CN112920288B - 一种具有降脂活性的黄大茶酸性多糖及其制备方法和应用 - Google Patents
一种具有降脂活性的黄大茶酸性多糖及其制备方法和应用 Download PDFInfo
- Publication number
- CN112920288B CN112920288B CN202110123087.1A CN202110123087A CN112920288B CN 112920288 B CN112920288 B CN 112920288B CN 202110123087 A CN202110123087 A CN 202110123087A CN 112920288 B CN112920288 B CN 112920288B
- Authority
- CN
- China
- Prior art keywords
- lytp
- yellow
- tea
- lipid
- polysaccharide
- Prior art date
- Legal status (The legal status 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 status listed.)
- Active
Links
- 229920001284 acidic polysaccharide Polymers 0.000 title claims abstract description 22
- 150000004805 acidic polysaccharides Chemical class 0.000 title claims abstract description 22
- 230000000694 effects Effects 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title abstract description 9
- 241001122767 Theaceae Species 0.000 title abstract 4
- 239000005642 Oleic acid Substances 0.000 claims abstract description 34
- 229920001282 polysaccharide Polymers 0.000 claims abstract description 27
- 239000005017 polysaccharide Substances 0.000 claims abstract description 27
- 150000004804 polysaccharides Chemical class 0.000 claims abstract description 27
- 210000004027 cell Anatomy 0.000 claims abstract description 24
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 claims abstract description 20
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 claims abstract description 20
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 claims abstract description 20
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 claims abstract description 20
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 claims abstract description 20
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 claims abstract description 20
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 13
- 239000004952 Polyamide Substances 0.000 claims abstract description 11
- 229920002647 polyamide Polymers 0.000 claims abstract description 11
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 claims abstract description 10
- 239000012528 membrane Substances 0.000 claims abstract description 10
- 150000002772 monosaccharides Chemical class 0.000 claims abstract description 9
- 239000002253 acid Substances 0.000 claims abstract description 8
- 238000004587 chromatography analysis Methods 0.000 claims abstract description 8
- 239000000203 mixture Substances 0.000 claims abstract description 8
- AEMOLEFTQBMNLQ-YMDCURPLSA-N D-galactopyranuronic acid Chemical compound OC1O[C@H](C(O)=O)[C@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-YMDCURPLSA-N 0.000 claims abstract description 7
- 239000008103 glucose Substances 0.000 claims abstract description 7
- 230000006372 lipid accumulation Effects 0.000 claims abstract description 6
- 210000005229 liver cell Anatomy 0.000 claims abstract description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 5
- 239000003814 drug Substances 0.000 claims abstract description 5
- 229940097043 glucuronic acid Drugs 0.000 claims abstract description 5
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 claims abstract description 4
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 claims abstract description 4
- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 claims abstract description 4
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 claims abstract description 4
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 claims abstract description 4
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 claims abstract description 4
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 claims abstract description 4
- 230000003544 deproteinization Effects 0.000 claims abstract description 4
- 229930182830 galactose Natural products 0.000 claims abstract description 4
- AEMOLEFTQBMNLQ-AQKNRBDQSA-N D-glucopyranuronic acid Chemical compound OC1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-AQKNRBDQSA-N 0.000 claims abstract description 3
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 claims abstract 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims abstract 2
- 244000269722 Thea sinensis Species 0.000 claims description 40
- 235000013616 tea Nutrition 0.000 claims description 35
- 235000020338 yellow tea Nutrition 0.000 claims description 20
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 15
- 240000001548 Camellia japonica Species 0.000 claims description 12
- 235000018597 common camellia Nutrition 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 12
- 238000002386 leaching Methods 0.000 claims description 10
- 238000001914 filtration Methods 0.000 claims description 8
- 235000006468 Thea sinensis Nutrition 0.000 claims description 7
- 238000005238 degreasing Methods 0.000 claims description 6
- 239000003480 eluent Substances 0.000 claims description 6
- 239000011344 liquid material Substances 0.000 claims description 6
- 239000008280 blood Substances 0.000 claims description 5
- 210000004369 blood Anatomy 0.000 claims description 5
- 150000002632 lipids Chemical class 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 5
- 239000012530 fluid Substances 0.000 claims description 4
- 230000001603 reducing effect Effects 0.000 claims description 4
- 238000004042 decolorization Methods 0.000 claims description 3
- 238000000502 dialysis Methods 0.000 claims description 3
- 238000000605 extraction Methods 0.000 claims description 3
- 239000000126 substance Substances 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 2
- 238000011068 loading method Methods 0.000 claims description 2
- 230000001376 precipitating effect Effects 0.000 claims description 2
- 238000010298 pulverizing process Methods 0.000 claims description 2
- 235000013402 health food Nutrition 0.000 claims 2
- 238000007605 air drying Methods 0.000 claims 1
- 239000000945 filler Substances 0.000 claims 1
- 235000013305 food Nutrition 0.000 abstract description 5
- 238000000926 separation method Methods 0.000 abstract description 5
- 238000005516 engineering process Methods 0.000 abstract description 3
- 230000036541 health Effects 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 14
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 10
- 239000000523 sample Substances 0.000 description 9
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 9
- 239000001963 growth medium Substances 0.000 description 7
- 239000004480 active ingredient Substances 0.000 description 5
- 235000012000 cholesterol Nutrition 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 4
- DATAGRPVKZEWHA-YFKPBYRVSA-N N(5)-ethyl-L-glutamine Chemical compound CCNC(=O)CC[C@H]([NH3+])C([O-])=O DATAGRPVKZEWHA-YFKPBYRVSA-N 0.000 description 4
- 239000002244 precipitate Substances 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 235000020279 black tea Nutrition 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 210000003494 hepatocyte Anatomy 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 230000007935 neutral effect Effects 0.000 description 3
- 150000008442 polyphenolic compounds Chemical class 0.000 description 3
- 235000013824 polyphenols Nutrition 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- UJOBWOGCFQCDNV-UHFFFAOYSA-N 9H-carbazole Chemical compound C1=CC=C2C3=CC=CC=C3NC2=C1 UJOBWOGCFQCDNV-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- NPGIHFRTRXVWOY-UHFFFAOYSA-N Oil red O Chemical compound Cc1ccc(C)c(c1)N=Nc1cc(C)c(cc1C)N=Nc1c(O)ccc2ccccc12 NPGIHFRTRXVWOY-UHFFFAOYSA-N 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- 230000003044 adaptive effect Effects 0.000 description 2
- 229930013930 alkaloid Natural products 0.000 description 2
- 150000003797 alkaloid derivatives Chemical class 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 235000009569 green tea Nutrition 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 235000020333 oolong tea Nutrition 0.000 description 2
- 238000012856 packing Methods 0.000 description 2
- 239000000049 pigment Substances 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000007873 sieving Methods 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 239000012086 standard solution Substances 0.000 description 2
- 229940026510 theanine Drugs 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- FNEHAOQZWPHONV-UHFFFAOYSA-N 9h-carbazole;sulfuric acid Chemical compound OS(O)(=O)=O.C1=CC=C2C3=CC=CC=C3NC2=C1 FNEHAOQZWPHONV-UHFFFAOYSA-N 0.000 description 1
- 235000019224 Camellia sinensis var Qingmao Nutrition 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 238000001212 derivatisation Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- UZIWRCBLXLKBID-UHFFFAOYSA-N dodecasodium sulfuric acid tetraborate Chemical compound [Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+].[O-]B([O-])[O-].[O-]B([O-])[O-].[O-]B([O-])[O-].[O-]B([O-])[O-].OS(O)(=O)=O UZIWRCBLXLKBID-UHFFFAOYSA-N 0.000 description 1
- 230000035622 drinking Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000000105 evaporative light scattering detection Methods 0.000 description 1
- 235000019225 fermented tea Nutrition 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000007407 health benefit Effects 0.000 description 1
- 239000005457 ice water Substances 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000019520 non-alcoholic beverage Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 235000020339 pu-erh tea Nutrition 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000002390 rotary evaporation Methods 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000820 toxicity test Toxicity 0.000 description 1
- 150000003626 triacylglycerols Chemical class 0.000 description 1
- 235000020334 white tea Nutrition 0.000 description 1
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
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
-
- 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
-
- 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
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Polymers & Plastics (AREA)
- Materials Engineering (AREA)
- Biochemistry (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Sustainable Development (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Diabetes (AREA)
- Hematology (AREA)
- Obesity (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明公开了一种具有降脂活性的黄大茶酸性多糖及其制备方法和应用，其中黄大茶酸性多糖组分LYTP‑2的分子量为4.57×104Da，糖醛酸含量在62％以上，其单糖组成及摩尔比为半乳糖醛酸:葡萄糖醛酸:阿拉伯糖:半乳糖:鼠李糖:葡萄糖:甘露糖＝69.45:1:10.41:10.13:9.78:6.03:1.36。本发明黄大茶酸性多糖组分LYTP‑2，采用聚酰胺色谱脱蛋白脱色与超滤膜分离多糖组分技术联用制得，具有显著的降脂功能，可明显降低油酸诱导HepG2细胞的脂质积累，且对正常肝细胞LO2无损害作用。LYTP‑2作为新型天然的高活性降脂多糖，可广泛用于保健食品与医药健康领域。
Description
技术领域
本发明属于保健食品领域，具体涉及一种具有降脂活性的黄大茶酸性多糖及其制备方法和应用。
背景技术
茶作为世界上最重要的非酒精饮料之一，因其独特的香气和滋味，受到越来越多消费者的喜爱。近年来，饮茶所带来的健康益处已成为食品科学和营养学倍受关注的话题。根据发酵的程度，茶可分为绿茶(未发酵的茶)、白茶(微发酵)、黄茶(部分发酵)、乌龙茶(半发酵茶)、红茶(全发酵茶)、黑茶(发酵后的茶)等六种类型。科学研究表明茶叶中的茶多酚、茶多糖、生物碱、茶氨酸等活性成分具有抗氧化、抗肿瘤、增强免疫、降血糖、血脂、减肥和抗辐射等多种功效。其中茶多糖作为茶的主要活性成分在普洱茶(黑茶)、绿茶、乌龙茶等茶类已有比较系统的研究与开发，而产自安徽皖西大别山一带的黄大茶虽然有其降血糖、降血脂活性的文献报道，但其活性成分的物质基础尚不清楚，黄大茶中茶多糖的主要活性组分及其理化性质和功效关系亟待明确。
发明内容
本发明旨在提供一种具有降脂活性的黄大茶酸性多糖及其制备方法和应用。
申请人对安徽省霍山县的黄大茶进行了活性成分筛选实验，证明其茶多糖组分的减肥降脂活性明显高于茶多酚、茶氨酸、生物碱等成分，进一步深入探究黄大茶的多糖活性组分时发现黄大茶多糖提取物成分非常复杂，尤其是含大量的蛋白(含量接近40％)和色素，而用常规的除多糖蛋白方法(Sevag试剂法)根本无法将其除尽。本发明通过聚酰胺色谱与超滤分离技术联用，不仅将黄大茶多糖中蛋白含量从38.98％降至6.0％以下，茶色素及多酚类成分全部除去，且获得了富含半乳糖醛酸的黄大茶多糖组分(LYTP-2)，降脂作用十分优异。迄今，未见相关文献报道。
本发明分离获得的黄大茶酸性多糖LYTP-2，其分子量为4.57×104Da，糖醛酸含量在62％以上，其单糖组成及摩尔比为半乳糖醛酸:葡萄糖醛酸:阿拉伯糖:半乳糖:鼠李糖:葡萄糖:甘露糖＝69.45:1:10.41:10.13:9.78:6.03:1.36。
本发明黄大茶酸性多糖能明显降低油酸诱导的HepG2细胞的脂质积累，LYTP-2在400μg/mL时，能将油酸诱导HepG2细胞内总胆固醇(TC)、甘油三酯(TG)含量降低58.99％、57.29％，且对正常肝细胞LO2无损害作用，具有很重要的学术研究价值和开发应用前景。
本发明具有降脂活性的黄大茶酸性多糖的制备方法，首先将黄大茶依次经粉碎、脱脂、提取、醇沉处理后，采用聚酰胺色谱方法脱色脱蛋白，制得黄大茶粗多糖LYTP；然后将制得的LYTP透析、冻干后进行超滤分离，分别过超滤膜截留分子量为1×104Da和1×105Da进行分级，将分子量在1×104至1×105Da之间的超滤液组分浓缩、冻干，制得黄大茶多糖LYTP-2。具体包括如下步骤：
步骤1：粉碎
将黄大茶粉碎后过80目筛，制得黄大茶粉；
步骤2：脱脂
将步骤1所得黄大茶粉1000g按照1g:10mL的液料比加入95％乙醇，70℃浸提2-3h，重复浸提2-3次，过滤后收集茶滤渣，晾干；
步骤3：提取
将步骤2所得茶滤渣以1g:20mL的液料比加入蒸馏水，90℃浸提2-3h，重复浸提3-4次，合并提取液；
步骤4：醇沉
将步骤3所得提取液浓缩至1000mL，加入4000mL的95％乙醇，4℃静置10-12h，5000rpm离心10-15min，收集沉淀物；
步骤5：聚酰胺色谱法脱蛋白/脱色
采用AKTApurifier TM10色谱仪(美国GE公司)，色谱条件为：色谱柱XK50/30(50mm×30cm)，色谱填料80-100目聚酰胺，样品为步骤4所得沉淀物水溶液，上样量为2/5柱体积，洗脱液为纯水，流速2-5mL/min，收集3-5个柱体积的洗脱液，浓缩，透析(透析袋截留分子量3.5×103Da，去除小分子物质)，冻干，制得黄大茶粗多糖LYTP；
步骤6：超滤分离多糖组分
超滤装置UltrareservoirTMMasterflex(美国PALL公司)，10K、100K和500K超滤膜包(PALL OmegaTM)，称取适量步骤5制得的LYTP，纯水溶解，0.22μ滤膜过滤上样，依次过10K、100K和500K膜包，收集膜截留液和透过液，得到分子量在1×104Da以下、1×104-1×105Da、1×105-5×105Da、5×105Da以上四个多糖组分，浓缩、冻干，分别制得黄大茶多糖LYTP-1、LYTP-2、LYTP-3和LYTP-4。
本发明黄大茶酸性多糖LYTP-2的应用，是用于制备具有降脂功能的药物或保健食品，可明显降低油酸诱导HepG2细胞的脂质积累，且对正常肝细胞LO2无损害作用。
本发明以油酸诱导的HepG2细胞和正常肝细胞LO2为研究对象，通过细胞实验证明黄大茶酸性多糖LYTP-2的降脂活性。
本发明使用的黄大茶原料产自安徽霍山县亨大茶叶有限公司。
本发明的有益效果体现在：
1、本发明得到的具有显著降脂活性黄大茶酸性多糖LYTP-2，HPLC检测其分子量为4.57×104Da，单糖组成及摩尔比为半乳糖醛酸:葡萄糖醛酸:阿拉伯糖:半乳糖:鼠李糖:葡萄糖:甘露糖＝69.45:1:10.41:10.13:9.78:6.03:1.36，半乳糖醛酸为其主要成分。
2、本发明采用聚酰胺色谱与超滤分离技术联用取代Sevag试剂法制取LYTP-2，方法简捷、绿色环保，适用于规模化制备，应用前景广阔。
3、本发明黄大茶酸性多糖LYTP-2具有显著的降脂功能，可明显降低油酸诱导HepG2细胞的脂质积累，且对正常肝细胞LO2无损害作用。LYTP-2作为新型天然的高活性降脂多糖，可广泛用于保健食品与医药健康领域。
附图说明
图1黄大茶酸性多糖LYTP-2相对分子质量的高效液相色谱图，色谱条件为安捷伦1260HPLC-ELSD，TSKgel G3000PWxL(7.8mm×30cm，7μm)。图1显示，黄大茶酸性多糖LYTP-2的出峰时间为6.417min，测得LYTP-2的分子量为4.57×104Da。
图2标准单糖组成的高效液相色谱图(*-溶剂峰，1-甘露糖，2-鼠李糖，3-葡萄糖醛酸，4-半乳糖醛酸，5-葡萄糖，6-半乳糖，7-阿拉伯糖，8-木糖，9-岩藻糖)。
图3黄大茶酸性多糖LYTP-2单糖组成的高效液相色谱图(*-溶剂峰，1-甘露糖，2-鼠李糖，3-葡萄糖醛酸，4-半乳糖醛酸，5-葡萄糖，6-半乳糖，7-阿拉伯糖)。
图4HepG2细胞的油红O染色图(对照组-未经油酸诱导，模型组-油酸诱导，LYTP-1-油酸+LYTP-1，LYTP-2-油酸+LYTP-2，LYTP-3-油酸+LYTP-3，LYTP-4-油酸+LYTP-4)。图4显示，与模型组相比，LYTP-2红色区域显著减少，说明LPYT-2明显降低油酸诱导HepG2细胞的脂质积累。
图5LYTP-1、LYTP-2、LYTP-3和LYTP-4对油酸诱导HepG2细胞总脂质的影响(对照组-未经油酸诱导，模型组-油酸诱导，LYTP-1-油酸+LYTP-1，LYTP-2-油酸+LYTP-2，LYTP-3-油酸+LYTP-3，LYTP-4-油酸+LYTP-4)。由图5可知，与模型组相比，LYTP-2组的总脂质显著降低，且明显优于其他组分。
图6LYTP-1、LYTP-2、LYTP-3和LYTP-4对油酸诱导HepG2细胞内总胆固醇(TC)、甘油三酯(TG)的影响(对照组-未经油酸诱导，模型组-油酸诱导，LYTP-1-油酸+LYTP-1，LYTP-2-油酸+LYTP-2，LYTP-3-油酸+LYTP-3，LYTP-4-油酸+LYTP-4)。图6显示，与模型组相比，LYTP-2组总胆固醇(TC)、甘油三酯(TG)分别降低58.99％、57.29％，而LYTP-1降低29.63％、18.12％，LYTP-3降低50.84％、46.02％，LYTP-4降低25.22％、29.34％，说明相较于其他组分，LYTP-2降低油酸诱导HepG2细胞内TC、TG效果最优。
图7LYTP-2对正常肝细胞LO2的毒性检测。由图7可知，与空白对照组相比，LYTP-2对LO2细胞无毒副作用。
具体实施方式
以下通过具体的实施例描述本发明的制备，降脂活性，所举实施例只用于解释本发明，并非用于限定本发明的保护范围。
实施例1：黄大茶酸性多糖LYTP-2的制备
1、粉碎：将黄大茶粉碎后过80目筛，制得黄大茶粉；
2、脱脂：将步骤1所得黄大茶粉1000g按照1:10的液料比(g/mL)加入95％乙醇，70℃浸提2-3h，重复浸提2-3次，过滤后收集茶滤渣，晾干；
3、提取：将步骤2所得的茶滤渣以1:20的液料比(g/mL)加入纯水，90℃浸提2-3h，重复浸提3-4次，合并提取液；
4、醇沉：将步骤3所得提取液浓缩至1000mL，加入4000mL的95％乙醇，4℃静置10-12h，5000rpm离心10-15min，收集沉淀物；
5、聚酰胺色谱法脱蛋白/脱色：采用AKTApurifier TM10色谱仪(美国GE公司)，色谱条件：色谱柱XK50×30(50mm×30cm)，色谱填料80-100目聚酰胺，样品为步骤4所得沉淀物水溶液，上样量为2/5柱体积，洗脱液为纯水，流速2-5mL/min，收集3-5个柱体积的洗脱液，浓缩，透析(透析袋截留分子量3.5×103Da，去除小分子物质)，冻干，制得黄大茶粗多糖LYTP；
6、超滤膜分离：超滤装置UltrareservoirTMMasterflex(美国PALL公司)，10K、100K和500K超滤膜包(PALL OmegaTM)，称取适量步骤5制得得的LYTP，纯水溶解，0.22μm滤膜过滤上样，依次过10K、100K和500K膜包，收集膜截留液和透过液，得到分子量在1×104Da以下、1×104-1×105Da、1×105-5×105Da、5×105Da以上四个多糖组分，浓缩、冻干后制得黄大茶多糖，分别命名为LYTP-1、LYTP-2、LYTP-3、LYTP-4。
实施例2：黄大茶酸性多糖LYTP-2的中性糖、糖醛酸和单糖组成测定
1、苯酚-硫酸法测定黄大茶酸性多糖LYTP-2的中性糖含量。精确吸取2mLLYTP-2溶液(50μg/mL)和葡萄糖标准溶液，加入1mL 6％苯酚和5mL浓硫酸，设置三组平行，490nm处测定吸光值，测得LYTP-2的中性糖含量为23％。
2、咔唑–硫酸法测定黄大茶酸性多糖LYTP-2的酸性糖醛酸含量。精确吸取1mLLYTP-2溶液(50μg/mL)和半乳糖醛酸标准溶液，冰水浴加入5mL 0.478％四硼酸钠-硫酸，沸水浴20min，冷却后加入0.2mL0.15％咔唑，混匀静置2h，设置三组平行，520nm处测定吸光值，测得LYTP-2的糖醛酸含量为62％以上。
3、LYTP-2的单糖组成分析采用酸水解-柱前PMP衍生化方法处理样品，高效液相色谱法分析测定。精确称取5mg LYTP-2溶于5mL的2M的三氟乙酸中，氮气封管，110℃水解8h。冷却至室温后与适量的甲醇混合，反复旋蒸至中性，加入1mL纯水，备用。
标准单糖和已水解的样品溶液中加入等量的0.3MNaOH溶液，混匀。随后取适量混合溶液于具塞玻璃试管中，加入等量的0.5MPMP-甲醇溶液，70℃水浴1h。取出冷却至室温，氯仿萃取3-4次，分离上清液。高效液相色谱检测，采用DAD检测器。色谱条件为：色谱柱
C18柱(5μm，4.6mm×250mm)；流动相A为乙腈，流动相B为0.05MKH2PO4；梯度洗脱的时间设置为0-5min、5-10min、10-30min、30-60min，流动相B的比例设置为83％-82％-81％-80％-83％；检测波长为245nm；柱温设置为30℃；流速为1mL/min；进样量为10μL。
实施例3：黄大茶多糖的降脂活性测定
利用油酸诱导的HepG2细胞和正常肝细胞LO2为模型，对黄大茶多糖组分LYTP-1、LYTP-2、LYTP-3、LYTP-4的降脂活性进行评价。
1、降脂活性的检测
(1)总脂质的检测：取生长状态良好的HepG2细胞，接种于48孔板，置于5％CO2培养箱中适应性培养24h后，吸除培养基，空白对照组加入新鲜培养基，模型组加入0.2mM油酸溶液，实验样品组分别加入0.2mM油酸及400μg/mLLYTP-1、LYTP-2、LYTP-3、LYTP-4溶液,继续培养24h，油红O染色，显微镜下观察拍照(见图4)。拍完照后，每孔加入200μL的异丙醇，测定总脂质含量，结果如图5所示。
(2)总胆固醇(TC)、甘油三酯(TG)的检测：取生长状态良好的HepG2细胞，接种于6孔板，置于5％CO2培养箱中适应性培养24h后，吸除培养基，空白对照组加入新鲜培养基，模型组加入0.2mM油酸溶液，实验样品组分别加入0.2mM油酸及400μg/mL LYTP-1、LYTP-2、LYTP-3、LYTP-4溶液，继续放入培养箱中培养24h后，收集、破碎细胞，测定细胞内总胆固醇、甘油三酯含量，结果如图6所示。
2、正常肝细胞LO2毒性的检测
取生长状态良好的LO2细胞，接种于96孔板，置于5％CO2培养箱在适应性培养24h，吸除培养基，空白对照组加入新鲜培养基，样品组依次加入100μg/mL、200μg/mL、400μg/mL、800μg/mL、1000μg/mLLYTP-2溶液，继续培养24h。然后吸除培养液，MTT法检测细胞存活率，结果如图7所示。
Claims (4)
1.一种具有降脂活性的黄大茶酸性多糖，其特征在于：
所述黄大茶酸性多糖为分子量4.57×104Da的多糖组分LYTP-2，糖醛酸含量在62％以上，其单糖组成及摩尔比为半乳糖醛酸:葡萄糖醛酸:阿拉伯糖:半乳糖:鼠李糖:葡萄糖:甘露糖＝69.45:1:10.41:10.13:9.78:6.03:1.36。
2.一种权利要求1所述的黄大茶酸性多糖的制备方法，其特征在于包括如下步骤：
(1)将黄大茶依次经粉碎、脱脂、提取、醇沉处理后，采用聚酰胺色谱方法脱色脱蛋白，制得黄大茶粗多糖LYTP；
所述脱脂是将粉碎后获得的黄大茶粉按照1g:10mL的液料比加入95％乙醇中，70℃浸提2-3h，重复浸提2-3次，过滤后收集茶滤渣，晾干；
所述提取是将脱脂后获得的茶滤渣以1g:20mL的液料比加入蒸馏水中，90℃浸提2-3h，重复浸提3-4次，合并提取液；
采用聚酰胺色谱方法脱色脱蛋白时，采用AKTApurifier TM 10色谱仪，色谱条件为：色谱柱XK50/30，色谱填料80-100目聚酰胺，上样量为2/5柱体积，洗脱液为纯水，流速2-5mL/min，收集3-5个柱体积的洗脱液，浓缩，透析去除小分子物质，透析袋截留分子量3.5×103Da；
(2)将(1)制得的LYTP透析、冻干后进行超滤分离，分别过超滤膜截留分子量为1×104Da和1×105Da进行分级，将分子量在1×104至1×105Da之间的截留液组分浓缩、冻干，制得黄大茶多糖LYTP-2。
3.一种权利要求1所述的黄大茶酸性多糖的应用，其特征在于：用于制备具有降脂功能的药物或保健食品。
4.根据权利要求3所述的应用，其特征在于：
所述药物或保健食品可显著降低油酸诱导HepG2细胞的脂质积累，且对正常肝细胞LO2无损害作用。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110123087.1A CN112920288B (zh) | 2021-01-29 | 2021-01-29 | 一种具有降脂活性的黄大茶酸性多糖及其制备方法和应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110123087.1A CN112920288B (zh) | 2021-01-29 | 2021-01-29 | 一种具有降脂活性的黄大茶酸性多糖及其制备方法和应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112920288A CN112920288A (zh) | 2021-06-08 |
| CN112920288B true CN112920288B (zh) | 2022-04-29 |
Family
ID=76168264
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110123087.1A Active CN112920288B (zh) | 2021-01-29 | 2021-01-29 | 一种具有降脂活性的黄大茶酸性多糖及其制备方法和应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112920288B (zh) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114835827B (zh) * | 2022-03-23 | 2023-03-28 | 安徽大学 | 一种超高压制取黄大茶多糖的方法及其应用 |
| CN114751997B (zh) * | 2022-04-25 | 2023-02-28 | 安徽农业大学 | 一种具有抗炎活性的黄大茶多糖及其制备方法和应用、抗炎药物组合物 |
| CN115466338B (zh) * | 2022-09-29 | 2023-08-29 | 安徽慧之诺医学生物科技有限公司 | 一种黄大茶多糖及其药物组合物和应用 |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101798356A (zh) * | 2010-04-08 | 2010-08-11 | 晋江市恒源科技开发有限公司 | 一种分离提取天然茶多糖的方法 |
| CN103450365A (zh) * | 2012-07-05 | 2013-12-18 | 陈小强 | 一种以高速逆流色谱技术纯化制备茶多糖复合物的方法 |
| CN105566402B (zh) * | 2015-12-29 | 2017-11-21 | 上海师范大学 | 一种综合提取茶香精、茶多糖和茶多酚的方法 |
| CN111793660A (zh) * | 2020-09-02 | 2020-10-20 | 安徽大学 | 一种具有降血糖作用的黄大茶多糖 |
-
2021
- 2021-01-29 CN CN202110123087.1A patent/CN112920288B/zh active Active
Non-Patent Citations (1)
| Title |
|---|
| Ultrasound irradiation alters the spatial structure and improves the antioxidant activity of the yellow tea polysaccharide;Haisong Wang 等;《Ultrasonics Sonochemistry》;20200922;第70卷;第105355页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112920288A (zh) | 2021-06-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112920288B (zh) | 一种具有降脂活性的黄大茶酸性多糖及其制备方法和应用 | |
| US10835552B2 (en) | Method for preparing linseed polysaccharide having antiviral activity and immunological activity, and use of the linseed polysaccharide | |
| CN103444943B (zh) | 以黑茶为原料综合提制茶褐素和茶多糖的方法 | |
| CN105566402B (zh) | 一种综合提取茶香精、茶多糖和茶多酚的方法 | |
| CN103965369B (zh) | 灵芝多糖、提取纯化方法及其作为烟草保润剂的应用 | |
| Meng et al. | Rapid screening and separation of active compounds against α-amylase from Toona sinensis by ligand fishing and high-speed counter-current chromatography | |
| CN102334574A (zh) | 一种从六堡茶中提取茶褐素粗品的方法 | |
| CN112010989B (zh) | 一种具有抗氧化活性的竹荪菌托多糖的制备方法 | |
| CN107056851A (zh) | 一种巴戟天总低聚糖的制备方法 | |
| CN112920287B (zh) | 具有免疫调节功效的阳春砂多糖及其制备方法和应用 | |
| CN103404651B (zh) | 一种低咖啡碱含量的速溶茶原料的制备技术 | |
| CN101307343A (zh) | 非酯型儿茶素制备方法 | |
| CN115028753B (zh) | 具有抗肿瘤功效的沙棘均一多糖及其分离纯化方法及应用 | |
| CN112794925B (zh) | 一种阳春砂多糖及其制备方法和应用 | |
| CN113105567B (zh) | 一种蝉拟青霉甘露聚糖及其制备和用途 | |
| CN105153320B (zh) | 一种海带多糖的提取方法 | |
| CN103450365A (zh) | 一种以高速逆流色谱技术纯化制备茶多糖复合物的方法 | |
| CN102875691B (zh) | 一种从茶渣中同步制备多种活性物质的方法 | |
| CN115368480B (zh) | 毛酸浆茎多糖及其制备方法与应用 | |
| CN104586910A (zh) | 雪菊提取物及其制备方法 | |
| CN115746157B (zh) | 一种美味扇菇多糖及其制备方法和用途 | |
| TWI752622B (zh) | 蘆薈萃取物及其製備方法 | |
| CN114917246B (zh) | 覆盆子多糖r2在制备抗肿瘤药物和抗炎制剂中的应用 | |
| CN111675771B (zh) | 一种北沙参多糖及其制备方法和应用 | |
| CN116731217B (zh) | 一种藤茶酸性多糖AGP-2a及其制备方法和在制备抗炎化妆品中的用途 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20240705 Address after: 673400 Daplo Tea Factory, Daplo Village Committee, Shangpa Town, Fugong County, Nujiang Lisu Autonomous Prefecture, Yunnan Province Patentee after: Fugong Yuanlue Dapuluo Tea Industry Co.,Ltd. Country or region after: China Address before: No. 111, Kowloon Road, Hefei, Anhui, Anhui Patentee before: ANHUI University Country or region before: China |