WO2022156615A9 - 一种富含egc的非酯型茶多酚的制备方法 - Google Patents

一种富含egc的非酯型茶多酚的制备方法 Download PDF

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WO2022156615A9
WO2022156615A9 PCT/CN2022/072230 CN2022072230W WO2022156615A9 WO 2022156615 A9 WO2022156615 A9 WO 2022156615A9 CN 2022072230 W CN2022072230 W CN 2022072230W WO 2022156615 A9 WO2022156615 A9 WO 2022156615A9
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egc
tea
ester
preparation
extraction
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WO2022156615A1 (zh
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任雪音
刘剑宏
曹后建
浦平南
刘德和
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江苏德和生物科技有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/82—Theaceae (Tea family), e.g. camellia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
    • C07D311/58—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
    • C07D311/60—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2
    • C07D311/62—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2 with oxygen atoms directly attached in position 3, e.g. anthocyanidins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
    • A61K31/353—3,4-Dihydrobenzopyrans, e.g. chroman, catechin
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07G—COMPOUNDS OF UNKNOWN CONSTITUTION
    • C07G99/00—Subject matter not provided for in other groups of this subclass
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/10—Preparation or pretreatment of starting material
    • A61K2236/15—Preparation or pretreatment of starting material involving mechanical treatment, e.g. chopping up, cutting or grinding
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/31—Extraction of the material involving untreated material, e.g. fruit juice or sap obtained from fresh plants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/35—Extraction with lipophilic solvents, e.g. Hexane or petrol ether
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization

Definitions

  • the invention belongs to the technical field of natural plant extract processing, and in particular relates to a preparation method of EGC-rich non-ester type tea polyphenols.
  • Tea polyphenol is a general term for catechins, flavonoids, phenolic acids and anthocyanins in tea. Its main function is anti-oxidation, and the elimination of oxygen free radicals is very obvious. It can effectively reduce the absorption of cholesterol in the large intestine, prevent and treat atherosclerosis, and also enhance the body's immunity, anti-tumor, anti-radiation, and anti-aging.
  • tea polyphenols Because of the unique efficacy of tea polyphenols, it has been widely used in the fields of medicine, health care products and food: in medicine, it has been used as a prevention and treatment drug for cardiovascular, liver, kidney and gastrointestinal diseases of the human body; in health care products, it has been used as an It is the main component of immunity, hypolipidemic, hypoglycemic and weight loss products. It is also used as the main component of sunscreens, shampoos and sterilizers. Adding tea polyphenols to oily foods can prevent food from spoiling and keep fresh.
  • tea polyphenols The main components of tea polyphenols are catechins, which are further divided into ester catechins and non-ester catechins. According to the different types of catechins in tea polyphenols, they are generally divided into ester teas. Polyphenols and non-ester tea polyphenols. There are certain differences between the two types of tea polyphenols in terms of taste, irritation to the stomach and stability (water solubility). The irritation of the mucous membrane is also stronger, and the stability of the product in water solubility is worse; the non-ester tea polyphenols are significantly better in terms of taste, irritation to the stomach and stability (water solubility). Ester catechins.
  • non-ester type tea polyphenols its application in food, food additives, and pharmaceutical industries has broader application prospects.
  • non-ester-type tea polyphenols contain a variety of non-ester-type catechin monomers, among which EGC has relatively high biological activity.
  • the current technology for preparing EGC-rich non-ester-type tea polyphenols is usually as follows: extracting tea polyphenols from tea leaves through an extraction solvent (at least one of water, ethanol, and ethyl acetate), filtering, concentrating, etc., adding some Exogenous hydrolase, etc. are hydrolyzed, and then the enzyme body is inactivated by high temperature, filtered and removed to obtain a non-ester type tea polyphenol solution, and then the product is purified by chromatography separation method, and finally non-ester type is obtained after drying. Tea polyphenol products.
  • an extraction solvent at least one of water, ethanol, and ethyl acetate
  • exogenous hydrolase is added, which is easy to cause product contamination, and improper handling can lead to the generation of allergens.
  • the chromatographic method is used to prepare the product, and the process is relatively complicated, which is not conducive to industrialized production. The loss of product during processing is also relatively large, and the yield is relatively low.
  • the purpose of the present invention is to provide a kind of preparation method of EGC-rich non-ester-type tea polyphenols, without adding any exogenous hydrolase, avoiding the pollution of exogenous hydrolase to the product, while the EGC content is high and the production process is simplified , The product yield is high, which is conducive to industrial production.
  • Tea pretreatment green tea leaves are used as raw materials, and the tea leaves are crushed and broken to obtain tea granules.
  • (6) mobile extraction-water washing extract the concentrated solution in step (5) with 1-2 times the volume of ethyl acetate, collect the ethyl acetate phase, and then concentrate the ethyl acetate phase to a solid content ratio of 2-10 %, carry out reverse-phase extraction with 0.2-1 times the volume of deionized water, collect the washed phase, and perform reverse-phase extraction with the same method for 1-5 times, combine the washing phases, and then use 1-2 times the volume of ethyl acetate for the washing phase. Extraction was performed and the ethyl acetate phase was collected.
  • step (9) Drying: the concentrated solution in step (8) is dried by spray-zero drying to obtain a non-ester type tea polyphenol product rich in EGC.
  • the 10-100 mesh sieve residue of the pretreated tea particles in step (1) is not more than 40%, that is, the pretreated tea particles in the range of 10-100 mesh are more than 60%.
  • the amount of deionized water added in step (2) is 1-5 times the volume of the pretreated tea leaves, the food-grade acid is citric acid, and the stirring speed is 50-300 r/min, so The enzymatic hydrolysis temperature is 30 ⁇ 50°C, and the enzymatic hydrolysis time is 4 ⁇ 12h.
  • the extraction temperature in step (3) is 60-90° C.
  • the amount of deionized water added in a single time is 6-10 times the quality of the tea raw material
  • the single extraction time is 20-60 minutes.
  • the cooling temperature in step (4) is 30-50° C., and the filtration precision is 100-300 mesh.
  • the concentration in step (5) is specifically: concentrating the filtrate in step (4) to 0.5-2.5Brix.
  • the ethyl acetate extraction temperature in step (6) is 30-60°C
  • the deionized water reverse-phase extraction temperature is 30-60°C.
  • the concentration in step (8) is specifically: vacuum concentration of the tea polyphenol aqueous solution in step (7) to 20-50Brix under the conditions of vacuum degree above -0.080MPa and temperature of 52-72°C.
  • the present invention utilizes the endogenous hydrolase contained in the tea itself to carry out enzymatic hydrolysis before soaking, and enzymatically hydrolyzes the ester-type catechins in the tea leaves into non-ester-type tea catechins (especially EGCG is converted into EGC), and then Extraction of the product; after the collected extract is processed, the product is purified and purified by the method of mobile extraction-water washing, and the purified product is concentrated and dried to obtain a non-ester type tea polyphenol product rich in EGC, tea
  • the polyphenol content is 90-99.5%
  • the EGC content is greater than 80%
  • the ester catechin EGCG+ECG content is within a controllable range (below 10%).
  • the endogenous hydrolase contained in the tea leaves is used in combination with the hydrolysis system to hydrolyze the ester-type tea polyphenols in the tea leaves to convert them to non-ester-type tea polyphenols to the greatest extent. Tea polyphenols can improve the content of non-ester tea polyphenol products and prevent the contamination of products by exogenous hydrolase.
  • the non-ester type tea polyphenol product rich in EGC obtained by the preparation method of the present invention the content of tea polyphenol is 90-99.5%, the content of EGC is greater than 80%, and the content of ester type catechin EGCG+ECG within the controllable range (below 10%).
  • a preparation method of EGC-rich non-ester type tea polyphenols comprising the following steps:
  • the green tea leaves are pulverized and wall-breaking, and the size of the treated tea leaves is more than 60% in the range of 10-100 mesh, that is, the 10-100 mesh sieve residue of the pretreated tea particles is not more than 40%. .
  • the content of tea polyphenols is 98.6% detected by UV spectrophotometer commonly used in the industry; the content of EGC is 81.3% by HPLC, and the content of ester catechin EGCG+ECG is 7.3%.
  • the green tea leaves are pulverized and wall-breaking, and the particle size of the treated tea leaves is more than 60% in the range of 10-100 mesh.
  • the content of tea polyphenols is 99.2% detected by UV spectrophotometer commonly used in the industry; the content of EGC is 83.0% by HPLC, and the content of ester catechin EGCG+ECG is 6.6%.
  • the green tea leaves are pulverized and wall-breaking, and the particle size of the treated tea leaves is more than 60% in the range of 10-100 mesh.
  • the extraction temperature is 50 ° C, and collect the ethyl acetate phase; after the ethyl acetate phase is concentrated to a solid content of 10%, use 0.8 times the volume of The deionized water is reversed-phase extraction (this is called washing), the washing temperature is 50 ° C, the washing phase is separated and collected, and then washed twice with the same method, and all the washing phases are collected and merged; and then extracted with 1.5 times the volume of ethyl acetate , the extraction temperature was controlled to be 50 °C, and the ethyl acetate phase was collected.
  • the content of tea polyphenols is 98.3% detected by UV spectrophotometer commonly used in the industry; the content of EGC is 80.8% by HPLC, and the content of ester catechin EGCG+ECG is 7.5%.
  • the present invention also includes other embodiments, and all technical solutions formed by equivalent transformation or equivalent replacement shall fall within the protection scope of the claims of the present invention.

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Abstract

本发明涉及一种富含EGC的非酯型茶多酚的制备方法，将经粉碎破壁的茶叶利用茶叶自身含量的内源性水解酶，将酶解后的茶叶浸提，浸提液经过滤、浓缩后通过移动萃取-水洗进行精制纯化，纯化后产品经过浓缩、干燥，得到富含EGC的非酯型茶多酚产品，茶多酚含量为90-99.5％，EGC的含量大于80％，酯型儿茶素EGCG+ECG的含量在10％以下。本发明的富含EGC的非酯型茶多酚的制备方法不添加任何外源性水解酶，避免外源性水解酶对产品的污染，同时EGC含量高、生产工艺简化、产品收率高，有利于工业化生产。

Description

一种富含EGC的非酯型茶多酚的制备方法 技术领域
本发明属于天然植物提取物加工技术领域，具体涉及一种富含EGC的非酯型茶多酚的制备方法。
背景技术
茶多酚是茶叶中儿茶素类、黄酮类、酚酸类和花色素类化合物的总称，其最主要的作用是抗氧化，对氧自由基的消除作用十分明显，同时茶多酚能够抑菌、杀菌，能有效降低大肠对胆固醇的吸收，防治动脉粥样硬化，还能增强机体免疫能力，抗肿瘤、抗辐射，延缓衰老等作用。因为茶多酚的独特功效，目前已被广泛应用于医药、保健品和食品领域：在医药上已经作为人体的心血管、肝、肾及肠胃病的防治药物；在保健品上已经作为提高人体免疫力、降血脂、降血糖及减肥等制品的主要成分，还作为防晒霜、洗发剂和杀菌消毒剂的主要成分；在油脂食品中加入茶多酚，可以防止食品蚝败和保鲜。
茶多酚最主要成份为儿茶素类物质，儿茶素类又分为酯型儿茶素和非酯型儿茶素，根据茶多酚中儿茶素的类型不同一般分为酯型茶多酚及非酯型茶多酚。两种类型的茶多酚在口感、对胃觉的刺激性及稳定性(水溶性)方面存在一定的差异，酯型茶多酚在对味蕾的刺激方面苦涩性更强，在对人体的胃粘膜的刺激性也更强，同时产品在水溶性中溶解的稳安性更差；而非酯型茶多酚，口感、对胃觉的刺激性及稳定性(水溶性)方面要明显优于酯型儿茶素。因此，非酯型茶多酚存在的这些优势，其在食品、食品添加剂、医药产业中的应用具有更加广阔的应用前景。且非酯型茶多酚含有多种非酯型儿茶素单体，其中EGC的生物活性相对较高。
目前制备富含EGC的非酯型茶多酚技术通常为：通过提取溶剂(水、乙醇、乙酸乙酯中至少一种)从茶叶中提取茶多酚，经过滤、浓缩等处理后，添加一些外源性的水解酶等进行水解，然后再通过高温灭活酶体，过滤去除，得到非酯型茶多酚溶液，再通过层析分离方法对产品进行纯化处理，最终干燥后得到非酯型茶多酚产品。
上述工艺中，加入了外源性水解酶，容易引起产品的污染，处理不好会导致过敏源的产生，同时采用层析方法进行产品的制备，工艺相对会比较复杂，不利于工业化生产，同时在加工过程中产品的损失也比较大，收率相对较低。
发明内容
本发明的目的是提供一种富含EGC的非酯型茶多酚的制备方法，不添加任何外源性水解酶，避免外源性水解酶对产品的污染，同时EGC含量高、生产工艺简化、产品收率高，有利于工业化生产。
本发明解决上述问题所采用的技术方案为：一种富含EGC的非酯型茶多酚的制备方法，包括以下步骤：
(1)茶叶预处理：采用绿茶茶叶为原料，将茶叶进行粉碎破壁处理，得到茶叶颗粒。
(2)内源酶酶解：将预处理后的茶叶颗粒加入到动态提取罐中，加入去离子水，并加入食品级酸调节pH为2.0-6.5，在搅拌条件下，茶叶自身的水解酶对茶叶中的酯型茶多酚进行酶解。
(3)浸提：将经酶解后的茶叶原料用去离子水浸提两次，浸提结束后过滤，合并两次滤液。
(4)过滤：将上述滤液冷却，再次过滤，收集滤液。
(5)浓缩：将步骤(4)中滤液浓缩。
(6)移动萃取-水洗：用1-2倍体积的乙酸乙酯对步骤(5)中浓缩液进行萃取，收集乙酸乙酯相，后将乙酸乙酯相浓缩至含固率为2-10％后，用0.2-1倍体积的去离子水进行反相萃取，收集水洗相，再同法反相萃取1-5次，合并水洗相，水洗相再用1-2倍体积的乙酸乙酯进行萃取，收集乙酸乙酯相。
(7)转溶：将步骤(6)中最终收集的乙酸乙酯相浓缩至浸膏，再加入去离子水进行溶解，得到茶多酚水溶液。
(8)浓缩：将步骤(7)中茶多酚水溶液真空浓缩。
(9)干燥：将步骤(8)中浓缩液通过喷零干燥方式进行干燥，得到富含EGC的非酯型茶多酚产品。
优选的，步骤(1)中所述预处理后的茶叶颗粒的10-100目筛余物不大于40％，即预处理后的茶叶颗粒在10-100目范围内的达60％以上。
优选的，步骤(2)中所述去离子水的加入量为预处理后的茶叶体积的1-5倍，所述食品级酸为柠檬酸，所述搅拌速度为50～300r/min，所述酶解温度为30～50℃，酶解时间为4～12h。
优选的，步骤(3)中所述浸提温度为60～90℃，单次去离子水加入量为茶叶原料质量的6～10倍，单次浸提时间为20-60min。
优选的，步骤(4)中所述冷却温度为30-50℃，所述过滤精度为100-300目。
优选的，步骤(5)中所述浓缩具体为：将步骤(4)中滤液浓缩至0.5-2.5Brix。
优选的，步骤(6)中所述乙酸乙酯萃取温度为30-60℃，所述去离子水反相萃取温度为30-60℃。
优选的，步骤(8)中所述浓缩具体为：将步骤(7)中茶多酚水溶液在真空度-0.080MPa以上、温度52-72℃条件下真空浓缩至20-50Brix。
本发明利用茶叶自身含量的内源性水解酶，在浸提前进行酶解，将茶叶中的酯型儿茶素酶解为非酯型茶儿素物质(特别是EGCG转化为EGC)，再进行产品的浸提；收集到的浸提液经处理后通过移动萃取-水洗的方法进行产品的精制纯化，纯化后产品经过浓缩、干燥工艺，得到富含EGC的非酯型茶多酚产品，茶多酚含量为90-99.5％，EGC的含量大于80％，酯型儿茶素EGCG+ECG的含量在可控范围内(10％以下)。
与现有技术相比，本发明的优点在于：
(1)不添加任何外源性水解酶，采用茶叶当中自身含有的内源性的水解酶结合水解体系，对茶叶中的酯型茶多酚进行水解，使其最大程度的转化为非酯型茶多酚，在提高非酯型茶多酚产品的含量同时杜绝了外源性水解酶对产品的污染。
(2)用移动萃取-水洗的方法进行产品纯化，不仅大大提高EGC含量，同时简化生产工艺，大大减少产品的损失，大大提高产品收率，有利于工业化生产。
(3)采用本发明制备方法所得到的富含EGC的非酯型茶多酚产品，茶多酚含量为90-99.5％，EGC的含量大于80％，酯型儿茶素EGCG+ECG的含量在可控范围内(10％以下)。
具体实施方式
以下结合实施例对本发明作进一步详细描述。
实施例1
一种富含EGC的非酯型茶多酚的制备方法，包括以下步骤：
(1)将绿茶茶叶进行粉碎破壁处理，处理后的茶叶颗粒大小在10-100目范围内的达60％以上，即预处理后的茶叶颗粒的10-100目筛余物不大于40％。
(2)将5.0Kg预处理好的茶叶加入到动态提取罐(带搅拌)中，加入4倍体积的去离子水，并用柠檬酸调节pH值为5.0，在转速100r/min、温度40℃条件下酶解6小时。
(3)后关闭搅拌，加入去离子水在80℃进行一次浸提，控制提取罐中茶叶与去离子水的质量比为1:8，浸提30min；一次浸提结束后，过滤出滤液，再加入同样比例的去离子水，同法浸提一次，浸提结束后过滤，合并两次滤液。
(4)将收集的滤液冷却至40℃，进一步用精度为200目的过滤器过滤并收集滤液。
(5)通过浓缩装置，将上述茶水浓缩至1.0Brix。
(6)再用1.2倍体积的乙酸乙酯对上述浓缩茶水进行萃取，萃取温度为40℃，收集乙酸乙酯相；将乙酸乙酯相浓缩至含固率为8％，用0.5倍体积的去离子水进行反相萃取(此称为水洗)，水洗温度为40℃，分离收集水洗相，再同法水洗3次，收集合并 所有水洗相；再用1.5倍体积的乙酸乙酯进行萃取，萃取温度为40℃，收集乙酸乙酯相。
(7)将收集的乙酸乙酯溶液浓缩至浸膏，再加入3L去离子水进行溶解，得到茶多酚水溶液。
(8)将茶多酚水溶液真空浓缩，控制真空度为-0.080MPa以上，温度52-72℃，将产品水溶液浓缩至35Brix。
(9)用喷零干燥方式对浓缩液进行干燥处理，收集得到305g粉剂产品。
产品采用行业内通用的紫外分光光度计检测茶多酚含量达98.6％；经HPLC检测，EGC含量为81.3％，酯型儿茶素EGCG+ECG含量为7.3％。
实施例2
(1)将绿茶茶叶进行粉碎破壁处理，处理后的茶叶颗粒大小在10-100目范围内的达60％以上。
(2)将5.0Kg预处理好的茶叶加入到动态提取罐(带搅拌)中，加入4倍体积的去离子水，并用柠檬酸调节pH值为6.0，在转速150r/min、温度45℃条件下酶解10小时。
(3)后关闭搅拌，加入去离子水在80℃进行一次浸提，控制提取罐中茶叶与去离子水的质量比为1:10，浸提30min；一次浸提结束后，过滤出滤液，再加入同样比例的去离子水，同法浸提一次，浸提结束后过滤，合并两次滤液。
(4)将收集的滤液冷却至40℃，进一步用精度为200目的过滤器过滤并收集滤液。
(5)通过浓缩装置，将上述茶水浓缩至1.0Brix。
(6)再用1.2倍体积的乙酸乙酯对上述浓缩茶水进行萃取，萃取温度为40℃，收集乙酸乙酯相；将乙酸乙酯相浓缩至含固率为8％，后用0.5倍体积的去离子水进行反相萃取(此称为水洗)，水洗温度为40℃，分离收集水洗相，再同法水洗3次，收集合并所有水洗相；再用1.5倍体积的乙酸乙酯进行萃取，萃取温度为40℃，收集乙酸乙酯相。
(7)将收集的乙酸乙酯相浓缩至浸膏，再加入3L去离子水进行溶解，得到茶多酚水溶液。
(8)将茶多酚水溶液真空浓缩，控制真空度为-0.080MPa以上，温度52-72℃，将产品水溶液浓缩至35Brix。
(9)用喷零干燥方式对浓缩液进行干燥处理，收集得到312g粉剂产品。
产品采用行业内通用的紫外分光光度计检测茶多酚含量达99.2％；经HPLC检测，EGC含量为83.0％，酯型儿茶素EGCG+ECG含量为6.6％。
实施例3
(1)将绿茶茶叶进行粉碎破壁处理，处理后的茶叶颗粒大小在10-100目范围内的达60％以上。
(2)将5.0Kg预处理好的茶叶加入到动态提取罐(带搅拌)中，加入4倍体积的去离子水，并用柠檬酸调节pH值为6.0，在转速100r/min、温度40℃条件下酶解8小时。
(3)后关闭搅拌，加入去离子水在80℃进行一次浸提，控制提取罐中茶叶与去离子水的质量比为1:8，浸提30min；一次浸提结束后，过滤出滤液，再加入同样比例的去离子水，同法浸提一次，浸提结束后过滤，合并两次滤液。
(4)将收集的滤液冷却至40℃，进一步用精度为200目的过滤器过滤并收集滤液。
(5)通过浓缩装置，将上述滤液浓缩至1.5Brix。
(6)再用1.5倍体积的乙酸乙酯对上述浓缩液进行萃取，萃取温度为50℃，收集乙酸乙酯相；将乙酸乙酯相浓缩至含固率为10％后，用0.8倍体积的去离子水进行反相萃取(此称为水洗)，水洗温度为50℃，分离收集水洗相，再同法水洗2次，收集合并所有水洗相；再用1.5倍体积的乙酸乙酯进行萃取，控制萃取温度为50℃，收集乙酸乙酯相。
(7)将收集的乙酸乙酯相浓缩至浸膏，再加入3L去离子水进行溶解，得到茶多酚水溶液。
(8)将茶多酚水溶液真空浓缩，控制真空度为-0.080MPa以上，温度52-72℃，将产品水溶液浓缩至40Brix。
(9)用喷零干燥方式对浓缩液进行干燥处理，收集得到303g粉剂产品。
产品采用行业内通用的紫外分光光度计检测茶多酚含量达98.3％；经HPLC检测，EGC含量为80.8％，酯型儿茶素EGCG+ECG含量为7.5％。
除上述实施例外，本发明还包括有其他实施方式，凡采用等同变换或者等效替换方式形成的技术方案，均应落入本发明权利要求的保护范围之内。

Claims (8)

  1. 一种富含EGC的非酯型茶多酚的制备方法，其特征在于：包括以下步骤：
    (1)茶叶预处理：采用绿茶茶叶为原料，将茶叶进行粉碎破壁处理，得到茶叶颗粒；
    (2)内源酶酶解：将预处理后的茶叶颗粒加入到动态提取罐中，加入去离子水，并加入食品级酸调节pH为2.0-6.5，在搅拌条件下，茶叶自身的水解酶对茶叶中的酯型茶多酚进行酶解；
    (3)浸提：将经酶解后的茶叶原料用去离子水浸提两次，浸提结束后过滤，合并两次滤液；
    (4)过滤：将上述滤液冷却，再次过滤，收集滤液；
    (5)浓缩：将步骤(4)中滤液浓缩；
    (6)移动萃取-水洗：用1-2倍体积的乙酸乙酯对步骤(5)中浓缩液进行萃取，收集乙酸乙酯相，后将乙酸乙酯相浓缩至含固率为2-10％后，用0.2-1倍体积的去离子水进行反相萃取，收集水洗相，再同法反相萃取1-5次，合并水洗相，水洗相再用1-2倍体积的乙酸乙酯进行萃取，收集乙酸乙酯相；
    (7)转溶：将步骤(6)中最终收集的乙酸乙酯相浓缩至浸膏，再加入去离子水进行溶解，得到茶多酚水溶液；
    (8)浓缩：将步骤(7)中茶多酚水溶液真空浓缩；
    (9)干燥：将步骤(8)中浓缩液通过喷零干燥方式进行干燥，得到富含EGC的非酯型茶多酚产品。
  2. 根据权利要求1所述的富含EGC的非酯型茶多酚的制备方法，其特征在于：步骤(1)中所述预处理后的茶叶颗粒的10-100目筛余物不大于40％。
  3. 根据权利要求1所述的富含EGC的非酯型茶多酚的制备方法，其特征在于：步骤(2)中所述去离子水的加入量为预处理后的茶叶体积的1-5倍，所述食品级酸为柠檬酸，所述搅拌速度为50～300r/min，所述酶解温度为30～50℃，酶解时间为4～12h。
  4. 根据权利要求1所述的富含EGC的非酯型茶多酚的制备方法，其特征在于：步骤(3)中所述浸提温度为60～90℃，单次去离子水加入量为茶叶原料质量的6～10倍，单次浸提时间为20-60min。
  5. 根据权利要求1所述的富含EGC的非酯型茶多酚的制备方法，其特征在于：步骤(4)中所述冷却温度为30-50℃，所述过滤精度为100-300目。
  6. 根据权利要求1所述的富含EGC的非酯型茶多酚的制备方法，其特征在于：步骤(5)中所述浓缩具体为：将步骤(4)中滤液浓缩至Brix 0.5-2.5。
  7. 根据权利要求1所述的富含EGC的非酯型茶多酚的制备方法，其特征在于：步 骤(6)中所述乙酸乙酯萃取温度为30-60℃，所述去离子水反相萃取温度为30-60℃。
  8. 根据权利要求1所述的富含EGC的非酯型茶多酚的制备方法，其特征在于：步骤(8)中所述浓缩具体为：将步骤(7)中茶多酚水溶液在真空度-0.080MPa以上、温度52-72℃条件下真空浓缩至Brix20-50。
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