201116272 六、發明說明： 【發明所屬之技術領域】 本發明係以可藉由糖醇抑制揮發性硫化合物產生爲特 徵之口臭去除劑者。 【先前技術】 在曰本以3萬人爲對象經由厚生省保健福祉動向調查 時，發現有7 0 %的人被發現口腔內有某種牙科之問題，其 中14.5%之人回答「對於口臭覺得有問題」（可以複數項 同時回答）’排名次於牙周病或生齲等相關之第4名，逐 年針對口臭之問題正提昇其關心。根據報告從口腔發出之 令人不適的臭味成份有揮發性硫化合物（V S C )、揮發性 氮化合物、低價脂肪酸等。此等中根據報告、官能試驗之 臭味強度、與口腔內所檢查出的濃度之間，具有極大之相 關連性者係VSC。自口腔內氣體中被檢測出之vsc係確認 有硫化氫、甲基硫醇、二甲基硫醚之三種爲各單獨、或混 合存在。其他物質雖可以自呼氣中檢驗出，但卻很少被檢 查出人之氣味臨界値以上的濃度。 口臭之主要原因的V S C係以口腔內細胞之殘骸或食物 中含硫氨基酸做爲基質，藉由口腔內之嫌氣性菌的代謝而 被產生者。尤其經由細菌之胱硫醚-β-合成酶、胱硫酸_ r . 裂合酶之關連、自牛胱胺酸會產生硫化氫、經由L-甲硫月安 酸-r-裂合酶之關連而自甲硫胺酸產生甲基硫醇。 vsc係不但爲惡臭物質，還具有強力之活體毒性。經 201116272 由報告獲知，藉由VSC會亢進具有變異功能之黏膜透過性 ，並且會促進纖維芽細胞之膠原合成阻礙、以及促進上皮 基底膜之損傷，而阻礙其合成。尤其硫化氫係在使用人白 血球之實驗中，顯示不但可增加活性氧之產生，另一方面 還會強烈地阻礙氧物歧化酶（Superoxide dismutase ( SOD ))，被報告VSC可能係具有發癌性者。爲此抑制口臭係 就維持口腔之健康而言乃具極重要之意義。 近年來針對口腔之維護，尤其就抗腐蝕之觀點，雖有 增加糖醇類的需要，但糖醇類對'口臭的影響卻仍未被解析 清楚。 在此乃對現今被常用於無糖口香糖的糖醇中之木糖醇 、麥芽糖醇、赤蘚糖醇及乳糖醇加以評估其對口臭有何種 影響，經由唾液培育試驗、菌代謝阻礙試驗進行評估。 有關糖醇類與口臭，減低消臭作用之先行技術有如下 之文獻。 專利文獻1係針對有關口香糖之發明者，將含有磷酸 二氫鈣、甘油磷酸鈣的組成物與膠體基質一起混練，製作 爲口香糖，經由10名評審員咀嚼，予以官能評估其對口臭 效果之速效性與持續性。結果使用配合6 0%木糖醇之口香 糖時，確認立即有效性者爲10/10人，確認具持續性者爲 7/10人。與之相比，將木糖醇改爲蔗糖60%配合時，確認 有速效性者爲5/10人，確認有持續性者爲3/10人。又，並 無記載咀嚼時間' 或確認速效性、持續性的具體時間。由 專利文獻1之發明詳細說明中的說明00 1 7中曾記載，含有 201116272 一種或二種以上選自木糖醇、山梨糖醇、赤蘚糖醇所成羣 之糖醇於口香糖中時’可以提高口臭除去效果，其配合量 係封全體之口香糖而g爲20〜85質量％，更以3〇〜80質量％ 爲宜。 專利文獻2係有關口腔用組成物者。揭示尤其可防止 、抑制由甲硫胺酸酶所引起之口臭的口腔用組成物。其申 請專利範圍第1項中記載其含有一種或二種以上選自甘露 醇、麥芽糖醇、山梨糖醇及此等之混合物所成羣，其製品 、製劑中通常以0 _ 1質量％以上’較佳以丨〜7 〇質量％之比例 配cu 。5式料液中添加牙銀P卜琳單細菌（P〇rphyr〇monas. gingivalis )之菌體懸濁液、甲硫胺酸，測定甲基硫醇之結 果，含有1質量％麥芽糖醇之試料液係與無添加者相比較， 得到可以阻礙20.0%甲硫胺酸酶活性之報告。 專利文獻3係有關口臭成份洗淨組成物及含其之口腔 用組成物、口香糖及口中清涼糖者。尤其爲有關可以洗淨 口腔內之吲哚、甲基吲哚、酚、對-甲酚等有氣味成份的 洗淨組成物、或口腔用組成物者。依申請專利範圍第4項 記載其含有一種或二種以上選自碳數4〜24之糖醇，申請專 利範圍第5項及發明之詳細說明00 1 9則記載由高度之口臭 成份洗淨效果而言，上述糖醇中尤以山梨糖醇、木糖醇、 乳糖醇、麥芽糖醇、異構麥芽糖（palatinit )爲宜之報告 。在使用於牙膏之實施例中則表示配合赤蘚糖醇、麥芽糖 醇者係對於具有口臭之試驗者而言，使用3 0分鐘後可以顯 著地減低其口臭（由3名專門評審員評估其官能）。 201116272 專利文獻4係揭示相轉變之口臭除去劑，尤其以甘油 一酸酯爲主基劑’含有聚合物、多元醇、口臭除去有效成 份及溶劑所成之口臭除去劑者。其申請專利範圍第1 4項中 揭示含有做爲除去口臭有效成份之非發酵性糖醇，於申請 專利範圍第16項記載做爲非發酵性糖醇之木糖醇、山梨糖 醇、赤藓糖醇、甘露醇、麥芽糖醇、乳糖醇、巴拉金糖醇 、巴拉金糖（palatinose)、寡糖。專利文獻4中曾以含有 木糖醇之比較例、實施例進行官能性評估以評價口臭除去 效果，惟並未確認木糖醇含量與官能性評估結果之相關性 〇 如上述，以往技術中係針對木糖醇、赤蘚糖醇、麥芽 糖醇、還包含實施例、認爲具有減低口臭之效果。惟針對 乳糖醇，雖在申請專利範圍中做爲組成物，有效成份記載 ，但未有實施例可以完全確認有具體之消臭效果。 先行技術文獻 〔專利文獻〕 〔專利文獻1〕特開2006-32545 5號公報 〔專利文獻2〕特開2003- 1 6045 8號公報 〔專利文獻3〕特開2004-203 8 7 2號公報 〔專利文獻4〕特表2009-500399 【發明內容】 〔發明欲解決之課題〕 本發明係以提供可藉由糖醇對揮發性硫化合物之產生 -8- 201116272 可具抑制做爲特徵之口臭除去劑爲目的者。 自口腔發出令人不舒服之臭味成份，根據報$胃vsc 、揮發性氮化合物、低級脂肪酸等。此等中根據報告， VSC係由官能性試驗之臭味強度、與自口腔內所檢查出之 濃度間具有極強烈之相關性。爲確認對糖醇之□臭所具影 響，經調查各種糖醇對VSC產生之影響，結果確認赤蘚糖 醇、乳糖醇、麥芽糖醇係pH非依賴性地可以抑制硫化氣、 甲基硫醇的產生。又，對於牙周病之主要原因菌的牙齦口卜 啉單胞菌（？〇^11丫1'011101133.§丨1^卜31丨3)則只需以1〇%濃度 之乳糖醇、麥芽糖醇即可以減少硫化氫之產生量至約 4 0〜6 0 % 〇 發明之效果 本發明係因可以顯示極顯著之抑制揮發性硫化合物的 產生作用，所以做爲含漱劑、牙膏、吸入劑、錠劑等製劑 ，又可做爲口香糖、糖果、糖果錠片、膠菓糖、餅乾、巧 克力等糖果、冰品、飲料等食品，日常予以利用、攝取， 對改善口臭及預防均極有效。 【實施方式】 實施本發明之形態 本發明之一實施形態係以糖醇爲有效成份之口臭除去 劑者。 本發明之另一實施形態係上述糖醇爲乳糖醇的如上述 ~ 9 - 201116272 記載之口臭除去劑者。 本發明之另一實施形態係以糖醇爲有效成份的口臭除 去劑所成含漱劑、牙膏劑、吸入劑及錠劑者。 本發明之又一實施形態係以糖醇爲有效成份之口臭除 去劑所成食品者。 本發明之又一實施形態係含有以糖醇爲有效成份之口 臭除去劑所成口香糖、糖果、糖果錠片、膠菓糖、餅乾、 巧克力等糖果、冰品、飲料等之食品者。 本發明之另一實施形態係以糖醇爲有效成份之可以對 於經由甲硫胺酸及牛胱胺酸代謝經路阻礙而產生揮發性硫 化合物時做爲其抑制劑者。 本發明之另一實施形態係上述糖醇爲乳糖醇、麥芽糖 醇之如上述所記載對產生揮發性硫化合物做爲抑制劑者。 以下藉由具體之實施例詳細說明本發明，惟本發明並 非可被此等所限制者。 (實施例1 ) 依以下進行糖醇類的唾液培養試驗。 1 _ 1 ·準備試料 做爲試料係使用木糖醇、赤蘚糖醇、乳糖醇、麥芽糖 醇、蔗糖。分別將此等溶解於脫離子水’使其成適當濃度 ，做爲試料溶液。 -10- 201116272 1 - 2 .唾液培養試驗 選出4名健康之成人（A、B、C、D，平均年齡32_0歲 )做爲被試驗者，測試係實施二連續。早上9:00〜9:30之間 ’自各被試驗者採取1 〇ml無刺激唾液（採取當天不吃早餐 ’亦不刷牙）’採取到之唾液保存於冰上面。在1.0 m 1無 刺激唾液中加入0.5 m 1試料溶液，於3 7 °C培養2 3小時。培 養23小時後’將目式料放在冰上。gc ( Gas Chromatography )分析前於3 7 °C振盪試料經丨5分鐘，以注射器取適當量之 (罐頭）頂隙氣體，進行GC分析。另外，中性條件下之試 驗係加入磷酸鉀緩衝液使其最後濃度爲20mM，維持反應 液於中性領域。 1 - 3 . G C分析 有關GC分析係全部均使用Agilent公司製的gC689〇。 分析营柱係使用HP-PL〇TQ(30mx0.53mmx40gm)、以初 期溫度：70°C/2.5min、昇溫：30°C/min，最後溫度：190 °C /3.5min、注入溫度：2〇〇。〇、檢驗器：FpD、檢出器溫 度：200 °C、流量：2〇ml/min之條件下進行分析。有關RT 係硫化氫爲1 · 1 min、甲基硫醇爲4 · 0m i n、二甲基—硫化物 爲5 · 6 m 1 η、—甲基二硫化物爲8 3 m丨η。針對全部試料以二 連續進行，算出平均値。 1 - 4 .結果 1 .蔗糖之效果[Technical Field] The present invention relates to a bad breath removing agent which is characterized in that a sugar alcohol can suppress the generation of a volatile sulfur compound. [Prior Art] When Sakamoto was surveyed by the Health and Welfare Department of the Ministry of Health and Welfare for 30,000 people, it was found that 70% of people were found to have some dental problems in their mouths, and 14.5% of them answered that "there is a feeling for bad breath." The problem (can be answered at the same time) 'The second place is ranked second in the period related to periodontal disease or oysters. The problem of bad breath is increasing its concern year by year. According to the report, the unpleasant odor components emitted from the oral cavity include volatile sulfur compounds (V S C ), volatile nitrogen compounds, and low-priced fatty acids. Among these, depending on the report, the odor intensity of the sensory test, and the concentration detected in the oral cavity, VSC is extremely relevant. The scsc detected from the gas in the oral cavity was confirmed to have three kinds of hydrogen sulfide, methyl mercaptan, and dimethyl sulfide, either alone or in combination. Other substances can be detected from exhalation, but they are rarely detected at concentrations above the threshold of human odor. The V S C, which is the main cause of bad breath, is caused by the residue of cells in the oral cavity or the sulfur-containing amino acid in the food as a substrate, and is produced by metabolism of anaerobic bacteria in the oral cavity. In particular, via cysteine-β-synthase of bacteria, sulphuric acid _ r. lyase, related to hydrogen sulfide from bovine cysteine, and related to L-methylsulfate-r-lyase Methyl mercaptan is produced from methionine. Vsc is not only a stench, but also has a strong living toxicity. According to the report, 201116272, VSC will break through the mucosal permeability of the mutated function, and will promote the collagen synthesis inhibition of the fibroblasts and promote the damage of the epithelial basement membrane, thereby hindering its synthesis. In particular, in the experiment using human white blood cells, hydrogen sulfide not only increases the production of active oxygen, but also strongly blocks the superoxide dismutase (SOD). It is reported that VSC may be cancerous. By. To suppress bad breath, it is of great importance to maintain the health of the mouth. In recent years, in view of oral maintenance, especially in terms of corrosion resistance, although there is a need to increase sugar alcohols, the effects of sugar alcohols on 'bad breath' have not been resolved. Here, the effects of xylitol, maltitol, erythritol and lactitol, which are commonly used in sugar alcohols of sugar-free chewing gum, on halitosis are evaluated by saliva cultivation test and bacterial metabolism inhibition test. Evaluation. The prior art for reducing the deodorization of sugar alcohols and bad breath has the following literature. Patent Document 1 relates to a chewing gum inventor, and a composition containing calcium dihydrogen phosphate or calcium glycerophosphate is kneaded together with a colloidal matrix to prepare a chewing gum, which is chewed by 10 panelists to evaluate its effectiveness against bad breath. Sex and persistence. As a result, when the chewing gum containing 60% xylitol was used, it was confirmed that the immediate effect was 10/10, and it was confirmed that the persistence was 7/10. In contrast, when xylitol was changed to 60% sucrose, 5/10 people were confirmed to have quick-acting properties, and 3/10 persons were confirmed to have persistence. Also, there is no specific time to describe the chewing time' or to confirm the quickness and sustainability. In the description of the detailed description of the invention of Patent Document 1, it is described in 00 1 7 that when one or two or more sugar alcohols selected from the group consisting of xylitol, sorbitol, and erythritol are used in the chewing gum, The bad breath removing effect can be improved, and the amount of the gum is a total of 20 to 85% by mass, more preferably 3 to 80% by mass. Patent Document 2 relates to an oral composition. An oral composition which particularly prevents and suppresses bad breath caused by methioninase is disclosed. In the first paragraph of the patent application, it is described that it contains one or two or more kinds selected from the group consisting of mannitol, maltitol, sorbitol, and the like, and the products and preparations thereof are usually 0 to 1% by mass or more. It is preferable to use cu in a ratio of 丨~7 〇 mass%. A sample suspension containing gram silver, P 〇rphyr〇monas. gingivalis, methionine, and a result of measuring methyl mercaptan, containing 1% by mass of maltitol The liquid system was reported to be able to block 20.0% methioninase activity compared to no adder. Patent Document 3 relates to a composition for cleaning a bad breath component, an oral composition containing the same, a chewing gum, and a refreshing sugar in the mouth. In particular, it relates to a cleaning composition or an oral composition which can wash odorous components such as hydrazine, methyl hydrazine, phenol, and p-cresol in the oral cavity. According to Item 4 of the patent application, it contains one or two or more kinds of sugar alcohols selected from carbon number 4 to 24, and the patent application scope 5 and the detailed description of the invention 00 1 9 describe the cleaning effect of the high-quality bad breath component. Among them, sorbitol, xylitol, lactitol, maltitol, and isomeric maltose (palatinit) are preferably reported among the above sugar alcohols. In the examples used in toothpaste, it is indicated that in combination with erythritol and maltitol, the tester with bad breath can significantly reduce the bad breath after 30 minutes (the evaluation is performed by three panelists) ). 201116272 Patent Document 4 discloses a phase change bad breath removing agent, particularly a monoglyceride-based base agent, which comprises a polymer, a polyhydric alcohol, an effective component for removing bad breath, and a bad breath removing agent. The non-fermentable sugar alcohol which is an effective component for removing bad breath is disclosed in Item 14 of the patent application scope, and xylitol, sorbitol, and red peony which are non-fermentable sugar alcohols described in Item 16 of the patent application. Sugar alcohol, mannitol, maltitol, lactitol, palatinitol, palatinose, oligosaccharides. In Patent Document 4, the functional evaluation of the comparative example and the example containing xylitol was carried out to evaluate the bad breath removing effect, but the correlation between the xylitol content and the functional evaluation result was not confirmed as described above, and the prior art was directed to Xylitol, erythritol, maltitol, and the examples are also considered to have an effect of reducing bad breath. However, for lactitol, although it is a composition and a valid ingredient in the scope of the patent application, no specific deodorizing effect can be completely confirmed by the examples. [Patent Document 1] [Patent Document 1] JP-A-2006-32545 5 (Patent Document 2) JP-A-2003- 1 6045 No. 8 (Patent Document 3) JP-A-2004-203 8 7 2 Patent Document 4] Special Table 2009-500399 [Disclosed from the Invention] [Problems to be Solved by the Invention] The present invention provides a bad breath removal characteristic which can be suppressed by the production of volatile sulfur compounds by sugar alcohols -8- 201116272 For the purpose of the agent. An unpleasant odor component is emitted from the mouth, according to the report of stomach vsc, volatile nitrogen compounds, lower fatty acids, and the like. According to the report, VSC has a strong correlation between the odor intensity of the functional test and the concentration detected from the oral cavity. In order to confirm the influence on the odor of sugar alcohols, the effects of various sugar alcohols on VSC were investigated, and it was confirmed that erythritol, lactitol, and maltitol can inhibit sulfur gas and methyl mercaptan independently without pH. The production. In addition, for the main cause of periodontal disease, the genus Porphyromonas gingivalis (?〇^11丫1'011101133.§丨1^卜31丨3) only needs 1% concentration of lactitol, maltose The alcohol can reduce the amount of hydrogen sulfide generated to about 40 to 60%. 效果 Effect of the invention The present invention is an anthraquinone-containing, toothpaste, and inhalant because it can exhibit a very significant inhibition of the production of volatile sulfur compounds. Preparations such as tablets and lozenges can also be used as chewing gum, candy, candy tablets, gum fructose, biscuits, chocolate and other confectionery, ice products, beverages, etc., which are used and ingested daily, and are extremely effective for improving bad breath and prevention. [Embodiment] Embodiments of the present invention One embodiment of the present invention is a bad breath removing agent containing a sugar alcohol as an active ingredient. According to another embodiment of the present invention, the above-mentioned sugar alcohol is lactose, and the bad breath removing agent according to the above-mentioned ~ 9 - 201116272 is used. Another embodiment of the present invention is a tanning agent, a toothpaste, an inhalant, and a lozenge which are formed by a bad breath removing agent containing a sugar alcohol as an active ingredient. Still another embodiment of the present invention is a food product obtained by using a bad breath removing agent containing a sugar alcohol as an active ingredient. Still another embodiment of the present invention is a food containing a chewing gum, a candy, a candy lozenge, a fructose, a biscuit, a chocolate, or the like, an ice product, a drink, or the like, which comprises a bad breath removing agent containing a sugar alcohol as an active ingredient. Another embodiment of the present invention is a sugar alcohol as an active ingredient which can be used as an inhibitor for the production of volatile sulfur compounds by methionine and bovine cysteine metabolism. According to still another embodiment of the present invention, the sugar alcohol is lactitol or maltitol as described above, and the volatile sulfur compound is produced as an inhibitor. The invention is illustrated in detail by the following examples, but the invention is not limited thereto. (Example 1) A saliva culture test of sugar alcohols was carried out as follows. 1 _ 1 ·Preparation of samples As a sample, xylitol, erythritol, lactitol, maltitol, and sucrose were used. These were dissolved in deionized water to make them into an appropriate concentration, and used as a sample solution. -10- 201116272 1 - 2 . Saliva culture test Four healthy adults (A, B, C, D, mean age 32_0 years old) were selected as subjects, and the test system was implemented in two consecutive sessions. Between 9:00 and 9:30 in the morning ‘ Take 1 〇ml of non-irritating saliva from each testee (take no breakfast on the day ‘and don’t brush your teeth)’ Take the saliva you have saved on the ice. A 0.5 m 1 sample solution was added to 1.0 m 1 of non-irritated saliva, and cultured at 37 ° C for 23 hours. After 23 hours of cultivation, the mesh was placed on ice. Gc (Gas Chromatography) The sample was shaken at 37 °C for 5 minutes before the analysis, and an appropriate amount of (canned) headspace gas was taken with a syringe for GC analysis. In addition, the test under neutral conditions was added to a potassium phosphate buffer to a final concentration of 20 mM, and the reaction solution was maintained in the neutral field. 1 - 3 . G C analysis All of the GC analysis systems used gC689® manufactured by Agilent. The analysis column system uses HP-PL〇TQ (30mx0.53mmx40gm), initial temperature: 70°C/2.5min, temperature rise: 30°C/min, final temperature: 190 °C /3.5min, injection temperature: 2〇 Hey. 〇, tester: FpD, detector temperature: 200 ° C, flow rate: 2 〇 ml / min for analysis. The RT-based hydrogen sulfide is 1 · 1 min, the methyl mercaptan is 4 · 0 m i n , the dimethyl-sulfide is 5 · 6 m 1 η, and the - methyl disulfide is 8 3 m 丨 η. The average enthalpy was calculated by continuously performing two samples for all the samples. 1 - 4. Results 1. Effect of sucrose
C -11 - 201116272 在上述唾液培養試驗中，添加蔗糖、使其最後濃度爲 0.057、0.114%。結果最後濃度爲0.057%時，pH値並未變 ，VSC之產生亦幾乎無任何變化，惟最後濃度爲〇.1 14%時 ，pH値爲5.6，偏向酸性，亦抑制硫化氫♦甲基硫醇之產 生，將此等結果示於圖1。 另外，在反應液中加入磷酸緩衝劑進行培養時，即使 蔗糖之最後濃度爲〇. 1 1 4%，反應液之pH値還可以保持於中 性附近，相反地，硫化氫•甲基硫醇之產生量反而增加。 由此可知，蔗糖之VSC抑制效果係受到pH値之影響者 。中性條件下，有關添加蔗糖可以使VSC之產生量提昇， 根據推測應爲蔗糖可以成爲口腔內細菌的營養來源，而可 以增加口臭原因菌所致。據測在沒有緩衝劑的條件下，雖 可藉由蔗糖增加口腔內細菌，但因蔗糖會被滋養化、乳酸 等被排出，降低pH値，會阻礙到甲硫胺酸酶活性、牛胱胺 酸代謝酶活性、或許口臭原因菌會被抑制其增殖而可以抑 制V S C之產生》 2.木糖醇之效果 有關木糖醇，在此於圖2示其以最後濃度1 4 3 %、5 7 % 濃度進行上述唾液保溫試驗之結果。使用木糖醇之此試驗 的全部評審員中針對硫化氫•甲基硫醇之產生量、及反應 液之pH値均未出現有極大之變化。 3.赤蘚糖醇之效果 -12- 201116272 有關赤蘚糖醇係如圖3所示’添加赤蘚糖醇亦對反應 液之p Η値有任何影響，硫化氫·甲基硫醇之產生量係依其 添加濃度之不同而被抑制。 4.乳糖醇之效果 有關乳糖醇係如圖4 - 1所示，會因添加乳糖醇而有反 應液之pH値不變者（4人中2人）、與會降低PH値者（4人 中2人）。可是每一個評審員均抑制了硫化氫•甲基硫醇 之產生量。 又，如圖4-2所示，中性條件中’經確認乳糖醇之效 果時，確認在中性條件下亦可抑制甲基硫醇產生。 5 .麥芽糖醇之效果 有關麥芽糖醇係有即使添加麥芽糖醇有反應液之PH値 不變之人（4人中1人），與pH値會降低（4人中3人）者。 如圖5 -1所示，全部評審員係即使添加麥芽糖醇亦幾乎未 見到硫化氫之產生量上的變化。另一方面，藉由添加麥芽 糖醇可抑制甲基硫醇產生。中性條件下係如圖5-2所示’ 雖可見到硫化氫產生量上昇之傾向，但甲基硫醇卻被抑制 其產生。 由以上結果可知，在試管內試驗系中獲知，木糖醇對 VSC之產生不會有不佳之影響。另，有關乳糖醇•麥芽糖 醇·赤蘚糖醇則獲知可以pH値非依賴性下抑制VSC之產生 。尤其有關乳糖醇係其抑制效果比赤蘚糖醇更強。 -13- 201116272 (實施例2 ) 依如下進行糖醇類的菌代謝阻礙試驗。 2-1.準備菌液 菌株係使用具核梭桿菌（F.nucleatum)與牙齦卟啉單 胞菌（P.gingivalis)。核梭桿菌係在含0.05 % L -牛腕胺酸 之3 %THB培養基中，嫌氣性地培養1天。培養1天後，確認 5 5 0nm時之吸光度爲0.8以上時，將5000rpm經4分鐘遠離之 上澄液丟棄。懸濁菌體於生理食.鹽水中，再次施行同樣之 操作，將所得菌體懸濁於2倍量原來菌液的生理食鹽水中 ，一邊冰冷同時供予試驗。 牙齦卟啉單胞菌係以TSB培養基（3%Tripticase豆肉湯 0.3%酵母、0.0005%高鐵血紅素、0.00005%維生素K)中 以嫌氣性下培養1天。培養1天後，確認5 50nm下之吸光度 爲1.4以上，以上述一樣之方法調製菌液。 2-2.甲硫胺酸代謝經路阻礙試驗C -11 - 201116272 In the above saliva culture test, sucrose was added to give a final concentration of 0.057 and 0.114%. At the final concentration of 0.057%, the pH値 did not change, and the VSC production did not change at all. However, when the final concentration was 〇.1 14%, the pH値 was 5.6, which was acidic, and also inhibited hydrogen sulfide. The production of alcohol is shown in Figure 1. Further, when a phosphate buffer is added to the reaction solution for cultivation, even if the final concentration of sucrose is 0.11%, the pH of the reaction solution can be maintained near neutral, and conversely, hydrogen sulfide/methyl mercaptan The amount of production increases. From this, it can be seen that the VSC inhibitory effect of sucrose is affected by pH 値. Under neutral conditions, the addition of sucrose can increase the production of VSC. It is speculated that sucrose can be a nutrient source for bacteria in the mouth and can increase the cause of bad breath. It has been measured that in the absence of a buffer, although sucrose can increase bacteria in the mouth, sucrose is sterilized, lactic acid, etc. are discharged, lowering pH 値, hindering methionase activity, bovine cystamine Acid metabolism enzyme activity, perhaps bad breath cause bacteria can be inhibited from proliferation and can inhibit the production of VSC. 2. The effect of xylitol is related to xylitol, which is shown in Fig. 2 as the final concentration of 143 %, 5 7 The % concentration was the result of the above saliva holding test. There was no significant change in the amount of hydrogen sulfide/methyl mercaptan produced and the pH of the reaction solution among all the judges of this test using xylitol. 3. Effect of erythritol-12- 201116272 About erythritol series As shown in Figure 3, 'addition of erythritol also has any effect on the p Η値 of the reaction solution, and the production of hydrogen sulfide·methyl thiol The amount is suppressed depending on the concentration to which it is added. 4. Effect of lactitol The lactose alcohol is shown in Figure 4-1. The pH of the reaction solution is unchanged due to the addition of lactitol (2 out of 4), and the decrease in PH (4 persons) 2 people). However, each reviewer suppressed the amount of hydrogen sulfide and methyl mercaptan produced. Further, as shown in Fig. 4-2, when the effect of lactitol was confirmed in the neutral condition, it was confirmed that methyl mercaptan production can be suppressed under neutral conditions. 5. Effect of maltitol The maltitol is a person (one of four persons) having a pH of the reaction solution even if maltitol is added, and the pH is lowered (three of four people). As shown in Fig. 5-1, all the reviewers showed little change in the amount of hydrogen sulfide produced even when maltitol was added. On the other hand, methyl mercaptan production can be suppressed by adding maltitol. Under neutral conditions, as shown in Figure 5-2, although the tendency of hydrogen sulfide production to increase is observed, methyl mercaptan is inhibited from being produced. From the above results, it was found that xylitol did not have a bad influence on the production of VSC in the in-tube test system. In addition, it is known that lactitol, maltitol, and erythritol inhibit the production of VSC at a pH-independent manner. In particular, the lactose alcohol has a stronger inhibitory effect than erythritol. -13-201116272 (Example 2) A bacterial metabolism inhibition test of sugar alcohols was carried out as follows. 2-1. Preparation of bacterial strains The strains used F. nucleatum and P. gingivalis. The Fusobacterium nucleatum strain was cultured in a 3% medium in THB medium containing 0.05% L-bovine glycosaminoglycan for 1 day. After 1 day of culture, when it was confirmed that the absorbance at 550 nm was 0.8 or more, the supernatant was separated from the supernatant at 5000 rpm for 4 minutes. The suspension cells were again subjected to the same operation in physiological saline solution, and the obtained cells were suspended in physiological saline solution of the original bacterial solution twice, and were subjected to ice-cooling while being supplied for the test. The P. gingivalis strain was cultured for 1 day under anaerobic conditions in TSB medium (3% Tripticase bean broth 0.3% yeast, 0.0005% methemoglobin, 0.00005% vitamin K). After 1 day of culture, it was confirmed that the absorbance at 5 50 nm was 1.4 or more, and the bacterial solution was prepared in the same manner as above. 2-2. Methionine Metabolism Test
在試管中放入2.47ml 0.1M磷酸緩衝液（pH値6.5)與 0.03 ml被試驗液。以混合氣體（氮：水：二氧化碳=8 : 1 :1 )取代頂部空間後，以矽栓密封、攪拌，在3 7 °C之水 浴中保溫。經5分鐘後以結核菌素注射器注入〇.2ml菌液， 經攪拌並保溫。經5分鐘後，以結核菌素注射器注入〇.3ml L-甲硫胺酸溶液（〇 · 5 % )、經攪拌，於3 7 °C保溫1 〇分鐘。 抽取5 00μ1頂部空間氣體，以GC分析測定甲基硫醇量。GC •14- 201116272 分析係如上述1-3—樣進行。 2-3 _牛胱胺酸代謝經路阻礙試驗 除了基質使用L-牛胱胺酸以外，其他則均與上述甲硫 胺酸代謝路阻礙試驗一樣之方法進行牛胱胺酸代謝經路阻 礙試驗。 2-4.結果 1.甲硫胺酸代謝經路阻礙試驗 確認糖醇及蔗糖對具核梭桿菌及牙齦卟啉單胞菌甲硫 胺酸代謝經路之阻礙活性的結果示於圖6-1之（a ) 、( b )。對於具核梭桿菌之甲硫胺酸代謝經路係正控制之氯化 鋅係最終lOOppm可以使甲基硫醇之產生量減少至40% (以 1 00%控制而言）。另一方面每一種糖醇均可以在最後1 0% 之濃度能夠控制甲基硫醇之產生量至8 0 %左右，雖不會很 強，但可確認具有甲硫胺酸代謝阻礙活性。另外，對牙齦 卟啉單胞菌的甲硫胺酸代謝經路則正控制之氯化鋅可以將 甲基硫醇之產生量控制至23 %爲止。又，乳糖醇、麥芽糖 醇雖不會很強，但仍可以抑制甲基硫醇之產生量至80〜90 % 左右，惟木糖醇、赤蘚糖醇則無法確認有明確之阻礙效果 2·牛胱胺酸之代謝經路阻礙試驗 對具核梭桿菌及牙齦卟啉單胞菌牛胱胺酸代謝經路， -15- 201116272 確認糖醇及蔗糖之阻礙活性結果示於圖6-2之（c ) 、 ( d )° 對於具核梭桿菌牛胱胺酸代謝經路係將木糖醇、赤蘚 糖醇、乳糖醇、麥芽糖醇以最後1 〇%濃度使硫化氫之產生 量減少至約60〜80%左右。有關蔗糖係將硫化氫之產生量抑 制至約40% »又，對牙齦卟啉單胞菌牛胱胺酸代謝經路係 使乳糖醇、麥芽糖醇、蔗糖爲最後濃度1 〇%，減少硫化氫 之產生量至約40〜60 %。另一方面，有關木糖醇、赤蘚糖醇 則未見到有明顯之效果。 由以上結果可知，對具核梭桿菌而言係木糖醇、赤蘚 糖醇、乳糖醇、麥芽糖醇係雖然不會很強，但仍可阻礙甲 硫胺酸、牛胱胺酸之二種的代謝經路。 另一方面，對牙齦卟啉單胞菌係只有乳糖醇與麥芽糖 醇可以強烈地阻礙牛胱胺酸之代謝經路。因此由這個現象 可知，糖醇之口臭抑制作用之一機制係有可能來自牛胱胺 酸·甲硫胺酸代謝阻礙效果的可能性。 以下製造含有本發明之口臭除去劑之含漱劑、牙膏、 口臭噴霧劑、錠劑、口香糖、糖果、糖果錠片、膠菓糖、 飲料。以下示此等之處方。又，藉由此等，本發明品之範 圍並不受此等所限制。 (實施例3 ) 依以下處方製造含漱劑。 -16- 201116272 乙醇 2.0重量％ 乳糖醇 10.0 香料 1.0 水 其餘 100.0 (實施例4 ) 依以下處方製造牙膏。 碳酸鈣 4 0.0重量％ 甘油 10.0 麥芽糖醇 20.0 羧甲基纖維素 2.0 月桂基硫酸納 2.0 香料 1 .0 糖精 0.1 Chlorohexigiline 0.0 1 水 其餘 100.0 (實施例5 ) 依以下處方製造口臭噴霧劑。 r~· -17- 201116272 乙醇 1 0.0重量％ 甘油 5.0 乳糖醇 10.0 麥芽糖醇 10.0 香料 0.05 著色劑 0.00 1 水 其餘 100.0 (實施例6 ) 依以下處方製造旋劑（trouche) 麥芽糖醇 7 2.3重量％ 木糖醇 20.0 ***膠 6.0 香料 1.0 單氟磷酸鈉 0.7 100.0 (實施例7 ) 依以下處方製造口香糖。 -18- 201116272 膠質料 2 0，0重量％ 麥芽糖醇 45.0 乳糖醇 33.0 香料 2.0 (實施例8 ) 100.0 依以下處方製造糖果。 麥芽糖醇 5 0.0重量％ 還原糖膠 34.0 檸檬酸 2.0 香料 0.2 水 其餘 (實施例9 ) 100.0 依以下處方製造糖果錠片。 麥芽糖醇 7 6 . 1重量％ 乳糖醇 19.0 蔗糖脂肪酸酯 0.2 香料 0.2 水 4.5 100.02.47 ml of 0.1 M phosphate buffer (pH 値 6.5) and 0.03 ml of the test solution were placed in a test tube. After replacing the headspace with a mixed gas (nitrogen: water: carbon dioxide = 8:1:1), it was sealed with a hydrazine plug, stirred, and incubated in a water bath at 37 °C. After 5 minutes, 2 ml of the bacterial solution was injected with a tuberculin syringe, stirred and kept warm. After 5 minutes, a solution of 3 ml of L-methionine solution (〇 · 5 %) was injected with a tuberculin syringe, stirred, and incubated at 37 ° C for 1 〇 minutes. A 500 11 headspace gas was taken and the amount of methyl mercaptan was determined by GC analysis. GC • 14- 201116272 The analysis was performed as described in 1-3 above. 2-3 _ bovine cysteic acid metabolism through the road obstruction test, except for the use of L-bovine cystine, the other methods are the same as the above-mentioned methionine metabolic pathway inhibition test for bovine cysteine metabolism pathway inhibition test . 2-4. Results 1. Methionine Metabolism The results of the obstruction activity of sugar alcohol and sucrose on the metabolic pathway of hemophilic acid of Fusobacterium nucleatum and P. gingivalis were shown in Figure 6 - 1 (a), (b). For the methionine metabolism of Fusobacterium nucleatum, the final zinc oxide of the zinc chloride system controlled by the path system can reduce the production of methyl mercaptan to 40% (in terms of 100% control). On the other hand, each of the sugar alcohols can control the amount of methyl mercaptan production to about 80% at the last 10% concentration, and although it is not strong, it can be confirmed to have a methionine metabolism inhibitory activity. In addition, zinc chloride, which is controlled by the methionine metabolism of P. gingivalis, can control the production of methyl mercaptan to 23%. Moreover, although lactitol and maltitol are not strong, the amount of methyl mercaptan can be suppressed to about 80 to 90%, but xylitol and erythritol cannot be confirmed to have a clear inhibitory effect. The metabolism of bovine cysteine was tested by the road obstruction test for the metabolism of Fusarium oxysporum and Porphyromonas gingivalis bovine cysteine, -15- 201116272. The results of inhibition of sugar alcohol and sucrose are shown in Figure 6-2. (c), (d)° Reduce the amount of hydrogen sulfide produced by the concentration of xylitol, erythritol, lactitol, and maltitol in the final concentration of serotonin About 60~80%. The sucrose system inhibits the production of hydrogen sulphide to about 40%. »In addition, the metabolism of carnosic acid to Porphyromonas gingivalis makes lactitol, maltitol and sucrose the final concentration of 1%, reducing hydrogen sulfide. The amount produced is about 40 to 60%. On the other hand, no significant effects were observed regarding xylitol or erythritol. From the above results, it can be seen that, for the genus Fusobacterium, xylitol, erythritol, lactitol, and maltitol are not strong, but they can still hinder methionine and bovine cysteine. Metabolic pathway. On the other hand, for the P. gingivalis strain, only lactitol and maltitol can strongly hinder the metabolism of bovine cysteine. Therefore, it is known from this phenomenon that one of the mechanisms of the bad breath inhibition of sugar alcohols is likely to be derived from the metabolic inhibitory effect of bovine cysteine and methionine. The gargle, toothpaste, bad breath spray, lozenge, chewing gum, candy, candy lozenge, gum fructose, and beverage containing the bad breath removing agent of the present invention are produced as follows. This is shown below. Further, the scope of the present invention is not limited by these. (Example 3) An expectorant was prepared according to the following prescription. -16- 201116272 Ethanol 2.0% by weight Lactitol 10.0 Perfume 1.0 Water The remaining 100.0 (Example 4) A toothpaste was prepared according to the following prescription. Calcium carbonate 4 0.0% by weight Glycerin 10.0 Maltitol 20.0 Carboxymethylcellulose 2.0 Sodium lauryl sulfate 2.0 Perfume 1 .0 Saccharin 0.1 Chlorohexigiline 0.0 1 Water The remaining 100.0 (Example 5) A bad breath spray was prepared according to the following formulation. r~· -17- 201116272 Ethanol 1 0.0% by weight Glycerin 5.0 Lactitol 10.0 Maltitol 10.0 Perfume 0.05 Colorant 0.00 1 Water remaining 100.0 (Example 6) Rotating agent (trouche) Maltitol 7 2.3 wt% wood Sugar alcohol 20.0 Acacia 6.0 Fragrance 1.0 Sodium monofluorophosphate 0.7 100.0 (Example 7) A chewing gum was prepared according to the following formulation. -18- 201116272 Glue 2 0,0 wt% Maltitol 45.0 Lactitol 33.0 Perfume 2.0 (Example 8) 100.0 Confectionery was made according to the following prescription. Maltitol 5 0.0% by weight Reducing gum 34.0 Citric acid 2.0 Perfume 0.2 Water Rest (Example 9) 100.0 A candy lozenge was prepared according to the following formula. Maltitol 7 6 . 1% by weight Lactitol 19.0 Sucrose fatty acid ester 0.2 Perfume 0.2 Water 4.5 100.0
C -19 - 201116272 (實施例1 〇) 依以下處方製造膠萬_。 明膠 6〇.〇重量％ 還原糖膠 2 1.40 麥芽糖醇 H.5 植物油脂 4.5 蘋果酸 2.0 香料 Λ π 1 〇〇. 〇 (實施例1 1 ) 依以下處方製造飲料。 橘子果汁 乳糖醇 檸檬酸 維生素C 香料 水 3 0. 〇重量％ 15.0 0.1 0.04 0.1 一其餘 100.0 產業上之可利用性 配合有本發明之糖醇的口臭去除抱丨 」吳太除劑係可適用於口香糖 糖果、糖果淀片4者，更可適用於除去口臭之特定保健 -20- 201116272 用食品者。 【圖式簡單說明】 圖1 :表示對產生蔗糖之VSC的影響圖。 圖2:表示對產生木搪醇之VSC的影響圖。 圖3:表示對產生赤蘚糖醇之VSC的影響圖。 圖4-1 :表示對產生乳糖醇之VSC的影響圖。 圖4-2 :表示對於在中性條件下產生乳糖醇之VSC的影 響圖。 圖5-1:表示對產生麥芽糖醇之VSC的影響圖° 圖5-2 :表示在中性條件下對產生麥芽糖醇之VSC的影 響圖。 圖6-1:表示糖醇類的菌代謝阻礙效果圖。 圖6-2 :表示糖醇類的菌代謝阻礙效果圖。 -21 -C -19 - 201116272 (Example 1 〇) The rubber _ is manufactured according to the following prescription. Gelatin 6 〇. 〇% by weight Reducing saccharide 2 1.40 Maltitol H.5 Vegetable oil 4.5 Malic acid 2.0 Perfume Λ π 1 〇〇. 〇 (Example 1 1 ) A beverage was prepared according to the following prescription. Orange juice, lactitol, citrate, vitamin C, perfume water, water, water, water, tea, water, tea, tea, tea, tea, tea, tea, tea, tea, tea, tea, tea, tea, tea, tea, tea, tea, tea, tea, tea, tea, tea, tea, tea, tea, tea Chewing gum candy, candy icing 4, can be more suitable for the removal of bad breath specific health -20- 201116272 food use. BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a graph showing the effect on VSC producing sucrose. Figure 2: shows the effect on the VSC producing xylitol. Figure 3: Diagram showing the effect on VSC producing erythritol. Figure 4-1: Diagram showing the effect on VSC producing lactitol. Figure 4-2: shows the effect of VSC on the production of lactitol under neutral conditions. Figure 5-1: Diagram showing the effect on VSC producing maltitol. Figure 5-2: Diagram showing the effect on VSC producing maltitol under neutral conditions. Fig. 6-1: Fig. 6-1 showing the effect of inhibition of the metabolism of sugar alcohols. Fig. 6-2 is a graph showing the effect of inhibition of the metabolism of sugar alcohols. -twenty one -