201114778 六、發明說明： 【發明所屬之技術領域】 本發明係關於從_與_栽培相關農業資材與其產物中萃取含 體物質之方法，及其可應用之相關領域。 【先前技術】 近年研究發現，多聽體具有許多生理活性，諸如調節免疫機能、抗炎 症、抗腫瘤活性、抗氧化、抗病毒、抗老化、清除自由基、解毒、降低膽 固醇等有心力效。由於多_之諸多有益功效，今常被做為有益添加物。 添加於食物、保健品中。常見之萃取多_的來源，大多從食、藥用葦類 的子實體或菌絲體中萃取取得，如：冬蟲夏草、靈芝等多種真_。 亦有研究指出可從經鶴培養走菌的培養基中萃取多醣體，如固態發酵培 養之基負、液態發酵培養之基質、太空包等培養基質。 萃取多_的方法，大多為熱水萃取法、鮮取法、酸驗萃取法等方 法；進-步可再藉由有機溶劑，如酒精等，進行沈殿純化。然而，該些萃 取法皆涉及高溫、高壓、高雜性之操作處理及設備，不但於製備過程有 工安之疑慮、且所需之產製設備，亦需有更高之維護等級，如耐高溫、耐 高壓、猶、祕^此外，若以酸驗萃取，將產生廢酸液、廢驗液，將 增加處理之難度，且亦有化學物質殘留之堪慮：若以鮮取法、熱水萃取 法’其所需加溫之水量極大’欲進行加溫時，將極費時、耗能、極具危險 性。此外，加熱大量之液體時’其熱穿透效率與熱均勻度將可能不足、不 均勻，皆將影響產品品質與萃取效率。 201114778 由上可知’上述製備多醣體之方法、來源仍具有諸多缺失，亟需加以改 良。本案發明人有鑑於此，乃玉思加以改良創新，研發完成本件「縣體、 其培養產物與培養紐巾萃取之含錄體物質、製備方法及其應用」。 【發明内容】 本發明之目的係在於提供-_便、環保、安全、具經濟效益之製備 含多膽體物質之方法、其產物及其各類應用。 • 本發明之次一目的係在於提供一種從、其培養產物與培養資材 (如：簟類子實體（細、益腳、菌絲體、已完成走菌之_培養基質或培 2液等)之再利财法、及其可應用之用途，如：可從中製備_酶體物 質等具經濟效益之主要產物、次級產物。 可達成上述發明目的之方法，包括有： 係從贿、_或已完歧菌之_培養基f等_農業資材與其產 物中，經磨碎 ' 高溫縫_、常溫萃取、·分料步驟後，萃取出含 鲁多種多醋體之物質。.該含多醣體物質可直接使用、或進一步加工後，作為 食品添加、保健品添加、醫藥品添加、或_補(如：作為純化某一種單 -多醋體之原物料、或作為製備改質之多醣體時的原物料)等適用用途。此 夕^於本發明製備過程中，所製得之次級產物，如：固態;慮摘、遽出廢 液等，可進一步回收再利用之。 【實施方式】 請參閱圖一所示，為本發明製備方法之流程圖。 本發明提供—種萃取含多賴物質㈣龄法，該方法包㈣下步驟： 201114778 步驟-.將摘資材與其產物’經壓磨破壞後可得一粗磨物，再將該粗磨 物以尚溫尚壓進行減菌； 步驟二：將步驟-之滅菌後粗磨物，加入、淋入或浸入常溫中性溶劑，在 室溫下麟混勻進行萃取，可得一萃取混合物； 步驟二.將步驟二之萃取混合物進行@液分離，即可製得含多醋體之粗萃 取液及固態濾出物。 其中於步驟-之破壞及滅菌步驟中，鋪紐與其產物係可由人工或 機械方式縣雌細麵、或魏適时式破壞之；其巾「高溫高壓滅菌」 係可由習狀高溫高壓滅、或其他_本發明之·方式，滅除其中 之有害微生物’確保後續製程之安全與潔淨。 其中於步驟二之萃取步驟令： a. 該常溫「中性溶劑」包含水、或可維持酸驗值為中性_6阳幻之 缓衝溶劑。 b. 在常溫下萃取，常溫包含介於代至35〇(：之溫度範圍，一般為a% 之室溫(正負差2〇C)。 c. 萃取過程中將粗磨物與中性溶劑混勻，係可由人力或機以娜 將之混勻。 丄萃取之大氣壓力可為常壓 '或可視需求調整。 e.萃取時間可介於10分鐘至12小時之間一般為3〇分鐘。 其中於步驟三之眺分離步射，該「醜分離」之方法包含但不限 於過渡、離心«關液分龄法。其中「磁」包含彻或結合不同孔 徑的慮網、渡布、絲、濾紙或細等適用濾材，或是卫業上常用之過渡 201114778 裝置如秒藻土磁等，渡除雜質而得到相對較純淨之慮液；「離心」包含利 用或結合—般離心機或連續式離心機，以獲得相對較純淨之上清液。 其中於步驟三可進-步絲萃取液以錢玉#加工方式，將之製成含 有多種多膽體之濃縮萃取液或乾燥粉劑，可供製成各種衍生製品(如^農縮 液體、粉末、濃縮氣態、噴霧等形式產品），或為特定多醣體之萃取原料來 源。6亥「加工方式」包含但不限於濃縮或嘴霧乾燥等適用本發明之方式。「濃 縮」泛指在食品或化料業上常用之濃縮方法，如加熱蒸發、減壓濃縮、 • 膜過渡、沉殺回溶、昇華或其他方法，濃縮比例可介於2倍錢。倍之間， —般以濃縮1G倍為佳。其巾若於步驟三將「粗萃取液」進H缩後可得 「濃縮萃取觀」及「渡出廢液」，該濃縮萃取濾液係指，相較於步驟二之 粗萃取液’單位濃度内具更高濃度之多膽體者：域出廢液包含減壓濃縮 之凝結水、含小分子廢液等’可回收再利用作為萃取溶劑、或其他適用之 用途。 其中於步驟三經固液分離後之固態渡出物，可如常回收作為堆肥、飼 _料、栽培介質、培養基質、嶋農業資材或燃料等應用。 本發明所稱「含多醣體物質」可包含經本發明方法製得之粗萃取液、 或其衍生物(如：濃縮萃取液、乾燥粉劑、固形物等形式），經測試確認該含 多醣體物質内富含多種多醣體，易於儲存與運輸，可以直接供使用者使用， 或亦可應用於各種食品、飼料或化妝品之製造。 前述多醣體測試包含多醣體含量測定、分子量測定等適用測試方法， "亥些方法包含苯酚-硫酸法、蒽酮(Anthr〇ne)反應法、高效能液相層析法 (HPLC)、凝膠滲逸層析(GPC)等方法。 201114778 本發明所述之「麵資材與其產物」包含但不限於各類富含多酶體之 摘資材與其產物，如：金針、舞益、白域、_、秀珍兹、 鮑魚益等之子實體(碰）’加工廢棄物如益腳、菌絲體、經過或已完成走菌 之基質。該「基質」包含各類適用菇類走菌之栽培基質、培養基質，如： 太空包、固態培養基、液態培養基等基質。 本發明係以下面的實施例予以示範闡明，但本發明不受下述實施例所 限制。此外’本發明所使用之材料、物有特別指明者，其餘皆為市售 易於取得之材料。 實施例金針菇多醣體萃取液的製備方法 新鮮金針賊腳5〇〇 g，先以手工或機器礙壓使益體細胞破碎，經 12PC高溫高壓滅菌後，加入〗，_址奶水於常溫下搜摔萃取分鐘， 使滅菌後之金針題_磨额水混合均勻。瓶合物㈣紗布過渡後， 以溏紙再次過渡，或聊啊離心後取上層液，得到粗萃取液·恤。 其中紗布可為八層紗布。 差JIM含蚩測宕及问牧率 取2 mL粗萃取液’經乾燥後秤重，最高可得到〇._5 g含多醜體固 形物1言之’即每·_鮮金針鶴腳可獲得含多雜固形物8.23g， 經換异回收率為1.65% (w/w，重量百分比）。 另取332 mL粗萃取液’經乾燥後科重，得到[心含多膽體固形物， 、。之即每5〇〇g新鮮金針紐腳可獲得含多酿體固形物3.心，經換算 回收率為〇_73% (w/w，重量百分比）。 201114778 -Λ 以獅·硫酿所製得含多__射含量，以葡萄糖 為標準品’結果如下所示： 凊參閱下表-及表二，多__物以相同比_溶後，平均㈣新 鮮金針兹益腳可沉殿0.27g多醣體。 表一以苯酴-硫酸法所測得之檢量線201114778 VI. Description of the Invention: TECHNICAL FIELD OF THE INVENTION The present invention relates to a method for extracting a body substance from ___ cultivation related agricultural materials and products thereof, and related fields in which it can be applied. [Prior Art] Recent studies have found that multi-hearing organisms have many physiological activities, such as regulating immune function, anti-inflammatory, anti-tumor activity, anti-oxidation, anti-virus, anti-aging, scavenging free radicals, detoxification, and reducing cholesterol. Due to the many beneficial effects, it is often used as a beneficial additive. Added to food and health products. The sources of common extractions are mostly extracted from the fruiting bodies or mycelium of food and medicinal mites, such as Cordyceps sinensis and Ganoderma lucidum. It has also been pointed out that polysaccharides can be extracted from the medium cultured by the crane, such as the base of solid-state fermentation, the substrate of liquid fermentation culture, and the culture medium of space pack. Most of the methods for extracting more _ are hot water extraction method, fresh extraction method, acid extraction method, etc.; further steps can be further purified by an organic solvent such as alcohol. However, these extraction methods involve high-temperature, high-pressure, high-hybrid operation processing and equipment, not only in the preparation process, but also in the production equipment, and also require a higher maintenance level, such as high temperature resistance. In addition, if it is extracted by acid, it will produce waste acid and waste liquid, which will increase the difficulty of treatment, and there are also chemical residues: if it is freshly taken, hot water is extracted. The law 'the amount of water it needs to warm is enormous' is extremely time consuming, energy intensive and extremely dangerous when it is desired to warm up. In addition, when heats a large amount of liquid, its heat penetration efficiency and heat uniformity may be insufficient and uneven, which will affect product quality and extraction efficiency. 201114778 It can be seen from the above that the above methods and sources for preparing polysaccharides still have many defects, and need to be improved. In view of this, the inventor of this case has made improvements and innovations, and has completed the research and development of this article "The recording body, preparation method and application of the county body, its culture products and culture towel extraction." SUMMARY OF THE INVENTION The object of the present invention is to provide a method for the preparation of a polycholinoid substance, a product thereof and various applications thereof, which are environmentally friendly, safe and economical. • The second object of the present invention is to provide a culture product and a culture material (for example, a scorpion fruit body (fine, beneficial foot, mycelium, finished _ culture medium or culture liquid 2, etc.) The re-profit method and its applicable applications, such as: the main products and secondary products that can be economically produced from the preparation of zymogen substances. The methods for achieving the above objects of the invention include: Or the _ medium f and so on _ agricultural materials and their products, after grinding 'high temperature seam _, normal temperature extraction, · material separation step, extract a variety of substances containing a variety of vinegar. The substance can be used directly, or further processed, as a food additive, a health supplement, a pharmaceutical addition, or a supplement (eg, as a raw material for purifying a single-polyphenol, or as a modified polysaccharide) In the preparation process of the present invention, the secondary products obtained, such as: solid; the waste, the waste liquid, etc., can be further recycled and reused. Please refer to Figure 1 for the invention. The present invention provides a method for extracting a medium containing a plurality of substances (four), and the method comprises the following steps: (4) The following steps: 201114778 Step--After crushing the material and its product, a coarse abrasive can be obtained after crushing, and then The coarse abrasive is sterilized by the temperature and pressure; Step 2: After the step-sterilization, the coarse abrasive is added, drenched or immersed in a normal temperature neutral solvent, and mixed at room temperature for extraction, thereby obtaining one Extracting the mixture; Step 2. The extraction mixture of the second step is subjected to @liquid separation to obtain a crude extract containing the polyacetate and the solid filtrate. Among the steps of the destruction and sterilization steps, the paving and its products It can be destroyed by artificial or mechanical method of female fine noodles or Wei Shishi; its towel "high temperature autoclaving" can be destroyed by high temperature and high pressure, or other methods of this invention, to eliminate harmful microorganisms in it' The process is safe and clean. The extraction step in step 2 is: a. The normal temperature "neutral solvent" contains water, or the acid can be maintained as a neutral _6 phantom buffer solvent. b. at room temperature Extraction, room temperature package Between generations up to 35 〇 (: temperature range, generally a% room temperature (positive and negative difference 2 〇 C). c. During the extraction process, the coarse grinding material is mixed with the neutral solvent, which can be manpower or machine Mix it. The atmospheric pressure of 丄 extraction can be adjusted to normal pressure' or visual demand. e. The extraction time can be between 10 minutes and 12 hours, usually 3 〇 minutes. The method of "ugly separation" includes, but is not limited to, transition, centrifugation, and liquid separation. Among them, "magnetic" includes a filter medium of different diameters, such as mesh, crossing cloth, silk, filter paper or fine, or The commonly used transition 201114778 device, such as second algae magnetic, etc., removes impurities to obtain relatively pure liquids; "centrifugation" involves the use or combination of a centrifuge or a continuous centrifuge to obtain relatively pure In the third step, the step-filament extract can be processed into a variety of multi-choledic concentrated extracts or dry powders by using Qianyu# processing method, which can be used to make various derivative products (such as Liquid, powder, concentrated gas, spray, etc.) Or extraction of raw materials for a specific source of the polysaccharide. 6Hai "Processing Method" includes, but is not limited to, a method of applying the present invention, such as concentration or mist drying. “Concentration” refers to the concentration methods commonly used in food or chemical industry, such as heating and evaporation, concentration under reduced pressure, • membrane transition, immersion, refining, sublimation or other methods. The concentration ratio can be between 2 times. Between times, it is better to concentrate 1G times. If the towel is in the third step, the "crude extract" is added to the H-shrinkage to obtain the "concentrated extraction view" and the "extracted waste liquid". The concentrated extract filtrate refers to the unit concentration of the crude extract as compared with the second step. Those with higher concentrations of biliary bodies: the effluent from the field contains condensed water concentrated under reduced pressure, and contains small molecule waste liquid, etc., which can be recycled and reused as an extraction solvent, or other suitable uses. The solid ferment after solid-liquid separation in step 3 can be recycled as a compost, feed, cultivation medium, culture medium, agricultural material or fuel. The "polysaccharide-containing substance" referred to in the present invention may comprise a crude extract obtained by the method of the present invention, or a derivative thereof (for example, a concentrated extract, a dry powder, a solid, etc.), and the polysaccharide-containing substance is confirmed by testing. It is rich in a variety of polysaccharides, easy to store and transport, can be used directly by users, or can be applied to the manufacture of various foods, feeds or cosmetics. The above polysaccharide test includes a suitable test method for determination of polysaccharide content, molecular weight measurement, etc., and some methods include phenol-sulfuric acid method, anthrone (Anthr〇ne) reaction method, high performance liquid chromatography (HPLC), and coagulation. Gel permeation chromatography (GPC) and other methods. 201114778 The "face material and its products" described in the present invention include, but are not limited to, various kinds of multi-enzyme-rich materials and products thereof, such as: gold needle, Wuyi, Baiyu, _, Xiuzhenz, abalone and the like ( Touch) 'Processing waste such as beneficial feet, mycelium, matrices that have passed or have been completed. The "matrix" comprises various cultivation substrates and culture substrates suitable for mushroom sterilization, such as: space capsules, solid medium, liquid medium and the like. The present invention is exemplified by the following examples, but the present invention is not limited by the following examples. Further, the materials and materials used in the present invention are specifically indicated, and the rest are commercially available materials which are easily available. EXAMPLES Preparation method of Flammulina velutipes polysaccharide extract Fresh golden needle thief foot 5〇〇g, firstly by manual or machine pressure to break the body cells, after 12PC high temperature autoclaving, add 〗 _, _ site milk at room temperature search After the extraction, the gold needles after sterilization are mixed uniformly. After the transition of the bottle (4) gauze, the paper is re-transitioned, or the supernatant is taken after centrifugation to obtain a crude extract/shirt. The gauze can be eight layers of gauze. Poor JIM contains 蚩 宕 问 问 问 问 问 问 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Containing 8.23 g of polyheterosolids, the recovery by conversion was 1.65% (w/w, weight percent). Another 332 mL of crude extract was taken to dry the body weight to obtain [heart containing multi-biliary solids, . That is, every 5 〇〇g fresh gold needles can obtain a multi-brew solids 3. Heart, and the converted recovery is 〇_73% (w/w, weight percentage). 201114778 -Λ The content of __ shots prepared with lion and sulphur is based on glucose. The results are as follows: 凊 Refer to the following table - and table 2, after __ things are dissolved in the same ratio _ (4) Fresh golden needles can be used to sink 0.27g polysaccharides. Table 1 Measured line measured by phenylhydrazine-sulfuric acid method
葡萄糖 490 run吸光值 3 平均值標準偏差 (g)/mL 1 2 0.001 0.077 0.058 0.002 0.220 0.214 0.003 0.365 0.373 0.004 0.508 0.495 0.005 0.543 0.562 0.006 0.335 0.334 0.007 0.636 0.685 0.008 0.746 0.744 0.009 1.023 1.014 0.010 0.963 0.952Glucose 490 run absorbance 3 mean standard deviation (g) / mL 1 2 0.001 0.077 0.058 0.002 0.220 0.214 0.003 0.365 0.373 0.004 0.508 0.495 0.005 0.543 0.562 0.006 0.335 0.334 0.007 0.636 0.685 0.008 0.746 0.744 0.009 1.023 1.014 0.010 0.963 0.952
〇^〇86~~0Ό74 〇]〇Ϊ4~ 0.189 0.208 〇.〇16 0.360 0.366 0.0〇7 0.377 0.460 0.072 0.516 0.540 〇.〇23 0.306 0.325 〇.〇16 0.691 0.671 0.030 0.708 0.733 〇〇21 1.002 1.013 〇.〇11 Q-938 0.95j_〇π13〇^〇86~~0Ό74 〇]〇Ϊ4~ 0.189 0.208 〇.〇16 0.360 0.366 0.0〇7 0.377 0.460 0.072 0.516 0.540 〇.〇23 0.306 0.325 〇.〇16 0.691 0.671 0.030 0.708 0.733 〇〇21 1.002 1.013 〇. 〇11 Q-938 0.95j_〇π13
表二以苯紛-硫酸法測量金針兹粗萃取液之多酿體含量 490 nm吸光值 萃取液批次Table 2 measures the multi-brew content of the crude extract of the golden needles by the benzene-sulfuric acid method. 490 nm absorbance value
A B C 平均 ----- 500 g新鮮菇腳 _平均值標準偏差之多醣含量㈦ 1.329 0.635 0.664 0.543 ~〇Ϊ86 — 0.455 0.102 0.694 0.417 0.298 〇2〇ί〇?Ι〇 0.692 0.605 0.654 0.650 0.27 2 因此，綜上所述，經本發明製備方法所得之粗萃取液富含多醣體約 佔多醣體固形重量的14%至19%。 201114778 本發明所提供之從菇類資材與其產物中萃取之含多醣體物質、製備方 法及其應用，與其他習用技術、產品相互比較時，更具有下列之優點. 1.本發明係以脑等農f諸與其產物作為原料，製備含多醋體物 質，其製借過程簡易、安全，且製備過程中無使用化學物質，因此，所妒 得之含多醣體物質，相較以往，使用者將無化學物質殘留之堪虞。 2. 本發明之細諸與其聽先行高溫高賊理，減除所有生物與可 能之病原微生物，有效確保後續製程之安全與潔淨。 3. 本發明於常溫下以中性溶劑萃取多酿體，取代習用之高溫高壓萃取 法，或熱水萃取法。伽有二為避免加熱大量賴時，其熱穿透效率 與熱均勻衫足崎響產；f與萃取效率；另_為避免製程中加溫量以 嘲計之液體，符合現今工業節能触、經濟與工安之要求。 4.本發明在常溫下以中性溶劑萃取多醣體，取代習用之利用酸驗的萃 取方法其優點為增加生產安全度，減少增設相關設備資本與生產成本、 及減）S邊萃取過程巾產生之舰液、祕液之產生，及後續之廢液處理。 • I仔之含多種多醣體物質，其應用形式廣，由於其無菌、 、’、他化子物貝殘留騎之可能’所製得者之含多醣體種類多，可直接供 使用者視，、而要使用之’如翻 '添加、物縣料 ' 塗佈、噴鮮各類適 用形式。 綜合上述，本發明之製備方法、所製備之含多_物質，相較以往， 本發明之製程將可實際為τ^ 為業s產顧，且崎得者顧形式廣、附加價 值高。 上列§羊細§兒明係針對本發明 之一可行貫施例之具體說明’惟該實施例 201114778 並非用、限制本I明之專利範圍，凡未脫離本發明技藝精神所為之等效實 施或變更，均銳含於本案之翻範圍中。 【圖式簡單說明】 圖一為本發明製備方法之流程圖。 【主要元件符號說明】 益 #»*> 【參考文獻】 L李志洲、揚海壽、閩鎖田。2003。金針菇多醣最佳提取工藝研究。寶雞 文理學院學報23: 38-41。ABC average ----- 500 g fresh mushroom feet _ average standard deviation of polysaccharide content (seven) 1.329 0.635 0.664 0.543 ~ 〇Ϊ86 — 0.455 0.102 0.694 0.417 0.298 〇2〇ί〇?Ι〇0.692 0.605 0.654 0.650 0.27 2 Therefore, In summary, the crude extract obtained by the preparation method of the present invention is rich in polysaccharides and accounts for about 14% to 19% of the solid weight of the polysaccharide. 201114778 The polysaccharide-containing material extracted from the mushroom material and the product thereof, the preparation method and the application thereof provided by the invention have the following advantages when compared with other conventional techniques and products. 1. The invention is based on the brain and the like. The agricultural products and their products are used as raw materials to prepare polyphenol-containing substances. The preparation process is simple and safe, and no chemical substances are used in the preparation process. Therefore, the polysaccharide-containing substances obtained by the user will be compared with the past. No chemical residue. 2. The invention is designed to reduce the safety and cleanliness of subsequent processes by eliminating the need for high temperature and high thief to reduce all biological and possible pathogenic microorganisms. 3. The present invention extracts a multi-brewed body at a normal temperature with a neutral solvent, replacing the conventional high temperature and high pressure extraction method or hot water extraction method. In order to avoid heating a large amount of heat, its heat penetration efficiency and heat uniformity are good; f and extraction efficiency; another _ to avoid the heating process in the process to ridicule the liquid, in line with today's industrial energy-saving touch, economy And the requirements of the work safety. 4. The invention extracts the polysaccharide body with a neutral solvent at normal temperature, and replaces the conventional extraction method using acid test, which has the advantages of increasing production safety, reducing the capital and production cost of the related equipment, and reducing the production of the S-side extraction process towel. The production of ship liquid, secret liquid, and subsequent waste liquid treatment. • I have a variety of polysaccharides, which have a wide range of applications. Because of their sterility, 'there is a possibility that the scorpion can be ridden, the polysaccharides can be used directly by the user. And to use the 'such as turn 'add, material county material' coating, spray various types of applicable forms. In summary, the preparation method of the present invention and the prepared multi-substance, compared with the prior art, the process of the present invention can be practically produced by τ^, and the sturdy person has a wide form and a high added value. The above is a detailed description of one of the possible embodiments of the present invention, which is not intended to limit the scope of the patents of the present invention, and is not equivalent to the equivalent of the spirit of the present invention. The changes are all in the scope of the case. BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a flow chart of a preparation method of the present invention. [Main component symbol description] Benefit #»*> [References] L Li Zhizhou, Yang Haishou, and Yusuotian. 2003. Study on the best extraction technology of Flammulina velutipes polysaccharides. Baoji University of Arts and Sciences, 23: 38-41.