WO2020000828A1 - 一种具有显著降血脂活性的龙须菜多糖及其制备方法与应用 - Google Patents

一种具有显著降血脂活性的龙须菜多糖及其制备方法与应用 Download PDF

Info

Publication number
WO2020000828A1
WO2020000828A1 PCT/CN2018/113239 CN2018113239W WO2020000828A1 WO 2020000828 A1 WO2020000828 A1 WO 2020000828A1 CN 2018113239 W CN2018113239 W CN 2018113239W WO 2020000828 A1 WO2020000828 A1 WO 2020000828A1
Authority
WO
WIPO (PCT)
Prior art keywords
polysaccharide
gracilaria
preparation
group
extraction
Prior art date
Application number
PCT/CN2018/113239
Other languages
English (en)
French (fr)
Inventor
游丽君
黄诗铭
李雄
赵振刚
李超
Original Assignee
华南理工大学
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 华南理工大学 filed Critical 华南理工大学
Priority to SG11202012298VA priority Critical patent/SG11202012298VA/en
Publication of WO2020000828A1 publication Critical patent/WO2020000828A1/zh

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
    • 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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • 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
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Definitions

  • the invention belongs to the field of deep processing of asparagus, and particularly relates to an asparagus polysaccharide having significant blood lipid lowering activity, and a preparation method and application thereof.
  • Graceria Lemaneofimrsi belongs to the red algae and Gracaialri plants. It is mainly produced in China, Guangdong, Hainan, Shandong and other places. It is rich in dietary fiber, vitamins and trace elements, has low fat content, and contains polysaccharides. Active ingredients such as phycobiliproteins and unsaturated fatty acids.
  • Longxue Cai has a long history of consumption in eastern China. Traditional Chinese medicine believes that Longxue Cai has the effect of "convenience, cure spleen and heat", and modern medical researchers believe that it has the ability to help digestion, degrease, clear the stomach, and stop bleeding. , The effect of blood pressure.
  • the clearance rate of DPPH radical, hydroxyl radical and superoxide ion radical is above 90% (patent application number 201611024913.2).
  • the Third Institute of Oceanography of the State Oceanic Administration discloses a Longan vegetable agarose oligosaccharide and a preparation method thereof, which can be applied to antioxidant and anti-ultraviolet health products and cosmetics (patent application number 201210347105.5); Polysaccharide extract of Gracilaria vulgaris, preparation method and application thereof, the obtained polysaccharide has good activity of inhibiting dipeptidyl peptidase IV, and the polysaccharide can be used in the research and development of blood glucose lowering drugs or health products (patent application number 201410252784.7) ; Shanghai Fisheries University has disclosed a method for preparing polysaccharides of Gracilaria radiata, and the obtained polysaccharide has a high proliferation effect on mouse lymphocytes (patent application number 200710044610.1); Shanghai Jiaotong University has disclosed the application of polysacc
  • the primary purpose of the present invention is to provide a Gracilaria vulgaris polysaccharide.
  • Another object of the present invention is to provide a method for preparing the above-mentioned polysaccharide of Gracilaria.
  • Yet another object of the present invention is to provide the application of the above-mentioned polysaccharide of Gracilaria.
  • a method for preparing a polysaccharide of Gracilaria with significant blood lipid lowering activity including the following steps:
  • step (3) ultrafiltration the extraction solution described in step (2), discarding the permeate, and collecting the retentate;
  • the sieving and sieving in step (1) is sieving through a 20-80 mesh sieve.
  • the pH value of the organic acid solution in step (2) is 2.0 to 4.0.
  • the organic acid solution in step (2) is a citric acid solution or a malic acid solution.
  • the mass of the organic acid solution in step (2) is 20 to 40 times the mass of the gracilis powder.
  • the parameters of the pre-homogenization process in step (2) are: 20-50 MPa, 1 to 3 minutes, and the number of homogenization times is 1 to 2 times;
  • control conditions of the dynamic high-pressure microjet extraction in step (2) are: the microjet pressure is 120-180MPa, the extraction temperature is 75-100 ° C, the extraction time is 80-160 minutes, and the number of extractions is 1-2 times .
  • the ultrafiltration in step (3) uses an ultrafiltration membrane with a cut-off molecular weight of 1 kDa.
  • the type of the ion exchange resin in step (4) is DEAE-fast flow or DEAE-52.
  • the concentrated solution obtained by the vacuum concentration in step (4) is 1/10 to 1/5 of the volume of the retentate.
  • the amount of ethanol used in step (5) accounts for 60% to 90% of the total volume of the concentrated solution after being mixed with ethanol.
  • the standing in step (5) is standing at 2 to 8 ° C for 8 to 14 hours.
  • the drying in step (6) is vacuum freeze drying or spray drying.
  • a gracilaria polysaccharide prepared by the above-mentioned preparation method has a molecular weight of 20-35 kDa and a sulfuric acid group content of 20-28 wt%.
  • the present invention has the following advantages and effects:
  • the present invention uses organic acid combined with dynamic high-pressure micro-jet technology to extract polysaccharides from Gracilaria pulcherrima.
  • the extraction rate is 3 to 5 times higher than that of traditional water extraction, which can greatly shorten the extraction time and reduce energy consumption.
  • the polysaccharide of Gracilaria vulgaris obtained by the present invention has better blood lipid lowering effect.
  • Figures 1a to 1e are oil red o-stained photographs (200 ⁇ ) of liver tissue of mice in the NC, HF, HF + AT, HF + LGP, and HF + HGP groups, respectively.
  • Figures 2a to 2e are HE stained photographs (400 ⁇ ) of liver tissue of mice in the NC, HF, HF + AT, HF + LGP, and HF + HGP groups, respectively.
  • An asparagus polysaccharide extract is prepared as follows:
  • step (3) ultra-filtration the extraction solution obtained in step (2) with a membrane having a molecular weight cut-off of 1 kDa, discarding the permeate, and collecting the retentate;
  • the obtained polysaccharide is freeze-dried to obtain the polysaccharide of Gracilaria pulcherrima, which is referred to as 1 # extract.
  • An asparagus polysaccharide extract is prepared as follows:
  • step (3) ultra-filtration the extraction solution obtained in step (2) with a membrane having a molecular weight cut-off of 1 kDa, discarding the permeate, and collecting the retentate;
  • the obtained polysaccharide is freeze-dried to obtain the polysaccharide of Gracilaria, which is referred to as 2 # extract.
  • An asparagus polysaccharide extract is prepared as follows:
  • step (3) ultra-filtration the extraction solution obtained in step (2) with a membrane having a molecular weight cut-off of 1 kDa, discarding the permeate, and collecting the retentate;
  • the obtained polysaccharide is freeze-dried to obtain the polysaccharide of Gracilaria pulcherrima, which is described as 3 # extract.
  • a polysaccharide of Gracilaria pulcherrima is prepared by a traditional water extraction method, and the preparation method is as follows:
  • step (3) ultrafiltration the final extraction solution obtained in step (2) with a membrane with a molecular weight cut-off of 1 kDa, discarding the permeate, and collecting the retentate;
  • polysaccharide extraction rate (%) (polysaccharide content (g) in the extract solution / raw material weight (g)) ⁇ 100.
  • Extract 1 # Extract 2 # Extract 3 # extract 4 # Extract Polysaccharide extraction rate (%) 23.65 ⁇ 0.27 19.80 ⁇ 0.62 20.31 ⁇ 0.80 5.46 ⁇ 0.52
  • a hypercholesterolemic model was produced by feeding high-cholesterol and high-fat male Kunming mice to evaluate the hypolipidemic effect of the polysaccharides of Gracilaria pulvinata prepared according to the aforementioned method.
  • statin liquid Take 7 atorvastatin calcium tablets (containing atorvastatin 140mg), suspend in 350mL of sterile normal saline, and formulate a concentration of 0.4mg / mL (based on atorvastatin content) ).
  • TC total cholesterol
  • TG triglyceride
  • LDL-C low density lipoprotein
  • HDL-C high density lipoprotein
  • FFA free fatty acid
  • model control group HF
  • atorvastatin intervention group HF + AT
  • low-dose dragongrass Polysaccharide Intervention Group HF + LGP
  • High-dose Gracilaria Intervention Group HF + HGP
  • Oral administration The normal control group (NC) and the model control group were perfused with sterile saline at 0.1 mL / 10 g daily, and the administration group was perfused with the corresponding drug in a volume of 0.1 mL / 10 g per day. A total of 42 days were administered. After 14, 28, and 42 days of administration, 6 mice in each group were randomly selected.
  • the blood was collected by eyeball blood collection. After the mice were sacrificed by cervical spine, the epididymal fat was weighed, and the liver was taken as Pathological analysis of tissue sections; centrifugation of serum within 2 hours of blood collection was performed according to the instructions of the test box, and the contents of TC, TG, LDL-C, HDL-C, and FFA were measured.
  • AI LDL-C / HDL-C
  • Figures 1a to 1e are oil red o-stained photographs (200 ⁇ ) of liver tissue of mice in the NC, HF, HF + AT, HF + LGP, and HF + HGP groups, respectively.
  • Figures 2a to 2e are HE stained photographs (400 ⁇ ) of liver tissue of mice in the NC, HF, HF + AT, HF + LGP, and HF + HGP groups, respectively.
  • the atorvastatin group and the low-dose group have a significant increase (p ⁇ 0.05), while the high-dose group does not. Significantly elevated effect (p> 0.05).
  • the low-dose polysaccharide group has a significant reducing effect (p ⁇ 0.05), while the atorvastatin group and the high-dose polysaccharide group have no significant effect. Reduction effect (p> 0.05).
  • the NC group has a normal liver morphology, and only a small number of hepatocytes contain fine dot-like orange-stained lipid droplets; the HF group has a diffuse, dark orange-red color.
  • the small piece of dark orange in the necrotic area indicates that a large number of liver cells contain large lipid droplets and the lipid droplets in the necrotic area are fused; the HF + AT group is diffusely spotted and droplet-like powder staining, compared with the model group, there are some lipid droplets.
  • the HF + LGP group was focally light orange, suggesting that the number of lipid droplets and the size of lipid droplets in the HF + LGP group were significantly reduced compared with the model group, and the fat-reducing effect was significant; the HF + HGP group showed nested tablets It is orange in color, and the lipid droplets are reduced and reduced compared with the model group, and the fat reduction effect is obvious.
  • liver cells in the NC group are regularly arranged, without steatosis, and there is no inflammation and necrosis;
  • the liver cells in the HF group are disorderly arranged, and the number of cells in the cytoplasm varies, and the size Various fatty vacuoles (arrow 1), severe fatty degeneration, severe hepatocyte watery degeneration (arrow 2), and focal infiltration of inflammatory cells (arrow 3), suggesting that the model group
  • the liver has a large amount of fat accumulation and moderate inflammation and necrosis;
  • the liver cells in the HF + AT group have a slightly regular arrangement with fatty vacuoles (arrow 1), hepatic water-like degeneration (arrow 2), and a small number of inflammatory cells (Arrow 3), suggesting that there is moderate steatosis and mild inflammation in the statin intervention group;
  • hepatocytes in the HF + LGP group are regularly arranged, no fat vacuoles
  • reagents Atorvastatin calcium tablets (Pfizer Pharmaceutical Co., Ltd., product batch number: S90890), alanine aminotransferase (GPT) kit (Nanjing Jiancheng Biotechnology Co., Ltd., production batch number: 20180420), aspartate aminotransferase (GOT) reagent Box (Nanjing Jiancheng Biotechnology Co., Ltd., production batch number: 20180420), Longsaurus polysaccharide (GLP) was made by our laboratory.
  • GLP Longsaurus polysaccharide
  • atorvastatin a positive control drug: take 7 atorvastatin calcium tablets (containing atorvastatin 140mg), suspend in 350mL of sterile normal saline, and prepare a concentration of 0.4mg / mL (using Atorvastatin content).
  • Modeling stage 102 Kunming mice, 22-24g, males, were randomly divided into two groups, one group of 24 was a normal control group and fed with ordinary feed; the other group of 78 was a hyperlipidemia model group ( (Hereinafter referred to as the model group), fed with high-cholesterol and high-fat diets, and fed freely with water. After 6 weeks of breeding, 6 mice were randomly selected from each group. After fasting for 8 hours, the blood was collected by eye removal and the serum was centrifuged.
  • model group hyperlipidemia model group
  • Total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and free fatty acid (FFA) kit instructions were used to measure TC, The contents of TG, LDL-C, HDL-C, and FFA are used to judge whether the modeling is successful or not according to the above indicators.
  • model control group HF
  • statin control group HF + AT
  • low-dose gracilaria polysaccharide group HF + LGP
  • high dose Gracilaria polysaccharide group HF + HGP
  • Oral administration The normal control group (NC) and the model control group were perfused with sterile saline at 0.1 mL / 10 g daily, and the administration group was perfused with the corresponding drug in a volume of 0.1 mL / 10 g per day. A total of 42 days were administered. After 14, 28, and 42 days of administration, 6 mice from each group were randomly selected.
  • GPT alanine aminotransferase
  • GOT aspartate aminotransferase
  • the high-fat mouse group had a significant increase (p ⁇ 0.01); compared with the model control group, the long-stemmed polysaccharides in the high-dose group and the high-dose group were all There was a significant decrease (p ⁇ 0.05), but compared with the atorvastatin group, there was no significant difference between the various groups of polysaccharides of A. radiata (p> 0.05).
  • the model control group had a very significant increase (p ⁇ 0.01); compared with the model control group, the atorvastatin group and the long-stemmed polysaccharide were low and high. Both the dose group had a significant reduction effect (p ⁇ 0.05); compared with the atorvastatin group, the low-dose polysaccharide group had a significant reduction effect (p ⁇ 0.05).
  • liver polysaccharides from A. radiata and atorvastatin show that there is a difference in the toxic and side effects of liver polysaccharides from A. radiata and atorvastatin.
  • a low dose (6.0 mg / mL polysaccharide content) is more safe than atorvastatin.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Polymers & Plastics (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Hematology (AREA)
  • Nutrition Science (AREA)
  • Diabetes (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Mycology (AREA)
  • Obesity (AREA)
  • Food Science & Technology (AREA)
  • Epidemiology (AREA)
  • Sustainable Development (AREA)
  • Biochemistry (AREA)
  • Materials Engineering (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

本发明公开了一种具有显著降血脂活性的龙须菜多糖及其制备方法与应用，属于龙须菜深加工领域。本发明所提取的龙须菜多糖的分子量为20～35kDa，硫酸基含量达20～28wt％。所述制备方法为将龙须菜干燥粉碎后，通过预均质、有机酸结合动态高压微射流技术提取、碱中和、超滤、离子交换树脂分离、浓缩和醇沉，从而制得龙须菜多糖。该龙须菜多糖的制备方法简单高效，适用于工业化生产，所得龙须菜多糖具有显著的降血脂活性，可用于制备降血脂药物或保健品。

Description

一种具有显著降血脂活性的龙须菜多糖及其制备方法与应用 技术领域
本发明属于龙须菜深加工领域，具体涉及一种具有显著降血脂活性的龙须菜多糖及其制备方法与应用。
背景技术
龙须菜(GracialriaLemaneofimrsi)属红藻门，江篱属(Gracaialri)植物，主要生产于我国广东、海南、山东等地，含有丰富的膳食纤维、维生素和微量元素，脂肪含量低，同时含有多糖、藻胆蛋白和不饱和脂肪酸等活性成分。龙须菜在中国东部地区有悠久的食用历史，传统中医认为龙须菜具有“利便、治瘿结热气”的功效，现代医学研究者则认为其具有助消化、解积腻、清肠胃、止血、降压的功效。
早期由于龙须菜产量稀少，因此国内对于龙须菜的研究报道主要集中在种植和加工方面，旨在提高其产量及繁殖率，而近年来关于龙须菜的研究重点逐渐由生产转移到了对其活性物质的提取及生理功能的研究方面，已见报道的有抗氧化活性、降血糖活性、免疫活性和抗肿瘤活性等。如朱益生公开了一种提高龙须菜萃取液的抗氧化成份的方法(专利申请号201010188414.3)；广东药科大学公开了一种龙须菜提取物及其制备方法和应用，该提取物对DPPH自由基、羟自由基、超氧离子自由基的清除率在90％以上(专利申请号201611024913.2)。国家海洋局第三海洋研究所公开了一种龙须菜琼胶寡糖及其制备方法，可应用于抗氧化、抗紫外线保健品和化妆品中(专利申请号201210347105.5)；华南理工大学公开了一种龙须菜多糖提取物及其制备方法与应用，所得多糖具有较好的抑制二肽基肽酶IV的活性，该多糖可应用于降血糖药物或保健品的研发中(专利申请号201410252784.7)；上海水产大学公开了一种龙须菜多糖的制备方法，所得多糖对小鼠淋巴细胞具有较高的增殖效果(专利申请号200710044610.1)；上海交通大学公开了一种龙须菜多糖的应用，该多糖可用于制备诱导细胞中的FADD和GADD基因的表达的抑制剂中的应用，在抗肿瘤药物或防癌保健品的制备中具有应用前景(专利申请号201610082951.7)。
发明内容
为了提高龙须菜的深加工技术，拓宽龙须菜的应用范围，本发明的首要目的在于提供一种龙须菜多糖。
本发明的另一目的在于提供上述龙须菜多糖的制备方法。
本发明的再一目的在于提供上述龙须菜多糖的应用。
本发明的目的通过下述技术方案实现。
一种具有显著降血脂活性的龙须菜多糖的制备方法，包含以下步骤：
(1)将龙须菜洗净、风干、粉碎过筛，得到龙须菜粉；
(2)向龙须菜粉中加入有机酸溶液，经过预均质处理后，结合动态高压微射流技术提取龙须菜粉，然后用碱中和，过滤，得到滤渣和提取液；
(3)将步骤(2)所述提取液进行超滤，舍弃透过液，收集截留液；
(4)用离子交换树脂分离所述截留液，取降血脂活性最高的组分进行真空浓缩，得浓缩液；
(5)将所述浓缩液与乙醇混合，静置，过滤得到多糖沉淀；
(6)将所述多糖沉淀进行干燥，得到所述龙须菜多糖。
优选的，步骤(1)所述粉碎过筛为粉碎后过20～80目筛。
优选的，步骤(2)所述有机酸溶液的pH值为2.0～4.0。
优选的，步骤(2)所述有机酸溶液为柠檬酸溶液或苹果酸溶液。
优选的，步骤(2)所述有机酸溶液的质量为所述龙须菜粉质量的20～40倍。
优选的，步骤(2)所述预均质处理的参数为：20～50MPa，1～3分钟，均质次数为1～2次；
优选的，步骤(2)所述动态高压微射流提取的控制条件为：微射流压力为120～180MPa，提取温度为75～100℃，提取时间为80～160分钟，提取次数为1～2次。
优选的，步骤(3)所述超滤采用截留分子量1kDa的超滤膜。
优选的，，步骤(4)所述离子交换树脂的型号为DEAE-fast flow或DEAE-52。
优选的，步骤(4)所述真空浓缩所得浓缩液为所述截留液体积的1/10～1/5。
优选的，步骤(5)所述乙醇的用量占所述浓缩液与乙醇混合后总体积的60％～90％，
优选的，步骤(5)所述静置是在2～8℃下静置8～14小时。
优选的，步骤(6)所述干燥为真空冷冻干燥或喷雾干燥。
由以上所述的制备方法制得的一种龙须菜多糖，所述龙须菜多糖的分子量为20～35kDa，硫酸基含量达20～28wt％。
以上所述的一种龙须菜多糖在制备降血脂药物或保健品中的应用。
相对于现有技术，本发明具有如下的优点及效果：
(1)本发明采用有机酸结合动态高压微射流技术提取龙须菜多糖，其提取率比传统水提 法提高3～5倍，可大幅缩短提取时间、降低能耗。
(2)本发明所得龙须菜多糖具有较好的降血脂功效。
附图说明
图1a～图1e分别为NC、HF、HF+AT、HF+LGP、HF+HGP组小鼠肝脏组织的油红o染色切片照片(200×)。
图2a～图2e分别为NC、HF、HF+AT、HF+LGP、HF+HGP组小鼠肝脏组织的HE染色切片照片(400×)。
具体实施方式
下面结合实施例对本发明作进一步详细的描述，但本发明的实施方式不限于此。
实施例1
一种龙须菜多糖提取物，其制备方法如下：
(1)龙须菜洗净、风干粉碎后过20目筛，取筛下粉末备用；
(2)取50g龙须菜粉置于容器中，与pH 2.0的柠檬酸按1:30的质量比混合均匀，30MPa预均质2次，每次均质2分钟，均质后，结合动态高压微射流技术提取，微射流压力125MPa，提取温度90℃，提取时间150分钟，提取2次，用NaOH中和至pH值为中性，过滤得滤渣和提取液；
(3)将步骤(2)所得的提取液用截留分子量1kDa的膜进行超滤，舍弃透过液，收集截留液；
(5)用离子交换树脂DEAE-Fast flow型号分离截留液，取降血脂活性最高的组分，真空浓缩至截留液体积的1/10，得浓缩液；
(6)在浓缩液中加入无水乙醇至乙醇终浓度为85wt％，混合均匀后，在6℃下静置12小时，过滤，除去上层清液部分，得多糖沉淀；
(7)将所得多糖沉淀冷冻干燥，得到所述龙须菜多糖，记为1#提取物。
实施例2
一种龙须菜多糖提取物，其制备方法如下：
(1)龙须菜洗净、风干、粉碎后过40目筛，取筛下粉末备用；
(2)取50g龙须菜粉置于容器中，与pH 3.5的柠檬酸按1:20的质量比混合均匀，25MPa预均质3次，每次均质1分钟，均质后，结合动态高压微射流技术提取，微射流压力160MPa，提取温度80℃，提取时间100分钟，提取1次，用NaOH中和至pH值为中性，过滤得滤渣和提取液；
(3)将步骤(2)所得的提取液用截留分子量1kDa的膜进行超滤，舍弃透过液，收集截留液；
(5)用离子交换树脂DEAE-52型号分离截留液，取降血脂活性最高的组分，真空浓缩至截留液体积的1/8，得浓缩液；
(6)在浓缩液中加入无水乙醇至乙醇终浓度为68wt％，混合均匀后，在4℃下静置10小时，过滤，除去上层清液部分，得多糖沉淀；
(7)将所得多糖沉淀冷冻干燥，得到所述龙须菜多糖，记为2#提取物。
实施例3
一种龙须菜多糖提取物，其制备方法如下：
(1)龙须菜洗净、风干、粉碎后过60目筛，取筛下粉末备用；
(2)取50g龙须菜粉置于容器中，与pH 3.0的柠檬酸按1:40的质量比混合均匀，45MPa预均质1次，每次均质3分钟，均质后，结合动态高压微射流技术提取，微射流压力148MPa，提取温度96℃，提取时间130分钟，提取1次，用NaOH中和至pH值为中性，过滤得滤渣和提取液；
(3)将步骤(2)所得的提取液用截留分子量1kDa的膜进行超滤，舍弃透过液，收集截留液；
(5)用离子交换树脂DEAE-Fast flow型号分离截留液，取降血脂活性最高的组分，真空浓缩至截留液体积的1/5，得浓缩液；
(6)在浓缩液中加入无水乙醇至乙醇终浓度为75wt％，混合均匀后，在2℃下静置8小时，过滤，除去上层清液部分，得多糖沉淀；
(7)将所得多糖沉淀冷冻干燥，得到所述龙须菜多糖，记为3#提取物。
对比例1传统水提法制备龙须菜多糖提取物
一种龙须菜多糖提取物，采用传统水提法进行制备，其制备方法如下：
(1)龙须菜洗净、风干、粉碎后过40目筛，取筛下粉末备用；
(2)取龙须菜粉于容器中煮沸120分钟后，过滤得滤渣和提取液；
(3)将步骤(2)所得的最终提取液用截留分子量1kDa的膜进行超滤，舍弃透过液，收集截留液；
(4)用离子交换树脂DEAE-Fast flow型号分离截留液，取降血脂活性最高的组分，真空浓缩至截留液体积的1/8，得浓缩液；
(5)在浓缩液中加入无水乙醇至乙醇终浓度为80wt％，混合均匀后，在0～4℃下静置12 小时，过滤，除去上层清液部分，得多糖沉淀；
(6)将所得多糖沉淀冷冻干燥，得到龙须菜多糖，记为4#提取物。
实验例1测定多糖提取率
测定实施例及对比例中各龙须菜多糖的多糖提取率。
采用苯酚-硫酸法测定1#～4#提取物中的多糖含量，计算多糖提取率，公式为：多糖提取率(％)＝(提取液中多糖含量(g)/原料重量(g))×100。
各龙须菜多糖提取物测定计算结果如表1所示：
表1
提取物 1#提取物 2#提取物 3#提取物 4#提取物
多糖提取率(％) 23.65±0.27 19.80±0.62 20.31±0.80 5.46±0.52
注：表内数据为平均值±标准偏差(n＝3)
从表1中数据可知，本发明所述方法的龙须菜多糖提取率明显高于传统水提法的多糖提取率。
实验例2龙须菜多糖的降血脂实验
本实验利用高胆固醇高脂饲料饲养雄性昆明小鼠造成高血脂模型，从而评价按前述方法制备得到的龙须菜多糖的降血脂作用。
1.实验材料
1.1药物及试剂：高胆固醇高脂饲料(蔗糖20％、猪油15％，胆固醇1.2％，胆酸钠0.2％、酪蛋白10％、磷酸氢钙0.6％、石粉0.4％、预混料0.4％，基础饲料52.2％；总能量4.4kcal/g；由广东省医学动物中心提供)，小鼠灌胃针管，一次性无菌注射器，离心管，无菌生理盐水，阿托伐他汀钙片(辉瑞制药有限公司，产品批号：S90890)，甘油三酯测试盒(南京建成生物科技有限公司，生产批号：20180420)，总胆固醇测试盒(南京建成生物科技有限公司，生产批号：20180420)，低密度脂蛋白测试盒(南京建成生物科技有限公司，生产批号：20180426)，高密度脂蛋白测试盒(南京建成生物科技有限公司，生产批号：20180426)，游离脂肪酸测试盒(南京建成生物科技有限公司，生产批号：20180420)龙须菜多糖(GLP)由本实验室按前述制备方法自制。
1.2实验主要仪器：低速离心机，酶标仪，移液枪。
1.3实验动物：昆明种小鼠，雄性，体重22～24g，购于广东省医学动物中心。
2.供试液样品
实施例1中制备的龙须菜多糖。
3.各供试样品液的配制：
3.1他汀药液的配制：取7片阿托伐他汀钙片(含阿托伐他汀140mg)，以350mL无菌生理盐水混悬，配制成浓度为0.4mg/mL(以阿托伐他汀含量计)的药液样品。
3.2低剂量龙须菜多糖药液的配制：精密称取3.57g龙须菜多糖。以350mL无菌生理盐水溶解，配制成多糖含量为6.0mg/mL的药液样品。
3.3高剂量龙须菜多糖药液的配制：精密称取13.37g龙须菜多糖。以350mL无菌生理盐水溶解，配制成多糖含量为22.5mg/mL的药液样品。
4.实验方法
4.1造模阶段：昆明种小鼠102只，22-24g，雄性，随机分成两组，一组24只，为正常对照组，以普通饲料喂养；另一组78只，为高血脂模型组(以下简称模型组)，以高胆固醇高脂饲料喂养，自由饮水摄食，饲养6周后，每组各随机选取6只小鼠，禁食不禁水8h后，采用摘眼球采血法，离心分离血清，按照试剂盒说明书操作，分别测定总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL-C)、高密度脂蛋白(HDL-C)、游离脂肪酸(FFA)的含量，根据上述指标判断造模是否成功。
4.2实验阶段：造模成功后，将模型组随机分为4组，每组18只，分别为：模型对照组(HF)、阿托伐他汀干预组(HF+AT)、低剂量龙须菜多糖干预组(HF+LGP)、高剂量龙须菜多糖干预组(HF+HGP)。灌胃给药。正常对照组(NC)和模型对照组每日按0.1mL/10g灌胃无菌生理盐水，给药组每日灌胃给予0.1mL/10g体积的相应药物。共给药42天。分别在给药14，28和42天后，每组随机选取6只小鼠禁食不禁水8h后，采用摘眼球采血法采血，脱颈椎处死小鼠后，取附睾周脂肪称重，取肝脏作组织切片病理学分析；采血2h内离心分离血清按照测试盒说明书操作，分别测定TC、TG、LDL-C、HDL-C、FFA的含量。
4.3数据处理与分析方法：以下实验数据均以“平均值±标准差”
Figure PCTCN2018113239-appb-000001
来表示。采用SPSS19.0进行one-way ANOVA进行单因素方差分析和差异显著性分析，方差之间无/有显著差异时以LSD/Dunnett's T3方法进行分析。
5.实验结果
5.1经测定，用于本实验例中的龙须菜多糖提取率为23.65±0.27wt％，经苯酚硫酸法检测得到多糖纯度为59.01±0.91wt％，经BCA试剂盒检测得到蛋白质含量仅为1.57±0.37wt％，硫酸基含量为22.85±0.70wt％，分子量为31.45±5.21kDa。
5.2龙须菜多糖对小鼠血清总胆固醇(TC)含量的影响(mmol/L)、甘油三酯(TG)含 量的影响(mmol/L)、低密度脂蛋白(LDL-C)含量的影响(mmol/L)、高密度脂蛋白(HDL-C)含量的影响(mmol/L)、游离脂肪酸(FFA)含量的影响(mmol/L)、动脉粥样硬化指数(AI)的影响(mmol/L)、附睾周脂重指数的影响(×10 -2g/g)
Figure PCTCN2018113239-appb-000002
见表2～表8。
表2
Figure PCTCN2018113239-appb-000003
注： *p＜0.05，高脂组与正常对照组比较； **p＜0.01，高脂组与正常对照组比较；不同字母表示高脂小鼠各组之间存在显著性差异(p＜0.05)。下同。
表3
Figure PCTCN2018113239-appb-000004
表4
Figure PCTCN2018113239-appb-000005
表5
Figure PCTCN2018113239-appb-000006
表6
Figure PCTCN2018113239-appb-000007
表7
Figure PCTCN2018113239-appb-000008
注：AI＝LDL-C/HDL-C
表8
Figure PCTCN2018113239-appb-000009
Figure PCTCN2018113239-appb-000010
图1a～图1e分别为NC、HF、HF+AT、HF+LGP、HF+HGP组小鼠肝脏组织的油红o染色切片照片(200×)。
图2a～图2e分别为NC、HF、HF+AT、HF+LGP、HF+HGP组小鼠肝脏组织的HE染色切片照片(400×)。
6.分析与讨论
从0d结果来看，模型与空白组相比，TC的浓度有显著性升高(p＜0.05)，而TG、LDL-C的浓度均有极显著性升高(p＜0.01)，HDL-C浓度分别有显著性降低(p＜0.05)和极显著性降低(p＜0.01)，该结果表明实验动物造模成功。在干预42d后TC浓度变化方面，与模型组相比，龙须菜多糖低、高剂量组，具有显著性差异(p＜0.05)，阿托伐他汀组虽有降低，但无显著性差异(p＞0.05)。在干预42d后TG浓度变化方面，与模型组相比，龙须菜多糖低、高剂量组，具有显著性差异(p＜0.05)。在干预14d后LDL-C浓度变化方面，与模型组相比，他汀组具有显著性降低作用(p＜0.05)，而龙须菜多糖低、高剂量组不具有显著性降低作用(p＞0.05)，随着干预时间增加直至42d，高脂小鼠的LDL-C浓度均有下降趋势，但各组之间无显著性差异(p＞0.05)。在干预42d后HDL-C浓度变化方面，与模型组相比，阿托伐他汀组和龙须菜低剂量组，具有显著性升高作用(p＜0.05)，龙须菜高剂量组不具有显著性升高作用(p＞0.05)。在干预42d后FFA浓度变化方面，与模型组相比，龙须菜多糖低剂量组具有显著性降低性作用(p＜0.05)，阿托伐他汀组和龙须菜高剂量组不具有显著性降低作用(p＞0.05)。在干预42d后血清动脉粥样硬化指数(AI＝LDL-C/HDL-C)变化方面，与模型组相比，龙须菜低、高剂量组具有显著性降低作用(p＜0.05)，阿托伐他汀组不具有显著性降低作用(p＞0.05)。在干预42d后附睾周脂重指数(附睾周脂肪重量/体重，×10 -2g/g)变化方面，与模型组相比，阿托伐他汀组、龙须菜低、高剂量组均具有显著性降低作用(p＜0.05)。
从图1a～e的各组小鼠肝脏组织油红o染色切片结果可见：NC组呈正常肝形态，仅有少量肝细胞含细点状橘染脂滴；HF组呈弥漫性滴状深橘红色，坏死区小片状深橘红色，提示大量肝细胞内含大脂滴且坏死处脂滴融合；HF+AT组呈弥漫点状、小滴状粉染，与模型组相比脂滴有所减小，但数量减少不明显；HF+LGP组呈灶性浅橘色，提示含脂滴的肝细胞数量及脂滴大小较模型组明显减少，减脂效果显著；HF+HGP组呈巢片性点状橘色，脂滴较模型组减少、减小，减脂效果明显。
从图2a～e的各组小鼠肝脏组织HE染色切片结果可见：NC组肝细胞排列规则，无脂肪变性，无炎症坏死情况；HF组肝细胞排列紊乱，胞质内可见数量不等，大小不一的脂肪空泡 (箭头标注1)，有重度脂肪变性情况出现，有重度肝细胞水样变性情况(箭头标注2)，同时有炎性细胞灶性浸润(箭头标注3)，提示模型组肝脏有大量脂肪积累，同时有中度炎症坏死情况；HF+AT组肝细胞排列稍有规则，有脂肪空泡(箭头标注1)、肝细胞水样变性(箭头标注2)和少量炎症细胞分布(箭头标注3)，提示他汀干预组有中度脂肪变性情况，同时有轻度炎症情况出现；HF+LGP组肝细胞排列规则，未见脂肪空泡，但有多处局灶性炎性细胞浸润，提示低剂量龙须菜多糖干预后脂肪积累情况有明显好转，但炎症有加重，可能处于机体修复后期；HF+HGP组肝细胞排列规则，有少量脂肪空泡(箭头标注1)，脂肪空泡体积较模型组的明显减小，有轻度脂肪变性情况，未见炎症出现，提示HF+HGP组肝脏脂肪积累情况较模型组有明显好转。
以上实验结果表明，低剂量(多糖含量为6.0mg/mL)的龙须菜多糖对高血脂小鼠的TC、TG、FFA和AI指数均有明显降低作用，并对HDL-C有显著性升高作用；高剂量(多糖含量为22.5mg/mL)的龙须菜多糖对高血脂小鼠的TC、TG和AI指数均有显著降低作用。综上，龙须菜多糖具有显著的降血脂功能，并能有效降低患动脉粥样硬化的风险。
实验例3龙须菜多糖与阿托伐他汀致肝损伤毒副作用比较
1.实验材料
1.1药品试剂：阿托伐他汀钙片(辉瑞制药有限公司，产品批号：S90890)，谷丙转氨酶(GPT)试剂盒(南京建成生物科技有限公司，生产批号：20180420)，谷草转氨酶(GOT)试剂盒(南京建成生物科技有限公司，生产批号：20180420)，龙须菜多糖(GLP)由本实验室自制。
1.2实验仪器：低速离心机，酶标仪，移液枪，
1.3实验动物：昆明种小鼠，雄性，体重22～24g，购于广东省医学动物中心。
2.实验样品的制备
实施例1中制备的龙须菜多糖。
3.实验样品的配制
3.1阳性对照药阿托伐他汀药液的配制：取7片阿托伐他汀钙片(含阿托伐他汀140mg)，以350mL无菌生理盐水混悬，配制成浓度为0.4mg/mL(以阿托伐他汀含量计)的药液样品。
3.2低剂量龙须菜多糖药液的配制：精密称取3.57g龙须菜多糖。以350mL无菌生理盐水溶解，配制成多糖含量为6.0mg/mL的药液样品。
3.3高剂量龙须菜多糖药液的配制：精密称取13.37g龙须菜多糖。以350mL无菌生理盐 水溶解，配制成多糖含量为22.5mg/mL的药液样品。
4.实验方法
4.1造模阶段：昆明种小鼠102只，22-24g，雄性，随机分成两组，一组24只，为正常对照组，以普通饲料喂养；另一组78只，为高血脂模型组(以下简称模型组)，以高胆固醇高脂饲料喂养，自由饮水摄食，饲养6周后，每组各随机选取6只小鼠，禁食不禁水8h后，采用摘眼球采血法，离心分离血清，按照总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)、游离脂肪酸(FFA)试剂盒说明书操作，分别测定TC、TG、LDL-C、HDL-C、FFA的含量，根据上述指标判断造模是否成功。
4.2实验阶段：造模成功后，将模型组随机分为4组，每组18只，分别为：模型对照组(HF)、他汀对照组(HF+AT)、低剂量龙须菜多糖组(HF+LGP)、高剂量龙须菜多糖组(HF+HGP)。灌胃给药。正常对照组(NC)和模型对照组每日按0.1mL/10g灌胃无菌生理盐水，给药组每日灌胃给予0.1mL/10g体积的相应药物。共给药42天。分别在给药14，28和42天后，每组随机选取6只小鼠禁食不禁水8h后，采用摘眼球采血法，离心分离血清，按照谷丙转氨酶(GPT)、谷草转氨酶(GOT)试剂盒说明书操作，分别测定GPT、GOT的活力。
4.3数据处理与分析方法：以下实验数据均以“平均值±标准差”
Figure PCTCN2018113239-appb-000011
来表示。采用SPSS19.0进行one-way ANOVA进行单因素方差分析和差异显著性分析，方差之间无/有显著差异时以LSD/Dunnett's T3方法进行分析。
5.实验结果
5.1经测定，用于本实验例中的龙须菜多糖提取率为23.65±0.27wt％，经苯酚硫酸法检测得到多糖纯度为59.01±0.91wt％，经BCA试剂盒检测得到蛋白质含量仅为1.57±0.37wt％，硫酸基含量为22.85±0.70wt％，分子量为31.45±5.21kDa。
5.2根据实验例2的实验结果，模型与空白组相比，TC的浓度有显著性升高(p＜0.05)，而TG、LDL-C的浓度均有极显著性升高(p＜0.01)，HDL-C浓度分别有显著性降低(p＜0.05)和极显著性降低(p＜0.01)，该结果表明实验动物造模成功。
5.3龙须菜多糖对小鼠血清谷丙转氨酶(GPT)含量的影响(U/L)
Figure PCTCN2018113239-appb-000012
龙须菜多糖对小鼠血清谷草转氨酶(GOT)含量的影响(U/L)
Figure PCTCN2018113239-appb-000013
见表9、表10。
表9
Figure PCTCN2018113239-appb-000014
*p＜0.05，高脂组与正常对照组比较； **p＜0.01，高脂组与正常对照组比较；不同字母表示高脂小鼠各组之间存在显著性差异(p＜0.05)。下同。
表10
Figure PCTCN2018113239-appb-000015
6.分析与讨论
在干预42d后GPT活力变化方面，与正常组相比，高脂小鼠组均具有极显著性升高(p＜0.01)；与模型对照组相比，龙须菜多糖低、高剂量组均有显著性降低(p＜0.05)，但与阿托伐他汀组相比，龙须菜多糖各组均无显著性差异(p＞0.05)。在干预42d后GOT活力变化方面，与正常组相比，模型对照组有极显著性升高(p＜0.01)；与模型对照组相比，阿托伐他汀组、龙须菜多糖低、高剂量组均具有显著性降低作用(p＜0.05)；与阿托伐他汀组相比，龙须菜多糖低剂量组具有显著降低作用(p＜0.05)。
以上结果表明，龙须菜多糖与阿托伐他汀在致肝损伤毒副作用方面存在差异，低剂量(多糖含量为6.0mg/mL)的龙须菜多糖比阿托伐他汀更安全。
上述实施例为本发明较佳的实施方式，但本发明的实施方式并不受上述实施例的限制，其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化，均应为等效的置换方式，都包含在本发明的保护范围之内。

Claims (10)

  1. 一种具有显著降血脂活性的龙须菜多糖的制备方法，其特征在于，包含以下步骤：
    (1)将龙须菜洗净、风干、粉碎过筛，得到龙须菜粉；
    (2)向龙须菜粉中加入有机酸溶液，经过预均质处理后，结合动态高压微射流技术提取龙须菜粉，然后用碱中和，过滤，得到滤渣和提取液；
    (3)将步骤(2)所述提取液进行超滤，舍弃透过液，收集截留液；
    (4)用离子交换树脂分离所述截留液，取降血脂活性最高的组分进行真空浓缩，得浓缩液；
    (5)将所述浓缩液与乙醇混合，静置，过滤得到多糖沉淀；
    (6)将所述多糖沉淀进行干燥，得到所述龙须菜多糖。
  2. 根据权利要求1所述的制备方法，其特征在于，步骤(1)所述粉碎过筛为粉碎后过20～80目筛。
  3. 根据权利要求1所述的制备方法，其特征在于，步骤(2)所述有机酸溶液的pH值为2.0～4.0；所述有机酸溶液为柠檬酸溶液或苹果酸溶液；所述有机酸溶液的质量为所述龙须菜粉质量的20～40倍。
  4. 根据权利要求1所述的制备方法，其特征在于，步骤(2)所述预均质处理的参数为：20～50MPa，1～3分钟，均质次数为1～2次；
    所述动态高压微射流提取的控制条件为：微射流压力为120～180MPa，提取温度为75～100℃，提取时间为80～160分钟，提取次数为1～2次。
  5. 根据权利要求1所述的制备方法，其特征在于，步骤(3)所述超滤采用截留分子量1kDa的超滤膜。
  6. 根据权利要求1所述的制备方法，其特征在于，步骤(4)所述离子交换树脂的型号为DEAE-fast flow或DEAE-52；所述真空浓缩所得浓缩液为所述截留液体积的1/10～1/5。
  7. 根据权利要求1所述的制备方法，其特征在于，步骤(5)所述乙醇的用量占所述浓缩液与乙醇混合后总体积的60％～90％；所述静置是在2～8℃下静置8～14小时。
  8. 根据权利要求1所述的制备方法，其特征在于，步骤(6)所述干燥为真空冷冻干燥或喷雾干燥。
  9. 由权利要求1-8任一项所述的制备方法制得的一种龙须菜多糖，其特征在于，所述龙须菜多糖的分子量为20～35kDa，硫酸基含量达20～28wt％。
  10. 权利要求9所述的一种龙须菜多糖在制备降血脂药物或保健品中的应用。
PCT/CN2018/113239 2018-06-28 2018-10-31 一种具有显著降血脂活性的龙须菜多糖及其制备方法与应用 WO2020000828A1 (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
SG11202012298VA SG11202012298VA (en) 2018-06-28 2018-10-31 Gracilaria lemaneiformis polysaccharide with significant hypolipidemic activity, preparation method thereof and use therefor

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201810686604.4 2018-06-28
CN201810686604.4A CN108586632B (zh) 2018-06-28 2018-06-28 一种具有显著降血脂活性的龙须菜多糖及其制备方法与应用

Publications (1)

Publication Number Publication Date
WO2020000828A1 true WO2020000828A1 (zh) 2020-01-02

Family

ID=63634813

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2018/113239 WO2020000828A1 (zh) 2018-06-28 2018-10-31 一种具有显著降血脂活性的龙须菜多糖及其制备方法与应用

Country Status (3)

Country Link
CN (1) CN108586632B (zh)
SG (1) SG11202012298VA (zh)
WO (1) WO2020000828A1 (zh)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108586632B (zh) * 2018-06-28 2021-08-06 华南理工大学 一种具有显著降血脂活性的龙须菜多糖及其制备方法与应用
CN110368400A (zh) * 2019-08-01 2019-10-25 大连工业大学 一种龙须菜多糖低聚物的制备方法及其应用

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101397346A (zh) * 2008-09-17 2009-04-01 上海海洋大学 一种具有免疫调节作用的龙须菜纯多糖的制备方法
CN104086664A (zh) * 2014-06-09 2014-10-08 华南理工大学 一种龙须菜多糖提取物及其制备方法与应用
CN104710541A (zh) * 2015-03-31 2015-06-17 广西还珠海洋生物科技有限公司 一种从海带中制备海带多糖的方法
CN106868072A (zh) * 2017-03-13 2017-06-20 珀莱雅化妆品股份有限公司 一种小分子量铁皮石斛多糖的制备方法
CN108586632A (zh) * 2018-06-28 2018-09-28 华南理工大学 一种具有显著降血脂活性的龙须菜多糖及其制备方法与应用

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101397346A (zh) * 2008-09-17 2009-04-01 上海海洋大学 一种具有免疫调节作用的龙须菜纯多糖的制备方法
CN104086664A (zh) * 2014-06-09 2014-10-08 华南理工大学 一种龙须菜多糖提取物及其制备方法与应用
CN104710541A (zh) * 2015-03-31 2015-06-17 广西还珠海洋生物科技有限公司 一种从海带中制备海带多糖的方法
CN106868072A (zh) * 2017-03-13 2017-06-20 珀莱雅化妆品股份有限公司 一种小分子量铁皮石斛多糖的制备方法
CN108586632A (zh) * 2018-06-28 2018-09-28 华南理工大学 一种具有显著降血脂活性的龙须菜多糖及其制备方法与应用

Also Published As

Publication number Publication date
CN108586632B (zh) 2021-08-06
CN108586632A (zh) 2018-09-28
SG11202012298VA (en) 2021-01-28

Similar Documents

Publication Publication Date Title
CN103768534B (zh) 一种具有抗肿瘤活性的中药组合物
WO2003099309A1 (en) Active fraction having anti-cancer and anti-metastasis isolated from acanthopanax species and fruits
CN103768152B (zh) 桂花苯乙醇苷提取物及其制备方法和用途
CN104284601B (zh) 用于促进类胰岛素生长因子分泌及骨骼生长的白何首乌提取物和续断提取物及其制造方法
CN101249259B (zh) 一种高含量、高活性口服云芝糖肽及其制备方法和应用
CN106084087A (zh) 一种瓜蒌多糖的制备方法
CN105639617A (zh) 一种茯苓的综合开发和应用的方法及其新用途
WO2020000828A1 (zh) 一种具有显著降血脂活性的龙须菜多糖及其制备方法与应用
CN107537028B (zh) 一种同时辅助降血糖及降血压的配方及其制备方法
CN106243239A (zh) 一种用于提高肝炎患者免疫力的可溶性小分子β‑1,3‑葡聚糖
CN105796587B (zh) 竹茹多糖在免疫调节、抗肿瘤中的应用
CN1814170A (zh) 一种治疗心血管疾病的药物滴丸及其制备方法
CN105777921A (zh) 一种黄芪多糖提取工艺及其用途
CN103342755A (zh) 枸杞多糖均一级份ⅳ及其制备方法和应用
CN102784247A (zh) 具有降血脂功能含壳寡糖的药物组合物
CN107137416B (zh) 一种防治非小细胞肺癌的药物组合物
CN104004110A (zh) 一种从瓜子金中提取的瓜子金多糖及其应用
CN103690573A (zh) 一种用于增强免疫力的天然药物组合物及其制备方法
CN108969580B (zh) 蓝布正总鞣质的制备方法及应用
CN108159146B (zh) 一种用于降血脂的中药组合物及制备方法
CN100589812C (zh) 冬虫夏草菌丝体抽提物的分离物及经口摄取用组合物
CN101804128B (zh) 一种治疗炎症性肠病的药物组合物及该组合物在制备治疗炎症性肠病药物中的用途
CN109294984A (zh) 一种体内高效扩增nk细胞的香菇多糖胶囊及其制备方法
CN104523851A (zh) 一种水红花子果皮总黄酮提取物的制备方法
CN108126006A (zh) 断血流口服制剂的制备工艺

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 18924909

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC (EPO FORM 1205A DATED 11/05/2021)

122 Ep: pct application non-entry in european phase

Ref document number: 18924909

Country of ref document: EP

Kind code of ref document: A1