TWI481351B - A yeast nutrition product and the preparation method thereof - Google Patents
A yeast nutrition product and the preparation method thereof Download PDFInfo
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Description
本發明涉及一種酵母營養食品及其製備方法。 The invention relates to a yeast nutritious food and a preparation method thereof.
酵母菌體是單細胞真核微生物,與香菇、木耳等同屬於菌藻類食物,其中含有較豐富的蛋白質和維生素、碳水化合物、脂肪、礦物質、核酸等多種營養成分。酵母菌中含有18種氨基酸,並且含人體必需的氨基酸,特別是在穀物蛋白中含量較少的賴氨酸,酵母菌體中含量較高,且氨基酸的比例接近聯合國糧農組織(FAO)推薦的較理想的氨基酸組成值,所以其營養價值較高。隨著世界人口的快速增長,食物缺乏日益顯著,尤其是蛋白質食品出現短缺。傳統的動植物蛋白質食品的生產受農業技術,及各類環境因數的限制,增長緩慢,難以滿足人類蛋白質消費的需求。酵母這種單細胞蛋白的出現開拓了獲取蛋白質的新領域。 Yeast cells are single-cell eukaryotic microorganisms, which are equivalent to mushrooms and fungus. They are rich in protein and vitamins, carbohydrates, fats, minerals, nucleic acids and other nutrients. Yeast contains 18 kinds of amino acids and contains essential amino acids, especially lysine which is less in cereal protein. The content of yeast is higher, and the ratio of amino acids is close to that recommended by FAO. The ideal amino acid composition value, so its nutritional value is higher. With the rapid growth of the world's population, food shortages are becoming more and more significant, especially in the shortage of protein foods. The production of traditional animal and plant protein foods is limited by agricultural technology and various environmental factors, and the growth is slow, which is difficult to meet the needs of human protein consumption. The emergence of this single-cell protein, yeast, opens up new areas of protein acquisition.
另外,酵母菌中含有豐富的維生素、礦物質以及多糖,能夠為人體提供更均衡的營養。並且酵母中脂肪含量很低,不含膽固醇,是低熱量、高營養的健康食品。 In addition, yeast is rich in vitamins, minerals and polysaccharides, which can provide a more balanced nutrition for the human body. And yeast has a low fat content and does not contain cholesterol. It is a low-calorie, high-nutrient health food.
由於酵母含有如此豐富的營養,因而使得用酵母來製備營養健康食品成為可能。目前,現有技術中以酵母為主要原料的營養食品僅包含少數幾種原料,酵母則僅採用酵母粉,並且製備工藝單一。例如專利“一種營養酵母食品及其製備方法”(200510015005.2),其所披露的營養酵母食品僅包括營養酵母 粉、全脂奶粉、β-環糊精、甜味劑、植脂末和其他添加劑,而製備該食品僅採用片劑生產工藝。因此,製備出包含多種營養成分、具有不同劑型、口感適合各種人群的酵母營養食品成為人們所關注的方向。 Since yeast contains such a rich nutrient, it is possible to prepare nutritious health foods using yeast. At present, the nutritious food with yeast as the main raw material in the prior art only contains a few raw materials, and the yeast uses only yeast powder, and the preparation process is single. For example, the patent "a nutritious yeast food and its preparation method" (200510015005.2), the nutrient yeast food disclosed therein only includes nutritional yeast Powder, whole milk powder, β-cyclodextrin, sweetener, non-dairy creamer and other additives, and the preparation of the food is only carried out using a tablet production process. Therefore, the preparation of yeast nutritious foods containing various nutrients, having different dosage forms, and having a mouthfeel suitable for various people has become a focus of attention.
為瞭解決現有技術中,酵母營養食品配方局限、製備工藝單一等問題,本發明提供了一種酵母營養食品及其製備方法。 In order to solve the problems in the prior art, the limitation of the yeast nutritious food formula, the single preparation process, and the like, the present invention provides a yeast nutritious food and a preparation method thereof.
本發明的一個方面,提供了一種酵母營養食品,該酵母營養食品,以酵母為主要原料,包括奶粉、麥芽糊精、乳糖、植脂末、碳酸鈣、澱粉、白砂糖以及矯味劑、香精,其特徵在於,所述酵母營養食品還包括B族維生素和富含微量元素的酵母,所述組分具有以下重量份:酵母5-97、奶粉1-20、植脂末1-50、碳酸鈣0.1-50、澱粉1-50、麥芽糊精0-50、乳糖0-50、白砂糖0-70、葉酸0-1、維生素B120-1維生素B10-10、 維生素B20-10、維生素A0-10、維生素C 0-20、富硒酵母0-10、富鋅酵母0-40、富鐵酵母0-10、富鉻酵母0-10、微晶纖維素0-20。 In one aspect of the invention, a yeast nutritious food is provided, which comprises yeast as a main raw material, including milk powder, maltodextrin, lactose, non-dairy creamer, calcium carbonate, starch, white sugar, flavoring agent, essence The yeast nutritious food further comprises a B vitamin and a micronutrient-rich yeast, the component having the following parts by weight: yeast 5-97, milk powder 1-20, non-dairy creamer 1-50, carbonic acid Calcium 0.1-50, starch 1-50, maltodextrin 0-50, lactose 0-50, white sugar 0-70, folic acid 0-1, vitamin B 12 0-1 vitamin B 1 0-10, vitamin B 2 0-10, vitamin A0-10, vitamin C 0-20, selenium-enriched yeast 0-10, zinc-rich yeast 0-40, iron-rich yeast 0-10, chromium-rich yeast 0-10, microcrystalline cellulose 0-20 .
根據本發明的酵母營養食品,其中,進一步包括具有下列重量份的組分:白砂糖1-70、葉酸0.00001-1、維生素B120.00001-1維生素B10.0001-10、維生素B20.0001-10、維生素A0.0001-10、維生素C 0.0001-20、富硒酵母0.0001-10、富鋅酵母1-40、富鐵酵母1-10、富鉻酵母0.0001-10、微晶纖維素1-20。 The yeast nutritious food according to the present invention, further comprising a component having the following parts by weight: white sugar 1-70, folic acid 0.00001-1, vitamin B 12 0.00001-1 vitamin B 1 0.0001-10, vitamin B 2 0.0001-10 , vitamin A0.0001-10, vitamin C 0.0001-20, selenium-enriched yeast 0.0001, zinc-rich yeast 1-40, iron-rich yeast 1-10, chromium-rich yeast 0.0001, microcrystalline cellulose 1-20.
根據本發明的酵母營養食品,其中,還包括以下重量份的組分: 脫水蔥片1-10、可哥粉1-20、果蔬粉1-20、芝麻1-50、奶茶粉1-20、豆奶粉1-20、燕麥片1-20。 A yeast nutritious food according to the present invention, which further comprises the following parts by weight: Dehydrated onion slices 1-10, Kege powder 1-20, fruit and vegetable powder 1-20, sesame 1-50, milk tea powder 1-20, soy milk powder 1-20, oatmeal 1-20.
根據本發明的酵母營養食品,其中,主要原料酵母可以是酵母粉或酵母乳。 A yeast nutritious food according to the present invention, wherein the main raw material yeast may be yeast powder or yeast milk.
根據本發明的酵母營養食品,其中,該酵母營養食品還包括富鐵酵母和富鉻酵母。 A yeast nutritious food according to the present invention, wherein the yeast nutritious food further comprises iron-rich yeast and chromium-rich yeast.
根據本發明的酵母營養食品,其中,富鐵酵母可以由是葡萄糖酸亞鐵、氨基酸螯合鐵、硫酸亞鐵、富馬酸亞鐵、乳酸亞鐵或它們任意的混合物取代;富鋅酵母可以由硫酸鋅、葡萄糖酸鋅、乳酸鋅、檸檬酸鋅、氨基酸螯合鋅或它們任意的混合物取代;富硒酵母可以由亞硒酸鈉取代;矯味劑是檸檬酸、蘋果酸。 The yeast nutritious food according to the present invention, wherein the iron-rich yeast may be substituted by ferrous gluconate, iron chelate iron, ferrous sulfate, ferrous fumarate, ferrous lactate or a mixture thereof; zinc-rich yeast may It is substituted by zinc sulfate, zinc gluconate, zinc lactate, zinc citrate, amino acid chelated zinc or any mixture thereof; the selenium-enriched yeast may be substituted by sodium selenite; the flavoring agent is citric acid, malic acid.
在本發明的酵母營養食品的一個具體實施方式中,其中,酵母營養食品包括以下重量份的組分:酵母5-97、奶粉1-20、植脂末1-50、碳酸鈣0.1-50、澱粉1-50、維生素C 0.1-0.3、白砂糖20-70、可哥粉1-20。 In a specific embodiment of the yeast nutritious food of the present invention, wherein the yeast nutritious food comprises the following components by weight: yeast 5-97, milk powder 1-20, non-dairy creamer 1-50, calcium carbonate 0.1-50, Starch 1-50, vitamin C 0.1-0.3, white sugar 20-70, Kog powder 1-20.
在本發明的酵母營養食品的一個具體實施方式中,其中,酵母營養食品包括以下重量份的組分:酵母5-97、奶粉1-20、植脂末1-50、碳酸鈣0.1-50、澱粉1-50、微晶纖維素1-20。 在本發明的酵母營養食品的一個具體實施方式中,其中,酵母營養食品包括以下重量份的組分:酵母5-95、奶粉1-20、植脂末1-50、碳酸鈣0.1-50、澱粉1-50、芝麻1-20、白砂糖1-20、麥芽糊精10-50。 In a specific embodiment of the yeast nutritious food of the present invention, wherein the yeast nutritious food comprises the following components by weight: yeast 5-97, milk powder 1-20, non-dairy creamer 1-50, calcium carbonate 0.1-50, Starch 1-50, microcrystalline cellulose 1-20. In a specific embodiment of the yeast nutritious food of the present invention, wherein the yeast nutritious food comprises the following components by weight: yeast 5-95, milk powder 1-20, non-dairy creamer 1-50, calcium carbonate 0.1-50, Starch 1-50, sesame 1-20, white sugar 1-20, maltodextrin 10-50.
在本發明的酵母營養食品的一個具體實施方式中,其中,酵母營養食品包括以下重量份的組分:酵母5-95、奶粉1-20、植脂末1-50、碳酸鈣0.1-50、澱粉1-50、白砂糖20-70、果蔬粉1-20、矯味劑1-20。 In a specific embodiment of the yeast nutritious food of the present invention, wherein the yeast nutritious food comprises the following components by weight: yeast 5-95, milk powder 1-20, non-dairy creamer 1-50, calcium carbonate 0.1-50, Starch 1-50, white sugar 20-70, fruit and vegetable powder 1-20, flavoring agent 1-20.
在本發明的酵母營養食品的一個具體實施方式中,其中,酵母營養食品包括以下重量份的組分:酵母5-95、奶粉1-20、植脂末1-50、碳酸鈣0.1-50、澱粉1-50、脫水蔥片1-10。 In a specific embodiment of the yeast nutritious food of the present invention, wherein the yeast nutritious food comprises the following components by weight: yeast 5-95, milk powder 1-20, non-dairy creamer 1-50, calcium carbonate 0.1-50, Starch 1-50, dehydrated onion slices 1-10.
在本發明的酵母營養食品的一個具體實施方式中,其中,酵母營養食品包括以下重量份的組分:酵母5-95、奶粉1-20、植脂末1-50、碳酸鈣0.1-50、澱粉1-50、葉酸0.05-0.1、維生素B120.0002、維生素A 0.1-0.3、維生素C 0.1-0.3、富鐵酵母1-10。 In a specific embodiment of the yeast nutritious food of the present invention, wherein the yeast nutritious food comprises the following components by weight: yeast 5-95, milk powder 1-20, non-dairy creamer 1-50, calcium carbonate 0.1-50, Starch 1-50, folic acid 0.05-0.1, vitamin B 12 0.0002, vitamin A 0.1-0.3, vitamin C 0.1-0.3, iron-rich yeast 1-10.
據本發明的酵母營養食品,其中,B族維生素是選自維生素B1、維生素B2、維生素B12、泛酸、煙酸組成的組中的任一種。 The yeast nutritious food of the present invention, wherein the B vitamin is any one selected from the group consisting of vitamin B 1 , vitamin B 2 , vitamin B 12 , pantothenic acid, and niacin.
在本發明的酵母營養食品的一個具體實施方式中,其中,酵母營養食品包括以下重量份的組分:酵母5-95、奶粉1-20、植脂末1-50、碳酸鈣0.1-50、澱粉1-50、富鋅酵母1-10、富硒酵母1-10、富鉻酵母1-10、富鐵酵母1-10。 In a specific embodiment of the yeast nutritious food of the present invention, wherein the yeast nutritious food comprises the following components by weight: yeast 5-95, milk powder 1-20, non-dairy creamer 1-50, calcium carbonate 0.1-50, Starch 1-50, zinc-rich yeast 1-10, selenium-enriched yeast 1-10, chromium-rich yeast 1-10, iron-rich yeast 1-10.
在本發明的酵母營養食品的一個具體實施方式中,其中, 酵母營養食品包括以下重量份的組分:酵母5-95,奶粉1-20、植脂末1-50、碳酸鈣0.1-50、澱粉1-50、奶茶粉、豆奶粉和燕麥片共60。 In a specific embodiment of the yeast nutritious food of the present invention, wherein The yeast nutritious food comprises the following components by weight: yeast 5-95, milk powder 1-20, non-dairy creamer 1-50, calcium carbonate 0.1-50, starch 1-50, milk tea powder, soy milk powder and oatmeal.
根據本發明的酵母營養食品,適合粉狀、雪片狀、片劑中的任何一種劑型。 The yeast nutritious food according to the present invention is suitable for any one of a powder form, a snow flake form, and a tablet form.
本發明的另一個方面,提供了根據本發明的酵母營養食品的製備方法。 In another aspect of the invention, there is provided a process for the preparation of a yeast nutritional food according to the invention.
根據本發明的用於製備酵母營養食品的方法,包括:將酵母和其他原料按照配方進行混合並加水攪拌均勻,然後噴霧乾燥並進行包裝以得到粉狀的酵母營養食品。 A method for preparing a yeast nutritious food according to the present invention comprises: mixing yeast and other raw materials according to a formula and uniformly stirring with water, followed by spray drying and packaging to obtain a powdery yeast nutritious food.
根據本發明的用於製備酵母營養食品的方法,包括:將酵母加水攪拌均勻,然後用滾筒乾燥機進行乾燥,其中乾燥溫度控制在180℃,乾燥後進行收集並用篩網過濾,再加入所述其他原輔料混合均勻,最後進行包裝,從而得到雪片狀的酵母營養食品。 The method for preparing a yeast nutritious food according to the present invention comprises: uniformly stirring the yeast with water, and then drying with a tumble dryer, wherein the drying temperature is controlled at 180 ° C, drying, collecting and filtering with a sieve, and then adding the The other raw materials are mixed evenly and finally packaged to obtain a snow flake yeast nutrition food.
根據本發明的用於製備酵母營養食品的方法,包括:將酵母和其他原料按照配方進行混合,然後通過制粒、乾燥、壓片等步驟製成片劑,最後經過包裝而獲得片劑的酵母營養食品。 The method for preparing a yeast nutritious food according to the present invention comprises: mixing yeast and other raw materials according to a formula, then forming a tablet by granulation, drying, tableting, etc., and finally obtaining a tablet yeast by packaging. Nutritious food.
與現有技術相比,本發明的酵母營養食品除了現有技術中已有的原料之外,還添加了維生素A、B族維生素、維生素C、富鐵酵母、富鋅酵母、富硒酵母、富鉻酵母等多種物質,提高了酵母食品的營養性,同時滿足了食用者對多種營養物質的需求;此外,本發明採用多種工藝進行酵母營養食品的製備,能夠獲得粉狀、雪片狀、片劑等不同劑型的酵母營養食品,開發 出口感適合不同人群服用的營養食品。 Compared with the prior art, the yeast nutritious food of the present invention adds vitamin A, B vitamins, vitamin C, iron-rich yeast, zinc-rich yeast, selenium-enriched yeast, and chromium-rich in addition to the raw materials existing in the prior art. A variety of substances, such as yeast, improve the nutritionality of the yeast food, while satisfying the needs of the consumer for a variety of nutrients; in addition, the present invention uses a variety of processes for the preparation of yeast nutritious foods, which can obtain powder, snow flakes, tablets Yeast nutritious foods of different dosage forms, development The sense of export is suitable for nutritious foods taken by different groups of people.
下文將列舉本發明的酵母營養食品的具體實施例。本領域中的普通技術人員應當明瞭,所列舉的具體實施例僅僅是為了例示的目的,而非為了限制本發明的範圍。 Specific examples of the yeast nutraceutical of the present invention will be enumerated below. It will be apparent to those skilled in the art that the specific embodiments are set forth herein for the purpose of illustration and description.
在本發明的實施例1中,酵母營養食品包括以下重量份的組分:酵母90、奶粉18、植脂末45、碳酸鈣5.3、澱粉50、富硒酵母2、富鋅酵母2、富鉻酵母2、葉酸0.1、維生素B10.1、維生素B20.1、維生素B120.001、富鐵酵母2。 In Example 1 of the present invention, the yeast nutritious food comprises the following components by weight: yeast 90, milk powder 18, non-dairy creamer 45, calcium carbonate 5.3, starch 50, selenium-enriched yeast 2, zinc-rich yeast 2, chromium-rich Yeast 2, folic acid 0.1, vitamin B 1 0.1, vitamin B 2 0.1, vitamin B 12 0.001, iron-rich yeast 2.
在本發明的實施例2中,酵母營養食品包括以下重量份的組分:酵母10、奶粉10、植脂末10、碳酸鈣10、澱粉17.6、富硒酵母8、富鋅酵母8、富鉻酵母8、富鐵酵母8、維生素B10.002、維生素B20.002、維生素B120.00005、白砂糖20、可哥粉10、麥芽糊精10。 In Example 2 of the present invention, the yeast nutritious food comprises the following components by weight: yeast 10, milk powder 10, non-dairy creamer 10, calcium carbonate 10, starch 17.6, selenium-enriched yeast 8, zinc-rich yeast 8, chromium-rich Yeast 8, iron-rich yeast 8, vitamin B 1 0.002, vitamin B 2 0.002, vitamin B 12 0.00005, white sugar 20, Kog powder 10, maltodextrin 10.
在本發明的實施例3中,酵母營養食品包括以下重量份的組分:酵母50、奶粉5、植脂末20、碳酸鈣40、澱粉30、富硒酵母5、富鋅酵母10、富鉻酵母5、葉酸0.05、維生素B15、維生素B25、維生素B120.001、白砂糖4.95。 In Example 3 of the present invention, the yeast nutritious food comprises the following components by weight: yeast 50, milk powder 5, non-dairy creamer 20, calcium carbonate 40, starch 30, selenium-enriched yeast 5, zinc-rich yeast 10, chromium-rich Yeast 5, folic acid 0.05, vitamin B 1 5, vitamin B 2 5, vitamin B 12 0.001, white sugar 4.95.
在本發明的實施例4中,酵母營養食品包括以下重量份的 組分:酵母5、奶粉10、植脂末30、碳酸鈣30、澱粉15、維生素B10.1、維生素B20.1、富鉻酵母5、果蔬粉4.8。 In Example 4 of the present invention, the yeast nutritious food comprises the following components by weight: yeast 5, milk powder 10, non-dairy creamer 30, calcium carbonate 30, starch 15, vitamin B 1 0.1, vitamin B 2 0.1, chromium-rich Yeast 5, fruit and vegetable powder 4.8.
在本發明的實施例5中,酵母營養食品包括以下重量份的組分:酵母97、奶粉1、植脂末1、碳酸鈣0.1、澱粉1、維生素B10.01、維生素B20.01、富硒酵母0.001。 In Example 5 of the present invention, the yeast nutritious food comprises the following components by weight: yeast 97, milk powder 1, non-dairy creamer 1, calcium carbonate 0.1, starch 1, vitamin B 1 0.01, vitamin B 2 0.01, selenium-enriched Yeast 0.001.
本發明的酵母營養食品適合包括粉狀、雪片狀、片劑的不同劑型,因而針對不同的劑型,本發明用於製備酵母營養食品的方法可包括以下三種: The yeast nutritious food of the present invention is suitable for different dosage forms including powder, snow flakes and tablets, and thus the method for preparing yeast nutritious food of the present invention may include the following three kinds for different dosage forms:
用於製備粉狀酵母營養食品的方法,包括:將酵母和其他原料按照配方進行混合並加水攪拌均勻,然後噴霧乾燥並進行包裝以得到粉狀酵母營養食品。 A method for preparing a powdery yeast nutritious food, comprising: mixing yeast and other raw materials according to a formula and uniformly stirring with water, then spray drying and packaging to obtain a powdery yeast nutritious food.
用於製備雪片狀酵母營養食品的方法,包括:將酵母加水攪拌均勻,然後用滾筒乾燥機進行乾燥,其中乾燥溫度控制在180℃,乾燥後進行收集並用篩網過濾,再加入所述其他原輔料混合均勻,最後進行包裝,從而得到雪片狀酵母營養食品。 The method for preparing a snow flake yeast nutrient food comprises: stirring the yeast water uniformly, and then drying with a tumble dryer, wherein the drying temperature is controlled at 180 ° C, drying, collecting, filtering with a sieve, and adding the other The raw materials are uniformly mixed and finally packaged to obtain a snow flake yeast nutritious food.
用於製備片劑酵母營養食品的方法,包括:將酵母和其他原料按照配方進行混合,然後通過制粒、乾燥、壓片等步驟製成片劑,最後經過包裝而獲得片劑酵母營養食品。片劑的製備可採用濕法制粒、一步制粒機制粒或直接壓片法。 The method for preparing a tablet yeast nutritious food comprises: mixing yeast and other raw materials according to a formula, then forming a tablet by granulating, drying, tableting, etc., and finally packaging to obtain a tablet yeast nutritious food. Tablets may be prepared by wet granulation, one-step granulation mechanism or direct compression.
通過功能實驗證明,本發明具有顯著的增強免疫力的功能,並且其作用比同類產品有所提高,具體實驗資料如下: Through functional experiments, the invention has significant immunity enhancing functions, and its effect is improved compared with similar products. The specific experimental data are as follows:
1 材料和方法 1 Materials and methods
1.1 受試物:本專利發明的酵母營養食品實施例1、2、3、 4、5(以下簡稱酵母營養食品A1、A2、A3、A4、A5)、專利“一種營養酵母食品及其製備方法”(200510015005.2)發明的酵母營養食品(以下簡稱酵母營養食品B,配方為:營養酵母粉18g、全脂奶粉44g、植脂末15g、蔗糖9g、果糖7.5g、β-環糊精2g可哥粉1.05g咖啡粉0.05g、可哥粉末香精3.4g)。成人推薦量為10g/d。按成人60kg體重計,即0.17g/kg.bw。按所需濃度配製成不同溶液灌胃。 1.1 Test substance: yeast nutritional food examples 1, 2, and 3 of the present invention 4, 5 (hereinafter referred to as yeast nutritional foods A1, A2, A3, A4, A5), the patent "a nutritional yeast food and its preparation method" (200510015005.2) invented yeast nutritional food (hereinafter referred to as yeast nutritional food B, the formula is: Nutritional yeast powder 18g, whole milk powder 44g, vegetable fat 15g, sucrose 9g, fructose 7.5g, β-cyclodextrin 2g Kog powder 1.05g coffee powder 0.05g, Kege powder flavor 3.4g). The recommended amount for adults is 10g/d. According to the adult 60kg body weight, namely 0.17g/kg.bw. Prepare different solutions at the required concentration.
1.2 試驗動物:清潔級BALB/c小鼠,雄性,200只,18-22g(湖北省實驗動物研究中心) 1.2 Test animals: clean grade BALB/c mice, male, 200, 18-22g (Hubei Experimental Animal Research Center)
1.3 分組、給藥:將小鼠隨機分為5個組:正常對照組、加酵母營養食品的A1、A2、A3、A4、A5組、加酵母營養食品B組。加酵母營養食品A1-A3的三個組和加酵母營養食品B組按30倍人體攝入量(5.0g/kg.bw)灌胃,灌胃體積為0.2ml/10g.bw。 1.3 Grouping and administration: The mice were randomly divided into 5 groups: normal control group, A1, A2, A3, A4, A5 group with yeast nutrition food, and yeast nutrition food group B. The three groups of yeast nutrition food A1-A3 and the yeast nutrition food group B were intragastrically administered with 30 times human body intake (5.0 g/kg.bw), and the gastric perfusion volume was 0.2 ml/10 g.bw.
1.4 主要儀器與試劑:5mm直徑打孔器、顯微鏡、微量血凝試驗板、離心機、酶標儀、96孔培養板、5%CO2培養箱、注射用墨汁、刀豆蛋白A(ConA)、MTT、DNFB、生理鹽水、雞紅細胞、Giemsa染液、YAC-1細胞、Hanks液、RPMI1640完全培養液、乳酸鋰、碘硝基氯化四氮唑(INT)、吩嗪二甲酯硫酸鹽(PMS)、NAD、Tris-HCl緩衝液 1.4 Main instruments and reagents: 5mm diameter puncher, microscope, micro blood coagulation test plate, centrifuge, microplate reader, 96-well culture plate, 5% CO2 incubator, ink for injection, concanavalin A (ConA), MTT, DNFB, saline, chicken red blood cells, Giemsa dye solution, YAC-1 cells, Hanks solution, RPMI1640 complete medium, lithium lactate, iodonitrotetrazolium chloride (INT), phenazine dimethyl sulfate ( PMS), NAD, Tris-HCl buffer
1.5 實驗方法 1.5 Experimental methods
1.5.1 碳廓清實驗和臟器/體重比值測定:實驗末期,小鼠稱重後,按體重從小鼠尾靜脈注入稀釋的印度墨汁(100mg/kg),注入墨汁後2、10min,分別從內眥靜脈叢取血 20μl,並立即將其加到2ml0.1% Na2CO3溶液中。用721分光光度計在600nm處測光密度值(OD),以Na2CO3溶液做空白對照。將小鼠處死,解剖動物,取出肝臟、脾臟及胸腺,用濾紙吸幹血跡後稱重。 1.5.1 Carbon clearance test and organ/body weight ratio measurement: At the end of the experiment, after the mice were weighed, the diluted Indian ink (100 mg/kg) was injected from the tail vein of the mouse according to the body weight, and the ink was injected 2, 10 min after the injection, respectively. 20 μl of blood was taken from the iliac venous plexus and immediately added to 2 ml of a 0.1% Na 2 CO 3 solution. The optical density value (OD) was measured at 600 nm using a 721 spectrophotometer, and the Na 2 CO 3 solution was used as a blank control. The mice were sacrificed, the animals were dissected, the liver, spleen and thymus were removed, and the blood was blotted with a filter paper and weighed.
1.5.2 遲髮型變態反應(DTH):小鼠連續灌胃30天后,每鼠腹部皮膚用脫毛劑進行脫毛處理,範圍約3cm×3cm,後用DNFB溶液50μl均勻塗抹致敏;5天后,再用DNFB溶液10μl均勻塗抹於小鼠右耳兩面,進行攻擊;24小時後,頸椎脫臼處死小鼠,剪下左右兩耳,用打孔器取下直徑5mm的耳片稱重。 1.5.2 delayed type hypersensitivity (DTH): After 30 days of continuous gavage in mice, each animal's abdominal skin was depilated with a depilatory agent, the range was about 3 cm × 3 cm, and then evenly sensitized with 50 μl of DNFB solution; after 5 days, 10 μl of DNFB solution was evenly applied to both sides of the right ear of the mouse to attack; after 24 hours, the mice were sacrificed by cervical dislocation, and the left and right ears were cut out, and the ear piece with a diameter of 5 mm was removed by a puncher and weighed.
1.5.3 小鼠腹腔巨噬細胞吞噬雞紅細胞實驗(半體內法):小鼠連續灌胃30天后,每鼠腹腔注射2%雞紅細胞懸液1ml。30min後,頸椎脫臼處死動物,仰位固定於蠟板上,正中剪開腹壁皮膚,經腹腔注入生理鹽水2ml,轉動鼠板1min,然後吸出腹腔洗液1ml,平均分滴於2片載玻片上,放入墊有濕紗布的搪瓷盒內,於37℃孵箱溫育30min。孵畢,在生理鹽水中漂洗,除去未貼片細胞。晾乾,以1:1丙酮甲醇溶液固定20min,4%(v/v)Giemsa-磷酸緩衝液染色3min,再用蒸餾水漂洗晾乾,於顯微鏡下計數巨噬細胞。 1.5.3 Mouse peritoneal macrophage phagocytosis chicken red blood cell experiment (half-in vivo method): After continuous gastric perfusion for 30 days, each mouse was intraperitoneally injected with 1 ml of chicken red blood cell suspension. After 30 minutes, the animals were sacrificed by cervical dislocation and fixed on the wax plate. The skin of the abdominal wall was cut in the middle, 2 ml of normal saline was injected into the abdominal cavity, the rat plate was rotated for 1 min, and then 1 ml of the peritoneal washing solution was aspirated, and the average was divided into 2 slides. Place in an enamel box with wet gauze and incubate for 30 min at 37 °C. After incubation, rinse in saline to remove unpatched cells. The cells were air-dried, fixed in 1:1 acetone-methanol solution for 20 min, stained with 4% (v/v) Giemsa-phosphate buffer for 3 min, rinsed with distilled water and dried, and the macrophages were counted under a microscope.
1.5.4 ConA誘導的小鼠脾淋巴細胞轉化實驗和小鼠NK細胞活性測定(乳酸脫氫酶LDH測定法):小鼠連續灌胃30天后,頸椎脫臼處死,無菌取脾,置於盛有適量無菌Hanks液的小平皿中,用鑷子輕輕將脾撕碎,經200目曬網過濾,製成單細胞懸液,將細胞懸液分為兩部分,分別用於ConA誘導的小鼠脾淋巴細胞轉化實驗和小鼠NK細胞活性測定。 1.5.4 ConA-induced spleen lymphocyte transformation assay and mouse NK cell activity assay (lactate dehydrogenase LDH assay): After 30 days of continuous gavage, the cervical vertebrae were sacrificed and the spleen was aseptically taken. In a small plate of appropriate amount of sterile Hanks solution, the spleen was gently shredded with tweezers, filtered through a 200 mesh net to prepare a single cell suspension, and the cell suspension was divided into two parts for ConA-induced mouse spleen. Lymphocyte transformation experiments and mouse NK cell activity assays.
1.5.4.1 ConA誘導的小鼠脾淋巴細胞轉化實驗:用Hanks液洗滌細胞懸液2次,每次離心10min(1000r/min),然後將細胞懸浮於1ml的完全培養液中,台盼藍染色計數活細胞數(在95%以上),調整細胞濃度為3×106個/ml。將每份細胞懸液分兩孔加入24孔培養板中,每孔1ml,一孔加75μlConA液,另一孔作為對照,置培養箱中培養72h。培養結束前4h,每孔吸去上清0.7ml,加入0.7ml不含血清的RPMI1640培養液,同時加入MTT50μl/孔,繼續培養4h。培養結束後,每孔加入1ml酸性異丙醇,混勻,溶解。然後分裝到96孔培養板,每孔做3個平行孔,用酶標儀以570nm波長測定光密度值。 1.5.4.1 ConA-induced spleen lymphocyte transformation in mice: Wash the cell suspension twice with Hanks solution, centrifuge for 10 min (1000 r/min), then suspend the cells in 1 ml of complete medium, trypan blue staining The number of viable cells (over 95%) was counted, and the cell concentration was adjusted to 3 × 10 6 /ml. Each cell suspension was added to a 24-well culture plate in two wells, 1 ml per well, one well was added with 75 μl of ConA solution, and the other well was used as a control, and cultured in an incubator for 72 hours. Four hours before the end of the culture, 0.7 ml of the supernatant was aspirated per well, 0.7 ml of serum-free RPMI1640 medium was added, and MTT 50 μl/well was added thereto, and the culture was continued for 4 hours. After the completion of the culture, 1 ml of acidic isopropanol was added to each well, and the mixture was mixed and dissolved. Then, it was dispensed into a 96-well culture plate, and three parallel holes were made per well, and the optical density value was measured by a microplate reader at a wavelength of 570 nm.
小鼠NK細胞活性測定(乳酸脫氫酶LDH測定法):試驗前24小時將靶細胞傳代培養,應用前以Hanks液洗3次,用RPMI1640完全培養液調整細胞濃度為4×105個/ml。用Hanks液洗滌細胞懸液2次,每次離心10min(1000r/min)。棄上清將細胞漿彈起,加入0.5ml滅菌水20s,裂解細胞後再加入0.5ml 2倍Hanks液及8ml Hanks液,離心10min(1000r/min),用1ml完全培養基重懸,用1%冰醋酸稀釋後計數,台盼藍染色計數活細胞數(在95%以上),調整細胞濃度為2×107個/ml。取靶細胞和效應細胞各100μl(效靶比50:1),加入96孔培養板中;靶細胞自然釋放孔加靶細胞和培養液各100μl、靶細胞最大釋放孔加靶細胞和1% NP40各100μl,上述各項均設三個複孔,於37℃、5% CO2培養箱中培養4小時,然後將培養板離心(1500r/min)5min,每孔吸取上清100μl置96孔培養板中,同時加入LDH基質液100μl,反應3min,每孔加入1mol/L的HCl30μl, 在酶標儀490nm處測定光密度值(OD)。 Determination of mouse NK cell activity (lactate dehydrogenase LDH assay): The target cells were subcultured 24 hours before the test, washed three times with Hanks before application, and adjusted to a cell concentration of 4×105 with RPMI1640 complete medium. Ml. The cell suspension was washed twice with Hanks solution and centrifuged for 10 min (1000 r/min) each time. Discard the supernatant and bounce the cytoplasm. Add 0.5 ml of sterilized water for 20 s. After lysing the cells, add 0.5 ml of 2 times Hanks solution and 8 ml of Hanks solution, centrifuge for 10 min (1000 r/min), resuspend with 1 ml of complete medium, and use 1%. After dilution with glacial acetic acid, the number of viable cells (over 95%) was counted by trypan blue staining, and the cell concentration was adjusted to 2×107 cells/ml. 100 μl of target cells and effector cells (50:1 target ratio) were added to 96-well culture plates; target cells were naturally released from wells plus 100 μl of target cells and culture medium, target cells were maximally released and target cells were added and 1% NP40 Each 100 μl, each of the above three sets of three wells, cultured in 37 ° C, 5% CO2 incubator for 4 hours, then the plate was centrifuged (1500r / min) for 5min, each well uptake 100μl 96-well culture plate In addition, 100 μl of LDH substrate solution was added at the same time, and the reaction was carried out for 3 min, and 30 μl of 1 mol/L HCl was added to each well. The optical density value (OD) was measured at 490 nm.
結果 result
2.1 兩種酵母營養食品對小鼠臟器/體重比的影響,結果見表1。 2.1 The effect of two yeast nutritious foods on the organ/body weight ratio of mice. The results are shown in Table 1.
兩種酵母營養食品脾臟/體重比值與對照組比較,P>0.05,無顯著差異;胸腺/體重比值與對照組比較,P<0.05,有顯著差異。 The spleen/weight ratio of the two yeast nutritious foods was compared with the control group, P>0.05, no significant difference; the thymus/body weight ratio was significantly higher than the control group, P<0.05.
2.2 兩種酵母營養食品對小鼠細胞免疫功能(小鼠脾淋巴細胞轉化實驗以及小鼠遲髮型變態反應(DTH))的影響,見表2。 2.2 The effects of two yeast nutritious foods on mouse cellular immune function (spleen lymphocyte transformation experiment and mouse delayed type hypersensitivity (DTH)) are shown in Table 2.
(1)與對照組相比P<0.01 (1) P<0.01 compared with the control group
(2)與對照組相比P<0.05 (2) P<0.05 compared with the control group
(3)酵母營養食品A1-A5組與酵母營養食品B組相比P<0.05 (3) Yeast nutritious food A1-A5 group compared with yeast nutritious food group B P<0.05
酵母營養食品A1-A5組淋巴細胞增殖實驗的OD差值與對照組相比P<0.01,有顯著性差異;耳腫程度與對照組相比也P<0.01,有顯著性差異,且酵母營養食品A1-A5組與酵母營養食品B組相比P<0.05,有顯著性差異。 The OD difference of lymphocyte proliferation test in yeast nutrition food A1-A5 group was significantly higher than that of the control group (P<0.01). The degree of ear swelling was also P<0.01 compared with the control group, and there was significant difference, and yeast nutrition. The food A1-A5 group had P<0.05 compared with the yeast nutritious food group B, and there was a significant difference.
兩種酵母營養食品對小鼠單核-巨噬細胞功能(小鼠碳廓清實驗、小鼠腹腔巨噬細胞吞噬雞紅細胞能力)的影響,見表3。 The effects of two yeast nutritious foods on mouse monocyte-macrophage function (mouse carbon clearance assay, mouse peritoneal macrophage phagocytosis of chicken red blood cells) are shown in Table 3.
(1)與對照組相比P<0.01 (1) P<0.01 compared with the control group
(2)與對照組相比P<0.05 (2) P<0.05 compared with the control group
(3)酵母營養食品A1-A5組與酵母營養食品B組相比P<0.05 (3) Yeast nutritious food A1-A5 group compared with yeast nutritious food group B P<0.05
酵母營養食品A1-A5組小鼠碳廓清實驗吞噬指數與對照組相比較,P<0.01,有極顯著差異;小鼠腹腔巨噬細胞吞噬雞紅細胞的吞噬率與對照組比較,P<0.01,有極顯著差異。並且, 酵母營養食品A1-A5組與酵母營養食品B組相比P<0.05,有顯著性差異。 Compared with the control group, the phagocytosis index of the mice in the A1-A5 group of yeast nutrition foods was significantly different from that of the control group. The phagocytosis rate of mouse peritoneal macrophages phagocytized chicken red blood cells was compared with the control group, P<0.01. There are significant differences. and, There was a significant difference between the yeast nutrition food group A1-A5 and the yeast nutrition food group B (P<0.05).
兩種酵母營養食品對小鼠NK細胞活性的影響,見表4。 The effects of two yeast nutritious foods on the activity of mouse NK cells are shown in Table 4.
(1)與對照組相比P<0.01 (1) P<0.01 compared with the control group
(2)與對照組相比P<0.05 (2) P<0.05 compared with the control group
(3)酵母營養食品A1-A5組與酵母營養食品B組相比P<0.01 (3) Yeast nutritious food A1-A5 group compared with yeast nutrient food group B P<0.01
酵母營養食品A1-A5組小鼠NK細胞活性與對照組相比較,P<0.01,有極顯著差異。並且,酵母營養食品A1-A5組與酵母營養食品B組相比P<0.01,有顯著性差異。 The activity of NK cells in the yeast nutrition food group A1-A5 was significantly higher than that in the control group (P<0.01). Moreover, the yeast nutrition food group A1-A5 had a significant difference compared with the yeast nutrient food group B (P<0.01).
結論 in conclusion
本實驗通過給小鼠灌以本發明的酵母營養食品(酵母營養食品A1-A5)和專利“一種營養酵母食品及其製備方法”(200510015005.2)發明的酵母營養食品(酵母營養食品B),並進行小鼠的細胞免疫功能實驗(小鼠脾淋巴細胞轉化實驗以及小鼠遲髮型變態反應(DTH))、單核-巨噬細胞功能實驗(小鼠碳廓清實驗、小鼠腹腔巨噬細胞吞噬雞紅細胞能力)、NK細 胞活性實驗。實驗結果表明本發明的酵母營養食品A1-A5比酵母營養食品B更能顯著提高小鼠的各項免疫功能。 In this experiment, a yeast nutrient food (yeast nutrient food B) invented by the yeast nutrient food of the present invention (yeast nutrient food A1-A5) and the patent "a nutritious yeast food and its preparation method" (200510015005.2) are implanted into the mouse, and Perform mouse cellular immune function experiments (mouse spleen lymphocyte transformation test and mouse delayed type allergy (DTH)), monocyte-macrophage function test (mouse carbon clearance test, mouse peritoneal macrophage phagocytosis) Chicken red blood cell capacity), NK fine Cell activity assay. The experimental results show that the yeast nutritional food A1-A5 of the present invention can significantly improve various immune functions of the mouse than the yeast nutritional food B.
以上,在特定的優選實施方式的基礎上對本發明進行了描述,但本領域中普通技術人員應當明瞭,在不偏離所附權利要求所限定的本發明的精神和範圍的情況下,可以增加多種變化和改動。 The invention has been described above on the basis of the specific preferred embodiments, but it will be apparent to those skilled in the art that various modifications can be made without departing from the spirit and scope of the invention as defined by the appended claims. Changes and changes.
Claims (10)
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TW098141533A TWI481351B (en) | 2009-12-04 | 2009-12-04 | A yeast nutrition product and the preparation method thereof |
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Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
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CN1927006A (en) * | 2005-09-07 | 2007-03-14 | 天津天士力制药股份有限公司 | Nutritious yeast foodstuff and its preparing process |
CN101569405A (en) * | 2008-04-29 | 2009-11-04 | 安琪酵母股份有限公司 | Yeast nutritive and health food |
CN101569414A (en) * | 2008-04-29 | 2009-11-04 | 安琪酵母股份有限公司 | Composition containing rich yeast selenium and yeast glucan |
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Publication number | Priority date | Publication date | Assignee | Title |
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CN1927006A (en) * | 2005-09-07 | 2007-03-14 | 天津天士力制药股份有限公司 | Nutritious yeast foodstuff and its preparing process |
CN101569405A (en) * | 2008-04-29 | 2009-11-04 | 安琪酵母股份有限公司 | Yeast nutritive and health food |
CN101569414A (en) * | 2008-04-29 | 2009-11-04 | 安琪酵母股份有限公司 | Composition containing rich yeast selenium and yeast glucan |
Non-Patent Citations (1)
Title |
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健康食品中常見的賦型劑-麥芽糊精,2007/06/01,網址;http://tw.myblog.yahoo.com/maysa73/article?mid=-2&prev=42&l=a&fid=14 * |
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