JP5600239B2 - Method for producing lignin extract and lignin extract - Google Patents

Description

  The present invention relates to an extract extracted from a material containing lignin under certain conditions and use thereof.

  The lignocellulosic material containing lignin and the black liquor that is the residue obtained by separating cellulose as the pulp raw material from the lignocellulosic material are not always used effectively, and the ratio of disposal as waste is not small. is there.

  On the other hand, techniques for effectively using lignin have also been developed. For example, a technique is known in which lignin in a lignocellulosic material is separated as a lignophenol derivative by reacting with an acid in the presence of phenol (Patent Documents 1 and 2). The lignophenol derivative has a polymer form in which lignin is appropriately reduced in molecular weight.

JP-A-2-233701 Japanese Patent Laid-Open No. 9-278904

  According to the methods described in Patent Documents 1 and 2, lignin having a complicated three-dimensional structure can be separated in a plant having a certain molecular weight in an easy-to-use form. However, in order to effectively use lignin, a method for obtaining a component derived from lignin from a lignin-containing material is not limited to this method, and it is desired that there are various aspects.

  Therefore, an object of the present invention is to provide a new method for producing a lignin extract and a lignin extract obtained thereby.

  The present inventor has found that the brownish extract obtained by crushing and pressing a lignin-containing material such as a lignocellulosic material in the presence of metal ions has various functions, and has completed the present invention. According to the present invention, the following means are provided.

  According to this invention, it is a manufacturing method of a lignin extract, Comprising: A manufacturing method provided with the process of making a metal ion and a lignin containing material contact, and extracting a lignin origin component from a lignin containing material is provided.

  The metal ions may be one or more selected from the group consisting of iron ions, copper ions, and cobalt ions. The lignin-containing material may include herbaceous foliage. Furthermore, the extraction step may be a step involving pressing or crushing of the lignin-containing material.

  According to the present invention, a lignin extract obtained by any of the above methods is also provided. According to this invention, the antiallergic agent which uses the said lignin extract as an active ingredient is also provided. Moreover, the wrinkle improving agent which uses the said lignin extract as an active ingredient is also provided. Furthermore, a stain improving agent comprising the above lignin extract as an active ingredient is also provided.

  The present invention relates to a method for producing a lignin extract and a lignin extract. The lignin extract obtained by the method of the present invention has an ultraviolet absorption effect, and has effects such as allergy symptom relief, wrinkle improvement and stain improvement. Hereinafter, embodiments of the present invention will be described in detail. In addition, although it is inference and does not constrain the present invention, it is considered that lignin reduced in molecular weight is obtained in a state stabilized with metal ions by the above production method. Such lignin is considered to be stabilized by its phenolic hydroxyl group, alcoholic hydroxyl group, etc. capturing metal ions to form a complex.

(Method for producing lignin extract)
(Lignin-containing material)
As a lignin containing material, the lignin contained in a plant structure or the derivative | guide_body of the said lignin should just be included. Examples of the lignin-containing material include plant-derived lignocellulose materials and materials obtained by chemically modifying the lignocellulose materials. Examples of plant-derived lignocellulosic materials include whole or parts of plants, processed products having woody parts, and the like. Plants include, but are not limited to, conifers, broad-leaved trees, eucalyptus, and other herbs, as well as rice, wheat, corn, sugarcane, kenaf, ghetto, moonfish, and other herbs and bamboo. . Non-use materials of plant-derived lignocellulose materials can also be used. Examples of non-use materials include thinned wood, pruned plant branches and leaves, and stalks and leaves after harvesting edible parts. Among these, herbaceous stems and leaves can be preferably used as the lignocellulosic material. Since these tissues are coarser than woody species, a lignin extract can be efficiently obtained. Moreover, a chlorophyll extract is also contained in a lignin extract by using the leaf part containing a chlorophyll.

  The lignocellulosic material is preferably crushed to some extent in consideration of extraction efficiency. The herbaceous lignocellulosic material is preferably cut appropriately, and the woody lignocellulosic material is preferably in the form of chips or powder.

  In addition to the lignophenol derivatives described in Patent Documents 1 and 2, as well as modified lignins such as sulfolignin, black liquor and tea gallate that are pulp extraction residues can be used as materials obtained by chemically modifying lignocellulosic materials. And vegetable food wastes. For example, the essential oil is extracted from Kumazasa by steam distillation, and the distilled water after separation of the essential oil contains polyphenols derived from Kumazasa lignin. Such a distillation residue can also be used as a lignin-containing material.

(Metal ions)
Examples of metal ions include transition metal ions and rare earth metal ions such as iron ions, copper ions, zinc ions, cobalt ions, nickel ions, europium ions, terbium ions, lanthanum ions, and neodymium ions. Among these, iron ions, copper ions, and cobalt ions can be preferably used. In particular, iron ions (Fe ³⁺ ) are most preferable from the viewpoints of suppressing the stability of the extract (anaerobic fermentation (rot)), cost, availability, and the like. Metal ions can be used alone or in combination of two or more. Copper ions and cobalt ions have a good green to blue appearance.

To make these metal ions exist, ferric oxide (Fe ₂ O ₃ ) can be used as iron ions, copper sulfate as copper ions, and cobalt sulfate as cobalt ions.

(Extraction process)
The extraction step of bringing the metal ions into contact with the lignin-containing material can be performed while appropriately mixing the lignin-containing material (by stirring or vibration) in a state where the metal ions can exist. The amount of metal ions present in the extraction process is not particularly limited. When crushed herbaceous foliage is used as a lignin-containing material, the extract is colored due to the presence of a sufficient amount of metal ions. For example, the iron ion is blackened. On the other hand, in the case of copper ions, it is possible to obtain an extract having a green color and excellent color tone. Therefore, it is possible to set an appropriate metal ion concentration by observing the appearance color of the extract, and therefore observing the appearance color of the extract. When iron ions are used as metal ions, typically, a solution of about 0.2 g / l or more and 1 g / l or less can be used as the ferric oxide solution.

  For example, the metal ion source in the extraction process may be rust (ferric oxide) inside an iron press used for extraction. The rusted iron press functions as a good iron ion supply source and can carry out a good extraction process.

  In order to carry out the extraction step in the presence of metal ions, it is preferred that there is an aqueous medium that can contain metal ions. In the extraction step, an aqueous medium such as water may be added separately in addition to the cell fluid and / or intercellular fluid supplied from the lignin-containing material itself. In particular, when a herbaceous lignocellulosic material is used, it is not always necessary to add an aqueous medium, but water or the like can be appropriately added in consideration of the concentration of the extract and extraction efficiency. On the other hand, when using woody lignocellulosic materials such as wood powder, it is preferable to use an appropriate amount of water.

  The extraction step is not particularly required to be heated, but may be heated to promote elution of lignin and promote binding with metal ions. In particular, when using copper ions, cobalt ions, or the like, it is preferable to perform an extraction step involving heating. For example, it is preferable to heat to about 80 ° C or higher and 100 ° C or lower. Further, although pressurization is not required, pressurization may be performed. The extraction step may be performed using, for example, a pressing and breaking device for producing paper.

  Prior to the extraction step or accompanying the extraction step, various operations for accelerating decomposition of lignin or promoting separation from cellulose can be performed. For example, lignophenol derivatization described in Patent Documents 1 and 2, explosion and steaming used for crushing and extraction of lignocellulose materials, oxidation treatment, reduction treatment, acid treatment, alkali treatment, enzyme treatment, microbial treatment, water and Examples include subcritical fluid processing of organic solvents, supercritical fluid processing, and the like. Each of these treatments may be performed on the lignin-containing material prior to the extraction step with metal ions, or may be performed in the extraction step.

Note that examples of the oxidizing agent for the oxidation treatment include ozone, Fe ³⁺ -HCl, alkali-H ₂ O ₂ , and Ce ⁴⁺ . Examples of the reducing agent for the reduction treatment include lithium tetrahydridoaluminate and sodium tetrahydroborate. Among these, sodium tetrahydroborate that selectively reduces a carbonyl group contained in lignin is preferably used. Examples of the alkali treatment chemical include various basic compounds such as sodium hydroxide, potassium hydroxide, ammonia, sodium carbonate, and sodium borate. As the chemical for acid treatment, sulfuric acid, hydrochloric acid, nitric acid, phosphoric acid, organic acid and the like are preferably used. As the enzyme treatment, a peroxidase that promotes fragmentation of lignin, a saccharifying enzyme that forms a glycoside, a glucuronyltransferase that oxidizes glucuron, and the like are preferably used. For microbial treatment, rot fungi and yeast are preferably used for fragmentation and modification of lignin because of ease of handling and safety.

  Further, when the lignin-containing material is immersed in an aqueous medium containing metal ions prior to the extraction step, an extract can be easily obtained. The immersion operation may employ a different method depending on the metal ion to be used. For example, in the case of iron ions, the lignin-containing material may be immersed in the metal salt solution used in the extraction step for about 1 to 24 hours and left to stand. In consideration of fermentation, it is preferable to stand at normal temperature or lower temperature. For example, when using copper ion or cobalt ion, it is preferable to promote capture of metal ions by lignin by heating to about 80 ° C. or more and 100 ° C. or less. The lignin-containing material after immersion may be subjected to an extraction step together with the immersion liquid, or may be performed only on the lignin-containing material separated from the immersion liquid.

  Although the time of the extraction process accompanied by mixing etc. is not specifically limited, For example, when obtaining a lignin extract from a lignocellulosic material, it should just be performed to such an extent that the restraint state of the cellulose by a lignin is substantially cancelled | released. Although it does not specifically limit, For example, it can implement in tens of minutes-about several hours. In the extraction step, mixing or the like can be stopped as necessary. Moreover, the extraction step may include adjusting the metal ion concentration by adding a metal salt solution as appropriate.

  The extract after the extraction process includes an extract under metal ions from lignin. In the extract, in addition to the case where a lignocellulase-based material is used as a lignin-containing material, polymer lignin may exist as a solid content. In this case, if necessary, the liquid portion can be separated as a lignin extract by a known solid-liquid separation technique such as centrifugation or filtration. Moreover, you may remove suitably the contaminating component in an extract according to the kind of used lignin containing material, and the use of a lignin extract.

  The extract obtained by the extraction process is usually colored. By adding an extract (typically an extract) extracted with citric acid from acids such as citric acid or other herbs to the extract, the color of the extract can be changed to a lighter color, Brightness can be improved. As such an extract for color tone adjustment, for example, a citric acid extract that exhibits a light pink to light purple color such as perilla leaves, hibiscus petals, cherry leaves, and the like can be used. What is necessary is just to set a citric acid concentration suitably. It is preferable to adjust the color tone at as low a temperature as possible.

(Lignin extract)
The lignin extract of the present invention is an extract of lignin in the presence of metal ions, and can be typically obtained by the above-described production method of the present invention. The lignin extract may be in a solid form such as liquid, slurry, paste or powder. For example, the extract obtained by the above method may be used as it is, or may be diluted or concentrated. Further, the extract may be dried and solidified by freeze drying or the like.

  It has been confirmed that the lignin extract can exhibit an ultraviolet absorbing action, an antiallergic action, a wrinkle improving action, a stain improving action and the like. The wrinkle improving action includes a wrinkle formation preventing action in addition to wrinkle improvement. The stain improving effect includes a stain formation preventing effect in addition to stain improving. In addition, the lignin extract has an anti-rot effect and is excellent in stability itself.

  The lignin extract can be used as an ultraviolet absorber based on the ultraviolet absorbing action. Ultraviolet absorbers can take various forms such as cosmetics, pharmaceuticals, and quasi-drugs that are external preparations for skin. Specifically, as cosmetics etc., sunscreen oil, sunscreen lotion, sunscreen cream, cream / milky lotion, lotion, perfume, funny, cosmetic oil, hair cosmetics, hair dye, perfume, powder, pack, It can be used as a foundation, powdered perfume, blusher, eyeliner, eye cream, eye shadow, mascara, eyebrow, nail cream, nail polish, lipstick, lip balm, soap, face wash, shampoo, rinse and the like. Examples of pharmaceuticals include aerosols, liquids, extracts, ointments, suspensions, patches, poultices, liniments, lotions and the like.

  The lignin extract can also be used as a wrinkle improving agent or a stain improving agent based on the wrinkle improving action. The wrinkle improving agent and the stain improving agent can take various forms as cosmetics, pharmaceuticals, quasi-drugs and the like for external use on the skin, like the ultraviolet absorber.

  Moreover, a lignin extract can be used as an antiallergic agent based on an antiallergic action. The antiallergic agent can take various forms such as cosmetics, quasi drugs, and pharmaceuticals. Specifically, cosmetic forms such as lotion, cream, milky lotion, funny, cosmetic oil, hair cosmetics, hair dye, paste perfume, powder, pack, foundation, powder perfume, blusher, eyeliner, eye cream, Eye shadow, mascara, eyebrow, lipstick, lip balm, soap, face wash, shampoo, rinse and the like. Examples of pharmaceuticals include aerosols, solutions, extracts, ointments, eye ointments, suspensions, patches, poultices, liniments, lotions and the like.

  The various preparations (external preparations for skin) containing the lignin extract of the present invention may further contain components having various effects other than the lignin extract. For example, moisturizer, keratin improving agent, keratolytic agent, antibiotics, skin permeation enhancer, blood circulation promoter, anti-inflammatory agent, cell activator, anti-inflammatory agent, analgesic agent, emollient, emollient, wound treatment agent And metabolism promoters.

  Various preparations containing the lignin extract of the present invention can contain various pharmaceutically acceptable ingredients in these preparations. For example, oil and fat components, phospholipids, UV absorbers, IR absorbers, emulsifiers, surfactants, antiseptics, stickers, antioxidants, whitening agents, vitamins, amino acids, hormones, peptides, bioactive plant extracts, Examples include fluorescent materials, pigments, dyes, fragrances, scrub agents, sequestering agents, binders, extenders, thickeners, saccharides, nutrient components, pH adjusters, chelating agents, and bactericides.

  In addition, about the addition amount of each component, the addition method, and the formulation method, it can follow a method well-known in this technical field. Those skilled in the art can easily prepare the various preparations containing the lignin extract based on the above general description and the specific disclosure of the examples, or by appropriately modifying or modifying them as necessary. Can be manufactured. The lignin extract content in the preparation is not particularly limited, and is appropriately selected according to the use of the preparation.

  Various preparations having the lignin extract of the present invention can obtain the above-mentioned various effects by external application to the skin or mucous membrane. The dosage of these preparations is not particularly limited. For example, after washing the lignin extract in the morning and evening, take an appropriate amount (for example, about 1 ml to 5 ml) according to the area to be applied, and prevent UV rays (face, neck, limbs, etc.), wrinkles, and spots. Can be applied to places, rashes caused by allergies, etc. In order to exert an antiallergic effect on the mucous membranes such as the nose and eyes, an appropriate amount may be applied to the mucous membrane of the nose and eyes and the surroundings.

  The present invention will be specifically described below with reference to examples. The materials, amounts used, ratios, processing details, processing procedures, and the like shown in the following examples can be changed as appropriate without departing from the spirit of the present invention. Therefore, the scope of the present invention should not be construed as being limited by the specific examples shown below.

(Preparation of lignin extract)
In this example, the crushed material of the shoot and leaf portion of the genus Ganyoga genus (Gentou (sannin)) is put into an iron crusher having rust as an iron ion source, and at room temperature without adding water To obtain a black-brown lignin extract. A brown supernatant obtained by removing the solid content of the extract by centrifugation was obtained. The obtained supernatant was diluted 3 times by adding 2 times the amount of water by volume. Moreover, it diluted 2 times with 1 time amount of water. In Examples 2, 4 and 5, the 3-fold diluted solution was used as the lignin extract, and in Example 3, the 2-fold diluted solution was used as the lignin extract. For the lignin extract (stock solution) prepared in this example, the gene mutagenicity test (reversion mutation test) was performed for three strains (Salmonella typhimurium TA100, Escherichia coli WP2uvrA strain, Salmonella yphimurium TA198 strain). As a result, mutagenicity was not observed. Moreover, when the irritation and sensitization by the Marzulli-maibach method were evaluated (50 test subjects), it was confirmed that there was no irritation and sensitization and hypoallergenicity.

(Wrinkle improvement test and antiallergic test)
Using the lignin extract (3-fold diluted solution) prepared in Example 1, the effect on human eye corner wrinkles and texture was evaluated by the following method. There were 19 subjects, and the test period was 3 months. In addition, the usage and dose were applied to the half-face twice / day (morning and evening), the lignin extract was applied, the eye corner replica was collected before use and after 3 months from use, and texture and wrinkle were analyzed and compared. . It should be noted that external preparations and cosmetics other than the lignin extract were not particularly limited, and those conventionally used or cosmetic oils (jojoba oil, squalane oil, etc.) were used.

  As for the analysis method, for wrinkles, a two-dimensional image analysis method using an oblique light proof by a replica that complies with the Cosmetic Functionality Evaluation Method Guidelines was used. The analysis items were the total number of wrinkles, the wrinkle area ratio, the maximum depth of maximum wrinkles, the average depth of maximum wrinkles, and the total average wrinkle depth. For the texture, a skin surface morphology analysis method using oblique illumination with a replica was used. The analysis items were the number of texture peaks (number of textures), texture distribution (texture irregularity), and texture spacing. These analysis items are defined as follows.

(1) Wrinkles Total number of wrinkles: Number of wrinkles = Number of binarized image extraction areas (N / cm ² )
Wrinkle area ratio (RWA) ^{* 1} : Total area of wrinkles in the vertical depth direction / measured area (1 cm ² ) × 100%
Maximum depth of maximum wrinkle ^{* 2} : Depth of the deepest part of all wrinkles (mm)
Maximum wrinkle average depth ^{* 2} : Average depth (mm) of the deepest wrinkle (1)
Total wrinkle average depth ^{* 2} : Average value of the depth of all wrinkles (mm)
* 1: Teruji Hayashi et al., Waging due to aging, sun exposure and cosmetics (Japan Cosmetic Engineers Journal, 27 (3), p355-373, 1993)
* 2: Anti-aging function evaluation committee, anti-wrinkle product evaluation guidelines for acquiring new effects (Journal of Japanese Cosmetics Society, 30 (4), p316-332, 2006)

(2) Number of texture peaks ^{* 3} : The number of textures (N / cm) and the number of textures in the length of 1 cm.
Texture distribution ^{* 3} : Standard deviation of texture interval (10/256 / mm), representing irregularity of texture.
Interval of texture ^{* 3} : Average value of distance from texture to next texture (10/256 / mm)
* 3: Toshiro Sone et al., Study on seasonal changes in skin surface morphology and skin roughness by image analysis (Journal of Japanese Cosmetic Science Society, 15 (2), p60-65, 1991)

  Table 1 shows the average values and standard deviations for these analysis items obtained before and after using the lignin extract. 1 to 3 show the results of oblique observation of each replica before and after use for three subjects.

  As shown in Table 1, no significant difference was observed before and after use for texture, but a significant difference in the total number of wrinkles and the wrinkle area ratio (Paired t-Test, * p <0.05, * * P <0.01) was observed. It was found that the total number of wrinkles is the number of wrinkles and has a tendency to reduce wrinkles. Further, the wrinkle area ratio is the total area of the wrinkles in the vertical depth direction, which indicates that the wrinkle depth has become shallow overall. In addition, as shown in FIGS. 1 to 3, the shadow of the replica (in the replica, the wrinkles are conversely mountain ranges) is remarkably small after use, and it is visually observed that the wrinkles are shallow. I was able to observe. It was also found that the tendency of such wrinkle depth to decrease tends to increase with deep wrinkles.

  In addition, symptom improvement was observed for hay fever patients (including allergic rhinitis) among the subjects, and for patients with atopic dermatitis in which a lignin extract was similarly applied to the affected area of atopic dermatitis, Reduction of itching was observed.

  From the above results, it was found that the lignin extract is effective in improving wrinkles. It was also found to be effective especially for deep wrinkles. It was also found that the lignin extract has antiallergic properties that alleviate and improve allergic symptoms. In particular, it was found that application to the face or the like is effective in improving hay fever and is effective in atopic dermatitis.

(Collagen production promoting effect test)
Using the lignin extract (2-fold diluted solution) prepared in Example 1, a collagen production promoting action test was performed by the following method. That is, human normal fibroblasts (Detroit 551) were cultured using MEM containing 10% FBS, 1% NEAA, 1 mmol / L sodium pyruvate, and then cells were collected by trypsin treatment. The collected cells were diluted with the above medium to a concentration of 2 × 10 ⁵ cells / ml and then seeded at 100 μl per well in a 96-well microplate and cultured overnight. After completion of the culture, the medium was removed, 150 μl of lignin extract dissolved in 0.5% FBS-containing MEM was added to each well, and cultured for 3 days. After culturing, the amount of collagen in each well was measured by ELISA. The publication of the collagen production promoting action was as follows. However, A is the amount of collagen when the test sample (lignin extract) is added, and B is the amount of collagen when the test sample is not added.
Collagen production promotion rate% = A / B × 100

  The collagen production promotion rate for the lignin extract was 859.2 ± 216.7 (200 μg / ml) and 188.5 ± 48.4 (50 μg / ml). This was measured at the same time, about 3 times or more of the extract obtained by extracting 50% ethanol with the distillation residue (distilled extract fraction, not including metal ions) from which essential oil was removed after steam distillation of moon peach leaves (monthly). Peach leaf extract 244.7 ± 67.4 (200 μg / ml), 112.1 ± 29.8 (50 μg / ml)).

  From the above, it was found that the lignin extract has a strong collagen production promoting action.

(Blemish improvement test)
Using the lignin extract (3-fold diluted solution) prepared in Example 1, the effect on facial stains (senile pigment spots) was evaluated by the following method. There were two test subjects, and the usage and dose were twice / day (morning and evening). The lignin extract was applied to the spot, and the spot was observed before use and after a certain period from use. In addition, the external preparations and cosmetics other than the lignin extract were not particularly limited, and the same conventional ones were used. The follow-up observation results are shown for one subject.

  In all subjects, the spots improved with time, and one person almost disappeared after 3 months of continuous use. In addition, as shown in FIG. 4, the other person improved considerably after about 8 months from the start of use, and further improved after 9 months (after about 17 months from the start of use). From the above results, it was clear that the lignin extract has a stain improving effect.

(Antiallergic test)
Using the lignin extract (3-fold diluted solution) prepared in Example 1, the antiallergic action was evaluated. The test subjects were two latex allergic patients, and the lignin extract was applied in an appropriate amount to the affected area when symptoms and symptoms occurred. As a result, itching was suppressed and improvement of allergic symptoms was confirmed for two subjects. From the above, it was found that the lignin extract has an antiallergic action that particularly suppresses early inflammation.

(Confirmation of components in lignin extract by GC-MS)
The lignin extract (stock solution) obtained in Example 1 was analyzed under general conditions using GC-MS. The results are shown in FIG.

  As shown in FIG. 5, various low molecular weight compounds considered to be derived from lignin could be confirmed in the lignin extract. From the above, it was found that lignin was stabilized by lowering the molecular weight by bringing a metal ion and a lignin-containing material into contact with each other, and a lignin extract having various functions was obtained.

It is a figure which shows the replica observation result which shows the wrinkle improvement tendency in 1 test subject among 3 test subjects. The observation result of the replica before use is shown on the left side, and the observation result of the replica at the same location is shown on the right side. It is a figure which shows the replica observation result which shows the wrinkle improvement tendency in another 1 person among 3 test subjects. The observation result of the replica before use is shown on the left side, and the observation result of the replica at the same location is shown on the right side. It is a figure which shows the replica observation result which shows the wrinkle improvement tendency in another one person among three test subjects. The observation result of the replica before use is shown on the left side, and the observation result of the replica at the same location is shown on the right side. It is a figure which shows the observation result which shows the spot improvement tendency in one test subject. The state before use is shown on the left side, the state of the same part after about 8 months has passed on the lower right side, and the state of the same part after about 17 months has passed since the start of use on the right side. It is a spectrum figure which shows the result of GC-MS.

Claims (10)

A method for producing a lignin extract,
In an iron crusher having rust inside, an extraction step of obtaining a colored extract by pressing and extracting a crushed material of lignocellulosic material that is a stem and leaf part of a plant belonging to herbs at room temperature ,
A manufacturing method comprising:   The manufacturing method according to claim 1, wherein the lignocellulosic material is alkali-treated prior to the extraction step. The manufacturing method of Claim 1 or 2 which processes the said lignocellulose material with the microorganisms containing a decaying fungus or yeast with the said extraction process.   Furthermore, the manufacturing method in any one of Claims 1-3 provided with the process of obtaining a clarified liquid by solid-liquid separation from the said extract.   Furthermore, the manufacturing method in any one of Claims 1-4 provided with the process of adding a citric acid or the citric acid extract of a plant to the said extract. The manufacturing method according to any one of claims 1 to 5 , wherein the plant is moon peach .   The lignin extract obtained by the manufacturing method in any one of Claims 1-6. The antiallergic agent which uses the extract obtained by the manufacturing method of Claim 6, or the extract obtained by diluting, concentrating or pulverizing the said extract as an active ingredient. The wrinkle improving agent which uses the extract obtained by the manufacturing method of Claim 6 or the extract obtained by diluting, concentrating or pulverizing the said extract as an active ingredient. The stain improving agent which uses as an active ingredient the extract obtained by the manufacturing method of Claim 6, or the extract obtained by diluting, concentrating or pulverizing the said extract .