WO2021098164A1 - 一种海带多酚的提取方法 - Google Patents

一种海带多酚的提取方法 Download PDF

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WO2021098164A1
WO2021098164A1 PCT/CN2020/089964 CN2020089964W WO2021098164A1 WO 2021098164 A1 WO2021098164 A1 WO 2021098164A1 CN 2020089964 W CN2020089964 W CN 2020089964W WO 2021098164 A1 WO2021098164 A1 WO 2021098164A1
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kelp
polyphenols
water
extracting
extraction
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PCT/CN2020/089964
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English (en)
French (fr)
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解万翠
杨锡洪
宋琳
车红霞
林鹏程
谢静雯
郁东兴
刘家鸣
刘永健
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青岛科技大学
山东省宏福慧科技有限公司
尚好科技有限公司
青岛智科检验检测有限公司
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Publication of WO2021098164A1 publication Critical patent/WO2021098164A1/zh
Priority to ZA2022/04640A priority Critical patent/ZA202204640B/en

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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
    • A61K36/02—Algae
    • A61K36/03—Phaeophycota or phaeophyta (brown algae), e.g. Fucus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants
    • 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
    • A61K2236/17—Preparation or pretreatment of starting material involving drying, e.g. sun-drying or wilting
    • 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
    • A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
    • 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
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying

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  • the invention relates to the technical field of plant extraction, in particular to a method for extracting kelp polyphenols.
  • seaweed phenolic compounds have strong biological activity and are a hot research topic in recent years. According to the molecular structure characteristics, seaweed phenolic compounds can be divided into simple phenolic compounds and polyphenolic compounds.
  • my country’s kelp resources are very rich. Kelp farming has become one of the pillar industries in China’s mariculture, with its output ranking first in the world. Kelp polyphenols have antioxidant, anti-tumor, antibacterial and chemical defense effects, and they have great development and utilization value.
  • kelp phenolic substances have problems such as easy oxidation, instability, complex extraction components, and low extraction rate, which restrict their industrial production and production.
  • the extraction of kelp polyphenols mainly uses dried, crushed and sieved kelp powder as raw materials, using a certain concentration of ethanol solution extraction method, but polyphenols in kelp usually form stable compounds with proteins and polysaccharides in the form of hydrogen bonds and hydrophobic bonds, resulting in the extraction rate Low.
  • Ultrasonic extraction, microwave extraction, metal ion precipitation extraction, supercritical CO 2 fluid extraction, biological enzymatic extraction, etc. are currently used.
  • Ultrasonic-assisted ethanol extraction is the most common method. This method can be used Ultrasonic oscillation loosens the structure of kelp, releases polyphenols, and improves extraction efficiency.
  • all current extraction methods deal with all kelp raw materials.
  • the effective concentration of polyphenols is low, many reagents need to be added, and the purity of polyphenols in the extract is low. Therefore, the technical problem of low polyphenol extraction rate is still not solved.
  • the antioxidant properties of kelp polyphenols are also relatively low, which restricts the preparation and application of kelp polyphenols as functional ingredients. Therefore, it is of great significance to explore new extraction methods of kelp polyphenols.
  • the present invention provides a method for extracting kelp polyphenols.
  • the present invention adopts the following technical solutions:
  • S 1 take the freeze-thaw method kelp frozen and thawed; preferably at -18 ⁇ -20 °C kelp frozen 6 ⁇ 12h, thawed at a temperature of 20-25 °C.
  • the dye layer was added an ethanol solution, after ultrasonic treatment combined filtrate and dried to give kelp polyphenol.
  • the content of kelp polyphenols in the kelp pigment layer is about 95-98%.
  • the inventor tried to extract kelp polyphenols directly from the kelp pigment, but they tried many methods and were successful. Later, after the inventor, after experimenting, he was surprised to find that the kelp was first After freezing and thawing, the kelp polyphenol extraction method provided by the present invention first uses a freeze-thaw method to process the kelp, freezes the kelp and then thawed, forming large ice crystals during the freezing process, destroying the kelp pigment layer structure, and the water flows out after thawing , To form a gap between the pigment layer and the kelp meat, which can make the subsequent pectinase fermentation better play its role.
  • the pectinase can fully contact the pectin in the pigment layer and degrade it, making the original kelp tighter.
  • the texture becomes looser and the pigment layer becomes looser, which facilitates detachment and destroys the structure of the kelp pigment layer.
  • the bound kelp polyphenols After pectinase degradation, the bound kelp polyphenols have been released, making the pigment layer easier to remove the pigment when ethanol is combined with ultrasonic extraction.
  • the kelp polyphenols in the layer are extracted, and the final amount of extraction solvent used is small, and the extraction rate can be significantly improved.
  • the fresh kelp can be directly cut into strips or pieces and then frozen; when the kelp is dried kelp, it is soaked with water, and the mass ratio of the dried kelp to water is 1:1 to 3, soaking The time is 1 ⁇ 3h. After washing, drain the water, cut into strips or pieces and freeze. When the kelp is fresh kelp, it can be washed directly, cut into strips or pieces, and then frozen.
  • the pectinase of the present invention is not specifically limited, and can be selected from those commonly used in the art.
  • the pectinase is selected from Aspergiluts niger.
  • the present invention can expand the culture of the pectinase so that the concentration of the pectinase bacteria solution can reach 10 6 ⁇ 10 8 CFU/mL, and the fresh kelp or rehydrated kelp with a mass of 1-5% of the bacteria Spray the kelp on the thawed kelp, rub it to make the bacteria better contact with the pectin, and ferment at 20-25°C for 30 ⁇ 90min; more preferably, the kelp and water after fermentation are mixed according to the mass ratio of 1:0.5-1 Stir.
  • the temperature is 20-40°C.
  • the invention adopts the freezing and thawing method to process the kelp. Now the kelp is frozen and then thawed. Large ice crystals are formed in the freezing process, which destroys the structure of the kelp pigment layer, so that the pigment layer is easy to be in the pigment layer when ethanol is combined with ultrasonic extraction. The kelp polyphenols are extracted. At this time, the ultrasonic time is relatively short and the heat is less. The temperature will not be too high due to the long-term ultrasonic treatment. The ultrasonic temperature can be maintained at 20-40 °C, while increasing the extraction rate, It also keeps the activity of kelp polyphenols to the greatest extent.
  • the kelp polyphenols mainly exist in the pigment layer.
  • the content of kelp polyphenols in the kelp pigment layer is about 95-98%.
  • the present invention uses the freezing and thawing method and the pectinase fermentation method to complete the better separation of the pigment layer and the kelp meat.
  • the kelp polyphenols in the pigment layer were extracted by alcohol extraction combined with ultrasonic. It can be seen from the above technical solutions that the extraction method of kelp polyphenols provided by the present invention reduces the consumption of 95% ethanol to 1/3 to 1/2 of the traditional extraction method, and the extraction rate is increased by 10% to 25%. After functional evaluation, The oxidation resistance is increased by about 10%, which promotes the industrial utilization of kelp polyphenols.
  • Fig. 1 is a graph showing the detection result of anti-oxidation performance of kelp polyphenols provided by the present invention
  • Example 2 is a comparison diagram of the antioxidant capacity of kelp polyphenols provided in Example 4 of the present invention.
  • the invention discloses a method for extracting kelp polyphenols, and those skilled in the art can learn from the content of this article and appropriately improve the process parameters. It should be particularly pointed out that all similar replacements and modifications are obvious to those skilled in the art, and they are all deemed to be included in the present invention.
  • the method and application of the present invention have been described through the preferred embodiments. It is obvious that relevant personnel can make changes or appropriate changes and combinations to the methods and applications described herein without departing from the content, spirit and scope of the present invention to achieve and Apply the technology of the present invention.
  • the data in Table 1 shows that the amount of ethanol used in Examples 1-3 is significantly less than the amount of ethanol in the comparative example, and the extraction rate of kelp polyphenols obtained by the preparation method provided in Examples 1-3 is significantly higher than that of the traditional process. .
  • the invention only extracts the pigment layer, after pectinase degradation, the bound kelp polyphenols are released, and then combined with ultrasonic treatment, the polyphenols can be extracted efficiently at a lower temperature and a shorter time.
  • Example 1 The kelp polyphenol obtained in Example 1 and the kelp polyphenol obtained in Comparative Example 1 were tested for their antioxidant properties, and the detection method was as follows:
  • Free radical scavenging rate (A 0 -(A i -A i0 ))/A 0 ⁇ 100% (2)
  • a i is the absorbance after mixing 1 mL of DPPH solution and 1 mL of sample solutions of different concentrations.
  • a i0 is the absorbance after mixing 1 mL of sample solution and 1 mL of methanol, and the background absorption of the sample to be tested.
  • a 0 is the absorbance of the mixture of 1 mL DPPH solution and 1 mL deionized water as a blank control.
  • a 0 is the absorbance of water instead of the sample;
  • a 1 is the absorbance of the sample;
  • a 2 is the absorbance of the sample and distilled water instead of H 2 O 2 .
  • the kelp is frozen and then thawed. Large ice crystals are formed during the freezing process, which destroys the structure of the kelp pigment layer. After pectinase degradation, the bound kelp polyphenols have been released, making the pigment layer re-release. It is easy to extract kelp polyphenols in the pigment layer when ethanol is combined with ultrasonic extraction. At this time, the ultrasonic time is relatively short, the heat is less, and the temperature will not be too high due to the long-term ultrasonic treatment. The ultrasonic temperature can be maintained at 20 -40°C, while increasing the extraction rate, it also keeps the activity of kelp polyphenols to the greatest extent.

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Abstract

本发明属于植物提取技术领域，公开了一种海带多酚的提取方法。该方法包括以下步骤：S 1采取冻融法将海带冷冻、解冻；S 2、解冻后的海带加入果胶酶菌种发酵液接种发酵，发酵后色素液体层；S 3、将所述色素层中加入乙醇溶液，超声波处理后过滤，滤液干燥得到海带多酚。该提取方法95％乙醇使用量减少，提取率得到提高，抗氧化性也得到了提高，推动了海带多酚的产业化利用。

Description

一种海带多酚的提取方法 技术领域
本发明涉及植物提取技术领域，特别涉及一种海带多酚的提取方法。
背景技术
海洋植物海藻中含有多酚，研究人员将从海藻中分离提纯的各种酚类化合物称作海藻酚类(algae polyphenols)。海洋酚类化合物具有强大的生物活性，乃是近年的研究热门，按照分子结构特征，海藻酚类化合物可以分为简单酚类化合物和多酚类化合物。我国的海带资源十分丰富，海带养殖已成为中国海水养殖的支柱产业之一，产量居世界首位，海带多酚具有抗氧化、抗肿瘤、抗菌以及化学防御等作用，具有极大的开发利用价值，但海带酚类物质存在易氧化、不稳定、提取组分复杂、提取率低等问题，制约了其工业化制备生产。
海带多酚提取主要以干燥、粉碎过筛的海带粉为原料，采用一定浓度乙醇溶液提取法，但多酚在海带体内通常与蛋白质、多糖以氢键和疏水键形式形成稳定化合物，造成提取率低，目前多采用超声波提取法、微波浸提法、金属离子沉淀萃取法、超临界CO 2流体萃取法、生物酶解提取法等，其中采用超声波辅助的乙醇提取法最为常见，该方法可以通过超声波振荡使海带结构松散，释放多酚物质，提升提取效率。但目前所有的提取方法都针对全部海带原料处理，多酚有效浓度低，需要添加试剂多，而且提取物中多酚纯度低等问题，故多酚提取率低的技术问题仍然没有解决，得到的海带多酚的抗氧化性能也相对较低，制约了海带多酚作为功能性配料的制备和应用，因此探索新的海带多酚的提取方法具有重要意义。
发明内容
为了解决现有技术中的从海带原料浆液提取海带多酚造成海带多酚提取率和抗氧化性能较低的问题，本发明提供了一种海带多酚的提取方法。
为了解决上述技术问题，本发明采用以下技术方案：
一种海带多酚的提取方法，具体保护以下步骤：
S 1、采取冻融法将海带冷冻、解冻；优选地，海带于-18～-20℃下冷冻6～12h，于20-25℃温度下解冻。
S 2、解冻后的海带加入果胶酶菌种发酵液接种发酵，发酵后加水搅拌、过滤得到滤渣色素层；
S 3、将所述色素层中加入乙醇溶液，联合超声波处理后过滤，滤液干燥得到海带多酚。
海带色素层中海带多酚的含量大约为95-98％，发明人试图从海带色素中直接提取海带多酚，但是尝试了很多方法均为成功，后来发明人经过试验，惊喜的发现先将海带冷冻解冻后， 本发明提供的海带多酚提取方法，首先采用冻融法对海带进行处理，将海带冷冻再解冻，在冷冻过程中形成大的冰晶，破坏了海带色素层结构，解冻后水分流出，在色素层与海带肉之间形成空隙，可使后续果胶酶发酵时更好的发挥其作用，果胶酶能够充分和色素层中的果胶接触并将其降解，使原来海带的紧密质构变松散，色素层更加松散，利于脱离，破坏了海带色素层结构，经过果胶酶降解，已将结合态的海带多酚释放出来，使得色素层再采用乙醇联合超声波提取时容易将色素层中的海带多酚提取出来，最终所用提取溶剂用量很少，同时可显著提高提取率。
本发明如果原料采用新鲜海带，可直接将新鲜海带切成条或块后冷冻；当所述海带为干海带时，加水浸泡，所述干海带与水的质量比为1:1～3，浸泡时间为1～3h，洗净后沥干水分，切成条或块后冷冻。当海带为鲜海带时，直接洗净、切成条或块后冷冻即可。
本发明所述的果胶酶没有具体限制，可选自本领域常用的，作为优选，所述果胶酶选自黑曲霉(Aspergiluts niger)。
作为优选，本发明可将所述果胶酶扩大培养，使果胶酶菌菌液浓度达到10 6～10 8CFU/mL，，将鲜海带或复水海带质量1-5％的所述菌液喷洒到解冻后的海带上，揉搓使菌液更好与果胶质接触，20-25℃温度下发酵30～90min；更优选地，发酵后海带和水按照质量比1:0.5-1混合搅拌。
向步骤S 3中得到的色素层加入40-80g 95％质量浓度乙醇，加水使乙醇溶液质量浓度达到40-70％，然后进行超声辅助处理，超声处理功率为500-2000W，时间为5-15min，温度为20-40℃。本发明采用冻融法对海带进行处理，现将海带冷冻，再解冻，在冷冻过程中形成大的冰晶，破坏了海带色素层结构，使得色素层再采用乙醇联合超声波提取时容易将色素层中的海带多酚提取出来，此时，超声的时间比较简短，放热少，不会因为长时间的超声处理导致温度过高，超声温度能够保持在20-40℃，在增加提取率的同时，又最大程度的保持了海带多酚的活性。
海带多酚主要存在于色素层，海带色素层中海带多酚的含量大约为95-98％，本发明采用冻融法、果胶酶发酵法完成了色素层和海带肉的较好分离，最后采用醇提法联合超声波对色素层中的海带多酚进行提取。由以上技术方案可知，本发明提供的海带多酚的提取方法，95％乙醇使用量减少至传统提取方法的1/3～1/2，提取率提高10％～25％，经功能性评价，抗氧化性提高了10％左右，推动了海带多酚的产业化利用。
附图说明
图1为本发明所提供的海带多酚的抗氧化性能检测结果图；
图2为本发明实施例4所提供的海带多酚抗氧化能力对比图。
具体实施方式
本发明公开了一种海带多酚的提取方法，本领域技术人员可以借鉴本文内容，适当改进工艺参数实现。特别需要指出的是，所有类似的替换和改动对本领域技术人员来说是显而易见的，它们都被视为包括在本发明当中。本发明的方法及应用已经通过较佳实施例进行了描述，相关人员明显能在不脱离本发明内容、精神和范围内对本文所述的方法和应用进行改动或适当变更与组合，来实现和应用本发明技术。
为了使本领域技术人员能够更好的理解本发明，下面结合具体实施方式对本发明作进一步的详细说明。
实施例1 海带多酚的提取
S 1采用冻融法，将鲜海带100g切成条在-18℃冷冻6h，再用20℃空气中解冻。
S 2将黑曲霉(Aspergiluts niger)扩大培养，使黑曲霉(Aspergiluts niger)菌液浓度达到10 6～10 8CFU/mL，按照1％鲜海带质量将所述菌液喷洒到解冻后的海带上，20℃温度下发酵30min，发酵后海带和水按照质量比1:0.5混合、搅拌揉搓，过滤得到滤渣色素层(形态可见图1)；
S 3将色素层加入40g 95％质量浓度乙醇中使乙醇的浓度达到40％，然后进行超声处理，超声处理功率为500W，时间为5min，温度为30℃，将乙醇提取后的提取液过滤，滤液为海带多酚粗提液，于-20℃预冷6h，冷冻干燥得到海带多酚粗品2.9g，海带多酚的提取率为2.9％。
Figure PCTCN2020089964-appb-000001
实施例2 海带多酚的提取
S 1将干海带加水浸泡，所述干海带与水的质量比为1:2，浸泡时间为2h，洗净后沥干水分，100g复水海带切成条后在-20℃冷冻12h，再用25℃空气中解冻。
S 2将黑曲霉(Aspergiluts niger)扩大培养，使黑曲霉(Aspergiluts niger)菌液浓度达到10 6～10 8CFU/mL，按照3％复水海带质量，将所述菌液喷洒到解冻后的海带上，25℃温度下发酵90min，发酵后海带和水按照质量比1:1混合、搅拌揉搓，过滤得到滤渣色素层(形态可见图1)；
S 3将色素层加入80g 95％质量浓度乙醇中使乙醇的浓度达到70％，然后进行超声处理，超声处理功率为2000W，时间为15min，温度为40℃，将乙醇提取后的提取液过滤，滤液为海带多酚粗提液，于-40℃预冷12h，冷冻干燥得到海带多酚粗品，海带多酚的提取率为3.2g，提取率3.2％。
实施例3 海带多酚的提取
S 1采用冻融法，将鲜海带切成条在-19℃冷冻9h，再用22℃空气中解冻。
S 2将黑曲霉(Aspergiluts niger)扩大培养，使黑曲霉(Aspergiluts niger)菌液浓度达到10 6～10 8CFU/mL，将所述菌液喷洒到解冻后的海带上，23℃温度下发酵60min，发酵后海带和水按照质量比1:0.8混合、搅拌揉搓，得到色素层(形态可见图1)；
S 3将色素层加入60g 95％质量浓度乙醇中使乙醇的浓度达到55％，然后进行超声处理，超声处理功率为1000W，时间为10min，温度为20℃，将乙醇提取后的提取液过滤，滤液为海带多酚粗提液，于-30℃预冷9h，冷冻干燥得到海带多酚粗品，海带多酚的提取率为3.0g，提取率3.0％。
对比例1 采用传统法提取海带多酚
称取100g鲜海带，干燥，粉碎，过40-60目筛，添加100-200g 95％乙醇，添加水至乙醇浓度50-70％，采用超声波处理，时间60-120min，温度升至50-60℃，将提取后的海带多酚提取液于-30℃预冷9h，冷冻干燥得到海带多酚粗品，进行多次提取，其提取率在2.0-2.5％之间。将其95％乙醇用量和海带多酚提取率与实施例1-3比较，比较结果如表1所示。
表1 不同提取方法下的海带多酚提取率
Figure PCTCN2020089964-appb-000002
表1数据显示，实施例1-3所用乙醇用量明显少于对比例的乙醇用量，实施例1-3提供的制备方法得到海带多酚的提取率明显高于传统工艺的海带多酚的提取率。本发明仅对色素层提取，经过果胶酶降解，已将结合态的海带多酚释放出来，再与超声波处理结合，在较低温度、较短时间内，可高效提取多酚。
实施例4 海带多酚抗氧化性能检测
对实施例1得到的海带多酚和对比例1得到的海带多酚进行抗氧化性能检测，检测方法如下：
(1)将二苯代苦味肼基自由基(α,α-diphenyl-β-picrylhydrazine,DPPH)与甲醇溶液混合呈紫色，在517nm最大吸收值。体系中有自由基清除能力化合物存在，溶液颜色由紫色变黄色，吸光度时数值变小。
自由基清除率＝(A 0-(A i-A i0))/A 0×100％       (2)
A i为1mL DPPH溶液和1mL不同浓度样品溶液混合后吸光度。A i0为1mL样品溶液和1mL甲醇混合后吸光度，待测样品本底吸收。A 0为1mL DPPH溶液和1mL去离子水混合吸光度，作为空白对照。
(2)羟基自由基清除
分别取50μL不同浓度的样品置于96孔板内，再加入50μL 0.75mmol/L邻二氮菲溶液；100μL 0.2mol/L pH值是7.4PBS溶液；50μL 0.75mmol/L FeSO 4溶液；50μL体积分数0.01％H 2O 2溶液，混匀后置于37℃水浴恒温30min，在波长536nm检测样品吸光度，以水作空白对照组，Vc作为阳性对照组。
清除率＝(1-(A 1-A 2)/A 0)％         (3)
A 0为水代替样品的吸光度；A 1为样品的吸光度；A 2为样品和蒸馏水替代H 2O 2的吸光度。
采用上述DPPH法和羟基自由基清除法测定海带多酚抗氧化能力，最终检测结果见图2，图2显示，传统方法得到的海带多酚抗氧化能力明显不如实施例方法得到的海带多酚抗氧化能力，其抗氧化性能提高了10％左右。对比例1采用传统提取方法是对整个海带提取，用试剂多，增加超声波辅助处理，所用时间长，产热多，温度高，使多酚部分氧化损失，生物活性低；本发明采用冻融法对海带进行处理，现将海带冷冻，再解冻，在冷冻过程中形成大的冰晶，破坏了海带色素层结构，经过果胶酶降解，已将结合态的海带多酚释放出来，使得色素层再采用乙醇联合超声波提取时容易将色素层中的海带多酚提取出来，此时，超声的时间比较简短，放热少，不会因为长时间的超声处理导致温度过高，超声温度能够保持在20-40℃，在增加提取率的同时，又最大程度的保持了海带多酚的活性。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。

Claims (8)

  1. 一种海带多酚的提取方法，其特征在于，包括以下步骤：
    S 1、采取冻融法将海带冷冻、解冻；
    S 2、解冻后的海带加入果胶酶菌种发酵液接种发酵，发酵后加水搅拌，过滤得到滤渣色素层；
    S 3、将所述色素层中加入乙醇溶液，超声波处理后过滤，滤液干燥得到海带多酚。
  2. 如权利要求1所述的海带多酚的提取方法，其特征在于，步骤S 1，海带于-18～-20℃下冷冻6～12h。
  3. 如权利要求1或2所述的海带多酚的提取方法，其特征在于，步骤S 1，于20-25℃温度下解冻。
  4. 如权利要求1所述的海带多酚的提取方法，其特征在于，所述海带为鲜海带时，将所述海带切成条或块后冷冻；所述海带为干海带时，加水浸泡得到复水后海带，所述干海带与水的质量比为1:1～3，浸泡时间为1～3h，洗净后沥干水分，切成条或块后冷冻。
  5. 如权利要求1所述的海带多酚的提取方法，其特征在于，所述果胶酶选自黑曲霉(Aspergiluts niger)。
  6. 如权利要求1、4或5所述的海带多酚的提取方法，其特征在于，将所述果胶酶扩大培养，使果胶酶菌菌液浓度达到10 6～10 8CFU/mL，将鲜海带或复水海带质量1-5％的所述菌液喷洒到解冻后的海带上，20-25℃温度下发酵30～90min。
  7. 如权利要求6所述的海带多酚的提取方法，其特征在于，发酵后海带和水按照质量比1:0.5-1混合，搅拌揉搓，过滤得到滤渣色素层。
  8. 如权利要求1所述的海带多酚的提取方法，其特征在于，步骤S 3中添加40-80g 95％质量浓度的乙醇，加水使最终乙醇容易的浓度达到40-70％，然后进行超声辅助处理，超声处理功率为500-2000W，时间为5-15min，温度为20-40℃。
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