JP2020130093A - Composition for promoting intestinal butyric acid production - Google Patents
Composition for promoting intestinal butyric acid production Download PDFInfo
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Abstract
Description
本発明は、腸内における酪酸の産生促進用組成物及びそれを用いた飲食品又は医薬品に関するものである。 The present invention relates to a composition for promoting the production of butyric acid in the intestine and a food or drink or a pharmaceutical product using the same.
腸内環境を整えることは疾病予防・健康維持において重要である。腸内細菌が産生する短鎖脂肪酸は上皮細胞のエネルギー源として利用されるのみならず、炎症の抑制や血糖の上昇抑制等様々な働きを有している。短鎖脂肪酸の産生には、水溶性食物繊維が重要であり、水溶性食物繊維としてフルクタンの一種であるイヌリンが知られている。
特許文献１には、イヌリンとスルホニル尿素等とを組み合わせた糖尿病治療のための相乗作用組成物が開示されている。
Preparing the intestinal environment is important for disease prevention and health maintenance. Short-chain fatty acids produced by intestinal bacteria are not only used as an energy source for epithelial cells, but also have various functions such as suppressing inflammation and suppressing the rise of blood glucose. Water-soluble dietary fiber is important for the production of short-chain fatty acids, and inulin, which is a kind of fructan, is known as a water-soluble dietary fiber.
特許文献１のように、イヌリンとその他の化合物の組み合わせに関する効果は徐々に報告されてきているが、イヌリンと他の食物繊維の組み合わせによる腸内細菌由来の短鎖脂肪酸産生に与える影響に関しては、いまだ十分に明らかにされていない。
本発明は、腸内の酪酸産生を促進するための組成物及びそれを含む飲食品又は医薬品を提供することを目的とする。
As in
An object of the present invention is to provide a composition for promoting butyric acid production in the intestine and a food or drink or a drug containing the same.
本発明者は、上記課題を解決すべく鋭意研究を重ねた結果、イヌリンとセルロースとを組み合わせることによって、腸内で酪酸が産生促進されることを見出し、本発明を完成させるに至った。即ち、本発明は以下の発明を包含する。 As a result of diligent research to solve the above problems, the present inventor has found that the combination of inulin and cellulose promotes the production of butyric acid in the intestine, and has completed the present invention. That is, the present invention includes the following inventions.
［１］イヌリンとセルロースとを含む腸内酪酸産生促進用組成物。
［２］［１］に記載の組成物を含む、飲食品。
［３］［１］に記載の組成物を含む、医薬品。
[1] A composition for promoting intestinal butyric acid production containing inulin and cellulose.
[2] A food or drink containing the composition according to [1].
[3] A pharmaceutical product containing the composition according to [1].
本発明によると、腸内の酪酸産生を促進するための組成物や、斯かる組成物を用いた飲食品又は医薬品が得られる。 According to the present invention, a composition for promoting butyric acid production in the intestine, and foods and drinks or pharmaceuticals using such a composition can be obtained.
本発明の腸内酪酸産生促進用組成物（以下「本発明の組成物」とも称す）は、イヌリンと、セルロースとを含むことを特徴とする。 The composition for promoting intestinal butyric acid production of the present invention (hereinafter, also referred to as "composition of the present invention") is characterized by containing inulin and cellulose.
イヌリンは、典型的には、プレバイオティクスとして重要な役割を果たし、人類を含め多くの動物はイヌリン加水分解酵素を持たないので、吸収されずに大腸に到達する。大腸中のミクロフローラ（菌叢）の中に、イヌリン加水分解酵素のイヌリナーゼを分泌する菌が含まれ、特に有益菌の代表、ビフィズス菌が知られている。よって、イヌリンを含んだ食物を摂取すると、ビフィズス菌等の有益菌が優先的に増加し有害菌の繁殖を抑える。また、イヌリンの発酵過程で乳酸等の有機酸が増加することから整腸効果を有する。 Inulin typically plays an important role as a prebiotic, and many animals, including humans, do not have inulin hydrolases and therefore reach the large intestine without being absorbed. Bifidobacterium, which is a representative of beneficial bacteria, is known to contain bacteria that secrete inulinase, an inulin hydrolase, in the microflora (bacterial flora) in the large intestine. Therefore, when food containing inulin is ingested, beneficial bacteria such as bifidobacteria increase preferentially and the growth of harmful bacteria is suppressed. In addition, it has an intestinal regulating effect because organic acids such as lactic acid increase in the fermentation process of inulin.
本発明では、イヌリンに更にセルロースを含むようにした組成物が、腸内における酪酸を産生促進する優位な効果が得られた。 In the present invention, the composition in which inulin further contains cellulose has a superior effect of promoting the production of butyric acid in the intestine.
本発明における「イヌリン」は、末端にグルコースをもつフルクトースの多糖ポリマーである、水溶性食物繊維として知られているフルクタンのうち、Ｄ−フルクトフラノースがβ２→１グリコシド結合で重合した直鎖状フルクタンである「直鎖イヌリン」を含む。また、本発明における「イヌリン」は、β２→１グリコシド結合及び少なくとも１つのβ２→６グリコシド結合を有する分岐状フルクタンである「分岐鎖イヌリン」も含む。 "Inulin" in the present invention is a linear fructan, which is a polysaccharide polymer of fructose having glucose at the terminal and is known as a water-soluble dietary fiber, in which D-fructanose is polymerized by β2 → 1 glycosidic bond. Includes the fructan "straight inulin". The "inulin" in the present invention also includes "branched chain inulin" which is a branched fructan having a β2 → 1 glycosidic bond and at least one β2 → 6 glycosidic bond.
直鎖イヌリンは、通常鎖長が２〜１００の範囲であり、好ましくは２〜６０の範囲であり、平均鎖長は通常５〜２０であり、イヌリン中の結合の少なくとも９０％がβ２→１グリコシド結合である。斯かる直鎖イヌリンは、チコリ由来のイヌリンの場合は、例えばFrutafit（登録商標）又はRaftiline（登録商標）を用いることができ、酵素合成によるイヌリンを用いる場合は、例えばフジＦＦ（登録商標）を用いることができる。 Linear inulin usually has a chain length in the range of 2 to 100, preferably in the range of 2 to 60, an average chain length of usually 5 to 20, and at least 90% of the bonds in inulin are β2 → 1. It is a glycosidic bond. As such a linear inulin, in the case of chicory-derived inulin, for example, Fruitafit (registered trademark) or Raftiline (registered trademark) can be used, and in the case of using enzymatically synthesized inulin, for example, Fuji FF (registered trademark). Can be used.
一方、分岐鎖イヌリンは、テキーラの原料である多肉質の植物であるアガベ（リュウゼツラン）に豊富に含まれることが知られている（J. Agric. Food Chem., (2003) 51 (27), pp 7835-7840、J. Agric. Food Chem., (2006) 54 (20), pp 7832-7839、等）。本発明における分岐鎖イヌリンは、特に限定されないが、例えば上記アガベから国際公開０７／１４２３０６号パンフレット等に記載の常法により得られ、典型的には、アガベの茎部分であるピーニャから細断、搾汁、濾過、精製、濃縮、粉末乾燥等を行った処理物に含まれる。このような処理物はアガベイヌリンとも称される。 On the other hand, branched-chain inulin is known to be abundant in agave (agave), a succulent plant that is a raw material for tequila (J. Agric. Food Chem., (2003) 51 (27), pp 7835-7840, J. Agric. Food Chem., (2006) 54 (20), pp 7832-7839, etc.). The branched-chain inulin in the present invention is not particularly limited, but is obtained from, for example, the above-mentioned agave by the conventional method described in Pamphlet No. 07/142306 of International Publication No. 07/142306, and is typically shredded from pinha, which is a stem portion of agave. It is contained in processed products that have been squeezed, filtered, refined, concentrated, and powder-dried. Such processed products are also referred to as agave inulin.
本発明におけるイヌリンは、粉末状、溶液状又はシロップ状等の種々の形態を採ることができ、粘性を増加させて食品で用いる結着剤として利用してもよい。 The inulin in the present invention can take various forms such as powder, solution or syrup, and may be used as a binder for use in foods by increasing the viscosity.
本発明におけるセルロースは、β-グルコース分子がグリコシド結合により直鎖状に重合した天然高分子であり、典型的には、野菜や果実等の食物から不溶性食物繊維として摂取することができる。斯かるセルロースは、セルロースを含む食物の形態として含んでいてもよいし、抽出・精製等したセルロース粉末の形態で含んでいてもよく、特に限定されない。 Cellulose in the present invention is a natural polymer in which β-glucose molecules are linearly polymerized by glycosidic bonds, and can be typically ingested as insoluble dietary fiber from foods such as vegetables and fruits. Such cellulose may be contained in the form of a food containing cellulose, or may be contained in the form of an extracted / purified cellulose powder, and is not particularly limited.
イヌリン及びセルロースの含有量は、所望の効果を発揮する有効量を含んでいればよく、限定されないが、総食物繊維を基準とした重量比として、イヌリン：セルロースが２５：７５〜７５：２５（％）程度であればよい。総量としては、粉末状のイヌリン及びセルロースを合計した重量が、ヒトに対して１日あたり、通常１〜５０ｇ程度であればよい。総食物繊維を定量する方法としては、ＡＯＡＣ９９１．４３に則った、Megazyme社のTotal Dietary Fiber Assay Kitを用いることができる。 The contents of inulin and cellulose may include an effective amount that exerts a desired effect, and are not limited, but inulin: cellulose is 25:75 to 75:25 (25:75 to 75:25) as a weight ratio based on total dietary fiber. %) It is sufficient. As the total amount, the total weight of powdered inulin and cellulose may be about 1 to 50 g per day for humans. As a method for quantifying the total dietary fiber, Megazyme's Total Dietary Fiber Assay Kit according to AOAC991.43 can be used.
本発明の腸内酪酸産生促進用組成物は、主にヒトの大腸内の腸内細菌によって消化発酵されることにより、その代謝物である短鎖脂肪酸の酪酸の産生を相乗的に促進することができる。一般に、ヒトの大腸内では、腸内細菌が食物繊維を発酵する際に短鎖脂肪酸を産生し、健康維持に欠かせない役割を果たしている。ヒトの場合、酢酸、プロピオン酸、酪酸の３種が代表的な短鎖脂肪酸として知られている。ヒトの体で短鎖脂肪酸が作られる部位は腸内細菌が多い大腸で、作られた短鎖脂肪酸は大腸から体内に吸収される。吸収された短鎖脂肪酸のうち、本発明で産生促進される酪酸は、大腸上皮細胞のエネルギー源として利用される。その他、腸は全身の免疫細胞のおよそ６０％が集中し、腸の免疫バランスの崩れ、特に過剰な免疫反応が全身に影響すると言われているが、酪酸には過剰な免疫反応を抑える制御性Ｔ細胞を増やす効果が示唆されている（Nature (2013) 504, pp 446-450）。また、短鎖脂肪酸自体は酸性の成分なので、短鎖脂肪酸ができると弱酸性の腸内環境になる。弱酸性であると悪玉菌の出す酵素の活性が抑えられるため、発がん性物質である二次胆汁酸や有害な腐敗産物ができにくくなり、整腸作用により腸内環境が健康に保たれる。 The composition for promoting intestinal butyric acid production of the present invention synergistically promotes the production of butyric acid, which is a metabolite of short-chain fatty acids, by being digested and fermented mainly by intestinal bacteria in the human large intestine. Can be done. Generally, in the human large intestine, intestinal bacteria produce short-chain fatty acids when fermenting dietary fiber, and play an essential role in maintaining health. In the case of humans, three types of acetic acid, propionic acid, and butyric acid are known as typical short-chain fatty acids. The site where short-chain fatty acids are produced in the human body is the large intestine, which is rich in intestinal bacteria, and the produced short-chain fatty acids are absorbed into the body from the large intestine. Of the absorbed short-chain fatty acids, butyric acid whose production is promoted in the present invention is used as an energy source for colonic epithelial cells. In addition, about 60% of the immune cells of the whole body are concentrated in the intestine, and it is said that the immune balance of the intestine is disturbed, especially an excessive immune reaction affects the whole body, but butyric acid has a regulatory ability to suppress the excessive immune reaction. The effect of increasing T cells has been suggested (Nature (2013) 504, pp 446-450). In addition, since short-chain fatty acids themselves are acidic components, the formation of short-chain fatty acids creates a weakly acidic intestinal environment. When it is weakly acidic, the activity of enzymes produced by bad bacteria is suppressed, so that secondary bile acids, which are carcinogens, and harmful putrefactive products are less likely to be produced, and the intestinal environment is maintained healthy by the intestinal regulation action.
本発明の組成物は、ヒト又は動物用の医薬品又は飲食品として利用することもできる。本明細書では、動物用の飲食品を飼料とも称す。 The composition of the present invention can also be used as a pharmaceutical product or food or drink for humans or animals. In this specification, food and drink for animals is also referred to as feed.
本発明における医薬品は、本発明の組成物を含有する腸内酪酸産生促進薬等であり得る。上記医薬品の剤型としては、例えば、錠剤、カプセル剤、顆粒剤、散剤、シロップ剤、ドライシロップ剤、液剤、懸濁剤等の経口剤、及び吸入剤、経皮製剤、坐剤等の経腸製剤、点滴剤、注射剤等の非経口剤が挙げられる。上記液剤、懸濁剤等の液体製剤は、服用直前に水又は他の適当な媒体に溶解又は懸濁する形であってもよく、上記錠剤及び顆粒剤は周知の方法でその表面をコーティングされていてもよい。また上記注射剤は、必要に応じて溶解補助剤を含む滅菌蒸留水又は滅菌生理食塩水の溶液であり得る。 The pharmaceutical product in the present invention may be an intestinal butyric acid production promoter or the like containing the composition of the present invention. Examples of the dosage form of the above-mentioned pharmaceuticals include oral preparations such as tablets, capsules, granules, powders, syrups, dry syrups, liquids and suspensions, and enteral preparations such as inhalants, transdermal preparations and suppositories. Parenteral preparations such as preparations, infusions, and injections can be mentioned. Liquid preparations such as the above liquids and suspensions may be dissolved or suspended in water or other suitable medium immediately before administration, and the above tablets and granules are coated on the surface by a well-known method. May be. Further, the injection may be a solution of sterile distilled water or sterile physiological saline containing a solubilizing agent, if necessary.
本発明における医薬品は、本発明の組成物に加えて、必要に応じて薬学的に許容される種々の担体、例えば賦形剤、安定化剤、その他の添加剤等を含有していてもよく、あるいは、さらに他の薬効成分、例えば各種ビタミン類、ミネラル類、生薬等を含有していてもよい。当該医薬品は、本発明の組成物に、上述の担体及び他の薬効成分を配合し、常法に従って製造することができる。 In addition to the composition of the present invention, the pharmaceutical product of the present invention may contain various pharmaceutically acceptable carriers such as excipients, stabilizers, other additives and the like, if necessary. Alternatively, it may contain other medicinal ingredients such as various vitamins, minerals, and crude drugs. The pharmaceutical product can be produced according to a conventional method by blending the above-mentioned carrier and other medicinal ingredients with the composition of the present invention.
本発明における飲食品又は飼料は、本発明の組成物を含み、場合によって、腸内酪酸産生促進の効果を企図して、その旨を表示した健康食品、機能性飲食品、特定保健用飲食品、病者用飲食品、家畜、競走馬、鑑賞動物等のための飼料、ペットフード等としてもあり得る。 The food or feed in the present invention contains the composition of the present invention, and in some cases, a health food, a functional food or a food or a food for specified health use, which is intended to have an effect of promoting intestinal butyric acid production and is labeled to that effect. It can also be used as food and drink for the sick, livestock, racehorses, feed for ornamental animals, pet food, and the like.
本発明における飲食品及び飼料の形態は特に制限されず、本発明の組成物を配合できる全ての形態が含まれる。例えば当該形態としては、固形、半固形又は液状であり得、あるいは、錠剤、チュアブル錠、粉剤、カプセル、顆粒、ドリンク、ゲル、シロップ、経管経腸栄養用流動食等の各種形態が挙げられる。 The forms of food and drink and feed in the present invention are not particularly limited, and all forms in which the composition of the present invention can be blended are included. For example, the form may be solid, semi-solid or liquid, or may include various forms such as tablets, chewable tablets, powders, capsules, granules, drinks, gels, syrups, and liquid foods for tube-intestinal nutrition. ..
具体的な飲食品の例としては、緑茶、ウーロン茶や紅茶等の茶飲料、コーヒー飲料、清涼飲料、ゼリー飲料、スポーツ飲料、乳飲料、炭酸飲料、果汁飲料、乳酸菌飲料、発酵乳飲料、粉末飲料、ココア飲料、アルコール飲料、精製水等の飲料、バター、ジャム、ふりかけ、マーガリン等のスプレッド類、マヨネーズ、ショートニング、クリーム、ドレッシング類、パン類、米飯類、麺類、パスタ、味噌汁、豆腐、牛乳、ヨーグルト、スープ又はソース類、ベーカリー類（パン、パイ、ケーキ、クッキー、ビスケット、クラッカー等）、菓子（ビスケットやクッキー類、チョコレート、キャンディ、ケーキ、アイスクリーム、チューインガム、タブレット等）、栄養補助食品（丸剤、錠剤、ゼリー剤又はカプセル剤等の形態を有するサプリメント、グラノーラ様シリアル、グラノーラ様スネークバー、シリアルバー）等が挙げられる。 Specific examples of foods and drinks include green tea, tea beverages such as oolong tea and tea, coffee beverages, soft beverages, jelly beverages, sports beverages, dairy beverages, carbonated beverages, fruit juice beverages, lactic acid bacteria beverages, fermented milk beverages, and powdered beverages. , Cocoa beverages, alcoholic beverages, purified water and other beverages, butter, jam, sprinkles, margarine and other spreads, mayonnaise, shortening, creams, dressings, breads, rice, noodles, pasta, miso soup, tofu, milk, Yogurt, soups or sauces, bakeries (bread, pies, cakes, cookies, biscuits, crackers, etc.), confectionery (biscuits, cookies, chocolate, candy, cakes, ice cream, chewing gum, tablets, etc.), nutritional supplements ( Supplements in the form of rounds, tablets, jellies or capsules, granola-like cereals, granola-like cookie bars, serial bars) and the like.
本発明における飼料は、上記飲食品とほぼ同様の組成や形態で利用できることから、本明細書における飲食品に関する記載は、飼料についても同様に当てはめることができる。 Since the feed in the present invention can be used in substantially the same composition and form as the above-mentioned food and drink, the description regarding the food and drink in the present specification can be applied to the feed as well.
本発明における飲食品及び飼料は、本発明の組成物に、飲食品や飼料の製造に用いられる他の飲食品素材、各種栄養素、各種ビタミン、ミネラル、アミノ酸、各種油脂、種々の添加剤（呈味成分、甘味料、有機酸等の酸味料、界面活性剤、ｐＨ調整剤、安定剤、酸化防止剤、色素、フレーバー等）等を配合して、常法に従って製造することができる。あるいは、通常食されている飲食品又は飼料に、本発明の組成物を配合することにより、本発明に係る飲食品又は飼料を製造することもできる。 The food and drink and feed in the present invention are the composition of the present invention, other food and drink materials used in the production of food and drink and feed, various nutrients, various vitamins, minerals, amino acids, various fats and oils, and various additives (presentation). It can be produced according to a conventional method by blending a taste component, a sweetener, an acidulant such as an organic acid, a surfactant, a pH adjuster, a stabilizer, an antioxidant, a pigment, a flavor, etc.). Alternatively, the food or drink or feed according to the present invention can be produced by blending the composition of the present invention with the food or food or feed that is normally eaten.
本発明における医薬品、飲食品又は飼料における本発明の組成物の含有量は、所望する腸内酪酸産生促進効果が得られる量であればよく、医薬の剤型や飲食品の形態、投与又は摂取する個体の種、症状、年齢、性別等に応じて適宜変更され得るが、粉末状のイヌリン及びセルロースを合計した重量が、ヒトに対して１日あたり、通常１〜５０ｇ程度であればよい。 The content of the composition of the present invention in the drug, food or drink or feed in the present invention may be any amount as long as the desired effect of promoting intestinal butyric acid production can be obtained, and the dosage form of the drug or the form, administration or ingestion of the food or drink The total weight of powdered inulin and cellulose may be about 1 to 50 g per day for humans, although it can be appropriately changed depending on the species, symptoms, age, sex, etc. of the individual.
以下に実施例及び比較例を挙げて本発明を具体的に説明するが、これにより本発明の範囲が限定されるものではない。 Hereinafter, the present invention will be specifically described with reference to Examples and Comparative Examples, but the scope of the present invention is not limited thereto.
［試料の調製］
試料として、セルロース（試料名称を「Ｃｅｌｌ」とする）３ｇ、イヌリン（試料名称を「ＩＱ」とする）３ｇ、セルロース１．５ｇ及びイヌリン１．５ｇ（同様に「Ｃｅｌｌ＋ＩＱ」とする）を調製した。セルロースは富士フイルム和光純薬から購入し、イヌリンはＦｒｕｔａｆｉｔ ＩＱ（登録商標）（Ｓｅｎｓｕｓ社）を使用した。
[Sample preparation]
As samples, 3 g of cellulose (referred to as "Cell"), 3 g of inulin (referred to as "IQ"), 1.5 g of cellulose and 1.5 g of inulin (also referred to as "Cell + IQ") were prepared. .. Cellulose was purchased from Fujifilm Wako Pure Chemical Industries, Ltd., and inulin used Frutafit IQ® (Sensus).
［糞便懸濁液の調製］
ヒト大腸内を模した腸内細菌叢を再現するために、豚（４−５月齢）の糞便を用いた人口腸管モデルを採用した。糞便懸濁液は、凍結保存の豚糞便に１０倍の生理食塩水を加え混合した後、ストレインバッグ（栄研化学）を用いて濾過したものを使用した。
[Preparation of fecal suspension]
In order to reproduce the intestinal flora that imitates the human large intestine, we adopted an artificial intestinal model using feces of pigs (4-5 months old). As the stool suspension, 10-fold physiological saline was added to cryopreserved pig stool, mixed, and then filtered using a strain bag (Eiken Chemical Co., Ltd.).
［Ｉｎ ｖｉｔｒｏ腸内発酵試験］
Ｉｎ ｖｉｔｒｏ腸内発酵試験には、Ｂｉｏ Ｊｒ．８培養装置（エイブル）を用いた。蒸留水８０ｍLを加えた培養槽、及び蒸留水１００ｍＬにＮｕｔｒｉｅｎｔ Ｂｒｏｔｈ（ＤｉｆｃｏＬａｂｏｒａｔｏｒｉｅｓ）４．８ｇを溶解させた溶液をそれぞれ１２１℃、１５分間滅菌した。滅菌・放冷後、培養槽へ糞便懸濁液を２０ｍＬ混和し、一晩培養を行った。培養装置の設定条件は、温度３７℃、撹拌子回転数４００ｒｐｍであった。培養槽内のｐＨが５．５を下回った場合にはアルカリ溶液（２Ｎ 水酸化ナトリウム）が自動的に滴下されるように設定した。また、培養槽内を嫌気状態に保つため、二酸化炭素ガスを充填した。
一晩培養後、Ｎｕｔｒｉｅｎｔ Ｂｒｏｔｈを溶解させた溶液２０ｍＬ及び試料を培養槽に添加した。４８時間培養後、培養液を採取し、遠心分離することで培養上清を得た。
[In vitro intestinal fermentation test]
For in vitro fermentation tests, Bio Jr. 8 Incubator (ABLE) was used. A culture tank containing 80 mL of distilled water and a solution prepared by dissolving 4.8 g of Nutrient Broth (DifcoLaboratories) in 100 mL of distilled water were sterilized at 121 ° C. for 15 minutes, respectively. After sterilization and allowing to cool, 20 mL of the fecal suspension was mixed in the culture tank and cultured overnight. The setting conditions of the incubator were a temperature of 37 ° C. and a stirrer rotation speed of 400 rpm. When the pH in the culture tank fell below 5.5, the alkaline solution (2N sodium hydroxide) was set to be automatically added dropwise. In addition, carbon dioxide gas was filled in order to keep the inside of the culture tank in an anaerobic state.
After culturing overnight, 20 mL of a solution in which Nutrient Bouillon was dissolved and a sample were added to the culture tank. After culturing for 48 hours, the culture broth was collected and centrifuged to obtain a culture supernatant.
［短鎖脂肪酸の測定］
短鎖脂肪酸の測定には、超高速液体クロマトグラフ（ＮｅｘｅｒａＸ２、島津製作所）を用い、分析条件は表１とした。培養上清４５０ｕＬに０．５Ｎ 過塩素酸を１ｍＬ添加し、５分間静置した後、遠心分離させ、得られた上清をＨＰＬＣ前処理フィルターで濾過したものを測定サンプルとした。酪酸の同定は測定サンプルと標準溶液の保持時間の比較によって、定量はピーク面積の比較によって行った。
[Measurement of short-chain fatty acids]
An ultra-high performance liquid chromatograph (NexeraX2, Shimadzu Corporation) was used for the measurement of short-chain fatty acids, and the analysis conditions are shown in Table 1. 1 mL of 0.5N perchloric acid was added to 450 uL of the culture supernatant, allowed to stand for 5 minutes, centrifuged, and the obtained supernatant was filtered through an HPLC pretreatment filter to obtain a measurement sample. Butyric acid was identified by comparing the retention time of the measurement sample and the standard solution, and quantified by comparing the peak area.
［実施例１］
上記の通り試験を行い、各試料添加群の培養液中酪酸量を測定したところ、図１及び表２の結果が得られた。ＩＱ添加群又はＣｅｌｌ添加群と比較して、Ｃｅｌｌ＋ＩＱ添加群では、培養液中酪酸量の有意な増加が認められた。
これらの組み合わせの相乗効果を、以下の計算式により算出した。
〈相乗効果〉＝ＸＹ／（（Ｘ／２）＋（Ｙ／２））
（式中、Ｘはセルロース（Ｃｅｌｌ）単体、Ｙはイヌリン（ＩＱ）単体の短鎖脂肪酸量であり、ＸＹは各試料を組み合わせて添加した場合の短鎖脂肪酸量である。上記［試料の調製］の通り、ＸＹとＸ又はＹは総食物繊維量を等しくしているため、Ｘ又はＹは、その単体の短鎖脂肪酸量を１／２にしている）
上記計算式から、Ｃｅｌｌ＋ＩＱ添加群の相乗効果は、表２中の各値より、２．３５／（（０．５７／２）＋（０．７５／２））にて算出したところ３．６倍となった。よって、Ｃｅｌｌ＋ＩＱにおける相乗効果が確認され、イヌリンとセルロースの組み合わせによって腸内細菌叢からの酪酸産生が促進されたことが示された。
[Example 1]
When the test was carried out as described above and the amount of butyric acid in the culture solution of each sample addition group was measured, the results shown in FIGS. 1 and 2 were obtained. A significant increase in the amount of butyric acid in the culture solution was observed in the Cell + IQ-added group as compared with the IQ-added group or the Cell-added group.
The synergistic effect of these combinations was calculated by the following formula.
<Synergistic effect> = XY / ((X / 2) + (Y / 2))
(In the formula, X is the amount of short-chain fatty acid of cellulose (Cell) alone, Y is the amount of short-chain fatty acid of inulin (IQ) alone, and XY is the amount of short-chain fatty acid when each sample is added in combination. ], Since XY and X or Y have the same total dietary fiber amount, X or Y halves the amount of short-chain fatty acids of the simple substance).
From the above formula, the synergistic effect of the Cell + IQ addition group was calculated as 2.35 / ((0.57 / 2) + (0.75 / 2)) from each value in Table 2, and it was 3.6. It has doubled. Therefore, a synergistic effect on Cell + IQ was confirmed, and it was shown that the combination of inulin and cellulose promoted butyric acid production from the intestinal flora.
Claims (3)
A pharmaceutical product comprising the composition according to claim 1.
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