TWI733980B - 一種具有增強免疫力功能的粉劑及其製備方法 - Google Patents
一種具有增強免疫力功能的粉劑及其製備方法 Download PDFInfo
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- TWI733980B TWI733980B TW107103385A TW107103385A TWI733980B TW I733980 B TWI733980 B TW I733980B TW 107103385 A TW107103385 A TW 107103385A TW 107103385 A TW107103385 A TW 107103385A TW I733980 B TWI733980 B TW I733980B
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- poria
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- licorice
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Abstract
本發明屬於保健品領域,公開了一種粉劑,係由菊粉、靈芝、薏苡仁、茯苓、白朮、菟絲子、黃精、麥冬、甘草、枸杞子、五味子和甜菊糖苷製備而成。本發明所述粉劑的原料全部來源於天然中草藥,無輔料添加,成分天然,標籤清潔,符合人們追求食品天然健康的理念趨勢,且服用劑量小,服食方便,可溶於水中服用,冷水可溶,吸收快。本發明所述粉劑製備方法簡單,適合大規模生產攜帶方便,製得的粉劑穩定性好,存放時間長,口感好,入口即化,冷水可溶。實驗表明本發明所述粉劑具有增強免疫力作用,效果顯著,可用於製備具有增強免疫力功能的保健食品。
Description
本發明屬於保健品領域,具體涉及一種具有增強免疫力功能的粉劑及其製備方法,尤其是以天然中草藥為主要原料製備的具有增強免疫力功能的粉劑及其製備方法。
免疫力是人體自身的防禦機制,是人體識別和消滅外來侵入的任何異物(病毒、細菌等)。處理衰老、損傷、死亡、變性的自身細胞以及識別和處理體內突變細胞和病毒感染細胞的能力。隨著現代生活節奏的加快、工作壓力的不斷加大、飲食不均衡、睡眠不足、運動不夠......都容易讓人免疫力低下。免疫力低下的身體易於被感染或患癌症,各種原因使免疫系統不能正常發揮保護作用,在此情況下,極易招致細菌、病毒、真菌等感染,因此免疫力低下最直接的表現就是容易生病,因經常患病,加重了機體的消耗,所以一般有體質虛弱、營養不良、精神萎靡、疲乏無力、食欲降低、睡眠障礙等表現。生病、打針、吃藥,便成了家常便飯,每次生病都要很長時間才能恢復,而且常常反復發作,如感冒反復發作、扁桃體炎反復發作、哮喘反復發作、支氣管炎反復發作、肺炎反復發作、腹瀉反復發作,長此以往會導致身體和智力發育不良,還易誘發重大疾病。
近年來人民生活水準明顯提高人們的消費觀念、健康觀念發生了較大變化。為規避不健康帶來的各種不利影響,使人們越來越重視營養保健品的使用。目前提高免疫力類保健食品主要以補充維生素和礦物質為主,大多以口服液的形態存在。口服液劑攜帶不便,穩定性差,存放時間短。片劑在壓片時大都需要加入澱粉、羥甲基纖維素等大劑量輔料,崩解時間長。同時對吞咽有困難的人群服用不方便,如老人和兒童。
本產品採用常規乾燥工藝製成粉劑,攜帶方便,穩定性好,存放時間
長,服用劑量小,可直接口服和溶於水中服用,吸收快,無輔料添加,成分天然,全部來源於植物草本,標籤清潔,符合人們追求食品天然健康的理念趨勢。
有鑑於此,本發明的目的在於針對習知技術存在的缺陷提供一種具有增強免疫力功能的粉劑,如保健粉劑及其製備方法。本發明所述粉劑以天然中草藥為主要原料製備,具有增強免疫力功能,不含輔料,成分天然。
為實現本發明的目的,本發明採用如下技術方案:一種具有增強免疫力功能的粉劑,由菊粉、靈芝、薏苡仁、茯苓、白朮、菟絲子、黃精、麥冬、甘草、枸杞子、五味子和甜菊糖苷製備而成。
進一步地,作為優選,所述菊粉、靈芝、薏苡仁、茯苓、白朮、菟絲子、黃精、麥冬、甘草、枸杞子、五味子和甜菊糖苷的質量比為(50~80):(10~30):(3~8):(2~8):(1~5):(1~5):(1~5):(0.5~1):(0.2~0.8):(0.2~0.8):(0.2~0.8):(0.2~0.8)。
在一些實施方案中,所述菊粉、靈芝、薏苡仁、茯苓、白朮、菟絲子、黃精、麥冬、甘草、枸杞子、五味子和甜菊糖苷的質量比為50:16:8:8:5:5:5:1:0.5:0.5:0.5:0.5。
在一些實施方案中,所述菊粉、靈芝、薏苡仁、茯苓、白朮、菟絲子、黃精、麥冬、甘草、枸杞子、五味子和甜菊糖苷的質量比為80:10:3:2:1:1:1:0.5:0.5:0.4:0.3:0.3。
在一些實施方案中,所述菊粉、靈芝、薏苡仁、茯苓、白朮、菟絲子、黃精、麥冬、甘草、枸杞子、五味子和甜菊糖苷的質量比為65:20:5:3:1.5:1.5:1:1:0.5:0.5:0.5:0.5。
本發明還提供了所述的粉劑的製備方法,將靈芝、薏苡仁、茯苓、白朮、菟絲子、黃精、麥冬、甘草、枸杞子和五味子加水提取,過濾後收集提取液,加入菊粉混合後乾燥,加入甜菊糖苷製粒,烘乾,整粒。
優選地,本發明所述的粉劑的製備方法中,所述加水煎煮提取為第一次加8-15倍的水,提取2-4小時,第二次加10-20倍的水提取2-5小時。
優選的,所述乾燥前還包括濃縮的步驟,所述濃縮為真空濃縮或反滲透濃縮。
進一步優選地,所述乾燥為噴霧乾燥、冷凍乾燥、帶式乾燥、微波乾燥、真空乾燥。
優選地,本發明所述製備方法中產品細微性控制在30至100目。
由上述技術方案可知,本發明提供了一種粉劑由菊粉、靈芝、薏苡仁、茯苓、白朮、菟絲子、黃精、麥冬、甘草、枸杞子、五味子和甜菊糖苷製備而成。本發明所述粉劑的原料全部來源於天然中草藥,無輔料添加,成分天然,標籤清潔,符合人們追求食品天然健康的理念趨勢,且服用劑量小,服食方便,可溶於水中服用,冷水可溶,吸收快。本發明所述粉劑製備方法簡單,適合大規模生產攜帶方便,製得的粉劑穩定性好,存放時間長。口感好,入口即化,冷水可溶。實驗表明本發明所述粉劑具有增強免疫力作用,效果顯著,可用於製備具有增強免疫力功能的保健食品。
為了更清楚地說明本發明實施例或習知技術中的技術方案,下面將對實施例或習知技術描述中所需要使用的附圖作簡單地介紹。
圖1示本發明所述保健粉劑的製備流程;圖2示受試樣品對ConA誘導淋巴細胞轉化的影響圖;圖3示受試樣品對抗體生成細胞的影響圖;圖4示受試樣品對NK細胞活性的影響圖;圖5示受試樣品對小鼠巨噬細胞吞噬率和吞噬指數的影響圖。
本發明公開了一種具有增強免疫力功能的粉劑及其製備方法。本領域具有通常知識者可以借鑒本文內容,適當改進工藝參數實現。特別需要指出的是,所有類似的替換和改動對本領域具有通常知識者來說是顯而易見的,它們都被視為包括在本發明。本發明的方法及產品已經通過較佳實施例進行了描述,相關具有通常知識者明顯能在不脫離本發明內容、精神和範圍內對本文所述的方法進行改動或適當變更與組合,來實現和應用本發明技術。
為了進一步理解本發明,下面將結合本發明實施例,對本發明實施例中的技術方案進行清楚、完整地描述,顯然,所描述的實施例僅僅是本發明一部分實施例,而不是全部的實施例。基於本發明中的實施例,本領域具有通常知識者在沒有做出創造性勞動前提下所獲得的所有其他實施例, 都屬於本發明保護的範圍。
如無特殊說明,本發明實施例中所涉及的試劑均為市售產品,均可以通過商業管道購買獲得。
配方:
製備方法:
將靈芝、薏苡仁、茯苓、白朮、菟絲子、黃精、麥冬、甘草、枸杞子、五味子投入提取罐,加入8-15倍純化水,煎煮提取2-4小時後過濾,再加入10-20倍純化水,煎煮提取2-5小時後過濾,合併兩次濾液,濃縮,然後加入菊粉進行噴霧乾燥或帶式乾燥或冷凍乾燥,得中草藥提取乾粉。將甜菊糖苷加入中草藥提取乾粉中,製粒,烘乾,整粒,產品細微性控制在30至100目。
配方:
製備方法:
將靈芝、薏苡仁、茯苓、白朮、菟絲子、黃精、麥冬、甘草、枸杞子、五味子投入提取罐,加入8-15倍純化水,煎煮提取2-4小時後過濾,再加入10-20倍純化水,煎煮提取2-5小時後過濾,合併兩次濾液,濃縮,然後加入菊粉進行噴霧乾燥或帶式乾燥或冷凍乾燥,得中草藥提取乾粉。將甜菊糖苷加入中草藥提取乾粉中,製粒,烘乾,整粒,產品細微性控制在30至100目。
配方:
製備方法:
將靈芝、薏苡仁、茯苓、白朮、菟絲子、黃精、麥冬、甘草、枸杞子、五味子投入提取罐,加入8-15倍純化水,煎煮提取2-4小時後過濾,再加入10-20倍純化水,煎煮提取2-5小時後過濾,合併兩次濾液,濃縮,然後加入菊粉進行噴霧乾燥或帶式乾燥或冷凍乾燥,得中草藥提取乾粉。將甜菊糖苷加入中草藥提取乾粉中,製粒,烘乾,整粒,產品細微性控制在30至100目。
增強免疫力功能判定分為細胞免疫功能、體液免疫功能、單核-巨噬細胞功能、NK細胞活性四個方面,只要任意兩個方面結果陽性,可判定受試樣品具有增強免疫力功能。
1、受試樣品:採用實施例3製得粉劑樣品。
2、試驗動物:近交系BALB/C小鼠,雄性,18-22g。購買後適應性飼養一周,按體重隨機分組,每組12只。
3、分組及受試樣品給予時間:
試驗動物共分4組,分別為3個劑量組和對照組(等量的蒸餾水)。3個劑量以人體推薦量的5倍(166.67mg/kg)、10倍(333.33mg/kg)和30倍(1000mg/kg)設置,每個劑量設三個重複。受試樣品給予時間為30天。
4、試驗方法及結果:
4.1 受試樣品對小試體重的影響
表1結果顯示,經口給予小鼠不同劑量供試樣品30天後,與對照組比 較,三個劑量組體重均無顯著差異(P>0.05),表明受試品安全性較好。
4.2 受試樣品對脾臟/體重比值的影響
表2結果顯示,經口給予小鼠不同劑量受試樣品30天後,與對照組比較,三個劑量組脾臟/體重比值均無顯著差異(P>0.05)。
4.3 ConA誘導的小鼠脾淋巴細胞轉化實驗(MTT法)
(1)脾細胞懸液製備:無菌取脾臟,Hanks液中放置,鑷子將脾臟剪碎,製成單個細胞懸液,過200目篩網過濾,Hanks液清洗2次,每次離心10分鐘(1000rpm/min),然後將細胞懸液懸浮於完全培養基長,台盼藍染色計算活細胞數後調整細胞密度為3×106個/mL。
(2)脾細胞增殖檢測:將每一份細胞懸液分成兩孔加入24孔培養板中,每孔1mL,一孔加入ConA溶液(終濃度7.5ug/mL),另一孔做對照。細胞培養箱內孵育培養72小時。MTT法檢測每孔的吸光度OD值。
用加ConA孔的OD值減去對照孔代表淋巴細胞的增殖能力,統計結果見圖2。
圖2結果顯示,經口給予小鼠不同劑量受試樣品30天後,與對照組相比,三個劑量組對ConA誘導淋巴細胞轉化均無顯著性差異(P>0.05),實驗結果陰性。
4.4 遲髮型變態反應(DTH)
二硝基氟苯誘導小鼠DTH(耳腫脹法)
(1)DNFB溶液(1%濃度的二硝基氟苯)配製;
(2)致敏:每只小鼠腹部用硫化鋇脫毛,範圍約3cm×3cm,用DNFB溶液50μL均勻塗抹致敏
(3)DTH產生與測定:5天後,用DNFB溶液均勻塗抹於小鼠右耳兩面)進行攻擊,攻擊後24小時頸椎脫臼處死,剪下左右耳殼,用打孔器取下直徑8mm的耳片,稱重。
結果判定:用左右耳重量之差表示DTH的程度,結果見表3。
表3結果顯示,與對照組相比,受試樣品三個劑量組左右耳重量差值均無顯著性差異(P>0.05),實驗結果陰性。
4.5 抗體生成細胞檢測(Jerne改良撥片法)
(1)脾細胞懸液製備:將SRBC免疫4-5天的小鼠頸椎脫臼處死,取出脾臟,製成脾細胞懸液,調整細胞密度為5×106個/mL。
(2)空斑測定:將表層培養基溶解後,45℃水浴,與等量的pH7.2的2倍濃度的Hanks溶液混合,分裝小試管,每管0.5mL。再向管內加50μL10%SRBC,20μL的脾細胞懸液,迅速混勻傾倒於已經刷上瓊脂糖薄層的玻片上,做平行片,待凝固後,將玻片水準扣放在玻片架凹槽內,繼續溫育1.5小時,統計溶血空斑數結果見圖3。
圖3結果顯示,經口給予小鼠不同劑量受試樣品30天後,與對照組相比,受試樣品可以提高脾臟細胞的抗體生成,其中高劑量組空斑數具有顯著性差異(P<0.05),實驗結果陽性。
4.6 NK細胞活性測定(乳酸脫氫酶測定法)
靶細胞的傳代(YAC-1細胞):實驗前將靶細胞進行傳代培養,調整細 胞狀態。
脾細胞懸液製備方法同4.3,台盼藍染色計數活細胞需要在95%以上。分離的脾細胞懸液調整濃度為2×107個/mL。
NK細胞活性檢測:取靶細胞和效應細胞各自100μL(效應細胞:靶細胞=50:1)加入到96孔培養板內。靶細胞自然釋放孔加靶細胞和培養液各100μL,靶細胞最大稀釋孔加靶細胞和1%NP40或2.5%Triton各100μL。上述各實驗組均設置3個複孔,細胞培養箱培養4個小時,然後將培養板進行低速離心5min,每孔吸取上清100微升轉至平底的96孔培養板內。利用LDH檢測試劑盒,490nm檢測每孔的OD值,每個實驗組取三個複孔的平均值,計算NK細胞活性,結果見圖4。其中NK細胞活性的計算公式如下:
NK細胞活性(%)=(反應孔OD值-自然釋放孔OD值)/(最大釋放孔OD值-自然釋放孔OD值)×100%。
圖4結果顯示,經口給予小鼠不同劑量受試樣品30天後,與對照組相比,受試樣品低劑量組NK細胞活性無顯著性差異(P>0.05),中、高兩個劑量組NK細胞活性均有顯著性差異(P<0.05),實驗結果陽性。
4.7 小鼠腹腔巨噬細胞吞噬棘紅細胞實驗(滴片法)
小鼠巨噬細胞的啟動:實驗前4天給每只小鼠腹腔注射2%壓積羊血紅細胞0.2mL。用頸椎脫臼法處死小鼠,腹腔注射加小牛血清的Hanks溶液4ml/只,輕輕按揉腹部20次,充分洗出腹腔巨噬細胞。然後腹腔壁剪開一個小口,用膠頭習慣吸取腹腔洗液2mL。用加樣器吸取0.5mL腹腔洗液加入到0.5mL1%雞血紅細胞懸液的試管內,混勻。吸取混合液0.5mL加到撥片的瓊脂圈內。放置細胞培養箱內孵育20min,孵育結束後迅速用生理鹽水將未貼壁的細胞洗掉,甲醛溶液固定1min,Giemsa染色15min。沖洗,晾乾,用40×顯微鏡統計吞噬率和吞噬指數。結果見圖5。油鏡下計數巨噬細胞,每張片計數100個,按照以下公式計算吞噬率和吞噬指數。
吞噬百分數(%)=吞噬雞血紅細胞的巨噬細胞數/計數的巨噬細胞數×100;吞噬指數=被吞噬的雞紅細胞總數/計數的巨噬細胞數
圖5結果顯示,經口給予小鼠不同劑量供試樣品30天後,與對照組相比,受試樣品低劑量組吞噬百分率及吞噬指數無顯著差異(P>0.05),中、 高劑量組吞噬百分率和吞噬指數均有顯著差異(P<0.05),實驗結果陽性。
5、結論
本實驗方案符合動物福利原則,實驗過程符合動物實驗條件要求,資料詳實,結果經統計學分析,得出結論如下:
5.1 小鼠脾淋巴細胞轉化實驗和遲髮型變態反應實驗結果均為陰性,判定細胞免疫功能試驗結果陰性。
5.2 抗體生成細胞檢測實驗結果為陽性,判定體液免疫功能試驗結果陽性。
5.3 小鼠腹腔巨噬細胞吞噬棘紅細胞實驗結果為陽性,判定單核-巨噬細胞功能試驗結果陽性。
5.4 乳酸脫氫酶測定實驗結果為陽性,判定NK細胞活性結果陽性。
在細胞免疫功能、體液免疫功能、單核-巨噬細胞功能及NK細胞活性四個方面測定中,本試驗結果顯示供試樣品在體液免疫功能中的抗體生成、腹腔巨噬細胞吞噬雞紅細胞、NK細胞活性中受檢樣品均可顯著提高各指標,且與對照組相比具有顯著性差異。
綜上所述,在本實驗室條件下,BALB/C小鼠分別經口灌胃連續給予蒸餾水、低、中、高不同劑量的實施例3製得粉劑溶液30天後,檢測結果表明實施例3製得粉劑具有增強免疫力功效。
實施例1和實施例2製得粉劑的效果與實施例3的效果相當。
Claims (7)
- 一種粉劑,其係由菊粉、靈芝、薏苡仁、茯苓、白朮、菟絲子、黃精、麥冬、甘草、枸杞子、五味子和甜菊糖苷製備而成,其中所述菊粉、靈芝、薏苡仁、茯苓、白朮、菟絲子、黃精、麥冬、甘草、枸杞子、五味子和甜菊糖苷的質量比為(50~80):(10~30):(3~8):(2~8):(1~5):(1~5):(1~5):(0.5~1):(0.2~0.8):(0.2~0.8):(0.2~0.8):(0.2~0.8);所述粉劑由如下方法製備:將靈芝、薏苡仁、茯苓、白朮、菟絲子、黃精、麥冬、甘草、枸杞子和五味子加水煎煮提取,過濾後收集提取液,加入菊粉混合後乾燥,加入甜菊糖苷製粒,烘乾,整粒;所述粉劑具有增強免疫的功能。
- 如請求項1所述的粉劑,其中所述菊粉、靈芝、薏苡仁、茯苓、白朮、菟絲子、黃精、麥冬、甘草、枸杞子、五味子和甜菊糖苷的質量比為50:16:8:8:5:5:5:1:0.5:0.5:0.5:0.5。
- 如請求項1所述的粉劑,其中所述菊粉、靈芝、薏苡仁、茯苓、白朮、菟絲子、黃精、麥冬、甘草、枸杞子、五味子和甜菊糖苷的質量比為80:10:3:2:1:1:1:0.5:0.5:0.4:0.3:0.3。
- 如請求項1所述的粉劑,其中所述菊粉、靈芝、薏苡仁、茯苓、白朮、菟絲子、黃精、麥冬、甘草、枸杞子、五味子和甜菊糖苷的質量比為65:20:5:3:1.5:1.5:1:1:0.5:0.5:0.5:0.5。
- 如請求項1所述的粉劑,其中所述方法中的加水煎煮提取為第一次加8-15倍的水,提取2-4小時,第二次加10-20倍的水提取2-5小時。
- 如請求項1所述的粉劑,其中所述方法中的乾燥之前還包括濃縮的步驟,所述濃縮為真空濃縮或反滲透濃縮。
- 如請求項1所述的粉劑,其中所述方法中的乾燥為噴霧乾燥、冷凍乾燥、帶式乾燥、微波乾燥、真空乾燥。
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