JP5425413B2 - Elastase inhibitor - Google Patents
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- JP5425413B2 JP5425413B2 JP2008136810A JP2008136810A JP5425413B2 JP 5425413 B2 JP5425413 B2 JP 5425413B2 JP 2008136810 A JP2008136810 A JP 2008136810A JP 2008136810 A JP2008136810 A JP 2008136810A JP 5425413 B2 JP5425413 B2 JP 5425413B2
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- 229940122858 Elastase inhibitor Drugs 0.000 title claims description 10
- 239000003602 elastase inhibitor Substances 0.000 title claims description 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 53
- 239000000284 extract Substances 0.000 claims description 27
- 244000131360 Morinda citrifolia Species 0.000 claims description 22
- 235000017524 noni Nutrition 0.000 claims description 22
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- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 1
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Description
本発明は、モリンダ・シトリフォリアの種子を含むエラスターゼ阻害剤に関するものであり、さらに種子の大部分を占めるオイル成分を除いた成分を得る抽出条件により見出せたエラスターゼ阻害剤・抗酸化作用を併せ持つエラスターゼ阻害剤、及びモリンダ・シトリフォリアの種子を粉砕することなく加熱抽出して得られるエキスを含むエラスターゼ阻害剤・抗酸化作用を併せ持つエラスターゼ阻害剤に関するものであり、新規にして、かつ、安全な化粧品、医薬品、医薬部外品、飲食品素材に関するものである。 The present invention relates to an elastase inhibitor containing Morinda citrifolia seeds, and further includes an elastase inhibitor / antioxidant inhibitor found by extraction conditions for obtaining components excluding oil components that occupy most of the seeds The present invention relates to an elastase inhibitor and an elastase inhibitor including an extract obtained by heating and extracting Morinda citrifolia seeds without crushing, and a novel and safe cosmetic and pharmaceutical product It relates to quasi-drugs and food and drink materials.
ノニ〔（学名：モリンダ・シトリフォリア（Ｍｏｒｉｎｄａ ｃｉｔｒｉｆｏｌｉａ）、和名：ヤエヤマアオキ）〕は、インドネシアのモルッカ諸島原産、アカネ科の熱帯性植物である。主に南太平洋のポリネシアの島々を中心に群生し、東南アジア、オーストラリア、インド、アフリカ、沖縄、小笠原諸島などの熱帯・亜熱帯地域に広く分布している。モリンダ・シトリフォリアは古くから熱帯・亜熱帯地域において食用、あるいは伝承医薬品として用いられ、根や茎、樹皮、葉、花、果実などの各部位が、高血圧、糖尿病、心臓病、疼痛、関節炎、喘息、マラリア熱、捻挫、火傷、虫歯、下痢、伝染病などの各種疾病の治療に利用されてきた。近年、それらの伝統的薬効を科学的に証明する研究が世界各国で進められている。 Noni [(Scientific name: Morinda citrifolia, Japanese name: Yaeyama Aoki)] is a tropical plant of the Rubiaceae native to the Molucca Islands in Indonesia. It is mainly clustered around Polynesian islands in the South Pacific, and is widely distributed in tropical and subtropical regions such as Southeast Asia, Australia, India, Africa, Okinawa and the Ogasawara Islands. Morinda citrifolia has long been used as an edible or traditional medicinal product in tropical and subtropical regions. It has been used to treat various diseases such as malaria fever, sprains, burns, tooth decay, diarrhea, and infectious diseases. In recent years, research that scientifically proves these traditional medicinal effects has been promoted in various countries around the world.
長時間の紫外線暴露は、しわの形成などの光老化をもたらすことがよく知られている。エラスチンは皮膚の弾力を保つ主要な真皮マトリックス成分のひとつであり、特に繰り返しの伸縮に関与する繊維性のタンパク質であるが、紫外線の影響でエラスターゼ酵素の活性が亢進すると、エラスチンの過剰な分解がおこり、しわが形成される。従って、安全で、かつ有効なエラスターゼ阻害剤は、美容上の問題を解決する手段として非常に有用であるといえる。 It is well known that prolonged UV exposure results in photoaging such as wrinkle formation. Elastin is one of the major dermal matrix components that maintain the elasticity of the skin, and is a fibrous protein that is particularly involved in repetitive stretching, but when elastase enzyme activity increases due to the influence of ultraviolet light, excessive degradation of elastin occurs. A wrinkle is formed. Therefore, it can be said that a safe and effective elastase inhibitor is very useful as a means for solving cosmetic problems.
また、過度の紫外線は活性酸素を過剰に増加させ、しみやそばかすなどの色素沈着の原因となるメラニンの産生を促進する。また、過度の活性酸素は炎症反応を増悪させ、その治癒を遅らせるほか、肌荒れや過敏肌などさまざまな肌トラブルの原因となる。従って、安全で、かつ有効な抗酸化剤は美容上の問題を解決する手段として非常に有用であるといえる。 In addition, excessive ultraviolet rays excessively increase active oxygen, and promote the production of melanin that causes pigmentation such as stains and freckles. Excessive active oxygen exacerbates the inflammatory response, delays healing, and causes various skin problems such as rough skin and sensitive skin. Therefore, it can be said that a safe and effective antioxidant is very useful as a means for solving cosmetic problems.
モリンダ・シトリフォリアに関しては、特開２０００−９５６６３号の請求項及び［００１２］にモリンダ・シトリフォリアのみならずアルトカルパス ラクーチャ等の植物の抽出物が美白剤、活性酵素消却剤、抗菌剤として有効であることが記載され、又、有効成分として用いられる植物抽出物は葉／枝・幹／樹皮／花／果実／根またはそれらの乾燥物から、適当な抽出溶媒を用いて、抽出することにより、調製される、ことが記載され、種子のことは記載されていないし、エラスターゼ阻害効果についての記載もない。 With respect to Morinda citrifolia, the claims of JP 2000-95663 and [0012] are effective not only for Morinda citrifolia but also for plant extracts such as altocarpa lacquer as whitening agents, active enzyme repellents and antibacterial agents. In addition, a plant extract used as an active ingredient is prepared by extracting from leaves / branches / stems / bark / flowers / fruits / roots or dried products thereof using an appropriate extraction solvent. The seeds are not described, and the elastase inhibitory effect is not described.
又、特開２００７−５５９６３号の［００１７］には、ヤエヤマアオキ（モリンダ・シトリフォリア）の果実は、果肉及び種子のうちいずれか一方又はその両方を含むものであるが、より抗酸化活性を高めることができるため、種子を除いた果肉部分のみを用いることが好ましい、ことが記載されている。果実には当然種子を含むので、種子を含んでも良いが、種子を除いた果肉の部分のみを用いることが好ましい、と解釈することが出来、種子についての効果を認めておらず、エラスターゼ阻害効果についての記載もない。 In addition, according to [0017] of JP-A-2007-55963, the fruit of Yaeyama Aoki (Morinda citrifolia) contains either one or both of pulp and seeds, but can further enhance the antioxidant activity. Therefore, it is described that it is preferable to use only the pulp part excluding seeds. Since the fruit naturally contains seeds, seeds may be included, but it can be interpreted that it is preferable to use only the part of the flesh excluding the seeds. There is no description about.
又、特開２００５−１４５９４５号の請求項１には、ヤエヤマアオキ（モリンダ・シトリフォリア）熟成果実発酵液からなる皮膚機能賦活化粧料組成物のことが記載されているが種子のことについては全く記載されていない。 Further, claim 1 of Japanese Patent Application Laid-Open No. 2005-145945 describes a skin function activating cosmetic composition comprising a fermented fruit juice of yamayamaoki (Morinda citrifolia), but it does not completely describe seeds. Not.
又、特表２００３−５０８５５４号の［０００６］には、ヤエヤマアオキ（モリンダ・シトリフォリア）種子から抽出されたエッセンシャルオイル、及びオイルを抽出する方法に関することが記載され、その抽出方法は［０００８］に記載のように種子を砕き、破砕し、薄片にし、切断し、することが記載されているが、種子がエラスターゼ阻害効果を有することは記載されていない。 [0006] of Japanese translations of PCT publication No. 2003-508554 describes an essential oil extracted from Yaeyama Aoki (Morinda citrifolia) seeds and a method for extracting the oil. The extraction method is described in [0008]. Thus, it is described that the seed is crushed, crushed, sliced, and cut, but it is not described that the seed has an elastase inhibitory effect.
又、特開２００４−３５２７０１号は［０００２］、［０００４］等に、モリンダ・シトリフォリアの果実の種子及び種皮が抗炎症作用、殺菌効果、保湿効果を有する入浴剤として利用出来ることが記載されているが、種子がエラスターゼ阻害効果を有することは記載されていない。 JP-A 2004-352701 describes in [0002], [0004], etc. that Morinda citrifolia fruit seeds and seed coats can be used as bathing agents having anti-inflammatory, bactericidal and moisturizing effects. However, it is not described that the seed has an elastase inhibitory effect.
このように、従来においてはモリンダ・シトリフォリアの種子がエラスターゼ阻害効果を有すること、又、その種子からオイル成分を除去した後の残渣を加熱抽出して得られたエキスがエラスターゼ阻害効果、抗酸化作用を併せ持つエラスターゼ阻害効果を有すること、又、その種子を粉砕することなく加熱抽出して、得られたエキスがエラスターゼ阻害効果、抗酸化作用を併せ持つエラスターゼ阻害効果を有することは全く知られておらず、本発明者らの鋭意研究の結果初めて発見されたものである。 Thus, in the past, Morinda citrifolia seeds have an elastase inhibitory effect, and the extract obtained by heating and extracting the residue after removing the oil component from the seeds has an elastase inhibitory effect, an antioxidant effect It is not known at all that it has an elastase inhibitory effect that also has an elastase inhibitory effect that has both an elastase inhibitory effect and an antioxidant effect by extracting the seeds by heating without crushing the seeds. It was discovered for the first time as a result of intensive studies by the present inventors.
本発明の目的は、モリンダ・シトリフォリア種子のエラスターゼ阻害活性を発現させる製法で、かつ、抗酸化作用を最大限に引き出すことのできる製法によって調製したエキスを有効成分として含有することで、しわの形成をはじめとする光老化の抑制、皮膚の美白、炎症の治療促進、肌荒れの予防・改善等に有用でかつ安全な、皮膚外用剤および内服剤を提供することである。 The object of the present invention is to produce wrinkles by containing, as an active ingredient, an extract prepared by a production method that exhibits the elastase inhibitory activity of Morinda citrifolia seeds and that can maximize the antioxidant effect. It is intended to provide a skin external preparation and an internal preparation that are useful and safe for suppression of photoaging, including whitening of the skin, promotion of treatment of inflammation, prevention and improvement of rough skin, and the like.
本発明者らは、鋭意研究を重ねた結果、モリンダ・シトリフォリアの種子がエラスターゼ阻害活性のあることを見出した。 As a result of intensive studies, the present inventors have found that Morinda citrifolia seeds have elastase inhibitory activity.
また、モリンダ・シトリフォリアの種子からオイル成分を除去した後の残渣を加熱抽出して得られたエキスに、オイル成分には期待できないエラスターゼ阻害活性、及び抗酸化作用を併せ持つエラスターゼ阻害活性を見出した。 Moreover, the elastase inhibitory activity which has the elastase inhibitory activity which cannot be expected to an oil component, and the elastase inhibitory activity to the extract obtained by heat-extracting the residue after removing an oil component from the seed of Morinda citrifolia was discovered.
さらに、その方法で調製したエキスには、オイル成分には期待できないエラスターゼ阻害活性、及び抗酸化作用を併せ持つエラスターゼ阻害活性が発現することを見出した。 Furthermore, it has been found that the extract prepared by the method expresses an elastase inhibitory activity that cannot be expected from an oil component and an elastase inhibitory activity that has both an antioxidant effect.
また、モリンダ・シトリフォリアの種子をあえて粉砕することなく含水アルコールで加熱抽出して得られたエキスに、粉砕後の種子を含水アルコールで静置抽出するという一般的に知られている抽出方法で調製したエキスには発現しないエラスターゼ阻害活性を見出した。なお、この際のアルコールとしては一価および多価アルコールが用いられ、前者としてはメタノール、エタノール、プロパノール、ブタノールが例示され、後者としてはエチレングリコール、プロピレングリコールが例示される。 In addition, Morinda citrifolia seeds are prepared by a generally known extraction method in which the seeds after pulverization are statically extracted with hydrous alcohol to the extract obtained by heat extraction with hydrous alcohol without pulverization. The elastase inhibitory activity which is not expressed in the extracted extract was found. In this case, monohydric and polyhydric alcohols are used as the alcohol. Examples of the former include methanol, ethanol, propanol, and butanol, and examples of the latter include ethylene glycol and propylene glycol.
さらに、その方法で調製したエキスは、一般的に知られている抽出方法で得られたエキスよりも極めて強い抗酸化活性が発現することを見出し、かつ抗酸化作用を併せ持つエラスターゼ阻害活性を見出した。 Furthermore, the extract prepared by that method was found to exhibit an extremely strong antioxidant activity than the extract obtained by a generally known extraction method, and also found an elastase inhibitory activity that also has an antioxidant effect. .
モリンダ・シトリフォリアの種子は前記のように粉砕しない場合にエラスターゼ阻害活性を見出すことが出来るのであるが、オイル成分を除去する場合はその種子を粉砕することが望ましい。粉砕することで効率よくオイル成分を除去できるからである。 Morinda citrifolia seeds can find elastase inhibitory activity when not pulverized as described above, but it is desirable to pulverize the seeds when removing the oil component. This is because the oil component can be efficiently removed by grinding.
また、本発明における製法によって得られた抽出液あるいはエキスから、カラムクロマトグラフィーなどを用いて精製した単一成分を用いることもできる。 Moreover, the single component refine | purified using the column chromatography etc. from the extract or extract obtained by the manufacturing method in this invention can also be used.
以下、実施例を挙げて本発明をさらに説明するが、本発明はこれら実施例に限定されるものではない。 EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated further, this invention is not limited to these Examples.
実験材料にはタヒチ産のモリンダ・シトリフォリア果実、および種子を使用した。 The experimental materials used were Morinda citrifolia fruits and seeds from Tahiti.
参考例
検体１；乾燥種子を粉砕後、１０倍量のヘキサンで１時間１回還流抽出し、ろ液を減圧下濃縮して溶媒を留去したものをヘキサン抽出エキスとした（収率；１０．７％）。
検体２；検体１の調製においてヘキサン抽出した後、即ちヘキサン抽出によりオイル成分を除去した後の残渣を再度エタノールで１時間１回還流抽出し、ろ液を減圧下濃縮して溶媒を留去したものをエタノール抽出エキスとした（収率；３．６％）。
検体３；乾燥種子を粉砕後、１０倍量の５０％エタノールで一晩静置抽出し、ろ液を減圧下濃縮してエタノールを除き、凍結乾燥したものを５０％エタノール静置抽出エキスとした（収率；３．５％）。
検体４；粉砕していない乾燥種子を１０倍量の５０％エタノールで２時間２回還流抽出し、ろ液を減圧下濃縮してエタノールを除き、凍結乾燥したものを５０%エタノール加熱抽出エキスとした（収率；５．０％）。
検体５；種子を除き、粉砕した乾燥果実を１０倍量の５０％エタノールで２時間２回還流抽出し、ろ液を減圧下濃縮してエタノールを除き、凍結乾燥したものを５０%エタノール加熱抽出エキスとした（収率；３３．０％）。
Reference sample 1: The dried seeds were pulverized and then refluxed and extracted once with 10 times the amount of hexane for 1 hour. The filtrate was concentrated under reduced pressure and the solvent was distilled off to obtain a hexane extract (yield: 10 .7%).
Specimen 2: After hexane extraction in the preparation of Specimen 1, that is, after removing the oil component by hexane extraction, the residue was reflux extracted once more with ethanol once an hour, and the filtrate was concentrated under reduced pressure to distill off the solvent. The product was used as an ethanol extract (yield: 3.6%).
Specimen 3: After pulverizing the dried seeds, the extract was statically extracted overnight with 10 times the amount of 50% ethanol, and the filtrate was concentrated under reduced pressure to remove ethanol and freeze-dried to obtain a 50% ethanol static extract. (Yield; 3.5%).
Specimen 4: Dry seed that was not crushed was refluxed twice with 10 times the amount of 50% ethanol for 2 hours, the filtrate was concentrated under reduced pressure to remove ethanol, and the freeze-dried extract was extracted with 50% ethanol heated extract. (Yield; 5.0%).
Specimen 5: The seeds were removed, and the pulverized dried fruits were reflux-extracted with 10 times the amount of 50% ethanol twice for 2 hours. An extract was obtained (yield; 33.0%).
試験例１
エラスターゼ阻害活性試験；Ｃａｉｎｅｌｌｉらの方法に準じて行った。すなわち、８ｍＭの基質（Ｎ-メトキシスクシニル−Ｌ−アラニル−Ｌ−アラニル−Ｌ−プロリル−Ｌ−バリン−ｐ−ニトロアニリド）（シグマ社製）／ジメチルスルフォキシド（ＤＭＳＯ）溶液を調製し、これを基質溶液とした。２５０ｍＵ／ｍＬの酵素エラスターゼ溶液を０．１Ｍ ヘペス（Ｈｅｐｅｓ）緩衝液（０．５ Ｍ ＮａＣｌ、１０％ ＤＭＳＯ、ｐＨ ７．８）を用いて調製した。被検体液５μＬと酵素エラスターゼ（シグマ社製）溶液９０μＬを混合し、４℃、１５分間プレインキュベーションした後、基質溶液５μＬを加え反応を開始した。３７℃、２０分間インキュベーションした後、吸光波長４０５ｎｍで吸光度を測定した。被検体、および陽性対照薬のフェニルメタンスルフォニルフルオリド（ＰＭＳＦ）（シグマ社製）はＤＭＳＯに溶解した。
Test example 1
Elastase inhibitory activity test; performed according to the method of Cainelli et al. That is, an 8 mM substrate (N-methoxysuccinyl-L-alanyl-L-alanyl-L-prolyl-L-valine-p-nitroanilide) (manufactured by Sigma) / dimethyl sulfoxide (DMSO) solution was prepared, This was used as a substrate solution. A 250 mU / mL enzyme elastase solution was prepared using 0.1 M Hepes buffer (0.5 M NaCl, 10% DMSO, pH 7.8). A sample solution (5 μL) and an enzyme elastase (Sigma) solution (90 μL) were mixed and preincubated at 4 ° C. for 15 minutes, and then the substrate solution (5 μL) was added to start the reaction. After incubation at 37 ° C. for 20 minutes, the absorbance was measured at an absorption wavelength of 405 nm. The subject and the positive control drug phenylmethanesulfonyl fluoride (PMSF) (manufactured by Sigma) were dissolved in DMSO.
表１ 検体４及び５のエラスターゼ阻害作用
＃；ｐ＜０．０１（対照（ｂ）との比較））
Table 1 Elastase inhibitory action of specimens 4 and 5
#; P <0.01 (comparison with control (b)))
表１に示したように、果実および種子の５０％エタノール抽出エキスのエラスターゼ阻害活性を検討した結果、種子エキス（検体４）にエラスターゼ阻害活性が認められたが、種子を除いた果実エキス（検体５）にはエラスターゼ阻害作用はまったく認められなかった。 As shown in Table 1, as a result of examining the elastase inhibitory activity of 50% ethanol extract of fruits and seeds, elastase inhibitory activity was found in the seed extract (sample 4), but the fruit extract excluding the seeds (sample) In 5), no elastase inhibitory action was observed.
表２に示したように、オイル成分であるヘキサン抽出エキス（検体１）にはエラスターゼ阻害活性はまったく認められなかった。一方、オイル成分をヘキサン抽出によって除去した残渣のエタノール抽出エキス（検体２）にはエラスターゼ阻害活性が認められた。
また、粉砕した種子を５０％エタノールで静置抽出して得られた５０％エタノール静置抽出エキス（検体３）はエラスターゼ阻害活性をまったく示さなかった。一方、種子をあえて粉砕せずに加熱抽出して得られた５０％エタノール加熱抽出エキス（検体４）にはエラスターゼ阻害活性が認められた。陽性対照薬として用いたＰＭＳＦにもエラスターゼ阻害活性が認められた。
As shown in Table 2, no elastase inhibitory activity was observed in the hexane extract (specimen 1), which is an oil component. On the other hand, an elastase inhibitory activity was observed in the residual ethanol extract (sample 2) from which the oil component was removed by hexane extraction.
In addition, the 50% ethanol stationary extract (sample 3) obtained by stationary extraction of the ground seeds with 50% ethanol did not show any elastase inhibitory activity. On the other hand, elastase inhibitory activity was observed in the 50% ethanol heat extract (sample 4) obtained by heat extraction without pulverizing the seeds. PMSF used as a positive control drug also showed elastase inhibitory activity.
試験例２
１、１−ジフェニル−２−ピクリルヒドラジル（ＤＰＰＨ）ラジカル捕捉活性試験；ＤＰＰＨラジカル捕捉能試験はＭａｒｓｄｅｎらの方法に従った。すなわち、０．５Ｍに調整した酢酸緩衝液（ｐＨ５．５）０．４ｍＬ、エタノール１．６ｍＬ、および被検体液２ｍＬを加えて混合した。その混合した液に０．５ｍＭ ＤＰＰＨ（和光純薬社製）／エタノール溶液１ｍＬを加え、暗所で３０分間室温放置後、吸光波長５２０ｎｍで吸光度を測定した。
Test example 2
1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity test; DPPH radical scavenging ability test was in accordance with the method of Marsden et al. That is, 0.4 mL of acetate buffer (pH 5.5) adjusted to 0.5 M, 1.6 mL of ethanol, and 2 mL of the sample solution were added and mixed. To the mixed solution, 1 mL of 0.5 mM DPPH (manufactured by Wako Pure Chemical Industries, Ltd.) / Ethanol solution was added and allowed to stand at room temperature in the dark for 30 minutes, and the absorbance was measured at an absorption wavelength of 520 nm.
表３に示したように、オイル成分であるヘキサン抽出エキス（検体１）はラジカル捕捉活性をまったく示さなかった。一方、オイル成分をヘキサン抽出によって除去した残渣のエタノール抽出エキス（検体２）には濃度依存的にラジカルを捕捉する活性が認められた。
また、種子をあえて粉砕せずに加熱抽出して得られた５０％エタノール加熱抽出エキス（検体４）は５、１０、２０μｇ／ｍＬの濃度で、粉砕した種子を５０％エタノールで静置抽出して得られた５０％エタノール静置抽出エキス（検体３）のそれよりも約４倍もの強いラジカル捕捉活性を示した。
As shown in Table 3, the hexane extract (sample 1), which is an oil component, showed no radical scavenging activity. On the other hand, the residue-extracted ethanol extract (specimen 2) from which the oil component was removed by hexane extraction was found to have an activity of scavenging radicals in a concentration-dependent manner.
In addition, 50% ethanol heat extract (sample 4) obtained by heat extraction without pulverizing the seeds was extracted at a concentration of 5, 10, 20 μg / mL, and the pulverized seeds were left to stand with 50% ethanol. The radical scavenging activity was about 4 times stronger than that of the 50% ethanol stationary extract (specimen 3) obtained.
試験例３
試験例１において、エタノールをイソプロピルアルコールおよびプロピレングリコールに置きかえて実験を行ったところ、いずれにおいても同様の結果を得た。
Test example 3
In Experiment 1, when ethanol was replaced with isopropyl alcohol and propylene glycol, the same results were obtained in all cases.
以下に皮膚外用剤（化粧水）及び内服剤（カプセル剤）の配合例及び製造例を示す。なお、％とあるのは重量％を示す。 Hereinafter, formulation examples and production examples of the external preparation for skin (skin lotion) and internal use (capsule) are shown. “%” Means “% by weight”.
検体４を用い、下記成分を所定量混合して24時間静置し、ろ過して化粧水とした。
検体４ １０％
幼若ハッサクエキス １０％
メチルパラベン １％
エタノール ４％
精製水 ７５％
Using specimen 4, the following components were mixed in predetermined amounts, allowed to stand for 24 hours, and filtered to obtain a lotion.
Sample 4 10%
Juvenile Hassaku Extract 10%
Methylparaben 1%
Ethanol 4%
Purified water 75%
検体４を用い、下記各成分を所定量加水混合して造粒し、６０℃で６時間乾燥したのち整粒し、常法に従いカプセル充填し、２５０ ｍｇのハードカプセル剤とした。
検体４ ２０％
幼若ウンシュウミカン末 ３５％
還元麦芽糖 ２５％
アスコルビン酸 ５％
結晶セルロース末 １５％
Using Sample 4, the following components were mixed with a predetermined amount of water, granulated, dried at 60 ° C. for 6 hours, granulated, filled in a capsule according to a conventional method, and a 250 mg hard capsule was prepared.
Sample 4 20%
Juvenile Unshu Mikan powder 35%
Reduced maltose 25%
Ascorbic acid 5%
Crystalline cellulose powder 15%
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