JP2008212136A - Post-fermented tea and method for producing the same - Google Patents
Post-fermented tea and method for producing the same Download PDFInfo
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本発明は、茶を発酵させて得られる、抗酸化作用を有する後発酵茶、その抽出物及びそれらを含有する皮膚外用剤組成物、浴用剤組成物、食品組成物等に関する。 The present invention relates to a post-fermented tea having an antioxidative effect obtained by fermenting tea, an extract thereof, a skin external preparation composition, a bath preparation composition, a food composition and the like containing them.
世界中で愛飲されている緑茶、烏龍茶、紅茶といった茶は、栄養摂取だけではなく、嗜好品としても飲用される。また、ストレス解消や気分転換等を目的としても飲用される。これは茶が、味だけでなく香りをも楽しめる飲料であることを意味する。例えば、紅茶は、発酵工程中に茶葉に含まれる香り成分が増加するため、その香りを堪能する飲料として位置付けられている。 Teas such as green tea, oolong tea, and black tea that are loved all over the world are used not only for nutrition but also as a luxury product. It is also used for the purpose of stress relief and mood change. This means that tea is a beverage that can enjoy not only taste but also aroma. For example, black tea is positioned as a beverage that enjoys the aroma because the aroma component contained in the tea leaf increases during the fermentation process.
茶は、製法の違いから次の４種類に大別される。緑茶に代表される不発酵茶、烏龍茶に代表される半発酵茶、紅茶に代表される完全発酵茶及び黒茶に代表される後発酵茶である。
不発酵茶は生茶葉を蒸熱又は炒熱処理して殺青した後、揉捻、乾燥したものであり、生茶葉自身の持つ酵素群は失われている。それゆえ、カテキン由来の独特の苦味を持つ。
一方、半発酵茶及び完全発酵茶は、殺青を行わず、揉捻して生茶葉自身の持つ酵素群の作用で発酵を行わせた後、加熱により酵素群を失活させて乾燥させたものであり、その発酵の程度により半発酵茶と完全発酵茶に分けられる。一般的に、緑茶と比較し、苦味が少なく、独特の風味を持つものが多い。
また、後発酵茶は、通常、不発酵茶を原料に自然環境の微生物を生育させ、製造されたものである。生育する微生物としては乳酸菌、真菌、酵母等が知られている（非特許文献１）が、発酵工程上、該微生物をコントロールすることは難しい。それゆえ、生育する微生物の種類、発酵期間等により、風味が異なる。
Tea is roughly divided into the following four types depending on the manufacturing method. These are non-fermented tea typified by green tea, semi-fermented tea typified by oolong tea, fully fermented tea typified by black tea, and post-fermented tea typified by black tea.
Unfermented tea is obtained by steaming or heat-treating fresh tea leaves and killing them, then twisting them and drying them. The enzyme group of fresh tea leaves themselves is lost. Therefore, it has a unique bitterness derived from catechin.
On the other hand, the semi-fermented tea and the completely fermented tea are those that are not killed, are twisted and fermented by the action of the enzyme group of the fresh tea leaves themselves, and then the enzyme group is deactivated by heating and dried. Yes, it can be divided into semi-fermented tea and completely fermented tea depending on the degree of fermentation. In general, it has less bitterness and has a unique flavor compared to green tea.
Further, post-fermented tea is usually produced by growing natural microorganisms from unfermented tea as a raw material. Lactic acid bacteria, fungi, yeasts and the like are known as growing microorganisms (Non-patent Document 1), but it is difficult to control the microorganisms during the fermentation process. Therefore, the flavor differs depending on the type of microorganisms that grow and the fermentation period.
また、茶に水を加えて微生物発酵処理する方法に関しては、これまで、例えば特許文献１及び２において、真菌類を用いた技術が開示されている。 In addition, with regard to a method for microbial fermentation treatment by adding water to tea, for example, Patent Documents 1 and 2 disclose techniques using fungi.
特許文献１には、麹菌によるスラリー発酵を行い、ガレート基を有するカテキン類を低減させ、没食子酸含有量を高めた後発酵茶飲料が記載されている。そして、この後発酵茶は粉砕した原料茶葉を水に懸濁させた状態で行うスラリー発酵で、かつこのスラリー発酵が、麹菌の酵素は活性を有するが麹菌は生育・増殖しない温度帯で行うことにより製造されている。 Patent Document 1 describes a post-fermented tea beverage in which slurry fermentation with koji molds is performed to reduce catechins having a gallate group and increase the gallic acid content. And after this fermented tea is slurry fermentation performed in a state where the crushed raw tea leaves are suspended in water, and this slurry fermentation is performed in a temperature range where the koji mold enzyme is active but koji mold does not grow or proliferate. It is manufactured by.
特許文献２には、β−１，３／１，６−グルカン、キチン、γ−アミノ酪酸等の機能や香気成分産生を目的に、茶葉を２５〜８０重量％の水分を含むように調整し、真菌類を接種し、生育させ、発酵機能茶を得ることが記載されている。 In Patent Document 2, tea leaves are adjusted to contain 25 to 80% by weight of water for the purpose of producing functions such as β-1,3 / 1,6-glucan, chitin, and γ-aminobutyric acid and aroma components. Inoculate fungi and grow to obtain a fermented functional tea.
緑茶は、一般的に、カテキン類が豊富で、抗酸化活性が高く、抗がん効果、動脈硬化抑制効果、血圧上昇抑制効果、抗糖尿病効果、肥満予防効果、抗アレルギー効果、抗菌・抗ウイルス効果等が知られている（非特許文献２）。しかし、緑茶の味はカテキン特有の渋みを持ち、食品として使用する場合に有効量を摂取することは非常に困難が伴う。また、化粧品素材として使用する場合も皮膚への刺激性がある。それゆえ、より抗酸化活性が高く、刺激性の少ない素材が求められていた。 Green tea is generally rich in catechins, has high antioxidant activity, anticancer effect, arteriosclerosis inhibitory effect, blood pressure increase inhibitory effect, antidiabetic effect, obesity preventive effect, antiallergic effect, antibacterial / antiviral Effects and the like are known (Non-Patent Document 2). However, the taste of green tea has an astringency peculiar to catechin, and it is very difficult to take an effective amount when used as food. Also, when used as a cosmetic material, it is irritating to the skin. Therefore, a material having higher antioxidant activity and less irritation has been demanded.
また、太陽光線に含まれる紫外線は、ヒトの皮膚組織、例えば表皮層、真皮層にまで到達し、これら組織を構成する基底細胞（角化細胞）やメラニン細胞、線維芽細胞に刺激を与えたり、細胞を破壊する事により、皮膚に紅斑を生じさせ、色素沈着や時に炎症、皮膚がんをも発生させる事がある（非特許文献３）。それゆえ、紫外線暴露の影響を抑制するために、紫外線吸収あるいは紫外線散乱作用を有するサンスクリーン剤や、紫外線暴露より発生するフリーラジカルを捕捉する抗酸化剤、紫外線暴露による皮膚組織のダメージを防止あるいは修復する皮膚損傷防止剤／修復剤等の開発が行われてきた。しかしながら、サンスクリーン剤は、皮膚刺激や光毒性の問題、あるいは効果が強いゆえに、人によっては接触性皮膚炎を誘発する場合もある。これらサンスクリーン剤は合成によって得られる高分子化合物や金属化合物であり、それゆえ、より安全性の高い、天然物から得られる紫外線防御剤が求められている。 In addition, ultraviolet rays contained in sunlight reach human skin tissues such as the epidermis layer and dermis layer, and stimulate basal cells (keratinocytes), melanocytes, and fibroblasts that constitute these tissues. By destroying cells, erythema is produced in the skin, and pigmentation, sometimes inflammation, and skin cancer may occur (Non-patent Document 3). Therefore, in order to suppress the effects of UV exposure, sunscreen agents that have UV absorption or UV scattering action, antioxidants that trap free radicals generated by UV exposure, and prevent skin tissue damage from UV exposure or Developments have been made in skin damage prevention / restoration agents and the like to be repaired. However, sunscreen agents may cause contact dermatitis depending on the person because of skin irritation, phototoxicity problems, or strong effects. These sunscreen agents are high molecular compounds and metal compounds obtained by synthesis, and therefore there is a need for UV protection agents obtained from natural products with higher safety.
天然物由来の紫外線防御剤としてはケルセチン配糖体（特許文献３）、アカハギンナンソウ抽出物（特許文献４）、マンゴスチン（特許文献５）、キク科植物抽出物（特許文献６）やモクレン科オガタマノキ属植物からの抽出物（特許文献７）が知られている。特許文献３は培養細胞に紫外線を照射することにより紫外線防御作用を証明したものであり、その他の特許文献４〜７は紫外線Ａ波、紫外線Ｂ波の吸収波長領域を持つことから紫外線防御作用を示している。 As a natural product-derived ultraviolet light protective agent, quercetin glycoside (Patent Document 3), red-billed extract (Patent Document 4), mangosteen (Patent Document 5), asteraceae plant extract (Patent Document 6) and magnolia An extract from a plant belonging to the genus Ogatamanoki (Patent Document 7) is known. Patent Document 3 proves the ultraviolet protective effect by irradiating the cultured cells with ultraviolet rays, and other Patent Documents 4 to 7 have the absorption wavelength region of the ultraviolet A wave and the ultraviolet B wave. Show.
後発酵茶に関しては、皮膚外用剤として用いることは知られておらず、ましてや紫外線防御作用についても全く知られていない。
上記の如く、緑茶は、カテキン類が豊富で、抗酸化活性が高く、種々の効果を有するが、カテキン特有の渋みや皮膚への刺激性があるゆえ、より抗酸化活性が高く、刺激性の少ない素材が求められていた。また、従来の紫外線吸収剤に用いられる素材の中には人によっては接触性皮膚炎の誘発や光毒性作用が見られるため、紫外線暴露に起因する皮膚障害を防止でき、より安全性の高い、天然由来の素材が求められていた。一方、現在市場にある碁石茶、阿波番茶、プーアル茶等の後発酵茶は、渋味がなく、刺激性もないが、自然環境中で、微生物を生育させ、製造しているので、製品の品質を安定させることが難しく、また抗酸化活性は低いものである。
従って、本発明の目的は、抗酸化活性が高く、渋味や刺激性が弱く、種々の化粧料、浴用剤や食品等への適用が可能な植物由来の新たな素材を提供することにある。また、当該植物由来の新たな素材の製造方法を提供することにある。
As described above, green tea is rich in catechins, has high antioxidant activity, and has various effects, but because of its astringent nature and irritation to the skin, it has higher antioxidant activity and irritation. Less material was required. In addition, some materials used in conventional UV absorbers can induce contact dermatitis and phototoxic effects in some people, thus preventing skin damage caused by UV exposure and higher safety. Naturally derived materials were sought. On the other hand, post-fermented teas such as Shizukuishi tea, Awaban tea, Pu'er tea, etc. currently on the market have no astringency and no irritation, but because they grow and produce microorganisms in the natural environment, It is difficult to stabilize the quality, and the antioxidant activity is low.
Accordingly, an object of the present invention is to provide a new plant-derived material that has high antioxidant activity, low astringency and irritation, and can be applied to various cosmetics, bath preparations, foods, and the like. . Moreover, it is providing the manufacturing method of the new raw material derived from the said plant.
そこで本発明者らは、抗酸化活性の弱い後発酵茶に着目し種々検討した結果、抗酸化活性の指標の一つであるＳＯＤ活性を指標にして、後発酵茶の製造条件を検討した。まず、微生物として糸状菌、乳酸菌、酵母等を用いて茶を発酵させ、ＳＯＤ活性を指標にスクリーニングしたところ、糸状菌による発酵が最もＳＯＤ活性を高めることを見出した。
次いで、該糸状菌を用いて、培養・発酵条件についても鋭意検討を重ねた結果、ある特定の適度な水分を茶に含浸させ、該糸状菌を接種し、生育させることで、ＳＯＤ活性が著しく高い後発酵茶が得られること、さらに当該発酵茶は渋味が弱く、かつ刺激性が低いため化粧料、浴用剤や食品等の素材として有用であることを見出した。また、当該後発酵茶又はその抽出物は紫外線暴露による皮膚障害を有意に防止する抗紫外線ストレス活性を有するため、紫外線防御剤として有用であることを見出した。
Therefore, the inventors of the present invention focused on post-fermented tea with weak antioxidant activity, and as a result, studied the production conditions of post-fermented tea using SOD activity, which is one of the indexes of antioxidant activity, as an index. First, tea was fermented using microorganisms such as filamentous fungi, lactic acid bacteria, yeast, etc., and screening was performed using SOD activity as an index. As a result, it was found that fermentation by filamentous fungi has the highest SOD activity.
Next, as a result of intensive studies on culture and fermentation conditions using the filamentous fungi, tea was impregnated with certain appropriate water, inoculated with the filamentous fungus, and grown. It has been found that a high post-fermented tea can be obtained, and that the fermented tea is useful as a material for cosmetics, bath preparations, foods and the like because of its weak astringency and low irritation. Moreover, since the said post-fermented tea or its extract has the anti-UV stress activity which prevents the skin damage by UV exposure significantly, it discovered that it was useful as a UV protective agent.
すなわち、本発明は以下の発明を提供するものである。
（１）Ｃａｍｅｌｌｉａ ｓｉｎｅｎｓｉｓ種由来の茶に、茶乾燥重量１００重量部に対して１１０重量部以上の水を加え、糸状菌を接種して生育せしめることにより得られる、スーパーオキシドジスムターゼ（Ｓｕｐｅｒｏｘｉｄｅ Ｄｉｓｍｕｔａｓｅ）活性を固形分換算で２，０００ｕｎｉｔｓ／ｇ以上を有する後発酵茶。
（２）Ｃａｍｅｌｌｉａ ｓｉｎｅｎｓｉｓ種由来の茶に、茶乾燥重量１００重量部に対して１１０重量部以上の水を加え、糸状菌を接種し、生育せしめることを特徴とする、スーパーオキシドジスムターゼ（Ｓｕｐｅｒｏｘｉｄｅ Ｄｉｓｍｕｔａｓｅ）活性を固形分換算で２，０００ｕｎｉｔｓ／ｇ以上を有する後発酵茶の製造方法。
（３）（１）記載の後発酵茶から、水、エタノール、含水エタノール、１，３−ブチレングリコール及び含水１，３−ブチレングリコールからなる群より選択される１種又は２種以上の抽出溶媒を用いて抽出した後発酵茶抽出物。
（４）（１）記載の後発酵茶から、水、エタノール、含水エタノール、１，３−ブチレングリコール及び含水１，３−ブチレングリコールからなる群より選択される１種又は２種以上の抽出溶媒を用いて抽出することを特徴とする後発酵茶抽出物の製造方法。
（５）（３）記載の後発酵茶抽出物を含有することを特徴とする皮膚外用剤組成物。
（６）（３）記載の後発酵茶抽出物を含有することを特徴とする浴用剤組成物。
（７）（１）記載の後発酵茶又は（３）記載の後発酵茶抽出物を含有することを特徴とする食品組成物。
（８）（３）記載の後発酵茶抽出物を含有することを特徴とする抗酸化剤。
（９）（１）記載の後発酵茶又は（３）の後発酵茶抽出物を含有することを特徴とする紫外線防御剤。
（１０）紫外線暴露に起因する皮膚障害を防止するものである（９）記載の紫外線防御剤。
That is, the present invention provides the following inventions.
(1) Superoxide dismutase (Superoxide Dismutase) activity obtained by adding 110 parts by weight or more of water to 100 parts by weight of tea dry weight from tea derived from Camellia sinensis and inoculating and growing the fungus. Is a post-fermented tea having 2,000 units / g or more in terms of solid content.
(2) Superoxide dismutase (Superoxide Dismutase), characterized in that 110 parts by weight or more of water is added to 100 parts by weight of tea dry weight, tea is derived from Camellia sinensis seeds, inoculated with filamentous fungi, and allowed to grow. A method for producing post-fermented tea having an activity of 2,000 units / g or more in terms of solid content.
(3) One or more extraction solvents selected from the group consisting of water, ethanol, hydrous ethanol, 1,3-butylene glycol, and hydrous 1,3-butylene glycol from the post-fermented tea described in (1) After fermentation using fermented tea extract.
(4) One or more extraction solvents selected from the group consisting of water, ethanol, hydrous ethanol, 1,3-butylene glycol, and hydrous 1,3-butylene glycol from the post-fermented tea described in (1) A method for producing a post-fermented tea extract, characterized by being extracted using
(5) A skin external preparation composition comprising the post-fermented tea extract according to (3).
(6) A post-fermented tea extract according to (3).
(7) A post-fermented tea according to (1) or a post-fermented tea extract according to (3).
(8) An antioxidant comprising the post-fermented tea extract described in (3).
(9) An ultraviolet protective agent comprising the post-fermented tea described in (1) or the post-fermented tea extract of (3).
(10) The ultraviolet protective agent according to (9), which prevents skin damage caused by ultraviolet exposure.
本発明の後発酵茶及びその抽出物は、優れた抗酸化活性を有し、渋味が弱く、かつ刺激性が少ない。また、当該後発酵茶及びその抽出物は、安全性が高く、紫外線暴露によって誘導される皮膚障害を予防・軽減する効果を有する。従って、当該後発酵茶及びその抽出物は、化粧料等の皮膚外用剤、浴用剤、食品等において抗酸化剤や紫外線防御剤として使用することができる。 The post-fermented tea and its extract of the present invention have excellent antioxidant activity, weak astringency and little irritation. Further, the post-fermented tea and its extract are highly safe and have the effect of preventing / reducing skin disorders induced by exposure to ultraviolet rays. Therefore, the post-fermented tea and its extract can be used as an antioxidant or UV protection agent in skin preparations such as cosmetics, bath preparations, foods and the like.
本発明の後発酵茶の原料としての茶は、Ｃａｍｅｌｌｉａ ｓｉｎｅｎｓｉｓ種であれば、特に品種等には限定されない。また、収穫直後の生茶葉、青殺後の茶葉、粗揉後の茶葉、揉捻後の茶葉、中揉後の茶葉、精揉後の茶葉、荒茶等の緑茶であっても、茶茎、茶花及びこれらの粉末品等の加工品等であっても、さらに半発酵茶、完全発酵茶も使用可能であり、原料となる茶は後発酵茶の用途により、適宜選択可能である。このうち、不発酵茶を用いることが、高いＳＯＤ活性を有する後発酵茶を得る点から好ましい。 If the tea as a raw material of post-fermented tea of this invention is a Camellia sinensis seed | species, it will not be specifically limited to a kind | species etc. In addition, fresh tea leaves immediately after harvest, tea leaves after green killing, tea leaves after rough koji, tea leaves after twisting, tea leaves after middle cocoon, tea leaves after brewing, green tea such as rough tea, Semi-fermented tea and completely fermented tea can also be used for tea flowers and processed products such as these powder products, and the raw tea can be appropriately selected depending on the use of post-fermented tea. Among these, it is preferable to use non-fermented tea from the viewpoint of obtaining post-fermented tea having high SOD activity.
これらの茶の発酵に用いられる微生物は、高いＳＯＤ活性を有する後発酵茶を得る点から糸状菌であるのが好ましい。糸状菌としては、醸造食品や発酵食品等の食品に使用可能な糸状菌の中から選抜することが可能である。例えば、Ａｓｐｅｒｇｉｌｌｕｓ属、Ｅｕｒｏｔｉｕｍ属、Ｒｈｉｚｏｐｕｓ属等を使用することができ、より好ましくはＡｓｐｅｒｇｉｌｌｕｓ ｏｒｙｚａｅ、Ａｓｐｅｒｇｉｌｌｕｓ ｎｉｇｅｒ、Ｅｕｒｏｔｉｕｍ ｃｒｉｓｔａｔｕｍ、Ｒｈｉｚｏｐｕｓ ｏｌｉｇｏｓｐｏｒｕｓ等が挙げられ、さらには該糸状菌から誘導される変異株であっても使用することができる。これらを単独で若しくはそれらを組合せて使用してもよい。これらの菌株は、ＩＦＯ、ＩＡＭ、ＡＴＣＣ、ＮＲＲＣ等の菌株分譲機関、日本醸造協会や市販の種菌株販売会社等から入手可能である。 The microorganism used for fermentation of these teas is preferably a filamentous fungus from the viewpoint of obtaining post-fermented tea having high SOD activity. As the filamentous fungus, it is possible to select from filamentous fungi that can be used for foods such as brewed foods and fermented foods. For example, Aspergillus genus, Eurotium genus, Rhizopus genus and the like can be used, more preferably Aspergillus oryzae, Aspergillus niger, Eurotium cristatum, Rhizopus oligosporus, etc. Can also be used. These may be used alone or in combination. These strains are available from strain distribution agencies such as IFO, IAM, ATCC, and NRRC, the Japan Brewing Association, and commercial seed strain sales companies.
本発明においては、Ｃａｍｅｌｌｉａ ｓｉｎｅｎｓｉｓ種由来の茶に、茶乾燥重量１００重量部に対して１１０重量部以上、好ましくは１５０重量部以上、より好ましくは２００重量部以上、最適には３００〜４００重量部の水を加える。ここでの水分量の調整は原料となる茶に、水を加え、馴染ませ、糸状菌が生育できる条件にすることを意味する。例えば、荒茶を使用した場合、該荒茶はすでに約３〜５重量％の水分を既に含んでおり、加える水分量は、茶が含んでいる水分を考慮して調整する必要がある。また、生茶葉のように水分を７０〜８０重量％含んでいるものについては、加水しなくても良い。上記の水分含量は、培養・発酵に用いる糸状菌が繁殖又は発酵可能な量であり、かつ本発明の高いＳＯＤ活性を有する後発酵茶を得るために重要である。該糸状菌を優先的に繁殖させることで、雑菌汚染をより防止することができる。該水分調整後に、殺菌工程を加えることが好ましい。該殺菌条件は、例えば、８０〜１００℃で３０〜６０分間、あるいは１００〜１２１℃で１５〜３０分間等の条件で加熱殺菌を行うことが挙げられる。該殺菌後は、冷却し、該茶を殺菌済み培養・発酵槽に移し、下記の植菌工程を行う。なお、該殺菌工程は、茶の水分量が変化しないように密閉系で行うのが好ましい。 In the present invention, tea derived from Camellia sinensis species is 110 parts by weight or more, preferably 150 parts by weight or more, more preferably 200 parts by weight or more, and most preferably 300 to 400 parts by weight with respect to 100 parts by weight of tea dry weight. Add water. The adjustment of the amount of water here means that water is added to tea used as a raw material so as to adjust to a condition that allows filamentous fungi to grow. For example, when crude tea is used, the crude tea already contains about 3 to 5% by weight of moisture, and the amount of moisture to be added needs to be adjusted in consideration of the moisture contained in the tea. Moreover, what contains 70 to 80 weight% of water | moisture content like fresh tea leaves does not need to be hydrated. The water content is important for obtaining the post-fermented tea having a high SOD activity according to the present invention, which is an amount capable of propagating or fermenting the filamentous fungi used for culture and fermentation. By preferentially propagating the filamentous fungus, contamination with various bacteria can be further prevented. It is preferable to add a sterilization step after the moisture adjustment. Examples of the sterilization conditions include heat sterilization at 80 to 100 ° C. for 30 to 60 minutes, or at 100 to 121 ° C. for 15 to 30 minutes. After the sterilization, the mixture is cooled, the tea is transferred to a sterilized culture / fermentor, and the following inoculation process is performed. The sterilization step is preferably performed in a closed system so that the moisture content of tea does not change.
水分含量を調整した茶に、糸状菌を接種し、一般に２０〜４０℃で３〜３０日間、好ましくは２５〜３７℃で３〜７日間培養・発酵する。ここで培養・発酵方法は、通常用いられる方法であれば、液体培養・発酵法、固体培養・発酵法等いずれの方法でも良い。次いで、殺菌工程を加えることが好ましい。該殺菌条件は、例えば、８０〜１００℃で３０〜６０分間、あるいは１００〜１２１℃で１５〜３０分間等の条件で加熱殺菌を行うこと、又は、８０〜１２０℃で加熱乾燥し該糸状菌を殺菌するとともに、水分含量を１０重量％以下、好ましくは５重量％以下にする。これにより、本発明のＳＯＤ活性の高い後発酵茶が製造される。 The tea whose moisture content has been adjusted is inoculated with filamentous fungi and is generally cultured and fermented at 20 to 40 ° C. for 3 to 30 days, preferably at 25 to 37 ° C. for 3 to 7 days. Here, the culture / fermentation method may be any method such as liquid culture / fermentation method or solid culture / fermentation method as long as it is a commonly used method. Then, it is preferable to add a sterilization process. The sterilization conditions include, for example, heat sterilization at 80 to 100 ° C. for 30 to 60 minutes, or 100 to 121 ° C. for 15 to 30 minutes, or heat drying at 80 to 120 ° C. Is sterilized, and the water content is 10% by weight or less, preferably 5% by weight or less. Thereby, the post-fermented tea with high SOD activity of the present invention is produced.
本発明の後発酵茶は、そのままの形態でも利用可能であるが、加熱、乾熱、マイクロ波等で殺菌を行い、さらに粉砕後、粉末、ペーストの形で、あるいは溶媒を用いて抽出し、後発酵茶抽出物として利用できる。さらに、当該後発酵茶抽出物を、スプレードライヤー、ドラムドライヤー、フリーズドライヤー、エアードライヤー等を用いて乾燥し、粉末化を行うことで、後発酵茶抽出物末とすることができる。また、必要に応じて造粒機等を用いて顆粒品とすることができる。 The post-fermented tea of the present invention can be used as it is, but is sterilized by heating, dry heat, microwave, etc., and further pulverized, extracted in the form of powder, paste, or using a solvent, It can be used as a post-fermented tea extract. Furthermore, the post-fermented tea extract can be made into a post-fermented tea extract powder by drying and pulverizing the post-fermented tea extract using a spray dryer, drum dryer, freeze dryer, air dryer or the like. Moreover, it can be set as a granule using a granulator etc. as needed.
上記抽出の際に用いる溶媒としては、水又は炭素数１〜６のアルコール類若しくはグリコール類を単独で若しくはこれら溶媒の２つ以上の組合せが使用でき、例えば水、エタノール、含水エタノール、１，３―ブチレングリコール、含水１，３−ブチレングリコール等が挙げられる。好ましくは含水エタノール若しくは含水１，３−ブチレングリコール、より好ましくは水とエタノールの５〜９５：９５〜５の混合物（ｖ／ｖ）、水と１，３−ブチレングリコールの５〜９５：９５〜５の混合物（ｖ／ｖ）が使用できる。
抽出条件としては、約２０〜７０℃で０．１〜５時間抽出し、ろ過を行い、得られたろ液を８０〜９５℃で３０分間の条件で加熱処理を行い冷却することが好ましい。
上記抽出物は、適宜、不活性な不純物を除去するため、例えば液々分液、固液分液、濾過膜、活性炭、吸着樹脂、イオン交換樹脂等の公知の分離・精製方法によって、更に精製してもよい。具体的には、抽出物末を上記抽出溶媒を用いて再度抽出することが挙げられる。
As the solvent used in the above extraction, water, alcohols having 1 to 6 carbon atoms or glycols can be used alone, or a combination of two or more of these solvents can be used. For example, water, ethanol, hydrous ethanol, 1, 3 -Butylene glycol, hydrous 1,3-butylene glycol, etc. are mentioned. Preferably water-containing ethanol or water-containing 1,3-butylene glycol, more preferably a mixture of water and ethanol 5 to 95: 95-5 (v / v), water and 1,3-butylene glycol 5 to 95:95 A mixture of 5 (v / v) can be used.
As extraction conditions, it is preferable to extract at about 20 to 70 ° C. for 0.1 to 5 hours, perform filtration, and cool the obtained filtrate by heating at 80 to 95 ° C. for 30 minutes.
The above extract is further purified by a known separation / purification method such as liquid-liquid separation, solid-liquid separation, filtration membrane, activated carbon, adsorption resin, ion exchange resin, etc., in order to remove inert impurities as appropriate. May be. Specifically, the extract powder may be extracted again using the extraction solvent.
上記の製造方法により得られた後発酵茶又はその抽出物は、抗酸化活性の一つの指標であるＳＯＤ活性を、後発酵茶固形分１ｇ当り２，０００ｕｎｉｔｓ以上、好ましくは８，０００ｕｎｉｔｓ以上、より好ましくは９，０００ｕｎｉｔｓ以上、さらに好ましくは９，０００〜５００，０００ｕｎｉｔｓ有する。例えば、原料としての茶に緑茶を、Ａｓｐｅｒｇｉｌｌｕｓ ｏｒｙｚａｅを用いて本発明の製造方法にて製造した後発酵茶は、ＳＯＤ活性を、後発酵茶固形分１ｇ当り９，０００ｕｎｉｔｓ以上を有している。 The post-fermented tea or extract thereof obtained by the above production method has an SOD activity that is one index of the antioxidant activity of 2,000 units or more, preferably 8,000 units or more, per 1 g of post-fermented tea solid content. Preferably, it has 9,000 units or more, more preferably 9,000 to 500,000 units. For example, post-fermented tea produced by the production method of the present invention using Aspergillus oryzae as a raw material tea has SOD activity of 9,000 units or more per 1 g of post-fermented tea solid content.
本発明の後発酵茶又はその抽出物は、後記実施例に示すように優れたＳＯＤ活性を有することから、皮膚外用剤組成物、浴用剤組成物、食品組成物等における抗酸化剤として有用である。また、本発明の後発酵茶又はその抽出物は抗酸化活性及び抗紫外線ストレス活性を有するとともに、皮膚刺激が少なく、渋味も弱いことから、皮膚外用剤又は浴用剤に応用した場合、肌の老化防止効果、抗炎症効果、抗アレルギー効果、紫外線防御効果、紫外線暴露に起因する皮膚障害の防止効果等の機能が、食品に応用した場合、内臓脂肪低減効果、血糖値上昇抑制効果、糖尿病抑制効果、血圧上昇抑制効果等の機能、紫外線暴露に起因する皮膚障害の防止効果が、さらに、消臭剤としての機能が期待できる。
ここで、抗紫外線ストレス活性とは、紫外線照射により生じる細胞傷害に対する抑制活性をいう。また、紫外線暴露に起因する障害としては、紫外線暴露により生じる種々の皮膚疾患、例えば、光線過敏症等が挙げられる。
Since the post-fermented tea or the extract of the present invention has an excellent SOD activity as shown in Examples below, it is useful as an antioxidant in skin external preparation compositions, bath preparation compositions, food compositions and the like. is there. Further, since the post-fermented tea or the extract thereof of the present invention has antioxidant activity and anti-ultraviolet stress activity, has little skin irritation and weak astringency, when applied to a skin external preparation or bath preparation, Functions such as anti-aging effects, anti-inflammatory effects, anti-allergic effects, UV protection effects, and skin damage prevention effects caused by UV exposure, when applied to food, visceral fat reduction effect, blood sugar level increase suppression effect, diabetes suppression A function such as an effect, a blood pressure increase suppressing effect, a skin damage prevention effect due to ultraviolet exposure, and a function as a deodorant can be expected.
Here, anti-ultraviolet stress activity refers to inhibitory activity against cell damage caused by ultraviolet irradiation. Examples of the disorder caused by ultraviolet exposure include various skin diseases caused by ultraviolet exposure, such as photosensitivity.
本発明でいう皮膚外用剤とは、皮膚に適用しうる剤形のものであって、クリーム、化粧水、乳液、パック料、美容液、ファンデーション、パウダー、リップクリーム等の化粧料、抗炎症剤、抗真菌剤、鎮痛剤、抗掻痒剤、ステロイド剤等を含む医薬品・医薬部外品、シャンプー、リンス等の洗浄剤等を指す。 The topical skin preparation as used in the present invention is a dosage form that can be applied to the skin, and cosmetics such as creams, lotions, emulsions, packs, essences, foundations, powders, lip balms, and anti-inflammatory agents. , Refers to pharmaceuticals and quasi drugs including antifungal agents, analgesics, antipruritic agents, steroids, etc., and shampoos, rinses and other cleaning agents.
皮膚外用剤組成物の形態としては特に限定されず、例えば、薬用又は／及び化粧品用の化粧水、乳液、クリーム、軟膏、ローション、オイル、パック等の基礎化粧料；洗顔料や皮膚洗浄料、マッサージ用剤、クレンジング用剤、除毛剤、脱毛剤、髭剃り処理料、アフターシェーブローション、プレショーブローション、シェービングクリーム、シャンプー剤、リンス剤、トリートメント剤、パーマネント剤、染毛料、整髪料、ヘアートニック剤、育毛・養毛料等が挙げられる。その他、消臭・防臭剤や衛生用品、衛生綿類、ウエットテッシュ等に応用できる。
これらの皮膚外用剤の調製にあたっては、水、界面活性剤、キレート剤、油剤、乳化剤、保湿剤、増粘剤、多価アルコール、アルコール類、美白剤、防腐剤、紫外線吸収剤、紫外線散乱剤、香料等を適宜配合することができる。
The form of the external preparation for skin is not particularly limited, and includes basic cosmetics such as skin lotions, emulsions, creams, ointments, lotions, oils, packs and the like for medicinal and / or cosmetics; Massage agent, cleansing agent, hair remover, hair remover, shaving treatment, after shave lotion, pre-show lotion, shaving cream, shampoo, rinse agent, treatment agent, permanent agent, hair dye, hair conditioner, hair art And agents, hair growth and hair nourishing agents. In addition, it can be applied to deodorants and deodorants, sanitary products, sanitary cotton, wet tissue, etc.
In preparing these external preparations for skin, water, surfactants, chelating agents, oils, emulsifiers, moisturizers, thickeners, polyhydric alcohols, alcohols, whitening agents, preservatives, UV absorbers, UV scattering agents A fragrance or the like can be appropriately blended.
浴用剤組成物の形態としては、粉末状、顆粒状、液状、ゲル状、錠剤状等が挙げられる。また、食品組成物の形態としては、アンプル状、カプセル状、丸剤状、錠剤状、粉末状、顆粒状、固形状、液状、ゲル状、気泡状等の他、各種食品中に配合することもできる。これらの組成物の調製にあたっては、賦形剤、結合剤、滑沢剤等を適宜配合することができる。 Examples of the bath composition include powder, granule, liquid, gel, and tablet. In addition, the form of the food composition includes ampoules, capsules, pills, tablets, powders, granules, solids, liquids, gels, bubbles, etc., and other food compositions. You can also. In preparing these compositions, excipients, binders, lubricants, and the like can be appropriately blended.
また、皮膚外用剤、浴用剤及び食品への本発明後発酵茶又はその抽出物の配合量は、固形分（乾燥重量）換算で０．０００１〜２０重量％、０．０１〜１０重量％が好ましく、特に０．０１〜５重量％が好ましい。 Moreover, the blending amount of the post-invention fermented tea or its extract into the skin external preparation, bath preparation and food is 0.0001 to 20% by weight, 0.01 to 10% by weight in terms of solid content (dry weight). Particularly preferred is 0.01 to 5% by weight.
以下、実施例を示して本発明をさらに詳細かつ具体的に説明するが、本発明は以下の例によって限定されるものではない。 Hereinafter, the present invention will be described in more detail and specifically with reference to examples, but the present invention is not limited to the following examples.
［試験例１］
緑茶（荒茶）５００ｇを１００重量部としてそれぞれ０重量部、５０重量部、７０重量部、８０重量部、１００重量部、２００重量部、３００重量部、３５０重量部、４００重量部、５００重量部、７００重量部の水を加え、混合後、密閉系で、８０℃で６０分間加熱殺菌を行った。３０℃まで冷却後、該緑茶葉を、予め殺菌済みの培養・発酵槽に移し、厚み５ｃｍになるように堆積し、Ａｓｐｅｒｇｉｌｌｕｓ ｏｒｙｚａｅ ＩＦＯ５２３８を植菌した。植菌後、３０℃で７日間培養した後、乾燥させ、後発酵茶を得た。該後発酵茶に５，０００ｍＬの５０％（体積／体積）エタノールを添加し、撹拌抽出を行った。ろ過により固液分離を行い、得られたろ液を固形分３０％（重量／体積）まで濃縮し、８０℃で３０分間加熱処理を行い冷却して、後発酵茶抽出物を得た。該後発酵茶抽出物に関して、ＳＯＤ Ａｓｓａｙ Ｋｉｔ−ＷＳＴ（同仁化学社製）を用いてＳＯＤ活性を測定し、後発酵茶固形分当たりに換算した。結果を表１に示す。
[Test Example 1]
0, 50, 70, 80, 100, 200, 300, 350, 400, and 500 parts by weight of 500 g of green tea (raw tea). And 700 parts by weight of water were added and mixed, and then heat-sterilized at 80 ° C. for 60 minutes in a closed system. After cooling to 30 ° C., the green tea leaves were transferred to a previously sterilized culture / fermentor, deposited to a thickness of 5 cm, and inoculated with Aspergillus oryzae IFO5238. After inoculation, the cells were cultured at 30 ° C. for 7 days and then dried to obtain post-fermented tea. After that, 5,000 mL of 50% (volume / volume) ethanol was added to the fermented tea, followed by extraction with stirring. Solid-liquid separation was performed by filtration, and the obtained filtrate was concentrated to a solid content of 30% (weight / volume), heated at 80 ° C. for 30 minutes and cooled to obtain a post-fermented tea extract. With respect to the post-fermented tea extract, SOD activity was measured using SOD Assay Kit-WST (manufactured by Dojin Chemical Co., Ltd.) and converted to the amount of post-fermented tea solids. The results are shown in Table 1.
ＳＯＤ活性の測定は以下の通り行った。まず、試料となる後発酵茶抽出物を蒸留水で希釈し、０．０７５ｍｇ／ｍＬ、０．０３７５ｍｇ／ｍＬ、０．００７５ｍｇ／ｍＬ、及び０．００３７５ｍｇ／ｍＬになるようにサンプル溶液を調製した。このサンプル溶液を９６ｗｅｌｌ プレートの各ｗｅｌｌに２０μＬずつ分注した。次に、ＳＯＤ Ａｓｓａｙ Ｋｉｔ−ＷＳＴに同封されているＴｅｃｈｎｉｃａｌ Ｉｎｆｏｒｍａｔｉｏｎに従い調製したＷＳＴ ｗｏｒｋｉｎｇ ｓｏｌｕｔｉｏｎを２００μＬずつ加えよく混合した。さらに、ＳＯＤ Ａｓｓａｙ Ｋｉｔ−ＷＳＴに付属のＴｅｃｈｎｉｃａｌ Ｉｎｆｏｒｍａｔｉｏｎに従い調整したＥｎｚｙｍｅ ｗｏｒｋｉｎｇ ｓｏｌｕｔｉｏｎを２０μＬずつマルチチャンネルピペットにて加え、プレートリーダー中で３７℃、２０分間インキュベートし、４５０ｎｍの波長で吸光度を測定した。尚、各々にはＥｎｚｙｍｅ ｗｏｒｋｉｎｇ ｓｏｌｕｔｉｏｎの代わりに付属のＤｉｌｕｔｉｏｎ ｂｕｆｆｅｒを用いたブランクを設けた。さらに、サンプル溶液の代わりに蒸留水を用いたコントロール及びそのブランクを設けた。測定した吸光度から、「スーパーオキシド」と「ＷＳＴ−１」から「ＷＳＴ−１ ｆｏｒｍａｚａｎ」が生成される反応の阻害率を求め、さらにサンプルのＩＣ50値（単位：ｇ／ｍＬ）を算出した。
また、サンプル溶液の代わりにＳＯＤ標準品を用い、ＳＯＤ濃度と阻害率の関数にて検量線を作成した。作成した検量線から、ＳＯＤのＩＣ50値（単位：ｕｎｉｔｓ／ｍＬ）を算出した。
上記の「サンプルのＩＣ50値」及び「ＳＯＤのＩＣ50値」からサンプルのＳＯＤ活性を算出した。計算式は以下の通りである。
（サンプルのＳＯＤ活性 単位：ｕｎｉｔｓ／ｇ）＝（ＳＯＤのＩＣ50値）÷（サンプルのＩＣ50値）
The measurement of SOD activity was performed as follows. First, a post-fermented tea extract serving as a sample was diluted with distilled water, and sample solutions were prepared to be 0.075 mg / mL, 0.0375 mg / mL, 0.0075 mg / mL, and 0.00375 mg / mL. . 20 μL of this sample solution was dispensed into each well of a 96-well plate. Next, 200 μL of WST working solution prepared in accordance with Technical Information enclosed in SOD Assay Kit-WST was added and mixed well. Furthermore, 20 μL of Enzyme working solution adjusted according to Technical Information attached to SOD Assay Kit-WST was added by a multichannel pipette, incubated at 37 ° C. for 20 minutes in a plate reader, and the absorbance was measured at a wavelength of 450 nm. In each case, a blank using an attached dilution buffer was provided instead of the enzyme working solution. Further, a control using distilled water instead of the sample solution and its blank were provided. From the measured absorbance, the inhibition rate of the reaction in which “WST-1 formatzan” was generated from “superoxide” and “WST-1” was determined, and the IC 50 value (unit: g / mL) of the sample was calculated.
In addition, a standard curve was prepared using a function of SOD concentration and inhibition rate, using an SOD standard product instead of the sample solution. An IC 50 value (unit: units / mL) of SOD was calculated from the prepared calibration curve.
It was calculated SOD activity of the sample from the "IC 50 value of the sample" and "IC 50 value of SOD" above. The calculation formula is as follows.
(Sample SOD activity units: units / g) = (IC 50 values of SOD) ÷ (IC 50 value of the sample)
表１より、水添加量２００〜５００重量部の範囲で高いＳＯＤ活性を、３００〜４００重量部でより高いＳＯＤ活性を示すことがわかる。 From Table 1, it can be seen that a high SOD activity is exhibited in the range of 200 to 500 parts by weight of water addition, and a higher SOD activity is exhibited in the range of 300 to 400 parts by weight.
また、得られた後発酵茶抽出物を皮膚に塗布したところほとんど刺激性がなかった。また、味も渋味が低く良好であった。 Moreover, when the obtained fermented tea extract was applied to the skin, there was almost no irritation. Also, the taste was good with low astringency.
また、同様の方法で、従来から飲用されている後発酵茶のＳＯＤ活性を測定した結果を
表２に示す。
Moreover, the result of having measured the SOD activity of the post-fermented tea conventionally drunk by the same method is shown in Table 2.
表２より、本発明の後発酵茶が、従来の後発酵茶に比べて高いＳＯＤ活性を有することがわかる。 From Table 2, it can be seen that the post-fermented tea of the present invention has a higher SOD activity than the conventional post-fermented tea.
［実施例１］
緑茶（荒茶）５００ｇに、１，７５０ｍＬの水を加え混合後、密閉系で、８０℃で６０分間加熱殺菌を行った。３０℃まで冷却後、該緑茶を、予め殺菌済みの培養・発酵槽に移し、厚み４ｃｍになるように堆積し、Ａｓｐｅｒｇｉｌｌｕｓ ｏｒｙｚａｅ ＩＦＯ５２３８を植菌した。植菌後、３０℃で７日間培養・発酵させた。培養・発酵完了後、送風乾燥機にて乾燥させ、本発明の後発酵茶４８６ｇを得た。次いで、該後発酵茶に、５，０００ｍＬの水を添加し、５０℃で１時間、撹拌抽出を行った。ろ過により固液分離を行い、得られたろ液を固形分３０％まで濃縮し、８０℃で３０分間加熱処理を行い冷却して、後発酵茶水抽出物を得た。得られた後発酵茶水抽出物をフリーズドライヤーにて乾燥することで、後発酵茶水抽出物末を得た（１０８ｇ）。得られた後発酵茶水抽出物末に関して、ＳＯＤ Ａｓｓａｙ Ｋｉｔ−ＷＳＴを用いＳＯＤ活性を測定した結果、後発酵茶固形分１ｇ当たり、２０，０６２ｕｎｉｔｓ（後発酵茶水抽出物末１ｇ当たり８０，２３８ｕｎｉｔｓ）であった。得られた抽出物末は、渋味もなく、苦味もなく、皮膚刺激性もなく、良好であった。
[Example 1]
1,750 mL of water was added to 500 g of green tea (raw tea) and mixed, and then heat-sterilized at 80 ° C. for 60 minutes in a closed system. After cooling to 30 ° C., the green tea was transferred to a previously sterilized culture / fermentor, deposited to a thickness of 4 cm, and Aspergillus oryzae IFO5238 was inoculated. After inoculation, the cells were cultured and fermented at 30 ° C. for 7 days. After completion of the culture and fermentation, the mixture was dried with a blow dryer to obtain 486 g of post-fermented tea of the present invention. Next, 5,000 mL of water was added to the post-fermented tea, and stirring extraction was performed at 50 ° C. for 1 hour. Solid-liquid separation was performed by filtration, and the obtained filtrate was concentrated to a solid content of 30%, followed by heat treatment at 80 ° C. for 30 minutes and cooling to obtain a post-fermented tea water extract. The obtained post-fermented tea water extract was dried with a freeze dryer to obtain a post-fermented tea water extract powder (108 g). As a result of measuring the SOD activity using the SOD Assay Kit-WST for the obtained post-fermented tea water extract powder, it was 20,062 units (80,238 units per gram of post-fermented tea water extract powder) per 1 g of the post-fermented tea solid content. there were. The obtained extract powder was good with no astringency, no bitterness and no skin irritation.
［実施例２］
緑茶（荒茶）５００ｇに、２，５００ｍＬの水を加え混合後、密閉系で、８０℃で６０分間加熱殺菌を行った。３０℃まで冷却後、該緑茶を、予め殺菌済みの培養・発酵槽に移し、厚み５ｃｍになるように堆積し、Ａｓｐｅｒｇｉｌｌｕｓ ｏｒｙｚａｅ ＩＦＯ５２３８を植菌した。植菌後、３０℃で７日間培養・発酵した。培養・発酵完了後、送風乾燥機にて乾燥させ、本発明の後発酵茶４９２ｇを得た。次いで、５，０００ｍＬの５０％エタノール（以下５０％ＥｔＯＨという）を添加し、５０℃で１時間、撹拌抽出を行った。ろ過により固液分離を行い、得られたろ液を固形分３０％まで濃縮し、８０℃で３０分間加熱処理を行い、冷却して後発酵茶５０％ＥｔＯＨ抽出物を得た。得られた後発酵茶５０％ＥｔＯＨ抽出物をフリーズドライヤーにて乾燥することで、後発酵茶５０％ＥｔＯＨ抽出物末を得た（１１０ｇ）。得られた後発酵茶５０％ＥｔＯＨ抽出物末に関して、ＳＯＤ Ａｓｓａｙ Ｋｉｔ−ＷＳＴを用いＳＯＤ活性を測定した結果、後発酵茶固形分１ｇ当たり、１９，８８４ｕｎｉｔｓ（後発酵茶５０％ＥｔＯＨ抽出物末１ｇ当たり９０，３８４ｕｎｉｔｓ）であった。得られた抽出物末は、渋味もなく、苦味もなく、皮膚刺激性もなく、良好であった。
[Example 2]
To 500 g of green tea (raw tea), 2,500 mL of water was added and mixed, and then heat-sterilized at 80 ° C. for 60 minutes in a closed system. After cooling to 30 ° C., the green tea was transferred to a previously sterilized culture / fermentor, deposited to a thickness of 5 cm, and Aspergillus oryzae IFO5238 was inoculated. After inoculation, the cells were cultured and fermented at 30 ° C. for 7 days. After completion of the culture and fermentation, the mixture was dried with an air dryer to obtain 492 g of post-fermented tea of the present invention. Subsequently, 5,000 mL of 50% ethanol (hereinafter referred to as 50% EtOH) was added, followed by extraction with stirring at 50 ° C. for 1 hour. Solid-liquid separation was performed by filtration, and the obtained filtrate was concentrated to a solid content of 30%, heat-treated at 80 ° C. for 30 minutes, and cooled to obtain a post-fermented tea 50% EtOH extract. The obtained post-fermented tea 50% EtOH extract was dried with a freeze dryer to obtain a post-fermented tea 50% EtOH extract powder (110 g). With respect to the obtained post-fermented tea 50% EtOH extract powder, SOD activity was measured using SOD Assay Kit-WST. As a result, 19,884 units (1 g of post-fermented tea 50% EtOH extract powder per 1 g of post-fermented tea solid content was measured. 90,384 units). The obtained extract powder was good with no astringency, no bitterness and no skin irritation.
［実施例３］
収穫した直後の生茶葉（水分含量：７８重量％）５００ｇを水にて洗浄後、よく水をきり、密閉系で、８０℃で６０分間加熱殺菌を行った。３０℃まで冷却後、該茶葉を、予め殺菌済みの培養・発酵槽に移し、厚み４ｃｍになるように堆積し、Ａｓｐｅｒｇｉｌｌｕｓ ｏｒｙｚａｅ ＩＦＯ５２３８を植菌した。植菌後、３０℃で７日間培養・発酵させた。培養・発酵完了後、送風乾燥機にて乾燥させ、本発明の後発酵茶９３ｇを得た。次いで、該後発酵茶に、１，０００ｍＬの水を添加し、撹拌抽出を行った。ろ過により固液分離を行い、得られたろ液を固形分３０％まで濃縮し、８０℃で３０分間加熱処理を行い、冷却して後発酵茶水抽出物を得た。得られた後発酵茶水抽出物をフリーズドライヤーにて乾燥することで、後発酵茶水抽出物末を得た（１８ｇ）。得られた後発酵茶水抽出物末に関して、ＳＯＤ Ａｓｓａｙ Ｋｉｔ−ＷＳＴを用いＳＯＤ活性を測定した結果、後発酵茶固形分１ｇ当たり、９，１００ｕｎｉｔｓ（後発酵茶水抽出物末１ｇ当たり３７，０００ｕｎｉｔｓ）であった。得られた抽出物末は、渋味もなく、苦味もなく、皮膚刺激性もなく、良好であった。
[Example 3]
Freshly harvested fresh tea leaves (water content: 78% by weight) 500 g were washed with water, thoroughly drained, and sterilized by heating at 80 ° C. for 60 minutes in a closed system. After cooling to 30 ° C., the tea leaves were transferred to a previously sterilized culture / fermentor, deposited to a thickness of 4 cm, and Aspergillus oryzae IFO5238 was inoculated. After inoculation, the cells were cultured and fermented at 30 ° C. for 7 days. After completion of the culture and fermentation, the mixture was dried with a blow dryer to obtain 93 g of post-fermented tea of the present invention. Next, 1,000 mL of water was added to the post-fermented tea, followed by extraction with stirring. Solid-liquid separation was performed by filtration, and the obtained filtrate was concentrated to a solid content of 30%, heat-treated at 80 ° C. for 30 minutes, and cooled to obtain a post-fermented tea water extract. The obtained post-fermented tea water extract was dried with a freeze dryer to obtain a post-fermented tea water extract powder (18 g). With respect to the obtained post-fermented tea water extract powder, SOD activity was measured using SOD Assay Kit-WST. there were. The obtained extract powder was good with no astringency, no bitterness and no skin irritation.
［実施例４］ 実施例２で得られた後発酵茶５０％ＥｔＯＨ抽出物末１００ｇに５０％１，３−ブチレングリコール（以下５０％ＢＧという）を５，０００ｇ添加し、５０℃で３０分間抽出を行った。次いでろ過を行いろ液を得た。得られたろ液について精密ろ過を行い、５０％ＢＧにて後発酵茶抽出物固形分１％になるように希釈し、後発酵茶５０％ＢＧ抽出物を得た。得られた後発酵茶５０％ＢＧ抽出物に関して、ＳＯＤ Ａｓｓａｙ Ｋｉｔ−ＷＳＴを用いＳＯＤ活性を測定した結果、抽出物１ｇ当たり８９０ｕｎｉｔｓ（固形分１ｇ当たり８９，０００ｕｎｉｔｓ）であった。得られた抽出物は、皮膚刺激性がなく、良好であった。 [Example 4] 5,000 g of 50% 1,3-butylene glycol (hereinafter referred to as 50% BG) was added to 100 g of the post-fermented tea 50% EtOH extract powder obtained in Example 2, and the mixture was heated at 50 ° C for 30 minutes. Extraction was performed. Next, filtration was performed to obtain a filtrate. The obtained filtrate was subjected to microfiltration and diluted with 50% BG so as to have a post-fermented tea extract solid content of 1% to obtain a post-fermented tea 50% BG extract. As a result of measuring SOD activity using SOD Assay Kit-WST for the obtained post-fermented tea 50% BG extract, it was 890 units per gram of the extract (89,000 units per gram of solid content). The obtained extract was good without skin irritation.
［比較例１］
緑茶（荒茶）５００ｇに、６，７５０ｍＬの水を添加し撹拌抽出を行った。ろ過により固液分離を行い、得られたろ液を固形分３０％（重量／体積）まで濃縮し、８０℃で３０分間加熱処理を行い冷却し、緑茶抽出物を得た。得られた緑茶抽出物をフリーズドライヤーにて乾燥することで、緑茶抽出物末を得た。得られた緑茶抽出物末に関して、ＳＯＤ Ａｓｓａｙ Ｋｉｔ−ＷＳＴを用いＳＯＤ活性を測定した結果、緑茶固形分１ｇ当たり、４，７６０ｕｎｉｔｓ（緑茶抽出物末１ｇ当たり１９，８００ｕｎｉｔｓ）であった。ただし、この抽出物は渋味、苦味が強く、皮膚刺激性も強いものであった。
[Comparative Example 1]
6,750 mL of water was added to 500 g of green tea (raw tea), and stirring extraction was performed. Solid-liquid separation was carried out by filtration, and the obtained filtrate was concentrated to a solid content of 30% (weight / volume), followed by heat treatment at 80 ° C. for 30 minutes and cooling to obtain a green tea extract. The obtained green tea extract was dried with a freeze dryer to obtain a green tea extract powder. As a result of measuring SOD activity using the SOD Assay Kit-WST for the obtained green tea extract powder, it was 4,760 units (19,800 units per 1 g of green tea extract powder) per 1 g of green tea solid content. However, this extract had strong astringency and bitterness and strong skin irritation.
［試験例２］抗紫外線ストレス試験
紫外線は皮膚を構成する細胞の中の一種である線維芽細胞にストレスを与える要因の一つである。そのストレスにより線維芽細胞にＤＮＡ損傷を起こし、８−ＯＨｄＧの生成量を増加させる。紫外線Ａ波（３２０ｎｍ〜４００ｎｍ）は皮膚の表皮を通過し線維芽細胞の存在する真皮まで到達するので、紫外線Ａ波をヒト正常線維芽細胞へ照射することにより細胞にストレスを与えることができる。
それゆえ、ヒト正常線維芽細胞を用いて、実施例２で得られた後発酵茶５０％ＥｔＯＨ抽出物末の抗紫外線ストレス活性を、デオキシリボ核酸（以下ＤＮＡという）損傷マーカーである８−ヒドロキシデオキシグアノシン（以下８−ＯＨｄＧという）を測定することにより検討した。
[Test Example 2] Anti-ultraviolet stress test Ultraviolet rays are one of the factors that give stress to fibroblasts, which are a kind of cells constituting the skin. The stress causes DNA damage to fibroblasts and increases the amount of 8-OHdG produced. Since the ultraviolet A wave (320 nm to 400 nm) passes through the epidermis of the skin and reaches the dermis where fibroblasts are present, the cells can be stressed by irradiating human normal fibroblasts with the ultraviolet A wave.
Therefore, using human normal fibroblasts, the anti-ultraviolet light stress activity of the post-fermented tea 50% EtOH extract powder obtained in Example 2 was determined by deoxyribonucleic acid (hereinafter referred to as DNA) damage marker, 8-hydroxydeoxy. Examination was carried out by measuring guanosine (hereinafter referred to as 8-OHdG).
ヒト正常線維芽細胞の培養条件は、以下の通りとした。
ヒト正常線維芽細胞は、１０％ウシ胎児血清（シグマ社製）（以下ＦＢＳという）添加ダルベッコ改変イーグル培地（ナカライテスク社製）（以下ＤＭＥＭという）を培地として使用し、ＣＯ2インキュベーター（５％ＣＯ2、３７℃）内で、週２回の培地交換を行い、培養した。
The culture conditions for human normal fibroblasts were as follows.
For human normal fibroblasts, Dulbecco's modified Eagle medium (Nacalai Tesque) (hereinafter referred to as DMEM) supplemented with 10% fetal bovine serum (Sigma) (hereinafter referred to as FBS) was used as the medium, and a CO 2 incubator (5% (CO 2 , 37 ° C.) The medium was changed twice a week and cultured.
サンプル溶液の調製は、以下の通り行った。
実施例２で得られた後発酵茶５０％ＥｔＯＨ抽出物末１００ｍｇを秤量し、１０ｍＬの増殖培地に完全に溶解させた。その後、０．２μｍ滅菌フィルターを用いて滅菌ろ過し、１％溶液とした。この１％溶液を増殖培地で希釈し、０．０００１％、及び０．００００５％のサンプル溶液を調製した。
The sample solution was prepared as follows.
100 mg of the post-fermented tea 50% EtOH extract powder obtained in Example 2 was weighed and completely dissolved in 10 mL of growth medium. Thereafter, the solution was sterilized by filtration using a 0.2 μm sterilization filter to obtain a 1% solution. This 1% solution was diluted with growth medium to prepare 0.0001% and 0.00005% sample solutions.
抗紫外線ストレス試験は、以下の通り行った。
ヒト正常線維芽細胞を１０％ＦＢＳ添加ＤＭＥＭで３×１０5ｃｅｌｌｓ／ｍＬに調製し、１０ｍＬを１０ｃｍ ｄｉｓｈに播種した（３×１０6ｃｅｌｌｓ／ｄｉｓｈ）。翌日、０．０００１％及び０．００００５％のサンプルを添加した１０％ＦＢＳ添加ＤＭＥＭ１０ｍＬと交換した。１時間培養後、紫外線Ａ波を７０分間照射した（紫外線Ａ波強度：１０Ｊ／ｃｍ2）。直ちに、培養上清を除去し、トリプシン処理により細胞をｄｉｓｈより剥し、ピペットにて細胞懸濁液を１．５ｍＬエッペンドルフチューブに移した。
回収した細胞から、ＤＮＡ Ｅｘｔｒａｃｔｏｒ ＷＢ Ｋｉｔ（和光純薬工業社製）を用いてＤＮＡを次のように抽出した。
まず、１．５ｍＬエッペンドルフチューブを遠心分離後、上清捨て、リン酸緩衝生理食塩水（以下ＰＢＳという）を加え混合し、遠心（３，０００ｇ、５分）後、上清を除去した。得られた細胞ペレットに、Ｌｙｓｉｓ Ｓｏｌｕｔｉｏｎ（ＤＮＡ Ｅｘｔｒａｃｔｏｒ ＷＢ Ｋｉｔに付属）を１ｍＬ加え、Ｖｏｒｔｅｘにて撹拌後、遠心（１０，０００ｇ、２０秒）し、上清を除去した。上記操作を２回繰り返した後、Ｅｎｚｙｍｅ Ｒｅａｃｔｉｏｎ Ｓｏｌｕｔｉｏｎ（ＤＮＡ Ｅｘｔｒａｃｔｏｒ ＷＢ Ｋｉｔに付属）を２００μＬとＰｒｏｔｅａｓｅ（ＤＮＡ Ｅｘｔｒａｃｔｏｒ ＷＢ Ｋｉｔに付属）を１０μＬを加えて混合し、３７℃で１時間インキュベートした。その後、Ｓｏｄｉｕｍ Ｉｏｄｉｄｅ Ｓｏｌｕｔｉｏｎ（ＤＮＡ Ｅｘｔｒａｃｔｏｒ ＷＢ Ｋｉｔに付属）を３００μＬ加えて混合し、更にイソプロピルアルコール（和光純薬工業社製）０．５ｍＬを加えて、白い綿状のＤＮＡが完全に見えてくるまで静置した。遠心（１０，０００ｇ、１０分）後、上清を除去し、得られたＤＮＡに洗浄液（Ａ）（ＤＮＡ Ｅｘｔｒａｃｔｏｒ ＷＢ Ｋｉｔに付属）を１ｍＬ加え混合し、遠心（１０，０００ｇ、５分）した後、上清を除去し、更に洗浄液（Ｂ）（ＤＮＡ Ｅｘｔｒａｃｔｏｒ ＷＢ Ｋｉｔに付属）を１ｍＬ加え混合し、遠心（１０，０００ｇ、５分）した後、上清を除去し、沈殿物（ＤＮＡ）を軽く減圧乾燥した後、５０μＬの滅菌超純水に溶解した（ＤＮＡ水溶液）。
次に、５０μＬのＤＮＡ水溶液に、２００ｍＭ酢酸ナトリウム（ｐＨ４．８）を５．５μＬ、１ｍｇ／ｍＬに超純粋にて濃度を調製したｎｕｃｌｅａｓｅ Ｐ１（和光純薬工業社製）水溶液を５．５μＬ（２ｕｎｉｔｓ）加え、３７℃で１時間インキュベートした。更に、Ｔｒｉｓ−ＨＣｌ（１Ｍ、ｐＨ７．４）を５．５μＬ、Ａｌｋａｌｉｎｅ ｐｈｏｓｐｈａｔａｓｅ（タカラ社製）を１μＬ（２ｕｎｉｔｓ）添加し、３７℃にて更に１時間インキュベートしＤＮＡを加水分解した。ＤＮＡ加水分解液の２６０ｎｍ、２８０ｎｍ、３２０ｎｍにおける吸光度を分光光度計で測定し、ＤＮＡ水溶液の濃度、純度をチェックした。
最後に、ＤＮＡ加水分解液中に含まれる８−ＯＨｄＧを、高感度８−ＯＨｄＧ ｃｈｅｃｋ（日本老化制御研究所製）を用いたＥＬＩＳＡにより測定した。
５０μＬのＤＮＡ加水分解水溶液（約２０〜３０μｇ）を、８−ＯＨｄＧを固相化しているマイクロプレート用モジュール（高感度８−ＯＨｄＧ ｃｈｅｃｋに付属）のウェルに分注した。続いて各ウェルに、８−ＯＨｄＧと特異的に反応するモノクローナル抗体である第一抗体溶液（高感度８−ＯＨｄＧ ｃｈｅｃｋに付属）を５０μＬずつ添加し、プレートを左右に振動させ、よく混合した。４℃で一晩反応させた後、ウェルから反応液を捨て、洗浄液（高感度８−ＯＨｄＧ ｃｈｅｃｋに付属）で洗浄した。次に、各ウェルにモノクローナル抗体と結合する酵素標識抗体である第二抗体溶液（高感度８−ＯＨｄＧ ｃｈｅｃｋに付属）を１００μＬずつ分注し、よく混合した。常温で１時間反応後、ウェルから反応液を捨て、洗浄液（高感度８−ＯＨｄＧ ｃｈｅｃｋに付属）で洗浄した。各ウェルに反応停止液（高感度８−ＯＨｄＧ ｃｈｅｃｋに付属）を１００μＬ加え、反応を停止させた。マイクロプレートリーダーを使用して４５０ｎｍにおける各ウェルの吸光度を測定した。
The anti-ultraviolet stress test was performed as follows.
Normal human fibroblasts were prepared to 3 × 10 5 cells / mL with 10% FBS-added DMEM, and 10 mL was seeded on 10 cm dishes (3 × 10 6 cells / dish). The next day, the sample was replaced with 10 mL of 10% FBS-added DMEM supplemented with 0.0001% and 0.00005% samples. After culturing for 1 hour, ultraviolet A wave was irradiated for 70 minutes (ultraviolet A wave intensity: 10 J / cm 2 ). Immediately, the culture supernatant was removed, the cells were detached from the dish by trypsin treatment, and the cell suspension was transferred to a 1.5 mL Eppendorf tube with a pipette.
From the collected cells, DNA was extracted as follows using DNA Extractor WB Kit (manufactured by Wako Pure Chemical Industries, Ltd.).
First, after centrifuging a 1.5 mL Eppendorf tube, the supernatant was discarded, and phosphate buffered saline (hereinafter referred to as PBS) was added and mixed. After centrifugation (3,000 g, 5 minutes), the supernatant was removed. To the obtained cell pellet, 1 mL of Lysis Solution (attached to DNA Extractor WB Kit) was added, stirred with Vortex, centrifuged (10,000 g, 20 seconds), and the supernatant was removed. After repeating the above operation twice, 200 μL of Enzyme Reaction Solution (attached to DNA Extractor WB Kit) and 10 μL of Protease (attached to DNA Extractor WB Kit) were added and mixed, and incubated at 37 ° C. for 1 hour. Then, 300 μL of Sodium Iodide Solution (attached to DNA Extractor WB Kit) is added and mixed, and then 0.5 mL of isopropyl alcohol (manufactured by Wako Pure Chemical Industries, Ltd.) is added until white cotton-like DNA is completely visible. Left to stand. After centrifugation (10,000 g, 10 minutes), the supernatant was removed, and 1 mL of the washing solution (A) (attached to DNA Extractor WB Kit) was added to the obtained DNA and mixed, followed by centrifugation (10,000 g, 5 minutes). Thereafter, the supernatant was removed, and 1 mL of the washing solution (B) (attached to DNA Extractor WB Kit) was added and mixed. After centrifugation (10,000 g, 5 minutes), the supernatant was removed, and the precipitate (DNA) After lightly drying under reduced pressure, it was dissolved in 50 μL of sterile ultrapure water (DNA aqueous solution).
Next, 5.5 μL of nuclease P1 (manufactured by Wako Pure Chemical Industries, Ltd.) aqueous solution whose concentration was adjusted to 5.5 μL and 1 mg / mL with 200 mM sodium acetate (pH 4.8) in an ultrapure state was added to 50 μL of the DNA aqueous solution. 2 units) and incubated at 37 ° C. for 1 hour. Furthermore, 5.5 μL of Tris-HCl (1M, pH 7.4) and 1 μL (2 units) of Alkaline phosphatase (manufactured by Takara) were added, and the mixture was further incubated at 37 ° C. for 1 hour to hydrolyze the DNA. The absorbance at 260 nm, 280 nm, and 320 nm of the DNA hydrolyzate was measured with a spectrophotometer to check the concentration and purity of the aqueous DNA solution.
Finally, 8-OHdG contained in the DNA hydrolyzate was measured by ELISA using a high-sensitivity 8-OHdG check (manufactured by Japan Aging Control Laboratories).
50 μL of an aqueous DNA hydrolysis solution (about 20 to 30 μg) was dispensed into the wells of a microplate module (attached to the high-sensitivity 8-OHdG check) on which 8-OHdG was immobilized. Subsequently, 50 μL of the first antibody solution (attached to the high-sensitivity 8-OHdG check), which is a monoclonal antibody that specifically reacts with 8-OHdG, was added to each well, and the plate was shaken left and right and mixed well. After reacting at 4 ° C. overnight, the reaction solution was discarded from the well and washed with a washing solution (attached to high-sensitivity 8-OHdG check). Next, 100 μL of a second antibody solution (attached to the high-sensitivity 8-OHdG check), which is an enzyme-labeled antibody that binds to the monoclonal antibody, was dispensed into each well, and mixed well. After reacting at room temperature for 1 hour, the reaction solution was discarded from the well and washed with a washing solution (attached to high-sensitivity 8-OHdG check). 100 μL of a reaction stop solution (attached to the high-sensitivity 8-OHdG check) was added to each well to stop the reaction. The absorbance of each well at 450 nm was measured using a microplate reader.
抗紫外線ストレス試験の結果は図１に示す。
サンプル溶液で培養後、紫外線Ａ波照射処理した細胞のＤＮＡにおける８−ＯＨｄＧ生成量を、サンプル溶液無添加で紫外線Ａ波照射処理した陰性コントロールのそれと比較したところ、サンプル溶液は紫外線Ａ波照射に対する抗紫外線ストレス活性を有していることが示唆された。特に、０．０００１％においては、サンプル無添加で紫外線Ａ波無照射のブランクと近い値となり、最も抗紫外線ストレス活性を示した。
以上より、後発酵茶抽出物は、紫外線Ａ波に対する抗紫外線ストレス活性があると判断した。
The results of the anti-ultraviolet stress test are shown in FIG.
The amount of 8-OHdG produced in the DNA of cells treated with ultraviolet A wave irradiation after incubation with the sample solution was compared with that of the negative control treated with ultraviolet A wave irradiation without addition of the sample solution. It was suggested that it has anti-ultraviolet stress activity. In particular, at 0.0001%, the value was close to that of the blank with no sample added and without UV A wave irradiation, and showed the most anti-UV stress activity.
From the above, it was determined that the post-fermented tea extract has anti-ultraviolet stress activity against ultraviolet A waves.
以下に上述の実施例品を用いた配合例を示すが、各々の配合例は各製品の製造における常法により製造したもので、配合量のみを記した。また、単位は％（重量／重量）で記す。 Although the compounding example using the above-mentioned Example goods is shown below, each compounding example was manufactured by the usual method in manufacture of each product, and only the compounding quantity was described. The unit is expressed in% (weight / weight).
［配合例１］皮膚外用剤組成物（ローション）
後発酵茶５０％ＢＧ抽出物（実施例４） ５．０
エタノール １５．０
ヒドロキシエチルセルロース ０．１
防腐剤 ０．１
精製水 ７９．８
[Formulation Example 1] Skin preparation for external use (lotion)
Post-fermented tea 50% BG extract (Example 4) 5.0
Ethanol 15.0
Hydroxyethyl cellulose 0.1
Preservative 0.1
Purified water 79.8
［配合例２］皮膚外用剤組成物（乳液）
後発酵茶５０％ＢＧ抽出物（実施例４） ５．０
ステアリン酸 ０．２
セタノール １．５
ワセリン ３．０
流動パラフイン ７．０
ＰＯＥ（１０）モノオレイン酸エステル １．５
酢酸トコフェロール ０．２
グリセリン ５．０
トリエタノールアミン ０．１
防腐剤 ０．１
精製水 ７６．４
[Formulation Example 2] skin external preparation composition (milky lotion)
Post-fermented tea 50% BG extract (Example 4) 5.0
Stearic acid 0.2
Cetanol 1.5
Vaseline 3.0
Flowing paraffin 7.0
POE (10) monooleate 1.5
Tocopherol acetate 0.2
Glycerin 5.0
Triethanolamine 0.1
Preservative 0.1
Purified water 76.4
［配合例３］皮膚外用剤組成物（透明化粧水）
後発酵茶５０％ＢＧ抽出物（実施例４） ５．０
１，３−ブチレングリコール ６．０
グリセリン ４．０
オレイルアルコール ０．１
ＰＯＥ（２０）ソルビタンモノラウリン酸エステル ０．５
ＰＯＥ（１５）ラウリルアルコールエーテル ０．５
エタノール １０．０
香料 適量
色剤 適量
防腐剤 適量
褐色防止剤 適量
緩衝剤 適量
精製水 １００とする残余
[Formulation Example 3] External preparation for skin (transparent lotion)
Post-fermented tea 50% BG extract (Example 4) 5.0
1,3-butylene glycol 6.0
Glycerin 4.0
Oleyl alcohol 0.1
POE (20) sorbitan monolaurate 0.5
POE (15) lauryl alcohol ether 0.5
Ethanol 10.0
Fragrance Appropriate amount Colorant Appropriate amount Preservative Appropriate amount Browning agent Appropriate amount Buffering agent Appropriate amount Purified
［配合例４］皮膚外用剤組成物（柔軟化粧水）
後発酵茶５０％ＢＧ抽出物（実施例４） ０．５
ソルビット ４．０
ジプロピレングリコール ６．０
ＰＥＧ １５００ ５．０
ＰＯＥ（２０）オレイルアルコールエーテル ０．５
メチルセルロース ０．２
クインスシード ０．１
エタノール １０．０
香料 適量
色剤 適量
防腐剤 適量
キレート剤 適量
褐色防止剤 適量
緩衝剤 適量
精製水 １００とする残余
[Formulation Example 4] External preparation for skin (soft skin lotion)
Post-fermented tea 50% BG extract (Example 4) 0.5
Sorbit 4.0
Dipropylene glycol 6.0
PEG 1500 5.0
POE (20) oleyl alcohol ether 0.5
Methylcellulose 0.2
Quince Seed 0.1
Ethanol 10.0
Fragrance Appropriate amount Colorant Appropriate amount Preservative Appropriate amount Chelating agent Appropriate amount Browning agent Appropriate amount Buffering agent Appropriate amount Purified
［配合例５］皮膚外用剤組成物（半透明マイクロエマルション化粧水）
後発酵茶５０％ＢＧ抽出物（実施例４） ５．０
１，３−ブチレングリコール ６．０
グリセリン ５．０
ＰＥＧ ４０００ ３．０
オリーブ油 ０．５
ＰＯＥ（２０）ソルビタンモノステアリン酸エステル １．５
ＰＯＥ（５）オレイルアルコールエーテル ０．３
エタノール １０．０
香料 適量
色剤 適量
防腐剤 適量
緩衝剤 適量
褐色防止剤 適量
精製水 １００とする残余
[Formulation Example 5] skin external preparation composition (translucent microemulsion lotion)
Post-fermented tea 50% BG extract (Example 4) 5.0
1,3-butylene glycol 6.0
Glycerin 5.0
PEG 4000 3.0
Olive oil 0.5
POE (20) sorbitan monostearate ester 1.5
POE (5) oleyl alcohol ether 0.3
Ethanol 10.0
Fragrance Appropriate amount Colorant Appropriate amount Preservative Appropriate amount Buffering agent Appropriate amount Browning agent Appropriate amount Purified
［配合例６］皮膚外用剤組成物（収斂化粧水）
後発酵茶５０％ＢＧ抽出物（実施例４） ５．０
ジプロピレングリコール １．０
ソルビット １．０
ＰＯＥ（２０）オレイルアルコールエーテル １．０
スルホ石炭酸亜鉛 ０．２
クエン酸 ０．１
エタノール １５．０
香料 適量
防腐剤 適量
緩衝剤 適量
色剤 適量
褐色防止剤 適量
精製水 １００とする残余
[Composition Example 6] Skin external preparation composition (astringent lotion)
Post-fermented tea 50% BG extract (Example 4) 5.0
Dipropylene glycol 1.0
Sorbit 1.0
POE (20) oleyl alcohol ether 1.0
Zinc sulfocoborate 0.2
Citric acid 0.1
Ethanol 15.0
Perfume Appropriate amount Preservative Appropriate amount Buffering agent Appropriate amount Colorant Appropriate amount Browning agent Appropriate amount Purified
［配合例７］皮膚外用剤組成物（エモリエントローション）
後発酵茶５０％ＢＧ抽出物（実施例４） ５．０
ステアリン酸 ２．０
セチルアルコール １．５
ワセリン ４．０
スクワラン ５．０
グリセロールトリ−２−エチルヘキサン酸エステル ２．０
ソルビタンモノオレイン酸エステル ２．０
ジプロピレングリコール ５．０
ＰＥＧ １５００ ３．０
トリエタノールアミン １．０
防腐剤 適量
香料 適量
精製水 １００とする残余
[Composition Example 7] External preparation for skin (emollient lotion)
Post-fermented tea 50% BG extract (Example 4) 5.0
Stearic acid 2.0
Cetyl alcohol 1.5
Vaseline 4.0
Squalane 5.0
Glycerol tri-2-ethylhexanoate 2.0
Sorbitan monooleate 2.0
Dipropylene glycol 5.0
PEG 1500 3.0
Triethanolamine 1.0
Preservative Appropriate amount Perfume Appropriate amount Purified
［配合例８］皮膚外用剤組成物（シャンプー）
後発酵茶５０％ＢＧ抽出物（実施例４） ５．０
ポリオキシエチレンラウリルエーテル硫酸ナトリウム ５．０
ポリオキシエチレンラウリルエーテル硫酸トリエタノールアミン
３．０
ヤシ油脂肪酸サルコシンナトリウム ２．０
ヤシ油脂肪酸アミドプロピルベタイン ２．０
２−アルキルーＮ―カルボキシメチルーＮ―ヒドロキシエチルイ
ミダゾリニウムベタイン ２．０
ポリオキシエチレンラウリルエーテル １．０
ジステアリン酸エチレングリコール ３．０
ヤシ油脂肪酸ジエタノールアミド １．０
ポリオキシエチレン・メチルポリシロキサン共重合体 １．０
メチルポリシロキサン ０．５
ジイソステアリン酸ポリグリセリル ２．０
混合植物抽出液（１０） ０．５
加水分解ケラチン末 ０．３
アスコルビン酸ナトリウム ０．１
ピロクトンオラミン ０．５
メチルパラベン ０．３
塩化ジメチルジアリルアンモニウム・アクリルアミド共重合体
０．１
塩化Ｏ−（２−ヒドロキシ−３−（トリメチルアンモニオ）プロピル）
ヒドロキシエチルセルロース ０．１
センサマーＣＩ−５０ ３．０
精製水 １００とする残余
[Formulation Example 8] Skin preparation for external use (shampoo)
Post-fermented tea 50% BG extract (Example 4) 5.0
Sodium polyoxyethylene lauryl ether sulfate 5.0
Polyoxyethylene lauryl ether sulfate triethanolamine
3.0
Coconut oil fatty acid sarcosine sodium 2.0
Palm oil fatty acid amidopropyl betaine 2.0
2-alkyl-N-carboxymethyl-N-hydroxyethylimidazolinium betaine 2.0
Polyoxyethylene lauryl ether 1.0
Ethylene glycol distearate 3.0
Palm oil fatty acid diethanolamide 1.0
Polyoxyethylene / methylpolysiloxane copolymer 1.0
Methyl polysiloxane 0.5
Polyglyceryl diisostearate 2.0
Mixed plant extract (10) 0.5
Hydrolyzed keratin powder 0.3
Sodium ascorbate 0.1
Piroctone Olamine 0.5
Methylparaben 0.3
Dimethyldiallylammonium chloride / acrylamide copolymer
0.1
O- (2-hydroxy-3- (trimethylammonio) propyl) chloride
Hydroxyethyl cellulose 0.1
Sensormar CI-50 3.0
Residue with 100 purified water
［配合例９］浴用剤組成物（粉末タイプ）
後発酵茶水抽出物末（実施例１） １．０
硫酸ナトリウム ４９．５
炭酸水素ナトリウム ４９．５
[Formulation 9] Bath agent composition (powder type)
Post-fermented tea water extract powder (Example 1) 1.0
Sodium sulfate 49.5
Sodium bicarbonate 49.5
［配合例１０］浴用剤組成物（錠剤タイプ）
後発酵茶水抽出物末（実施例１） １．０
硫酸ナトリウム ４４．６
炭酸水素ナトリウム １４．７
コハク酸 ２１．７
滑沢剤 適量
色剤 適量
香料 適量
[Formulation Example 10] Bath composition (tablet type)
Post-fermented tea water extract powder (Example 1) 1.0
Sodium sulfate 44.6
Sodium bicarbonate 14.7
Succinic acid 21.7
Lubricant appropriate amount Colorant appropriate amount Fragrance appropriate amount
［配合例１１］食品組成物（あめ）
後発酵茶水抽出物末（実施例１） ０．２
砂糖 ５０．０
水飴 ３３．０
クエン酸 ２．０
香料 ０．２
水 １４．６
[Composition Example 11] Food composition (candy)
Post-fermented tea water extract powder (Example 1) 0.2
Sugar 50.0
Minamata 33.0
Citric acid 2.0
Fragrance 0.2
Water 14.6
［配合例１２］食品組成物（茶飲料（１））
後発酵茶水抽出物末（実施例１） ０．３
緑茶抽出物 ０．７
ビタミンＣ ０．１
水 ９８．９
[Composition Example 12] Food composition (tea beverage (1))
Post-fermented tea water extract powder (Example 1) 0.3
Green tea extract 0.7
Vitamin C 0.1
Water 98.9
［配合例１３］食品組成物（茶飲料（２））
後発酵茶水抽出物末（実施例１） １．０
ビタミンＣ ０．１
水 ９８．９
[Formulation Example 13] Food composition (tea beverage (2))
Post-fermented tea water extract powder (Example 1) 1.0
Vitamin C 0.1
Water 98.9
Claims (10)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2007157494A JP2008212136A (en) | 2006-06-20 | 2007-06-14 | Post-fermented tea and method for producing the same |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2006170622 | 2006-06-20 | ||
| JP2007029601 | 2007-02-08 | ||
| JP2007157494A JP2008212136A (en) | 2006-06-20 | 2007-06-14 | Post-fermented tea and method for producing the same |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JP2008212136A true JP2008212136A (en) | 2008-09-18 |
Family
ID=39832959
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2007157494A Pending JP2008212136A (en) | 2006-06-20 | 2007-06-14 | Post-fermented tea and method for producing the same |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2008212136A (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120100087A1 (en) * | 2010-10-25 | 2012-04-26 | Hui Kyoung Chang | Whitening cosmetic composition containing green tea extract |
| JP2014043406A (en) * | 2012-08-24 | 2014-03-13 | Gunma Univ | Agent having blood pressure elevation-suppressing action from post-fermented tea, and drink or food product and medicament comprising the same |
| JP2014050356A (en) * | 2012-09-07 | 2014-03-20 | Nippon Menaade Keshohin Kk | Differentiation inducer from stem cell to ectodermal cell |
| JP2017178898A (en) * | 2016-03-31 | 2017-10-05 | 株式会社ナリス化粧品 | Inhibitor for inhibiting skin cell growth inhibition |
| WO2019066261A1 (en) * | 2017-09-29 | 2019-04-04 | (주)아모레퍼시픽 | Antioxidizing, antiaging, or anti-inflammatory composition for strengthening skin barrier and caring for skin cell damage caused by fine dust including fermented tea extract |
| WO2019131274A1 (en) * | 2017-12-26 | 2019-07-04 | 国立大学法人 筑波大学 | Method for producing fermentation product derived from green tea extract, and koji fermentation product derived from green tea extract |
| CN112057377A (en) * | 2020-07-21 | 2020-12-11 | 黄山学院 | Fermented mulberry tea nerve-soothing mask and preparation method thereof |
| CN112868843A (en) * | 2021-01-29 | 2021-06-01 | 陕西科技大学 | Probiotic Fuzhuan tea beverage and preparation method thereof |
-
2007
- 2007-06-14 JP JP2007157494A patent/JP2008212136A/en active Pending
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120100087A1 (en) * | 2010-10-25 | 2012-04-26 | Hui Kyoung Chang | Whitening cosmetic composition containing green tea extract |
| JP2012092100A (en) * | 2010-10-25 | 2012-05-17 | Amorepacific Corp | Skin-whitening cosmetic composition containing green tea extract |
| US8545905B2 (en) * | 2010-10-25 | 2013-10-01 | Amorepacific Corporation | Whitening cosmetic composition containing green tea extract |
| JP2014043406A (en) * | 2012-08-24 | 2014-03-13 | Gunma Univ | Agent having blood pressure elevation-suppressing action from post-fermented tea, and drink or food product and medicament comprising the same |
| JP2014050356A (en) * | 2012-09-07 | 2014-03-20 | Nippon Menaade Keshohin Kk | Differentiation inducer from stem cell to ectodermal cell |
| JP2017178898A (en) * | 2016-03-31 | 2017-10-05 | 株式会社ナリス化粧品 | Inhibitor for inhibiting skin cell growth inhibition |
| WO2019066261A1 (en) * | 2017-09-29 | 2019-04-04 | (주)아모레퍼시픽 | Antioxidizing, antiaging, or anti-inflammatory composition for strengthening skin barrier and caring for skin cell damage caused by fine dust including fermented tea extract |
| CN111432797A (en) * | 2017-09-29 | 2020-07-17 | 株式会社爱茉莉太平洋 | Composition containing fermented tea extract for protecting skin cell damage caused by dust, enhancing skin barrier, and resisting oxidation, aging or inflammation |
| CN111432797B (en) * | 2017-09-29 | 2023-11-28 | 株式会社爱茉莉太平洋 | Composition for caring skin cell injury, enhancing skin barrier, and resisting oxidation, aging or inflammation containing fermented tea extract |
| AU2018341302B2 (en) * | 2017-09-29 | 2023-12-21 | Amorepacific Corporation | Antioxidizing, antiaging, or anti-inflammatory composition for strengthening skin barrier and caring for skin cell damage caused by fine dust including fermented tea extract |
| WO2019131274A1 (en) * | 2017-12-26 | 2019-07-04 | 国立大学法人 筑波大学 | Method for producing fermentation product derived from green tea extract, and koji fermentation product derived from green tea extract |
| CN112057377A (en) * | 2020-07-21 | 2020-12-11 | 黄山学院 | Fermented mulberry tea nerve-soothing mask and preparation method thereof |
| CN112868843A (en) * | 2021-01-29 | 2021-06-01 | 陕西科技大学 | Probiotic Fuzhuan tea beverage and preparation method thereof |
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