WO2021248601A1 - 以多聚体多酚为主的荔枝皮提取物在制备α-淀粉酶抑制剂中的应用 - Google Patents

以多聚体多酚为主的荔枝皮提取物在制备α-淀粉酶抑制剂中的应用 Download PDF

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WO2021248601A1
WO2021248601A1 PCT/CN2020/100763 CN2020100763W WO2021248601A1 WO 2021248601 A1 WO2021248601 A1 WO 2021248601A1 CN 2020100763 W CN2020100763 W CN 2020100763W WO 2021248601 A1 WO2021248601 A1 WO 2021248601A1
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preparation
peel extract
lychee
solution
polyphenols
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French (fr)
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杨子明
张利
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广西壮族自治区中国科学院广西植物研究所
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Priority to US17/002,760 priority Critical patent/US20210379135A1/en
Publication of WO2021248601A1 publication Critical patent/WO2021248601A1/zh

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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)
    • A61K36/77—Sapindaceae (Soapberry family), e.g. lychee or soapberry
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A01N65/08—Magnoliopsida [dicotyledons]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • 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
    • A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
    • 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
    • A61K2236/55—Liquid-liquid separation; Phase separation
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
    • Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
    • Y02P60/87—Re-use of by-products of food processing for fodder production

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  • the invention relates to the field of biotechnology, and more specifically to the application of a litchi bark extract mainly composed of polymer polyphenols in the preparation of ⁇ -amylase inhibitors.
  • Alpha-amylase inhibitors have a wide range of uses: (1) It can be used as drugs, health products or food for obesity and diabetes prevention: Starch is the main component of carbohydrates in the diet. It is hydrolyzed by saliva and pancreatic amylase to produce maltose. It is absorbed in the intestine in the form of glucose. Alpha-amylase inhibitors in the gastrointestinal tract can inhibit the activity of amylase in the gastrointestinal tract, slow down the digestion and absorption of the main carbohydrate starch in food, thereby inhibiting the increase in blood sugar concentration, and can effectively match the diet of diabetic patients treat. For obese patients, it can reduce the conversion of sugar to fat, delay intestinal emptying, and increase fat consumption to reduce weight.
  • Alpha-amylase inhibitor exerts weight loss and hypoglycemic effects by inhibiting amylase, and is excreted through the gastrointestinal tract, so it does not need to enter the blood circulatory system and does not act on the brain center. It does not suppress appetite while losing weight and reducing blood sugar. , There are no side effects even when the dosage is high, which is in line with the weight loss principles of the World Health Organization. (2) It can be used as a new insect-resistant and insecticidal biological pesticide: the ⁇ -amylase in insects decomposes starch, glycogen and other substances into maltose, glucose and fructose that can be directly absorbed, and participates in energy metabolism in the body.
  • Alpha-amylase inhibitors inhibit the activity of alpha-amylase and reduce the generalization of carbohydrates in insects, thereby inhibiting the growth and development of insects, and even causing insects to die. They can be used as new insect-resistant and insecticidal biological pesticides. (3) It can prevent dental caries: ⁇ -amylase inhibitors can also prevent the saccharification of starchy foods in the oral cavity and reduce the tartar formed by sugars, thereby preventing dental caries.
  • ⁇ -amylase inhibitors can increase the residual sugars in the intestinal tract, which is beneficial to the survival of the beneficial intestinal flora, and the residual sugars are fermented to produce lactic acid to make the intestinal tract
  • the environment tends to be acidic, which promotes the dissolution and absorption of calcium in food, resulting in the prevention and treatment of osteoporosis.
  • Litchi is the fruit of an evergreen plant in the Sapindaceae family. It is mainly produced in Guangdong, Guangxi, Fujian and other places in my country. At present, the annual production of litchi in my country is about 1.6 million tons, accounting for more than 80% of the total production of litchi in the world. About 20% of the peel waste of fresh fruit quality is produced during the deep processing of litchi. As a waste of bulk agricultural products, the peel of litchi has not been effectively developed and utilized.
  • the present invention provides the application of a litchi bark extract based on polymer polyphenols in the preparation of ⁇ -amylase inhibitors.
  • litchi bark extract mainly composed of polymer polyphenols in the preparation of ⁇ -amylase inhibitors.
  • the preparation method of the litchi bark extract mainly composed of polymer polyphenols is as follows:
  • the medicinal solution was purified by XDA-7 macroporous resin by column chromatography and eluted with a volume fraction of 60% ethanol aqueous solution to obtain the extract eluent; the column chromatography purification conditions were: loading liquid The mass concentration is 3.5mg/mL, the pH value of the drug solution is 4.5, the sample loading rate is 4BV/h; the flow rate of the eluent is 6BV/h;
  • the extract eluent was concentrated under reduced pressure under the conditions of a vacuum degree of 95kPa, a heating bath temperature of 40°C, and a cooling water temperature of 15°C, and spray-dried the obtained reflux solution at a feed rate of 51mL/min ,
  • the inlet air temperature is 160°C
  • the exhaust air temperature is 60°C
  • the concentration of the material liquid is 22%
  • the rotation speed of the atomizer is 21000r/min to obtain the lychee peel extract.
  • the litchi bark extract mainly composed of polymer polyphenols is used in the preparation of medicines for weight loss, blood sugar reduction, dental caries prevention, and osteoporosis prevention and treatment.
  • the application of the litchi bark extract mainly composed of polymer polyphenols in the preparation of biological pesticides is mainly composed of polymer polyphenols.
  • the specific application method of the litchi bark extract based on polymer polyphenols is: take the litchi bark extract based on polymer polyphenols, add or not add auxiliary materials, and prepare various preparations according to conventional methods; It can be made into existing conventional dosage forms such as capsules, tablets, granules or water solvents.
  • the present disclosure provides the application of a litchi peel extract based on polymer polyphenols in the preparation of ⁇ -amylase inhibitors.
  • the litchi peel contains a large amount of Polyphenols, polyphenols have multiple biological activities such as anti-oxidation, anti-tumor, prevention of cardiovascular disease, delaying aging, etc.; Enzymes have a good inhibitory effect, and ⁇ -amylase is used in weight loss and blood sugar reduction, biological pesticides, prevention of dental caries, prevention and treatment of osteoporosis.
  • Figure 1 is a diagram showing the content ratio of each polar part of the lychee peel extract of the present invention
  • Fig. 2 is a graph showing the inhibition curve of amylase by litchi bark extracts with different concentrations of polymer polyphenols.
  • the medicinal solution was purified by XDA-7 macroporous resin by column chromatography and eluted with a 60% ethanol aqueous solution with a volume fraction of 60% to obtain the extract eluate; the column chromatography purification conditions were: the mass concentration of the sample solution was 3.5mg/mL, the pH value of the drug solution is 4.5, and the sample loading rate is 4BV/h; the flow rate of the eluent is 6BV/h; Concentrate under reduced pressure at a cooling water temperature of 15°C, and spray-dry the obtained reflux liquid at a feed rate of 51mL/min, an inlet air temperature of 160°C, an exhaust air temperature of 60°C, and a liquid concentration of 22%
  • the Folin-phenol colorimetric method was used to determine the polyphenol content of 77.8%.
  • Example 1 The lychee bark extract obtained in Example 1 was studied for component separation, and the specific operations were as follows:
  • the litchi bark extract obtained in Example 1 was extracted step by step with n-hexane, ethyl acetate/anhydrous ether (1:1) to obtain the n-hexane phase and ethyl acetate/anhydrous ether of the litchi bark extract
  • the total phenol content of the three phases was determined separately for the three phases and the water phase. The results showed that the total phenol content of the water phase was the highest, followed by the total phenol content of the ethyl acetate/anhydrous ether phase, and the n-hexane phase was the smallest ( Figure 1).
  • the phenolic substances contained in the water phase are multimers of polyphenols, while the n-hexane phase and ethyl acetate/anhydrous ether phase are oligomers and monomers of polyphenols. Therefore, the lychee peel extract is mainly composed of polyphenols, followed by oligomers and monomers, of which polymer polyphenols account for about 79%.
  • Example 3 Inhibitory effect of litchi bark extract based on polymer polyphenols on ⁇ -amylase
  • disodium hydrogen phosphate take 35.81g of Na 2 HPO 4 ⁇ 12H 2 O and add 500ml of distilled water, fully dissolve and store;
  • DNS developer Dissolve 10.5g of sodium hydroxide with appropriate amount of water, and dissolve 91.0g of sodium potassium tartrate with appropriate amount of water. If necessary, dissolve with proper heating to fully dissolve the above two solutions. Then mix them evenly; then weigh 3.15g of DNS (3,5-dinitrosalicylic acid) into the above solution, and heat to completely dissolve it. After cooling, add 2.5ml of phenol, and finally add 2.5g of anhydrous sodium sulfite, stir well and add distilled water to make 500ml solution, and it can be used after 3 days.

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Abstract

以多聚体多酚为主的荔枝皮提取物在制备α-淀粉酶抑制剂中的应用，属于生物技术领域。以多聚体多酚为主的荔枝皮提取物对α-淀粉酶有很好的抑制作用，而α-淀粉酶在减肥降糖、生物农药、预防龋齿、预防和治疗骨质疏松症中都有应用。

Description

以多聚体多酚为主的荔枝皮提取物在制备α-淀粉酶抑制剂中的应用 技术领域
本发明涉及生物技术领域，更具体的说是涉及以多聚体多酚为主的荔枝皮提取物在制备α-淀粉酶抑制剂中的应用。
背景技术
α-淀粉酶抑制剂的用途十分广泛：(1)可以作为肥胖和糖尿病防治的药物、保健品或者食品：淀粉是饮食中的碳水化合物的主要组成，经唾液、胰淀粉酶水解生成麦芽糖，再以葡萄糖的形式在肠道内吸收。α-淀粉酶抑制剂在胃肠道中可以抑制胃肠道内淀粉酶的活性，使食物中主要的碳水化合物淀粉的消化吸收减慢，从而抑制血糖浓度的升高，可以有效地配合糖尿病人的饮食治疗。对于肥胖患者，可减少糖向脂肪转化，延缓肠道的排空，增加脂肪的消耗以减少体重。α-淀粉酶抑制剂通过对淀粉酶的抑制作用而发挥减肥、降糖效应，并经胃肠道排出体外，因此不必进入血液循环系统，不作用于大脑中枢，减肥降糖的同时不抑制食欲，使用剂量很高时也无副作用，符合世界卫生组织的减肥原则。(2)可以作为新型抗虫、杀虫生物农药：昆虫体内的α-淀粉酶将淀粉、糖原等物质分解成可以直接吸收的麦芽糖、葡萄糖和果糖等，参与体内的能量代谢。α-淀粉酶抑制剂抑制α-淀粉酶的活性，会降低昆虫体内糖类的通化作用，从而抑制昆虫的生长发育，甚至导致昆虫死亡，可以作为新型抗虫、杀虫生物农药。(3)可以预防龋齿：α-淀粉酶抑制剂还能阻止口腔中淀粉类食物的糖化，减少由糖类形成的齿垢，从而预防龋齿。(4)可以预防和治疗骨质疏松症：α-淀粉酶抑制剂可以使肠道内残留的糖类增加，有利于肠道有益菌群的生存，并将残留的糖发酵产生乳酸，使肠道趋于酸性环境，从而促使食物中钙的溶解和吸收，产生预防和治疗骨质疏松症的效果。
荔枝为无患子科常绿植物的果实，主产于我国广东、广西、福建等地，目前我国荔枝年产量约160万吨，占世界荔枝总产量80％以上。在荔枝的深加工过程中产生了约20％鲜果质量的果皮废弃物，荔枝果皮作为大宗农产品废弃 物至今未能较好地得到有效开发和利用。
因此，提供以多聚体多酚为主的荔枝皮提取物在制备α-淀粉酶抑制剂中的应用是本领域技术人员亟需解决的问题。
发明内容
有鉴于此，本发明提供了以多聚体多酚为主的荔枝皮提取物在制备α-淀粉酶抑制剂中的应用。
为了实现上述目的，本发明采用如下技术方案：
以多聚体多酚为主的荔枝皮提取物在制备α-淀粉酶抑制剂中的应用。
进一步，所述以多聚体多酚为主的荔枝皮提取物的制备方法如下：
(1)将新鲜荔枝果皮和8倍体积浓度80％乙醇水溶液在室温条件下提取2次，每次浸泡7d，过滤，合并滤液；
(2)将上述滤液在3500rpm条件下离心10min，收集上清液，得到药液；
(3)将药液过XDA-7大孔树脂进行柱层析纯化，用体积分数为60％乙醇水溶液洗脱，得到提取物洗脱液；所述柱层析纯化的条件为：上样液质量浓度为3.5mg/mL，药液的pH值为4.5，上样速率为4BV/h；洗脱液的流速为6BV/h；
(4)将所述提取物洗脱液在真空度95kPa，加热浴温度40℃，冷却水温度15℃的条件下进行减压浓缩，将得到的回流液进行喷雾干燥，进料速率51mL/min，进风温度160℃，排风温度60℃，料液浓度22％，雾化器转速21000r/min，得到荔枝皮提取物。
进一步，所述以多聚体多酚为主的荔枝皮提取物在制备减肥、降血糖、预防龋齿、预防和治疗骨质疏松症药物中的应用。
进一步，所述以多聚体多酚为主的荔枝皮提取物在制备生物农药中的应用。
以多聚体多酚为主的荔枝皮提取物的具体应用方法为：取以多聚体多酚为主的荔枝皮提取物，加入或不加入辅料，按常规方法制备成各种制剂；具体可以制成胶囊剂、片剂、颗粒剂或水溶剂等现有常规剂型。
经由上述的技术方案可知，与现有技术相比，本发明公开提供了以多聚体多酚为主的荔枝皮提取物在制备α-淀粉酶抑制剂中的应用，荔枝果皮中含有大量的多酚类化合物，多酚类物质具有抗氧化、抗肿瘤、预防心血管疾病、延缓衰老等多种生物活性；本发明研究证明以多聚体多酚为主的荔枝皮提取物对α-淀粉酶有很好的抑制作用，α-淀粉酶在减肥降糖、生物农药、预防龋齿、预防和治疗骨质疏松症中都有应用。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案，下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图仅仅是本发明的实施例，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据提供的附图获得其他的附图。
图1附图为本发明荔枝皮提取物各极性部分含量比例图；
图2附图为本发明不同浓度的以多聚体多酚为主的荔枝皮提取物对淀粉酶的抑制曲线。
具体实施方式
下面将结合本发明实施例中的附图，对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。
实施例1
荔枝皮提取物的制备方法，具体步骤如下：
将新鲜荔枝果皮和8倍体积浓度80％乙醇水溶液在室温条件下提取2次，每次浸泡7d，过滤，合并滤液；将上述滤液在3500rpm条件下离心10min，收集上清液，得到药液；将药液过XDA-7大孔树脂进行柱层析纯化，用体积分数为60％乙醇水溶液洗脱，得到提取物洗脱液；所述柱层析纯化的条件为：上样液质量浓度为3.5mg/mL，药液的pH值为4.5，上样速率为4BV/h；洗 脱液的流速为6BV/h；将所述提取物洗脱液在真空度95kPa，加热浴温度40℃，冷却水温度15℃的条件下进行减压浓缩，将得到的回流液进行喷雾干燥，进料速率51mL/min，进风温度160℃，排风温度60℃，料液浓度22％，雾化器转速21000r/min，得到荔枝皮提取物。
采用Folin-酚比色法进行测定，测得多酚含量为77.8％。
实施例2
对实施例1获得的荔枝皮提取物进行成分分离研究，具体操作如下：
取实施例1获得的荔枝皮提取物用正己烷、乙酸乙酯/无水乙醚(1:1)分步萃取，得到荔枝皮提取物浸提液的正己烷相、乙酸乙酯/无水乙醚相和水相，分别测定此三相中总酚的含量，结果表明水相的总酚含量最高，乙酸乙脂/无水乙醚相的总酚含量次之，正己烷相最小(图1)。水相中含有的酚类物质为多酚的多聚体，而正己烷相以及乙酸乙酯/无水乙醚相则为多酚的低聚体和单体。因此，荔枝皮提取物主要以多酚的多聚体为主，其次是低聚体和单体，其中多聚体多酚约占79％。
实施例3以多聚体多酚为主的荔枝皮提取物对α-淀粉酶的抑制作用
(一)配制试验试剂：
(1)pH＝7.0的0.01mol/L的PBS溶液的配制：
加0.2mol/L磷酸氢二钠61.0ml，加0.2mol/L磷酸二氢钠39.0ml，然后加0.8％氯化钠溶液至1L，既可以得到pH＝7.0的0.01mol/L PBS缓冲液。
0.2mol/L磷酸氢二钠：取Na 2HPO 4·12H 2O 35.81g加500ml蒸馏水，充分溶解后保存；
0.2mol/L磷酸二氢钠：取NaH 2PO 4·2H 2O 15.6g加500ml蒸馏水，充分溶解后保存；
0.8％氯化钠的配制：取NaCl 4g加500ml蒸馏水，充分溶解后保存。
(2)0.5％的淀粉溶液配制：取可溶性淀粉200mg溶于40ml pH＝7.0的0.01mol/L PBS缓冲液，加热使其溶解，充分溶解后保存。
(3)α-淀粉酶溶液的配制：取α-淀粉酶加入pH＝7.0的0.01mol/L PBS缓冲液，使其配制成10μg/ml的溶液，充分溶解后保存。
(4)DNS显色剂的配制：取10.5g氢氧化钠加适量的水溶解，另取91.0g的酒石酸钾钠加适量的水溶解，必要时可适当加热溶解，把以上两种溶液充 分溶解后混合均匀在一起；再称取3.15g的DNS(3,5-二硝基水杨酸)加入上述溶液中，加热使其完全溶解。冷却后加入2.5ml的苯酚，最后再加入2.5g的无水亚硫酸钠充分搅拌均匀后加蒸馏水使其配成500ml溶液，放置3天后即可使用。
(5)以多聚体多酚为主的荔枝皮提取物溶液的配置：将以多聚体多酚为主的荔枝皮提取物溶于pH＝7.0的0.01mol/L PBS缓冲液中，配成各种浓度的以多聚体多酚为主的荔枝皮提取物溶液。
(二)以多聚体多酚为主的荔枝皮提取物对α-淀粉酶抑制活性的测定
空白组：先加入0.9ml的pH＝7.0的0.01mol/L PBS缓冲液，再加入0.1mlα-淀粉酶溶液混合均匀，置于37℃恒温箱中水浴孵育10分钟；加入0.2ml的淀粉溶液启动反应，于37℃恒温箱中孵育10分钟，最后再加入1ml的DNS显色剂混合均匀后，放入95℃恒温箱中孵育5分钟；流水冷却，用半自动生化分析仪在波长为540nm的条件下测定OD值。
测定组：先加入0.8ml的pH＝7.0的0.01mol/L PBS缓冲液，再加入0.1ml不同浓度的以多聚体多酚为主的荔枝皮提取物溶液和0.1mlα-淀粉酶溶液混合均匀，置于37℃恒温箱中水浴孵育10分钟；加入0.2ml的淀粉溶液启动反应，于37℃恒温箱中孵育10分钟，最后再加入1ml的DNS显色剂混合均匀后，放入95℃恒温箱中孵育5分钟；流水冷却，用半自动生化分析仪在波长为540nm的条件下测定OD值。
对照组：先加入0.8ml的pH＝7.0的0.01mol/L PBS缓冲液，再加入0.1ml不同浓度的以多聚体多酚为主的荔枝皮提取物溶液和0.1ml pH＝7.0的0.01mol/L PBS缓冲液混合均匀，置于37℃恒温箱中水浴孵育10分钟；加入0.2ml的淀粉溶液启动反应，于37℃恒温箱中孵育10分钟，最后再加入1ml的DNS显色剂混合均匀后，放入95℃恒温箱中孵育5分钟；流水冷却，用半自动生化分析仪在波长为540nm的条件下测定OD值。
计算不同浓度的以多聚体多酚为主的荔枝皮提取物溶液对α-淀粉酶的抑制率，结果见图2。图2结果显示，当提取物的浓度不断升高时对α-淀粉酶的抑制效果也在不断的提高，以多聚体多酚为主的荔枝皮提取物在浓度为151.5μg/ml时对α-淀粉酶的抑制率达到80.2％，由此可以看出以多聚体多酚为主的荔枝皮提取物对α-淀粉酶有很好的抑制作用。经SPSS17.0计算软件计算 得出，以多聚体多酚为主的荔枝皮提取物对α-淀粉酶的半数抑制浓度(IC 50)为62.2μg/ml。
对所公开的实施例的上述说明，使本领域专业技术人员能够实现或使用本发明。对这些实施例的多种修改对本领域的专业技术人员来说将是显而易见的，本文中所定义的一般原理可以在不脱离本发明的精神或范围的情况下，在其它实施例中实现。因此，本发明将不会被限制于本文所示的这些实施例，而是要符合与本文所公开的原理和新颖特点相一致的最宽的范围。

Claims (4)

  1. 以多聚体多酚为主的荔枝皮提取物在制备α-淀粉酶抑制剂中的应用。
  2. 根据权利要求1所述的以多聚体多酚为主的荔枝皮提取物在制备α-淀粉酶抑制剂中的应用，其特征在于，所述以多聚体多酚为主的荔枝皮提取物的制备方法如下：
    (1)将新鲜荔枝果皮和8倍体积浓度80％乙醇水溶液在室温条件下提取2次，每次浸泡7d，过滤，合并滤液；
    (2)将上述滤液在3500rpm条件下离心10min，收集上清液，得到药液；
    (3)将药液过XDA-7大孔树脂进行柱层析纯化，用体积分数为60％乙醇水溶液洗脱，得到提取物洗脱液；所述柱层析纯化的条件为：上样液质量浓度为3.5mg/mL，药液的pH值为4.5，上样速率为4BV/h；洗脱液的流速为6BV/h；
    (4)将所述提取物洗脱液在真空度95kPa，加热浴温度40℃，冷却水温度15℃的条件下进行减压浓缩，将得到的回流液进行喷雾干燥，进料速率51mL/min，进风温度160℃，排风温度60℃，料液浓度22％，雾化器转速21000r/min，得到荔枝皮提取物。
  3. 根据权利要求1所述的以多聚体多酚为主的荔枝皮提取物在制备α-淀粉酶抑制剂中的应用，其特征在于，所述以多聚体多酚为主的荔枝皮提取物在制备减肥、降血糖、预防龋齿、预防和治疗骨质疏松症药物中的应用。
  4. 根据权利要求1所述的以多聚体多酚为主的荔枝皮提取物在制备α-淀粉酶抑制剂中的应用，其特征在于，所述以多聚体多酚为主的荔枝皮提取物在制备生物农药中的应用。
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