a high level of storage stability.

[0016] A first subject of the present invention is therefore an agent for the non-oxidative coloring of keratinic fibers, in particular human hair, comprising i) at least one indican-containing component which has been pretreated at a temperature in the range from 60 °C to 200 °C, furthermore ii) an enzyme mixture comprising at least one endo-1,4-beta-glucanase (EC 3.2.1.4), at least one beta-glucosidase (EC 3.2.1.21) and at least one exo-1,4-beta-D-glucanase (EC 3.2.1.91), furthermore iii) cysteine, furthermore iv) isatin and v) water.

[0017] According to the invention, non-oxidative coloring means that the coloring takes place without the addition of oxidizing agents other than atmospheric oxygen.

[0018] The indigo precursor or indirubin precursor indican occurs naturally in plants. Indican is a glucoside of indoxyl and occurs naturally in some plants known as indigo plants, such as Isatis tinctoria (woad) or Indigofera tinctoria. Synonyms for indican are 3-indoxyl-ß-D-glucopyranoside, indoxyl-ß-D-glucoside, 3-(ß-D-glucosido)indole, or plant indican.

[0019] Before indican is converted into the dyes indigo or indirubin, it must be cleaved into indoxyl and the glucoside residue. This cleavage preferably occurs enzymatically. Cleavage should occur in situ, i.e., shortly before application to the keratin fibers to be dyed.

[0020] Accordingly, the indican-containing component, which has been pretreated at a temperature in the range of 60 °C to 200 °C, and the enzyme mixture ii) according to the invention should be physically separated from each other until use.

[0021] A further subject of the present invention is therefore a kit for the non-oxidative coloring of keratinic fibers, in particular human hair, with natural dyes, which comprises the following compartments which are physically separate from one another: i) a composition (Ind) which contains at least one indican-containing component which has been pretreated at a temperature in the range from 60 °C to 200 °C, and ii) an aqueous composition (E) which contains an enzyme mixture comprising at least one endo-1,4-beta-glucanase (EC 3.2.1.4), at least one beta-glucosidase (EC 3.2.1.21) and at least one exo-1,4-beta-D-glucanase (EC 3.2.1.91), iii) optionally at least one further compartment, and which is characterized in that cysteine ​​is contained in at least one of the compartments and isatin is contained in at least one of the compartments.

[0022] "Non-oxidative" is understood according to the invention to mean that no oxidizing agents other than atmospheric oxygen are used in the agents, kits, and methods according to the invention. In particular, the oxidizing agents hydrogen peroxide, persulfates, perbromates, percarbonates, perborates, and percarbamides are excluded. Atmospheric oxygen does not constitute an oxidizing agent in the context of the invention.

[0023] Preferred agents and kits for non-oxidative dyeing according to the invention are characterized in that the indican-containing component i) is selected from comminuted parts of an indigo or indirubin-producing plant and from indican-containing extracts of an indigo or indirubin-producing plant, as well as mixtures of these indican-containing components i).

[0024] Particularly preferred agents and kits for non-oxidative coloring according to the invention are characterized in that the indican-containing component i) is selected from indican-containing extracts of an indigo or indirubin-producing plant.

[0025] Further agents and kits for non-oxidative coloring preferred according to the invention are characterized in that the extraction agent used to obtain the at least one indican-containing extract from a plant producing indigo or indirubin is selected from water, furthermore C1-C4 alkanols and C2-C4 polyols, in particular ethanol, isopropanol, n-propanol, ethylene glycol, 1,2-propanediol, glycerol and 1,3-butylene glycol, and mixtures of these extraction agents, preferably selected from water, ethanol, mixtures of water and at least one C1-C4 alkanol, mixtures of water and at least one C2-C4 polyol and water / ethanol mixtures, particularly preferably selected from water, ethanol and water / ethanol mixtures.

[0026] Preferred agents and kits for non-oxidative dyeing according to the invention are characterized in that the indigo or indirubin producing plant(s) is / are selected from at least one species of the following genera:

[0027] - Indigofera, in particular Indigofera tinctoria, Indigo suffruticosa, Indigofera articulata, Indigofera arrecta, Indigofera heterantha “Gerardiana”, Indigofera argentea or Indigofera longiracemosa;

[0028] - Isatis, especially Isatis tinctoria (woad);

[0029] - Persicaria, especially Persicaria tinctoria (dyer's knotweed);

[0030] - Wrightia, especially Wrightia tinctoria;

[0031] - Calanthe, especially Calanthe veratrifolia; and

[0032] - Baphicacanthus cusia, synonym Strobilanthes cusia.

[0033] Particularly preferred plants according to the invention which produce indigo or indirubin are selected from the genus Indigofera and in particular from Indigofera tinctoria.

[0034] The plants of the aforementioned genera and species contain no or only small amounts of indigo or indirubin. Instead, the plants contain indican, which, after enzymatic cleavage of the glucoside residue, is a precursor to indigo. The parts of the aforementioned plant genera and species that are rich in indican and also contain indigo and indirubin are the leaves of these plants. To obtain the indican-containing component i) according to the invention, the plant leaves of the aforementioned plant genera and species are dried and ground and used as plant powder.

[0035] Plant powders preferred according to the invention have a particle size of less than 500 pm, particularly preferably in the range of 120 pm - 200 pm, extremely preferably in the range of 150 pm - 180 pm. Further plant powders preferred according to the invention have a bulk density in the range of 0.20 - 0.60 g / cm 3 , particularly preferably in the range of 0.25 - 0.40 g / cm 3 Other plant powders preferred according to the invention have, based on their weight, a moisture content of less than 10% by weight, preferably 0.1-6.0% by weight, particularly preferably 0.2-3.0% by weight. The moisture content is determined after 3 hours of drying at 105°C.

[0036] According to the invention, it is preferred that the powder of the indigo- or indirubin-producing plant, based on its weight, consists of at least 50 wt.%, preferably at least 80 wt.%, particularly preferably more than 80-100 wt.%, of the leaves of the plant. Further preferred embodiments according to the invention are characterized in that the powder of the indigo- or indirubin-producing plant has a particle size of less than 500 pm, particularly preferably in the range of 120 pm-200 pm, extremely preferably in the range of 150 pm-180 pm, and also a bulk density in the range of 0.20-0.60 g / cm 3 , particularly preferably in the range of 0.25 - 0.40 g / cm 3 and, based on its weight, has a moisture content of less than 10 wt.%, preferably 0.1 - 6.0 wt.%, particularly preferably 0.2 - 3.0 wt.%.

[0037] Further agents and kits preferred according to the invention are characterized in that the powder of the indigo or indirubin-producing plant contains indican in an amount of 2-5 wt.%, preferably 2.5-4.5 wt.%, particularly preferably 3-4.1 wt.%, based on the weight of the powder, wherein the indican content is converted to pure indican.

[0038] Deactivation of the natural enzyme content in the indican-containing component i)

[0039] Further agents and kits for non-oxidative coloring preferred according to the invention are characterized in that the pretreatment of the at least one indican-containing component i) is carried out at a temperature in the range from 60 °C to 200 °C by heating, in particular by heating in a solvent, further by treatment with steam or by dry heating, preferably for a period of 1 to 120 minutes, particularly preferably 2 to 60 minutes, extraordinarily preferably 5 to 10 minutes.

[0040] Option 1 : Indican-containing component i) is an extract

[0041] Particularly preferred agents and kits for non-oxidative coloring according to the invention are characterized in that the indican-containing component i) is selected from at least one indican-containing extract of an indigo or indirubin-producing plant.

[0042] Extraordinarily preferred agents and kits for non-oxidative coloring according to the invention are characterized in that the indican-containing component i) is selected from at least one indican-containing extract of an indigo or indirubin-producing plant, wherein the extraction agent used to obtain the at least one indican-containing extract from an indigo or indirubin-producing plant is selected from water, furthermore C1-C4-alkanols and C2-C4-polyols, in particular ethanol, isopropanol, n-propanol, ethylene glycol, 1,2-propanediol, glycerol and 1,3-butylene glycol, and mixtures of these extraction agents, preferably selected from water, ethanol, mixtures of water and at least one C1-C4-alkanol, mixtures of water and at least one C2-C4-polyol and water / ethanol mixtures, particularly preferably selected from water, ethanol and Water / ethanol mixtures.

[0043] The indican can be extracted using these aforementioned polar solvents. Since the extraction can be easily carried out at a temperature in the range of 60°C to 200°C by heating the solvent, this preferred procedure not only achieves the isolation of the indican but also the simultaneous deactivation of the plant's own enzymes. The deactivation of the plant's own enzymes is desirable according to the invention, because it was surprisingly discovered that the resulting coloration can be significantly improved by replacing the plant's own enzymes, which, as a natural product, are subject to numerous qualitative and quantitative fluctuations in composition, with a standardized enzyme mixture.

[0044] In a further preferred embodiment of the invention, the extracted indican is not separated from the solvent after completion of the extraction procedure and separation of the solid plant parts, for example by filtration or decanting, but is used directly as indican-containing component i), optionally after concentration by evaporation of the solvent or after dilution, wherein optionally further cosmetic auxiliaries, such as preservatives, thickeners, perfumes, pH regulators or hair conditioners, can be added.

[0045] In another, likewise preferred embodiment of the invention, the extracted indican is also separated from the solvent after completion of the extraction procedure and separation of the solid plant parts, for example by filtration or decantation, for example by freeze-drying or by distilling off the solvent, so that the indican-containing component i) is obtained in powder form. To produce the colorant preferred according to the invention, the powdered indican-containing component i) can be taken up in water and then mixed with the enzyme mixture and optionally additional water. In a further preferred embodiment, to produce the colorant preferred according to the invention, the powdered indican-containing component i) can be mixed with the enzyme mixture and with water.

[0046] Option 2: dried and crushed plant parts of an indiqo-producing plant as indican-containing component i)

[0047] According to the invention, dried and crushed plant parts of an indigo-producing plant which have been pretreated at a temperature in the range of 60°C to 200°C can also be used as indican-containing component i).

[0048] The heat pretreatment of this embodiment of the invention is characterized in that the pretreatment of the at least one indican-containing component i) is carried out at a temperature in the range from 60 °C to 200 °C by heating, in particular by heating without solvent, further by treatment with steam or by dry heating, preferably for a period of 1 to 120 minutes, particularly preferably 2 to 60 minutes, extraordinarily preferably 5 to 10 minutes.

[0049] Further colorants preferred according to the invention are characterized in that as at least one indican-containing component i) dried and comminuted parts of at least one indigo or indirubin-producing plant, which have been pretreated at 60 - 200°C, are contained in a total amount of 1 - 80 wt.%, preferably 2 - 50 wt.%, particularly preferably 3 - 25 wt.%, extraordinarily preferably 4 - 15 wt.%, further extraordinarily preferably 5 - 10 wt.%, further extraordinarily preferably 6 - 7 wt.%, based on the weight of the agent.

[0050] Further colorants preferred according to the invention are characterized in that as at least one indican-containing component i) powdered leaves of Indigofera tinctoria are contained in an amount of 1-80 wt.%, preferably 2-50 wt.%, particularly preferably 3-25 wt.%, extraordinarily preferably 4-15 wt.%, further extraordinarily preferably 5-10 wt.%, further extraordinarily preferably 6-7 wt.%, based on the weight of the agent.

[0051] Preferred non-oxidative coloring agents according to the invention are characterized in that the indican-containing component i) contains, based on its weight, 0.01-6 wt.% indican. Preferred indican-containing components i) contain, based on their weight, 0.02-5 wt.%, preferably 0.05-4.5 wt.%, particularly preferably 0.1-4 wt.%, extraordinarily preferably 0.2-3.9 wt.% indican. Depending on the type of indican-containing component i), different subranges from the aforementioned broad ranges of indican quantities may be particularly preferred.

[0052] If the indican-containing component i) is dried and crushed plant parts of an indigo-producing plant, its indican content, in each case based on its weight, is preferably 1 - 6 wt.%, particularly preferably 2 - 5 wt.%, particularly preferably 3 - 4.5 wt.%, extremely preferably 3.5 - 4.1 wt.%.

[0053] If the indican-containing component i) is the solvent-containing extract from dried plant parts of an indigo-producing plant, its indican content, in each case based on its weight, is preferably 0.01 - 2 wt.%, particularly preferably 0.02 - 1 wt.%, particularly preferably 0.1 - 0.5 wt.%, extremely preferably 0.2 - 0.4 wt.%.

[0054] Coloring agents and coloring processes preferred according to the invention are characterized in that cysteine ​​is present or applied to the keratin fibers in an amount of 0.1 - 5.0 wt.%, preferably 0.5 - 3.0 wt.%, particularly preferably 0.8 - 2.0 wt.%, extraordinarily preferably 1.0 - 1.5 wt.%, based on the weight of the ready-to-use coloring agent.

[0055] According to the invention, the ready-to-use colorant is understood to mean the agent obtained by mixing all components contained in the compartments of the coloring kit according to the invention and which is intended and used for immediate application to the keratin fibers. The application of the ready-to-use colorant according to the invention or preferred according to the invention to the keratin fibers preferably takes place within 1 second to 30 minutes, particularly preferably within 10 seconds to 15 minutes, after completion of the preparation of the ready-to-use colorant, i.e., immediately or without delay after the preparation of the ready-to-use colorant.

[0056] Cysteine, as a chiral amino acid, possesses a stereogenic center and can occur in mirror-image form, namely in the form of L-cysteine ​​and D-cysteine. Both L-cysteine ​​and D-cysteine, as well as mixtures thereof, are encompassed by the present invention. Within the scope of the present invention, both possible enantiomers can therefore be used equally as specific compounds or as mixtures thereof, particularly as racemates. However, it is particularly advantageous according to the invention to use the naturally occurring isomer form, in this case L-cysteine.

[0057] Extraordinarily preferred coloring agents and coloring methods according to the invention are characterized in that L-cysteine ​​is present or applied to the keratin fibers in an amount of 0.1 - 5.0 wt.%, preferably 0.5 - 3.0 wt.%, particularly preferably 0.8 - 2.0 wt.%, extraordinarily preferably 1.0 - 1.5 wt.%, based on the weight of the ready-to-use coloring agent.

[0058] An aqueous solution of cysteine ​​reacts acidically. A 1 wt% solution of L-cysteine ​​in deionized water has a pH of 6.49, measured at 20°C. Cysteine ​​is therefore not an alkalizing agent.

[0059] Further coloring agents and coloring processes preferred according to the invention are characterized in that isatin is present in an amount of 0.01 - 1.0 wt.%, preferably 0.05 - 0.5 wt.%, particularly preferably 0.08 - 0.3 wt.%, extraordinarily preferably 0.1 - 0.2 wt.%, based on the weight of the ready-to-use coloring agent, or is applied to the keratin fibers.

[0060] For the effects according to the invention, it has proven particularly advantageous if cysteine ​​is contained in a molar excess to isatin or is applied to the keratin fibers.

[0061] Further preferred coloring agents and coloring processes according to the invention are therefore characterized in that cysteine ​​is contained in a molar excess to isatin or is applied to the keratin fibers.

[0062] Particularly preferred dyeing agents and dyeing methods according to the invention are characterized in that the molar ratio of cysteine ​​to isatin is 1.5 to 20, preferably 3 to 18, particularly preferably 5 to 15, extremely preferably 10 to 13.

[0063] The dyes according to the invention also contain water. The water essentially serves as a solvent for the enzymes, the at least one indican, cysteine, and isatin. If dried and ground plant parts of an indigo- or indirubin-producing plant, which have previously been subjected to heat treatment, are used as the indican-containing component i), these are preferably dispersed in water to extract the indican and make it accessible for reaction to indigo or indirubin.

[0064] If the indican-containing component i) is the solvent-containing extract from dried parts of an indigo or indirubin-producing plant, it preferably contains water as the solvent.

[0065] Water in the ready-to-use dye

[0066] Ready-to-use colorants preferred according to the invention are characterized in that they contain, in each case based on their weight, water in an amount of 19.9 - 95.0 wt.%, preferably 30.0 - 90.0 wt.%, particularly preferably 50.0 - 88.0 wt.%, extraordinarily preferably 60.0 - 85.0 wt.%.

[0067] Water in the indican-containing extract

[0068] If the indican-containing component i) is the solvent-containing extract from dried plant parts of an indigo-producing plant, it preferably contains water in an amount of 0.5 - 99.5 wt.%, more preferably 10.0 - 95.0 wt.%, particularly preferably 30.0 - 90.0 wt.%, extraordinarily preferably 50.0 - 85.0 wt.%, more extraordinarily preferably 65 - 80 wt.%, based on its weight.

[0069] Water in the indican-containing plant powder dispersion

[0070] If dried and comminuted plant parts of an indigo or indirubin-producing plant which have previously been subjected to a heat treatment are used as indican-containing component i), these are preferably dispersed in water, this dispersion preferably containing water in an amount of 15.0 - 99.5 wt.%, more preferably 30.0 - 95.0 wt.%, particularly preferably 50.0 - 90.0 wt.%, extraordinarily preferably 60.0 - 85.0 wt.%, more extraordinarily preferably 70 - 80 wt.%, based on its weight.

[0071] Water in the indican-containing dry extract dispersion

[0072] If the dried extract from the plant parts of an indigo-producing plant is used as the indican-containing component i), it is preferably dispersed or dissolved in water, wherein this dispersion or solution, in each case based on its weight, preferably contains water in an amount of 15.0-99.5 wt.%, more preferably 30.0-95.0 wt.%, particularly preferably 50.0-90.0 wt.%, extraordinarily preferably 60.0-85.0 wt.%, more extraordinarily preferably 70-80 wt.%. Water in the composition (E) containing the enzyme mixture ii)

[0073] The enzyme mixture ii) used according to the invention, comprising at least one endo-1,4-beta-glucanase (EC 3.2.1.4), at least one beta-glucosidase (EC 3.2.1.21), and at least one exo-1,4-beta-D-glucanase (EC 3.2.1.91), can preferably be dispersed or dissolved in water before being mixed with the indican-containing component i) to form a colorant according to the invention. This enzyme dispersion preferably contains, in each case based on its weight, water in an amount of 15.0-99.5 wt.%, more preferably 30.0-95.0 wt.%, particularly preferably 50.0-90.0 wt.%, extraordinarily preferably 60.0-80.0 wt.%, further extraordinarily preferably 64-75 wt.%.

[0074] Water as a component of the optional compartments

[0075] The water content of the colorant according to the invention can also be influenced by at least one further component which is contained in a compartment physically separate from the other components and which is neither different from the indican-containing component i) nor from the enzyme mixture ii) used according to the invention.

[0076] The cysteine ​​used according to the invention can be contained in at least one of the compartments selected from the indican-containing extract, the indican-containing plant powder dispersion, the composition (E) containing the enzyme mixture ii) and a further, optional compartment which, based on its weight, preferably additionally contains 0.05 - 99.9 wt.% water.

[0077] The isatin used according to the invention can be contained in at least one of the compartments selected from the indican-containing extract, the indican-containing plant powder dispersion, the composition (E) containing the enzyme mixture ii) and a further, optional compartment which, based on its weight, preferably additionally contains 0.05 - 99.9 wt.% water.

[0078] In order to save packaging material, it is preferred according to the invention that the cysteine ​​and the isatin are contained in the compartment with the indican-containing component.

[0079] The coloring agent according to the invention is further characterized in that it contains an enzyme mixture comprising

[0080] - at least one endo-1,4-beta-glucanase (EC 3.2.1.4),

[0081] - at least one beta-glucosidase (EC 3.2.1 .21) and

[0082] - contains at least one exo-1,4-beta-D-glucanase (EC 3.2.1.91).

[0083] Surprisingly, it was discovered that using an enzyme mixture according to the invention, dyeings can be achieved in which the final color result is achieved shortly after the completion of the actual dyeing process and does not change over time. However, using an indican-containing component with its natural enzyme content results in a very unstable dyeing result, with the color shifting significantly within two weeks of dyeing.

[0084] The enzymes contained in the enzyme mixture ii) used according to the invention are selected from a group of enzymes generally referred to as cellulases.

[0085] The term "cellulase," as used herein, refers to enzymes that catalyze the hydrolysis of 1,4-beta-D-glucoside bonds present in cellulose (cellobiose) and / or lichenin and / or beta-D-glucans. Cellulases are often also capable of hydrolyzing the 1,4-bonds in beta-D-glucans, which possess 1,3-bonds in addition to 1,4-bonds. Cellulases are capable of cleaving cellulose into beta-glucose. Consequently, cellulases act particularly on cellulose-containing or cellulose-derivative-containing residues and catalyze their hydrolysis. The decisive factor as to whether an enzyme is a cellulase within the scope of the invention is its ability to hydrolyze 1,4-beta-D-glucoside bonds in cellulose.

[0086] The term "cellulase activity" is defined here as an enzyme that catalyzes the hydrolysis of 1,4-beta-D-glucoside bonds in beta-1,4-glucan (cellulose). Cellulose activity is measured using a standard method, e.g., as follows: Cellulases release glucose from CMC (carboxymethylcellulose). Samples are incubated with a substrate (1.25 wt% CMC) under defined reaction conditions (100 mM sodium phosphate buffer pH 7.5, 40°C, 15 min). Reaction with p-hydroxybenzoic acid hydrazide (PAHBAH) in the presence of bismuth produces a yellow dye that can be determined photometrically at 410 nm. An alkaline pH value is required during the color reaction. The amount of sugar released corresponding to the color is a measure of enzyme activity (Lever, Anal. Biochem., 1972, 47 & 1977, 81).

[0087] Cellulases can be divided into three categories:

[0088] 1 . Endoglucanase (EC 3.2.1 .4), also known as endo-1 ,4-beta-glucanase, beta-1 ,4-glucanase, avicelase, beta-1 ,4-endoglucanhydrolase, endo-1 ,4-beta-D-glucanohydrolase, carboxymethylcellulase or celludextrinase;

[0089] 2. Cellulose-1,4-beta-cellobiosidase (non-reducing end) (EC 3.2.1.91), also known as exoglucanase, 1,4-beta-cellobiohydrolase, 4-beta-D-glucan cellobiohydrolase (non-reducing end), avicelase, exo-1,4-beta-D-glucanase or exocellobiohydrolase, releases cellobiose from the non-reducing ends of the ß-D-glucan chains by hydrolysis of the (1->4)-beta-D-glucosidic bonds in cellulose and cellotetraose;

[0090] 3. Beta-glucosidase (EC 3.2.1.21), also known as cellobiase, beta-D-glucoside glucohydrolase, amygdalase, or gentobiase, hydrolyzes terminal, non-reducing beta-D-glucosyl groups, releasing beta-D-glucose. Cellulases preferably used according to the invention are not derived from plants, but rather from bacteria or fungi. For the production of cellulases particularly preferably used according to the invention, fungi selected from Aspergillus spp., in particular Aspergillus niger, Aspergillus niger SOI017, Aspergillus niger AS 3.4309, Aspergillus niger HDF 05, Aspergillus oryzae, Aspergillus strain SA 58, Aspergillus terreus EMOO 6-4, as well as mixtures of Aspergillus niger and Aspergillus oryzae, and further selected from Trichoderma reesei, are used. Chemically modified or protein-engineered mutants are included.

[0091] Examples of cellulases with endo-1,4-glucanase activity (EC 3.2.1.4) are described in WO 2002 / 099091, e.g. those with a sequence of at least 97% identity to the amino acid sequence of positions 1 to 773 of SEQ ID NO:2 of WO 2002 / 099091.

[0092] A preferred embodiment of the invention is characterized in that the agent for non-oxidative coloring contains an enzyme mixture comprising at least one endo-1,4-beta-glucanase (EC 3.2.1.4) in a total amount of 20-35 wt.%, at least one beta-glucosidase (EC 3.2.1.21) in a total amount of 15-25 wt.% and at least one exo-1,4-beta-D-glucanase (EC 3.2.1.91) in a total amount of 25-35 wt.%, wherein the amounts refer to the total weight of the enzyme mixture ii).

[0093] In a further preferred embodiment of the invention, in addition to the at least one endo-1,4-beta-glucanase (EC 3.2.1.4), the at least one beta-glucosidase (EC 3.2.1.21) and the at least one exo-1,4-beta-D-glucanase (EC 3.2.1.91), at least one further enzyme with cellulase activity is additionally contained, selected from exo-1,4-beta-glucosidase (EC 3.2.1.74), cellulose-1,4-beta-cellobiosidase (EC 3.2.1.176) and oligooxyloglucan-reducing end-specific cellobiohydrolase (EC 3.2.1.150) as well as mixtures of these enzymes.

[0094] A further particularly preferred embodiment of the invention is characterized in that the agent for non-oxidative coloring comprises an enzyme mixture comprising at least one endo-1,4-beta-glucanase (EC 3.2.1.4) in a total amount of 20-35 wt.%, at least one beta-glucosidase (EC 3.2.1.21) in a total amount of 15-25 wt.%, and at least one exo-1,4-beta-D-glucanase (EC 3.2.1.91) in a total amount of 25-35 wt.%, as well as at least one further enzyme with cellulase activity, selected from exo-1,4-beta-glucosidase (EC 3.2.1.74), cellulose-1,4-beta-cellobiosidase (EC 3.2.1.176) and oligooxyloglucan-reducing end-specific Cellobiohydrolase (EC 3.2.1.150) and mixtures of these enzymes in a total amount which, together with the amounts of the other enzymes, amounts to 100% by weight, the amounts being based on the total weight of the enzyme mixture ii).A further particularly preferred embodiment of the invention is characterized in that the agent for non-oxidative coloring contains, based on its weight, an enzyme mixture with cellulase activity which contains at least one endo-1,4-beta-glucanase (EC 3.2.1.4), at least one beta-glucosidase (EC 3.2.1.21) and at least one exo-1,4-beta-D-glucanase (EC 3.2.1.91) in a total amount of 0.0001 - 1 wt.%, preferably 0.001 - 0.1 wt.%, more preferably 0.002 - 0.05 wt.%, particularly preferably 0.003 - 0.04 wt.%, extraordinarily preferably 0.01 - 0.03 wt.%, more extraordinarily preferably 0.015 - 0.02 wt.%.

[0095] The precursor to the dyes indigo and indirubin is indican. Indican, which occurs naturally in plants, is a glycoside, more precisely a glucoside, which is cleaved into indoxyl and the glycoside residue before the indican is converted into the dye indigo or indirubin. This cleavage occurs preferably enzymatically. The cleavage should take place in situ, i.e., shortly before application to the keratin fibers to be dyed.

[0096] The above-discussed agent for non-oxidative coloring of keratin fibers is therefore preferably commercially marketed as a kit comprising at least two components (Ind) and (E), which are physically separate from each other.

[0097] A further subject of the present invention is therefore a kit for the non-oxidative coloring of keratinic fibers, in particular human hair, with natural dyes, which comprises the following compartments which are physically separate from one another: i) a composition (Ind) which contains at least one indican-containing component which has been pretreated at a temperature in the range from 60 °C to 200 °C, and ii) an aqueous composition (E) which contains an enzyme mixture comprising at least one endo-1,4-beta-glucanase (EC 3.2.1.4), at least one beta-glucosidase (EC 3.2.1.21) and at least one exo-1,4-beta-D-glucanase (EC 3.2.1.91), iii) optionally at least one further compartment, characterized in that cysteine ​​is contained in at least one of the compartments and isatin is contained in at least one of the compartments.

[0098] The enzyme mixture ii) used according to the invention, comprising at least one endo-1,4-beta-glucanase (EC 3.2.1.4), at least one beta-glucosidase (EC 3.2.1.21) and at least one exo-1,4-beta-D-glucanase (EC 3.2.1.91), can preferably be dispersed or dissolved in water before it is mixed with the indican-containing component i), the cysteine ​​and the isatin to form a ready-to-use colorant according to the invention. This enzyme dispersion, which represents component (E) of the kit according to the invention, preferably contains, based on its weight, water in an amount of 15.0-99.5% by weight, more preferably 30.0-95.0% by weight, particularly preferably 50.0-90.0% by weight, extraordinarily preferably 60.0-80.0% by weight, and further extraordinarily preferably 64-75% by weight. The enzyme active substance content in the aqueous composition (E) is preferably, based on its weight, 1-70% by weight, particularly preferably 5-50% by weight.-%, furthermore particularly preferably 10 - 35 wt.%, extraordinarily preferably 15 - 25 wt.%, further extraordinarily preferably 18 - 22 wt.%.

[0099] Further colorants preferred according to the invention are characterized in that saponins are contained in a total amount of zero to less than 0.01 wt.%, preferably from zero to 0.005 wt.%, particularly preferably from zero to 0.001 wt.%, in each case based on the weight of the ready-to-use colorant.

[0100] With regard to further preferred embodiments of the dyeing kits according to the invention, what has been said regarding the dyeing agents according to the invention applies mutatis mutandis.

[0101] The present invention further provides a process for the non-oxidative coloring of keratinic fibers, in particular human hair, which comprises the following process steps in the specified order: a) Providing a composition (Ind) which contains at least one indican-containing component which has been pretreated at a temperature in the range from 60 °C to 200 °C, and providing an aqueous composition (E) which contains an enzyme mixture comprising at least one endo-1,4-beta-glucanase (EC 3.2.1.4), at least one beta-glucosidase (EC 3.2.1.21) and at least one exo-1,4-beta-D-glucanase (EC 3.2.1.91), and optionally providing at least one further component in a physically separate compartment, wherein at least one of the compositions (Ind) or (E) or at least one optional component contains cysteine ​​and at least one of the compositions (Ind) or (E) or at least one optional component contains isatin, b) mixing the compositions mentioned above under a) with one another to produce a ready-to-use colorant, c) applying the ready-to-use colorant obtained in step b) to the keratin fibers, d) leaving to act for a time of 30 seconds to 60 minutes, preferably 5 to 45 minutes, particularly preferably 15 to 30 minutes, e) rinsing the keratin fibers with water, f) optionally drying the keratin fibers.

[0102] The ready-to-use dye obtained in step b) is applied to the keratin fibers. The ready-to-use dye obtained in step b) is preferably applied to the keratin fibers within 1 second to 30 minutes, particularly preferably within 10 seconds to 15 minutes, after completion of process step b), i.e., immediately after the preparation of the ready-to-use dye.

[0103] Surprisingly, it was found that the temporal development of the coloring result on the keratin fibers can be further accelerated if the ready-to-use coloring agent according to the invention is allowed to act under the influence of heat, for example by means of a heat lamp or a drying hood or a hair dryer.

[0104] A particularly preferred embodiment of the dyeing process according to the invention is therefore characterized in that the ready-to-use dyeing agent obtained in step b) is allowed to act under the influence of heat, preferably at a temperature of 25 - 60 °C, particularly preferably at a temperature of 30 - 50 °C, extremely preferably at a temperature of 35 - 40 °C.

[0105] In conventional dyes and dyeing processes using natural dyes, compounds, especially salts, of metals other than alkali metals are often used to improve the adhesion of the natural dye to the keratin fibers. Such compounds are also referred to as "mordants." For example, US20040055096A1 discloses a pretreatment or posttreatment of keratin fibers with metals or metal salts selected from aluminum, iron, zinc, nickel, calcium, and magnesium. Other mordants include transition metals and lanthanides, as well as their salts and complexes.

[0106] The content of metal ions other than alkali metal ions, which is usually present in the water used, e.g., in tap water or city water, is not taken into account here. For example, tap water can contain an average of 2 mg of copper ions per liter, or approximately 0.0002% by weight of copper ions. Tap water contains approximately 20–120 mg of calcium ions per liter, or approximately 0.002–0.012% by weight of calcium ions. Tap water contains approximately 2–25 mg of magnesium ions per liter, or approximately 0.0002–0.0025% by weight of magnesium ions. At such low concentrations, no perceptible mordant effect occurs. For the purposes of the present invention, the agents according to the invention are "free of mordants" if the mordant(s) is / are present at a maximum concentration that is naturally customary for the ingredients used, such as water, enzymes, and plant parts.

[0107] The dyeing agents, dyeing kits and dyeing processes according to the invention can dispense with the use of such mordants.

[0108] Preferred colorants and coloring processes according to the invention are therefore characterized by being free of mordants. In order to preserve the hair-protecting potential of natural dyes, preferred processes according to the invention are limited to those processes in which the keratin fibers were not treated with a mordant within a period of up to 7 days before and up to 7 days after the application of a colorant according to the invention. Further preferred colorants, coloring kits, and coloring processes according to the invention are characterized by being free of hydrogen peroxide. Further preferred colorants, coloring kits, and coloring processes according to the invention are characterized by being free of mordants and hydrogen peroxide.

[0109] With regard to further preferred embodiments of the dyeing processes according to the invention, what has been said regarding the dyeing agents and dyeing kits according to the invention applies mutatis mutandis.

[0110] The keratin fibers to be colored, to which the ready-to-use colorant prepared in step b) by mixing the compositions provided in step a) is applied in process step c), are preferably dry.

[0111] In order to preserve the hair-protecting potential of natural dyes, the claimed method is preferably limited to those methods in which the keratin fibers have not been treated with an oxidizing agent within a period of up to 7 days prior to the application of the coloring agent according to the invention.

[0112] Oxidizing agents commonly used in hair cosmetics, but which are not intended to be used in hair treatments according to the invention, even as pretreatments, include hydrogen peroxide, persulfates, perbromates, percarbonates, perborates, and percarbamides. The oxygen contained in ambient air does not constitute an oxidizing agent in the context of the invention.

[0113] In order to preserve the hair-protecting potential of natural dyes, preferred methods according to the invention are limited to those methods in which the keratin fibers have not been treated with a keratin-reducing compound within a period of up to 7 days prior to the application of a coloring agent according to the invention.

[0114] The keratin fibers are preferably dried after rinsing out the ready-to-use colorant. Drying can be done without actively applying heat. However, drying can also be done with the addition of heat at a temperature of 25–120°C, particularly preferably at a temperature of 30–80°C, and most preferably at a temperature of 35–60°C. Heat is preferably applied using a heat lamp, a drying rod, a hair dryer, a flat iron, or a hair dryer.

[0115] A further feature of the dyeing process according to the invention is that, after application to the keratin fibers, the ready-to-use dye is allowed to act there for a period of 30 seconds to 60 minutes, preferably 5 to 45 minutes, particularly preferably 20 to 35 minutes, and extremely preferably 25 to 30 minutes. After the dyeing time has elapsed, the keratin fibers are rinsed with water to wash out the dye.

[0116] Optionally, the keratin fibers can be dried after this rinsing step. This can be done with an absorbent cloth, such as a towel. The towel-dried keratin fibers can also be partially or completely dried with a hairdryer or other heat source. It is also possible to let the keratin fibers air dry.

[0117] Further dyeing processes preferred according to the invention are characterized in that no oxidation dye precursors are used in them. Typical oxidation dye precursors are p-aminophenol, 4-amino-3-methylphenol, 4-amino-2-aminomethylphenol, 4-amino-2-(1,2-dihydroxyethyl)phenol, 4-amino-2-(diethylaminomethyl)phenol, 2-(2,5-diaminophenyl)ethanol, 2-(1,2-dihydroxyethyl)-p-phenylenediamine, N,N-bis-(2-hydroxyethyl)-p-phenylenediamine, N-(4-amino-3-methylphenyl)-N-[3-(1H-imidazol-1-yl)propyl]amine, N,N'-bis-(2-hydroxyethyl)-N,N'-bis-(4-aminophenyl)-1,3-diamino-propan-2-ol, Bis-(2-hydroxy-5-aminophenyl)methane, 1,3-bis-(2,5-diaminophen- oxy)propan-2-ol, N,N'-bis-(4-aminophenyl)-1,4-diazacycloheptane, 1,10-bis-(2,5-diaminophenyl)-1,4,7,10-tetraoxadecane, 2,4,5,6-tetraaminopyrimidine, 4-Hydroxy-2,5,6-triaminopyrimidine, 2-hydroxy-4,5,6-triaminopyrimidine, 2,3-diamino-6,7-dihydro-1H,5H-pyrazolo-[1,2-a]-pyrazol-1-one, 3-aminophenol, 5-amino-2-methylphenol,3-Amino-2-chlor-6-methylphenol, 2-Hydroxy-4-aminophenoxy- ethanol, 5-Amino-4-chlor-2-methylphenol, 5-(2-Hydroxyethyl)-amino-2-methylphenol, 2,4-Dichlor-3- aminophenol, 2-Aminophenol, 3-Phenylendiamin, 2-(2,4-Diaminophenoxy)ethanol, 1 ,3-Bis(2,4-di- aminophenoxy)propan, 1-Methoxy-2-amino-4-(2-hydroxyethylamino)benzol, 1 ,3-Bis(2,4-diamino- phenyl)propan, 2, 6-Bis(2'-hydroxyethylamino)-1 -methylbenzol, 2-({3-[(2-Hydroxyethyl)amino]-4- methoxy-5-methylphenyl}amino)ethanol, 2-({3-[(2-Hydroxyethyl)amino]-2-methoxy-5-methyl- phenyl}amino)ethanol, 2-({3-[(2-Hydroxyethyl)amino]-4,5-dimethylphenyl}amino)ethanol, 2-[3-Mor- pholin-4-ylphenyl)amino]ethanol, 3-Amino-4-(2-methoxyethoxy)-5-methylphenylamin, 1-Amino-3- bis-(2-hydroxyethyl)aminobenzol, Resorcin, 2-Methylresorcin, 4-Chlorresorcin, 1 ,2,4-Trihydroxy- benzol, 2-Amino-3-hydroxypyridin, 3-Amino-2-methylamino-6-methoxypyridin, 2,6-Dihydroxy-3,4-di- methylpyridin, 3,5-Diamino-2,6-dimethoxypyridin, 1-Phenyl-3-methylpyrazol-5-on, 1-Naphthol,1,5-dihydroxynaphthalene, 2,7-dihydroxynaphthalene, 1,7-dihydroxynaphthalene, 1,8-dihydroxynaphthalene, 2-(1,3-benzodioxole-5-ylamino)ethanol (1-beta-hydroxyethyl-3,4-methylenedioxyaniline), 4-hydroxyindole, 6-hydroxyindole, 7-Hydroxyindole, 4-Hydroxyindoline, 6-Hydroxyindoline and 7-Hydroxyindoline.

[0118] The compositions (Ind) and (E) used according to the invention and preferably according to the invention, and the at least one optional component in the separate compartment, can optionally contain, independently of one another, further additives that can optimize their application properties. Preferred additives are, in particular, thickeners, which ensure that the inventive or preferred ready-to-use colorant produced from the compositions (Ind) and (E) and optionally further optional components in separate compartments adheres better to the keratin fibers during application.

[0119] Compositions (Ind) and (E) used particularly preferably according to the invention contain - independently of one another - at least one or more hydrophilic thickeners, which are preferably selected from polysaccharides other than cellulose, which may be chemically and / or physically modified. Compounds from the group of polysaccharides are particularly preferred as hydrophilic thickeners according to the invention, since the basic structures of the polysaccharides are of natural origin and biodegradable. Preferred hydrophilic polysaccharide thickeners are selected from xanthan gum, alginic acids (and their corresponding physiologically acceptable salts, the alginates), agar agar (with the polysaccharide agarose present in agar agar as the main constituent), starch fractions and starch derivatives such as amylose, amylopectin and dextrins, karaya gum, locust bean gum, gum arabic, pectins, dextrans and guar gum, and mixtures thereof.

[0120] In preferred embodiments, xanthan gum is included as a hydrophilic thickener with a view to reliable viscosity adjustment and residue-free application to keratin fibers and the scalp.

[0121] Compositions (Ind) and (E) which are particularly preferably used according to the invention contain, in each case based on the weight, independently of one another, at least one hydrophilic thickener in a total amount of from 0.1 to 5% by weight, preferably from 0.5 to 4% by weight, more preferably from 1 to 3.5% by weight and most preferably from 1.2 to 2% by weight.

[0122] In a further preferred embodiment of the present invention, the compositions (Ind) and (E) used according to the invention contain, in each case based on their weight, independently of one another, 0.1 to 3 wt.%, preferably 0.5 to 2.5 wt.%, more preferably 1.2 to 2.0 wt.%, of xanthan gum.

[0123] Compositions (Ind) and (E) used particularly preferably according to the invention contain, independently of one another, at least one organic solvent having a phenyl group in the molecule. This solvent is preferably selected from phenoxyethanol, benzyl alcohol, and mixtures thereof. Surprisingly, it has been found that such aromatic solvents can have a positive effect on the dyeing results of the dyeing process according to the invention; this has been observed particularly with aromatic solvents.

[0124] In a further preferred embodiment of the present invention, the compositions (Ind) and (E) used according to the invention contain, in each case based on their weight, independently of one another, 0.1 to 3% by weight, preferably 0.5 to 2.5% by weight, more preferably 0.8 to 1.0% by weight, of at least one organic solvent which has a phenyl group in the molecule. In a further preferred embodiment of the present invention, the compositions (Ind) and (E) used according to the invention contain, in each case based on their weight, independently of one another, 0.1 to 3% by weight, preferably 0.5 to 2.5% by weight, more preferably 0.8 to 1.0% by weight, of at least one organic solvent selected from phenoxyethanol, benzyl alcohol and mixtures thereof.

[0125] Compositions (Ind) and (E) which are particularly preferably used according to the invention contain, independently of one another, at least one aliphatic solvent selected from C1-C4 alkanols and C2-C4 polyols, in particular selected from ethanol, isopropanol, n-propanol, ethylene glycol, 1,2-propanediol, glycerol and 1,3-butylene glycol, and mixtures of these solvents, but only in a total amount of 0.01-8% by weight, preferably 0.1-6% by weight, particularly preferably 0.5-4% by weight, in each case based on the weight of the compositions (Ind) or (E).

[0126] Other compositions (Ind) and (E) which are particularly preferably used according to the invention contain, independently of one another, no aliphatic solvent selected from C1-C4 alkanols and C2-C4 polyols.

[0127] The above statements regarding compositions (Ind) and (E) also apply mutatis mutandis to the at least one optional component in the at least one optional physically separate compartment.

[0128] Powders from the leaves of Indigofera tinctoria, which are preferably used according to the invention, react approximately neutrally in aqueous dispersion, with slight deviations between a slightly acidic and a slightly alkaline pH range. A 5 wt. % dispersion of ground leaves of Indigofera tinctoria in deionized water has a pH of 6.6 to 7.4, preferably 6.8 to 7.2, particularly preferably 6.9 to 7.1, each measured at 20°C.

[0129] Ready-to-use colorants preferred according to the invention have a pH of 2.0 to less than 10.0, preferably from 4.0 to 9.0, particularly preferably from 5.0 to less than 8.0, extraordinarily preferably from 6.6 to less than 7.5, in each case measured at 20°C.

[0130] In a preferred embodiment of the dyeing process according to the invention, the desired pH is adjusted using an acid or a base. Preferred acids are selected from citric acid, lactic acid, gluconic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, glutaric acid, galactaric acid (mucic acid), tartaric acid, malic acid, sulfuric acid and phosphoric acid, as well as mixtures of these acids. Preferred bases are sodium hydroxide, potassium hydroxide, arginine, lysine, monoethanolamine, triethanolamine, 2-amino-2-methylpropan-1-ol, and mixtures of these bases. In a further preferred embodiment of the dyeing process according to the invention, the desired pH is adjusted using a buffer system selected from a mixture of a medium-strength or weak acid with its conjugate or corresponding base (or the respective salt) and a mixture of a medium-strength or weak base with its conjugate orcorresponding acid.

[0131] According to the invention, preferred suitable corresponding acid-base pairs are those which stabilize the compositions used according to the invention in the pH range from 2.0 to less than 10.0, preferably from 4.0 to 9.0, particularly preferably from 5.0 to less than 8.0, extraordinarily preferably from 6.6 to less than 7.5, in each case measured at 20°C.

[0132] Particularly preferred buffer systems according to the invention are selected from

[0133] - Ammonia / ammonium salt mixtures, wherein the ammonium salt is preferably selected from ammonium chloride, ammonium bromide, ammonium hydrogen sulfate, ammonium sulfate, ammonium dihydrogen phosphate, diammonium hydrogen phosphate, ammonium phosphate, ammonium bicarbonate, ammonium carbonate, ammonium nitrate, ammonium acetate, ammonium glycolate, ammonium gluconate, ammonium tartrate, ammonium lactate, and mixtures of these ammonium salts, particularly preferably selected from ammonium chloride, ammonium hydrogen sulfate, ammonium sulfate, ammonium dihydrogen phosphate, diammonium hydrogen phosphate, ammonium phosphate, ammonium bicarbonate, and ammonium carbonate, extremely preferably selected from ammonium chloride,

[0134] - mixtures of hydrogen phosphate and dihydrogen phosphate, in particular the alkali metal salts of hydrogen phosphate and dihydrogen phosphate, particularly preferably the sodium salts and / or the potassium salts of hydrogen phosphate and dihydrogen phosphate,

[0135] - mixtures of alkali metal bicarbonate with alkali metal carbonate, in particular mixtures of sodium or potassium bicarbonate with sodium or potassium carbonate,

[0136] - mixtures of citric acid and its salts, in particular the alkali metal citrates, in particular the sodium salts, in particular trisodium citrate,

[0137] - mixtures of tartaric acid and its salts, in particular alkali metal tartrates, in particular potassium salts, in particular potassium hydrogen tartrate,

[0138] - mixtures of phthalic acid and its salts, in particular potassium salts, in particular potassium hydrogen phthalate,

[0139] - mixtures of lactic acid and its salts, in particular lactic acid / sodium lactate mixtures,

[0140] - mixtures of gluconic acid and its salts, in particular gluconic acid / sodium gluconate mixtures,

[0141] - mixtures of succinic acid and its salts, in particular the sodium salts, in particular sodium hydrogen succinate and disodium succinate, and - mixtures of malic acid and its salts, in particular the sodium salts, in particular sodium hydrogen malate and disodium malate, and

[0142] - Ammonia / ammonium salt mixtures, wherein the ammonium salt is preferably selected from ammonium chloride, ammonium bromide, ammonium hydrogen sulfate, ammonium sulfate, ammonium dihydrogen phosphate, diammonium hydrogen phosphate, ammonium phosphate, ammonium bicarbonate, ammonium carbonate, ammonium nitrate, ammonium acetate, ammonium glycolate, ammonium gluconate, ammonium tartrate, ammonium lactate, and mixtures of these ammonium salts, particularly preferably selected from ammonium chloride, ammonium hydrogen sulfate, ammonium sulfate, ammonium dihydrogen phosphate, diammonium hydrogen phosphate, ammonium phosphate, ammonium bicarbonate, and ammonium carbonate, extremely preferably selected from ammonium chloride.

[0143] Other buffer systems, such as acetic acid / sodium acetate, are also suitable in principle according to the invention. However, due to the vinegar odor, such a buffer is not acceptable for the production of a commercial cosmetic product.

[0144] In principle, other buffer systems are also suitable.

[0145] The specialist can obtain the appropriate weights of buffer salts to adjust the desired pH value from the relevant manuals.

[0146] Examples of implementation

[0147] The exemplary embodiments presented below are intended to explain the subject matter of the invention in more detail without limiting it thereto.

[0148] The dyeing for this invention was carried out on strands of buffalo belly hair (brush body tied, approx. 8 cm of free hair).

[0149] Preparation of the indican-containing component i) (Composition (Ind)

[0150] 95 ml of demineralized water was heated to approximately 95 °C. Subsequently, 5 grams of ground leaves of Indigofera tinctoria L* were added (5 wt% of the total dispersion) and stirred for 5 minutes at approximately 95 °C.

[0151] * The following raw materials were used:

[0152] "C30-Indigo Leaves Powder" (Indigofera tinctoria / _.); powdered leaves of Indigofera tinctoria, pH of the 5 wt.% dispersion in water: 6.8 to 7.2 (20°C) or "Indigo Powder" (Indigofera tinctoria L ), powdered leaves of Indigofera tinctoria, pH of the 5 wt.% dispersion in water: 6.8 to 7.2 (20°C)

[0153] The dispersion was then cooled to room temperature (20 °C) and filtered (filter: Whatman 10340810, viscose, pore size 100 pL, approx. 5 to 10 min, vacuum pump at approx. 500 mbar).

[0154] The filtrate was a brown, opaque solution without precipitate, with a pH of approximately 7.7 (20 °C). This indican-containing component i) is referred to below as "Composition (Ind)".

[0155] Treating hair strands with composition (Ind) does not produce color under standard dyeing conditions (50 ml of composition (Ind) per 1 g of hair strand, dyeing for 30 minutes at room temperature (20 °C) while stirring, rinsing for 30 seconds under running deionized water, combing 20 times at 20 °C, drying with a hairdryer, distance: d = 10 cm, temperature: 80 ± 5 °C, combing 20 times).

[0156] Coloring according to the invention

[0157] The aqueous composition (E) according to the invention, which contains an enzyme mixture (ii) according to the invention comprising at least one endo-1,4-beta-glucanase (EC 3.2.1.4), at least one beta-glucosidase (EC 3.2.1.21), and at least one exo-1,4-beta-D-glucanase (EC 3.2.1.91), was purchased from Novozymes. This aqueous composition (E) is referred to below as the "cellulase blend."

[0158] The enzyme mixture ii) in the “Cellulase Blend” from Novozymes comprised at least one endo-1,4-beta-glucanase (EC 3.2.1.4) in a total amount of 20-35 wt%, at least one beta-glucosidase (EC 3.2.1.21) in a total amount of 15-25 wt%, and at least one exo-1,4-beta-D-glucanase (EC 3.2.1.91) in a total amount of 25-35 wt%, as well as at least one further enzyme with cellulase activity in a total amount which, together with the amounts of the other enzymes, makes up 100 wt%, whereby the amounts refer to the total weight of the enzyme mixture ii).

[0159] The enzyme active substance content in the raw material used, the Novozymes "Cellulase Blend," which was available as an aqueous composition (E), was 190 pg / μl, or 19 wt.%. Furthermore, the "Cellulase Blend" contained, based on its weight, 66 wt.% water and 15 wt.% of a sucrose / D-glucose mixture.

[0160] To prepare the dye composition according to the invention, 0.5 g of L-cysteine ​​and 0.05 g of isatin were added to 49.4 ml of the inventive composition (Ind) and dissolved. This dispersion was heated to approximately 37°C on a stir plate while stirring. Upon reaching this temperature, 0.05 ml of the above-described "Cellulase Blend" from Novozymes was added. The resulting dye composition according to the invention had a total enzyme content of 0.019 wt.% based on its weight. This ready-to-use dye had a pH of 7.0, measured at 20°C.

[0161] A strand of buffalo belly hair (tied in a round, approx. 8 cm of free hair) was placed in this ready-to-use dye, which was warm to 37 °C, for 30 minutes and stirred at 37 °C.

[0162] The liquor ratio, i.e. the weight of dye per gram of hair, was 50 g of dye per 1 g of hair (50:1).

[0163] After the dyeing process, the hair strands were rinsed under running deionized water for 30 seconds and combed 20 times (20 °C).

[0164] The hair strands were then dried with a commercially available hair dryer at a defined distance (d = 10 cm) and a defined temperature (T = 80 ± 5 °C) with 20 combs.

[0165] Determination of staining results using spectrophotometric measurements

[0166] All colorimetric measurements were performed using the Spectraflash SF 600 colorimetric device from Datacolor.

[0167] The color difference, also known as dE or AE, can be easily determined colorimetrically using a colorimeter that measures colors in the L*, a*, b* color space, for example a Datacolor Spectraflash SF 600 colorimeter.

[0168] The L*,a*,b* color space refers to the CIELAB color space. The L value represents the brightness of the color (black-white axis); the higher the L value, the brighter the color. The a value represents the red-green axis of the system; the higher this value, the more the color is shifted toward red. The b value represents the yellow-blue axis of the system; the higher this value, the more the color is shifted toward yellow.

[0169] The color shift AE, i.e. the color difference between two (hair) colors, for each of which an L*, a*, b* value combination has been determined, is calculated according to the following formula:

[0170] AE = (AL 2 + Aa 2 + From 2 ) 05

[0171] The larger the value for AE, the more pronounced the color difference.

[0172] A D65 illuminant and a diffuse / 8° optical configuration were used for the spectrophotometer measurements. The spectral reflectance data for each sample from 380 nm to 700 nm were converted to colorimetric data using DCI Color software. Reflectance measurements were determined for each hair sample, with the average of four measurements recorded.

[0173] The color difference (AE) between the uncolored strand and the colored strand was calculated according to the following formula:

[0174] Lv, av, bv: colorimetric values ​​for dyed strands

[0175] Ln, an, bn: Colorimetric values ​​for undyed strands

[0176] In order to further understand the temporal development of the final color, measurements were taken on day 0 directly after coloring and 14 days after coloring and compared.

[0177] On day 0, immediately after coloring, the strands exhibited an intense magenta hue. The change in the shade of this strand over the two weeks following coloring became even more consistent (AE of 11.6) (see Table 1).

[0178] The differences between the strands dyed according to the invention and the strands dyed analogously with the composition (Ind) and cellulase blend, but without cysteine ​​and isatin, directly after dyeing (day 0) and 14 days after dyeing are shown in Table 1 (mean values).

[0179] Table 1: Comparison of staining with heat-treated composition (Ind) + reactivation by cellulase addition, with and without cysteine / isatin combination 0 days and 14 days after staining

[0180]

[0181] The conversion of the L a b values ​​into Pantone values ​​and color names was carried out using the website https: / / www.e-paint.co.uk / convert-lab.asp.

[0182] The addition of cysteine ​​and isatin to the indican-containing solution, which has been activated with cellulase, results in an intense magenta tone (frenzied red (7421 U, Pantone U)) visible on the hair fiber after coloring, rather than a blue-turquoise / blue-gray tone (see Table 1). The change in the color tone of this strand in the two weeks following coloring becomes even more uniform overall, but is negligible (AE of approximately 1 to 3).

[0183] In the above-described dyeing according to the invention, in contrast to the comparison dyeing without cysteine ​​and without isatin, the magenta tone can be observed on the hair strand directly after treatment.

[0184] Influence of dyeing temperature

[0185] The same coloring, performed at 20°C, shows a pinkish tint immediately after treatment. This was not quantifiable due to residual moisture. However, an irregular, stronger red-magenta tone developed during subsequent blow-drying (temperature at 5 cm from the hair dryer: 96°C, at 15 cm from the hair dryer: 79°C). The color difference between coloring at 37°C followed by blow-drying and coloring at 20°C followed by blow-drying was AE = 28.0.

[0186] The differences between the control strand and the strands with the addition of cysteine / isatin immediately after staining at 20 °C (day 0) are shown below (mean values). Table 2: L*, a*, b*, AE values ​​of the indican / cellulase stains with cysteine / isatin and

[0187] Indican / cellulase alone, dyeing at 20 °C and blow-drying

[0188] Influence of the pH value of the dye on the color result

[0189] The following shows how the pH value of the ready-to-use dye affects the color result.

[0190] The dyes are prepared and the highlights are dyed according to the process described above. In the inventive, ready-to-use dye consisting of indican solution, cellulase blend, 1 wt.% cysteine, and 0.1 wt.% isatin in water, a pH of 7.0 is spontaneously established. The pH of other inventive, ready-to-use dyes was adjusted to 5, 8, and 10 with sodium hydroxide solution or hydrochloric acid, each measured at 20°C. The dyeing was carried out at 37°C. Following dyeing, all highlights were rinsed and dried according to the usual protocol (see above). The color results were compared with those achieved at the "spontaneous" pH of 7.0. The dye dispersion with a pH of 10 is not in accordance with the invention (V-1).

[0191] Table 3: L*, a*, b*, AE values ​​and color designations of indican / cellulase stains with cysteine ​​and isatin at 37 °C and different pH values ​​directly after staining (day 0) It can be observed that the color intensity and magenta tone increase at pH 5. In a more alkaline environment, the a* value, which represents the red intensity, decreases significantly. At pH 10 (not according to the invention), the desired color is not visible on the hair fiber.

[0192] The data presented suggest that the dye according to the invention produces the darkest, most intense, and most reddish color at pH 5. The dye in the slightly alkaline range (pH 8) comes close to the color result with the spontaneous pH value (7.4) but shows a weaker pink tone (ΔE = 18.2). At a pH of 10, the dye does not achieve a color result comparable to the spontaneously occurring pH value of 7.0 (ΔE = 37.9).

[0193] By initially deactivating the enzyme activity present in indican-containing, indigo- or indirubin-producing plant parts and subsequently adding a selected enzyme mixture with cellulase activity shortly before application in combination with cysteine ​​and isatin, the staining performance of the initially non-staining composition (Ind) can be restored. Furthermore, an immediate staining effect is achieved that does not change noticeably within 14 days.

[0194] Influence of the reducing agent

[0195] The following shows how replacing cysteine ​​with another reducing agent affects the color result.

[0196] The dyes were prepared and the strands were dyed according to the process described above. In addition to the inventive, ready-to-use dye consisting of an indican-containing composition (Ind), a cellulase blend, 1 wt.% cysteine, and 0.1 wt.% isatin in water, a comparative dye (V-2) consisting of an indican-containing composition (Ind), a cellulase blend, 1 wt.% ascorbic acid, and 0.1 wt.% isatin in water was prepared. One batch was dyed on strands of hair at 20°C and another batch at 37°C. Both strands were subsequently rinsed and dried according to the usual protocol (see above). The pH of the dispersion with the indican-containing composition (Ind), cellulase blend, ascorbic acid, and isatin was spontaneously in the range of 4.3–4.7 (measured at 20 °C)—that is, without the addition of additional pH adjusters. The color results obtained are summarized in Tables 4 and 5.Table 4: Comparison between treatment according to the invention with indican / cellulase and cysteine ​​and isatin at 37 °C and non-treatment according to the invention with indican / cellulase and ascorbic acid and isatin at 37 °C directly after rinsing and blow-drying (L*, a*, b* values ​​are shown).

[0197] Using ascorbic acid instead of cysteine ​​results in a slightly less dark, redder, and more yellowish color when stained at 37 °C. The two color results differ by AE = 14.3.

[0198] Table 5: Comparison between treatment according to the invention with indican / cellulase and cysteine ​​and isatin at 20 °C and non-treatment according to the invention with indican / cellulase and ascorbic acid and isatin at 20 °C immediately after rinsing and blow-drying (L*, a*, b* values ​​are shown)

[0199] The difference between the color results according to the invention and those not according to the invention is smaller when dyed at 20°C than when dyed at 37°C: AE = 5.5 instead of 14.3. The strand treated with cysteine ​​(according to the invention) is slightly lighter, less red, and less yellowish than the strand not dyed according to the invention.

Claims

Patent claims 1. Agent for the non-oxidative coloring of keratinic fibers, in particular human hair, containing i) at least one indican-containing component which has been pretreated at a temperature in the range from 60 °C to 200 °C, furthermore ii) an enzyme mixture comprising at least one endo-1,4-beta-glucanase (EC 3.2.1.4), at least one beta-glucosidase (EC 3.2.1.21) and at least one exo-1,4-beta-D-glucanase (EC 3.2.1.91), furthermore iii) cysteine, furthermore iv) isatin and v) water.

2. Agent for non-oxidative coloring according to claim 1, characterized in that the indican-containing component i) is selected from comminuted parts of an indigo or indirubin-producing plant and from indican-containing extracts of an indigo or indirubin-producing plant and mixtures of these indican-containing components i).

3. Agent for non-oxidative coloring according to claim 2, characterized in that the extraction agent used to obtain the at least one indican-containing extract from a plant producing indigo or indirubin is selected from water, furthermore C1-C4 alkanols and C2-C4 polyols, in particular ethanol, isopropanol, n-propanol, ethylene glycol, 1,2-propanediol, glycerol and 1,3-butylene glycol, and mixtures of these extraction agents, in particular mixtures of water and at least one C1-C4 alkanol, mixtures of water and at least one C2-C4 polyol, particularly preferably water / ethanol mixtures.

4. Agent according to one of claims 2 or 3, characterized in that the indigo or indirubin producing plant(s) is / are selected from at least one species of the following genera: - Indigofera, in particular Indigofera tinctoria, Indigo suffruticosa, Indigofera articulata, Indigofera arrecta, Indigofera heterantha “Gerardiana”, Indigofera argentea or Indigofera longi-racemosa; - Isatis, especially Isatis tinctoria (woad); - Persicaria, especially Persicaria tinctoria (dyer's knotweed); - Wrightia, especially Wrightia tinctoria; - Calanthe, especially Calanthe veratrifolia; and - Baphicacanthus cusia, synonym Strobilanthes cusia, wherein the indigo or indirubin producing plant(s) is / are preferably selected from the genus Indigofera and in particular from Indigofera tinctoria.

5. Agent according to one of claims 1 to 4, characterized in that the indican-containing component i) contains, based on its weight, 0.01 - 6 wt.% indican.

6. Agent according to one of claims 1 to 5, characterized in that the pretreatment of the at least one indican-containing component i) is carried out at a temperature in the range from 60 °C to 200 °C by heating, in particular by heating in a solvent, further by treatment with steam or by dry heating, preferably for a period of 1 to 120 minutes, particularly preferably 2 to 60 minutes, extraordinarily preferably 5 to 10 minutes.

7. Agent according to one of claims 2 to 6, characterized in that the extracted indican is separated from the solvent after completion of the extraction procedure and separation of the solid plant parts, so that the indican-containing component i) is in powder form.

8. Agent according to one of claims 1 to 7, characterized in that the enzyme mixture ii) contains at least one endo-1,4-beta-glucanase (EC 3.2.1.4) in a total amount of 20 - 35 wt.%, at least one beta-glucosidase (EC 3.2.1.21) in a total amount of 15 - 25 wt.%, and at least one exo-1,4-beta-D-glucanase (EC 3.2.1.91) in a total amount of 25 - 35 wt.%, wherein the amounts refer to the total weight of the enzyme mixture ii).

9. Agent according to one of claims 1 - 8, characterized in that it contains, based on its weight, an enzyme mixture with cellulase activity which contains at least one endo-1,4-beta-glucanase (EC 3.2.1.4), at least one beta-glucosidase (EC 3.2.1.21) and at least one exo-1,4-beta-D-glucanase (EC 3.2.1.91) in a total amount of 0.0001 - 1 wt.%, preferably 0.001 - 0.1 wt.%, further preferably 0.002 - 0.05 wt.%, particularly preferably 0.003 - 0.04 wt.%, extraordinarily preferably 0.01 - 0.03 wt.%.

10. Agent for non-oxidative coloring according to one of claims 1 to 9, characterized in that cysteine ​​is contained in an amount of 0.1 - 5.0 wt.%, preferably 0.5 - 3.0 wt.%, particularly preferably 0.8 - 2.0 wt.%, extraordinarily preferably 1.0 - 1.5 wt.%, based on the weight of the agent.

11. Agent according to one of claims 1-10, characterized in that isatin is present in an amount of 0.01-1.0 wt.%, preferably 0.05-0.5 wt.%, particularly preferably 0.08-0.3 wt.%. %, particularly preferably 0.1 - 0.2 wt.%, based on the weight of the agent.

12. Agent according to one of claims 1-11, characterized in that it contains, based on its weight, 19.9-95.0 wt.%, preferably 30.0-90.0 wt.%, particularly preferably 50.0-88.0 wt.%, extraordinarily preferably 60.0-85.0 wt.%, of water.

13. Agent according to one of claims 1-12, characterized by a pH value of 2.0 to less than 10.0, preferably from 4.0 to 9.0, particularly preferably from 5.0 to less than 8.0, extraordinarily preferably from 6.7 to less than 7.5, in each case measured at 20 °C.

14. Kit for the non-oxidative coloring of keratinic fibers, in particular human hair, with natural dyes, comprising the following compartments which are physically separate from one another: i) a composition (Ind) containing at least one indican-containing component which has been pretreated at a temperature in the range of 60°C to 200°C, and ii) an aqueous composition (E) containing an enzyme mixture comprising at least one endo-1,4-beta-glucanase (EC 3.2.1.4), at least one beta-glucosidase (EC 3.2.1.21) and at least one exo-1,4-beta-D-glucanase (EC 3.2.1.91), iii) optionally at least one further compartment, characterized in that cysteine ​​is contained in at least one of the compartments and isatin is contained in at least one of the compartments, further characterized in that the compositions i) (Ind) and ii) (E) as described in any one of claims 1-13.

15. A method for the non-oxidative coloring of keratin fibers, in particular human hair, comprising the following process steps in the specified order: a) Providing a composition (Ind) containing at least one indican-containing component which has been pretreated at a temperature in the range of 60°C to 200°C, and providing an aqueous composition (E) containing an enzyme mixture comprising at least one endo-1,4-beta-glucanase (EC 3.2.1.4), at least one beta-glucosidase (EC 3.2.1.21) and at least one exo-1,4-beta-D-glucanase (EC 3.2.1.91), and optionally providing at least one further component in a physically separate compartment, wherein in at least one of the compositions (Ind) or (E) or at least one optional component cysteine ​​and in at least one of the compositions (Ind) or (E) or at least one optional component isatin, b) mixing the compositions mentioned above under a) with one another to produce a ready-to-use colorant, c) applying the ready-to-use colorant obtained in step b) to the keratin fibers, d) leaving it to act for a time of 30 seconds to 60 minutes, preferably 5 to 45 minutes, particularly preferably 15 to 30 minutes, e) rinsing the keratin fibers with water, f) optionally drying the keratin fibers, characterized in that the ready-to-use colorant obtained in step b) is an agent according to one of claims 1 - 13, wherein the leaving to act of the ready-to-use colorant obtained in step b) preferably takes place under the influence of heat, particularly preferably at a temperature of 25 - 60 °C, further particularly preferably at a temperature of 30 - 50 °C, extremely preferably at a temperature of 35 - 40 °C.

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