CN106588664A - Water-soluble caffeoyl structural grease, and preparation method and application thereof - Google Patents
Water-soluble caffeoyl structural grease, and preparation method and application thereof Download PDFInfo
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- CN106588664A CN106588664A CN201611010656.7A CN201611010656A CN106588664A CN 106588664 A CN106588664 A CN 106588664A CN 201611010656 A CN201611010656 A CN 201611010656A CN 106588664 A CN106588664 A CN 106588664A
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- water soluble
- soluble coffee
- caffeoyl
- structure grease
- grease
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- 239000004519 grease Substances 0.000 title claims abstract description 43
- 238000002360 preparation method Methods 0.000 title abstract description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 43
- 125000005456 glyceride group Chemical group 0.000 claims abstract description 17
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 claims abstract description 13
- 238000006243 chemical reaction Methods 0.000 claims abstract description 12
- 230000002255 enzymatic effect Effects 0.000 claims abstract description 12
- 238000001308 synthesis method Methods 0.000 claims abstract description 11
- 239000000126 substance Substances 0.000 claims abstract description 8
- 239000002537 cosmetic Substances 0.000 claims abstract description 5
- 235000013305 food Nutrition 0.000 claims abstract description 4
- 239000002253 acid Substances 0.000 claims description 46
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 31
- 235000021539 instant coffee Nutrition 0.000 claims description 30
- 235000016213 coffee Nutrition 0.000 claims description 21
- 235000013353 coffee beverage Nutrition 0.000 claims description 21
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 15
- 238000000605 extraction Methods 0.000 claims description 11
- 239000004367 Lipase Substances 0.000 claims description 10
- 108010048733 Lipozyme Proteins 0.000 claims description 9
- FCCDDURTIIUXBY-UHFFFAOYSA-N lipoamide Chemical compound NC(=O)CCCCC1CCSS1 FCCDDURTIIUXBY-UHFFFAOYSA-N 0.000 claims description 9
- 239000000706 filtrate Substances 0.000 claims description 8
- 238000001914 filtration Methods 0.000 claims description 8
- 238000013019 agitation Methods 0.000 claims description 7
- 239000007788 liquid Substances 0.000 claims description 7
- 238000001816 cooling Methods 0.000 claims description 6
- 238000004090 dissolution Methods 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 5
- 239000000376 reactant Substances 0.000 claims description 5
- 238000002390 rotary evaporation Methods 0.000 claims description 5
- 238000002425 crystallisation Methods 0.000 claims description 4
- 238000003756 stirring Methods 0.000 claims description 4
- 108010084311 Novozyme 435 Proteins 0.000 claims description 2
- 108090001060 Lipase Proteins 0.000 claims 1
- 102000004882 Lipase Human genes 0.000 claims 1
- XYIBRDXRRQCHLP-UHFFFAOYSA-N ethyl acetoacetate Chemical compound CCOC(=O)CC(C)=O XYIBRDXRRQCHLP-UHFFFAOYSA-N 0.000 claims 1
- 235000019421 lipase Nutrition 0.000 claims 1
- 239000007787 solid Substances 0.000 claims 1
- QAIPRVGONGVQAS-DUXPYHPUSA-N trans-caffeic acid Chemical compound OC(=O)\C=C\C1=CC=C(O)C(O)=C1 QAIPRVGONGVQAS-DUXPYHPUSA-N 0.000 abstract description 22
- 235000004883 caffeic acid Nutrition 0.000 abstract description 11
- ACEAELOMUCBPJP-UHFFFAOYSA-N (E)-3,4,5-trihydroxycinnamic acid Natural products OC(=O)C=CC1=CC(O)=C(O)C(O)=C1 ACEAELOMUCBPJP-UHFFFAOYSA-N 0.000 abstract description 10
- 229940074360 caffeic acid Drugs 0.000 abstract description 10
- QAIPRVGONGVQAS-UHFFFAOYSA-N cis-caffeic acid Natural products OC(=O)C=CC1=CC=C(O)C(O)=C1 QAIPRVGONGVQAS-UHFFFAOYSA-N 0.000 abstract description 10
- 230000003078 antioxidant effect Effects 0.000 abstract description 8
- 239000003963 antioxidant agent Substances 0.000 abstract description 6
- 238000000034 method Methods 0.000 abstract description 5
- 108090000790 Enzymes Proteins 0.000 abstract description 4
- 102000004190 Enzymes Human genes 0.000 abstract description 4
- 235000013336 milk Nutrition 0.000 abstract description 2
- 239000008267 milk Substances 0.000 abstract description 2
- 210000004080 milk Anatomy 0.000 abstract description 2
- LDVVTQMJQSCDMK-UHFFFAOYSA-N 1,3-dihydroxypropan-2-yl formate Chemical compound OCC(CO)OC=O LDVVTQMJQSCDMK-UHFFFAOYSA-N 0.000 abstract 4
- 230000007547 defect Effects 0.000 abstract 1
- 230000001766 physiological effect Effects 0.000 abstract 1
- 240000007154 Coffea arabica Species 0.000 description 18
- 239000000523 sample Substances 0.000 description 11
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 230000002902 bimodal effect Effects 0.000 description 7
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 7
- 238000002835 absorbance Methods 0.000 description 6
- 239000012071 phase Substances 0.000 description 6
- 230000006837 decompression Effects 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 239000013078 crystal Substances 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 3
- 238000005160 1H NMR spectroscopy Methods 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 150000003254 radicals Chemical class 0.000 description 3
- 230000002000 scavenging effect Effects 0.000 description 3
- -1 small molecule polyphenol Chemical class 0.000 description 3
- 239000012224 working solution Substances 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 2
- OAKJQQAXSVQMHS-UHFFFAOYSA-N Hydrazine Chemical compound NN OAKJQQAXSVQMHS-UHFFFAOYSA-N 0.000 description 2
- 108091000080 Phosphotransferase Proteins 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 235000009508 confectionery Nutrition 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 239000007791 liquid phase Substances 0.000 description 2
- 230000005311 nuclear magnetism Effects 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 150000007965 phenolic acids Chemical class 0.000 description 2
- 102000020233 phosphotransferase Human genes 0.000 description 2
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 102000004022 Protein-Tyrosine Kinases Human genes 0.000 description 1
- 108090000412 Protein-Tyrosine Kinases Proteins 0.000 description 1
- 125000002252 acyl group Chemical group 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 208000026106 cerebrovascular disease Diseases 0.000 description 1
- 150000001793 charged compounds Chemical class 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 201000006549 dyspepsia Diseases 0.000 description 1
- 239000008157 edible vegetable oil Substances 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 238000000132 electrospray ionisation Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 239000002932 luster Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000005809 transesterification reaction Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C69/00—Esters of carboxylic acids; Esters of carbonic or haloformic acids
- C07C69/66—Esters of carboxylic acids having esterified carboxylic groups bound to acyclic carbon atoms and having any of the groups OH, O—metal, —CHO, keto, ether, acyloxy, groups, groups, or in the acid moiety
- C07C69/73—Esters of carboxylic acids having esterified carboxylic groups bound to acyclic carbon atoms and having any of the groups OH, O—metal, —CHO, keto, ether, acyloxy, groups, groups, or in the acid moiety of unsaturated acids
- C07C69/732—Esters of carboxylic acids having esterified carboxylic groups bound to acyclic carbon atoms and having any of the groups OH, O—metal, —CHO, keto, ether, acyloxy, groups, groups, or in the acid moiety of unsaturated acids of unsaturated hydroxy carboxylic acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/37—Esters of carboxylic acids
- A61K8/375—Esters of carboxylic acids the alcohol moiety containing more than one hydroxy group
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/62—Carboxylic acid esters
-
- 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
Abstract
The invention belongs to the field of structural grease, and discloses water-soluble caffeoyl structural grease, and a preparation method and application thereof, in particular application thereof as an antioxidant. The chemical name of the water-soluble caffeoyl structural grease is 1-caffeoyl monoglyceride. The chemical structure is as shown in Formula (1). The invention also provides an enzymatic synthesis method. According to the method, an exchange reaction between caffeoyl acetate and glyceride is catalyzed by enzymes through a solvent-free system to prepare the 1-caffeoyl monoglyceride, so that the preparation of water-soluble caffeoyl derivatives is realized. The 1-caffeoyl monoglyceride is a new functional phenololipoid, has multiple physiological activities of caffeic acid, has higher antioxidant activity in both a water-soluble system and an alcohol-soluble system, and overcomes the defect of poor water solubility of the caffeic acid. Meanwhile, the 1-caffeoyl monoglyceride, serving as a potential natural water-soluble antioxidant substance, can be applied to the fields of emulsified food such as milk and functional grease and cosmetics, and has great economical and social values.
Description
Technical field
The invention belongs to Structure grease field, more particularly to a kind of water soluble coffee acid Structure grease and preparation method thereof with
Using especially as the application of antioxidant.
Background technology
In recent years, gradually stepping up with people's quality of life and living standard, Structure grease as functional grease, by
Gradually favored by people, such as a sweet ester can use a part for oil as feature emulsifying agent, DAG as family
Edible oil is substituted, the effect with fat-reducing.With going deep into for research, constantly there is new Structure grease to enter the life of people.Phenol
Fat contains one or more phenolic acid molecules as the new Structure grease of a class, on its molecular structure, and which not only has fatty acid
Trophism, the also physiologically active with phenolic acid, are gradually paid close attention to by research worker.
Caffeic acid is a kind of small molecule polyphenol, with various physiologically actives such as scavenging hydroxyl, DPPH, ABTS etc. freely
Base, suppresses Fyn Kinase activities, mitigates the skin canceration that UVB causes, and suppresses the sub- kinases of albumen and tyrosine-kinase enzyme activity, reduces
Melanic generation, suppresses the growth and breeding of various bacteria, blood fat reducing and blood sugar effect, prevention cardiovascular and cerebrovascular disease etc..
Caffeic acid is relatively low in the dissolubility of aqueous solution systems, limits its application in hydrophilic system, therefore, prepare coffee
The soluble derivative of acid can expand caffeinic range of application, and water soluble coffee acid Structure grease not only solves coffee
Low solubility problem of the acid in aqueous environment, and which is used as the new phenol fat of a class, in functional food, medicine and cosmetics neck
Domain is with a wide range of applications.
The content of the invention
In order to overcome the solubility that above-mentioned prior art caffeic acid itself is present with not enough, the primary and foremost purpose of the present invention
It is that a kind of water soluble coffee acid Structure grease is provided.
Another object of the present invention is to provide a kind of preparation method of above-mentioned water soluble coffee acid Structure grease.
Still a further object of the present invention is to provide above-mentioned water soluble coffee acid Structure grease as Natural Antioxidants in breast
The application in the field such as relieving dyspepsia product and cosmetics.
The purpose of the present invention is realized by following proposal：
A kind of water soluble coffee acid Structure grease, its chemical name is the mono- coffee acid glycerides (1-MCG) of 1-, chemical constitution
As shown in following formula ()：
The invention provides a kind of enzymatic synthesis method of above-mentioned water soluble coffee acid Structure grease, its reaction equation
For：
Specifically include following steps：
Caffeic acetate is mixed with glycerol, heated and stirred, is lowered the temperature, add immobilized-lipase, isothermal reaction to separate,
Obtain water soluble coffee acid Structure grease.
Caffeic acetate used is 1 with the mol ratio of glycerol:3～1:30.
The consumption of the immobilized-lipase is the 2～12% of caffeic acetate and glycerol gross mass.
Described immobilized-lipase can be Novozym435, Lipozyme 435, Lipozyme TL IM and
At least one in Lipozyme RM IM.
The temperature of the heated and stirred is preferably 75～85 DEG C, and the heated and stirred is used to make caffeic acetate fill with glycerol
Divide mixing, preferably stir 20～30min.
The cooling is preferably decreased to 55～75 DEG C.It is preferred that stirring after cooling after 20～30min makes system temperature constant again
Add enzyme.
The time of the isothermal reaction is preferably 0.5～48h of stirring reaction.
The separation preferably includes following steps：Water dissolution reactant mixture, sucking filtration is added to be extracted to filtrate with dichloromethane
Obtain upper layer of extraction liquid, crystallisation by cooling after vacuum concentrated by rotary evaporation.
Gained crystal is preferably by water washing and drying carries out purification.
The enzymatic synthesis method of the present invention, is prepared with glyceride transesterification reaction by solvent-free system enzymatic caffeic acetate
The mono- coffee acid glycerides of 1-, realize the preparation of water soluble coffee acid derivative.In addition, the mono- coffee acid glycerides of 1- are a kind of new
Phenol fat, be functional grease, with caffeinic various physiologically actives, which is respectively provided with stronger in water solublity and alcohol-soluble system
Antioxidant activity, overcomes the weakness of caffeic acid poorly water-soluble, will further expand caffeinic application, while the mono- coffees of 1-
Coffee acid glyceride can be applied to food (especially as antioxidant as a kind of potential natural Water-soluble antioxidant material
It is the emulsified foods such as milk, functional grease) and the field such as cosmetics, with huge economy and social value.
The present invention is had the following advantages and beneficial effect relative to prior art：Preparation method of the present invention is using without molten
Agent system enzymatic reaction, relative to prior art, reaction yield is high, the response time is short, and Product Byproduct is few, reaction condition temperature
With, enzyme not easy in inactivation, and the product for producing has more preferable quality at aspects such as color and luster, local flavors, using immobilized-lipase, can
To realize recycling and reusing for enzyme, so as to reduce production cost.
Description of the drawings
Fig. 1 is the liquid phase figure of the sample for reacting 0h (A) and 12h (B).
Fig. 2 is the mass spectrum of the mono- coffee acid glycerides of 1-.
Fig. 3 is the mono- coffee acid glycerides of 1-1H-NMR (A) and13C-NMR (B) figure.
Fig. 4 be the mono- coffee acid glycerides (1-MCG) of 1-, caffeic acetate (EC), caffeic acid (CA) DPPH free radicals it is clear
Except the influence curve of vigor.
What Fig. 5 was represented be the mono- coffee acid glycerides (1-MCG) of 1-, caffeic acetate (EC), caffeic acid (CA) ABTS from
The influence curve of vigor is removed by base.
Specific embodiment
With reference to embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are not limited
In this.
Reagent used in the following example can be obtained from commercial channel.
Embodiment 1：The preparation of water soluble coffee acid Structure grease
The caffeic acetate of 3mmol with 9mmol glycerol in reactor, 80 DEG C, 300rpm magnetic agitation 30min, afterwards
After 65 DEG C of magnetic agitation 20min, 6% (w/w) immobilized-lipase Lipozyme TL IM are added to react 48h at 65 DEG C.With
40mL distills water dissolution reactant mixture, and decompression sucking filtration obtains filtrate, extracts filtrate, extraction 3～5 with 40mL dichloromethane every time
It is secondary, upper layer of extraction liquid is collected, 50 DEG C of vacuum concentrated by rotary evaporations obtain about 10mL concentrated solutions crystallisation by cooling in 4 DEG C of refrigerators, afterwards
Decompression sucking filtration obtains white crystal, and is washed with cold distilled water more than 3 times, is dried, obtains water solublity under room temperature low moisture environments
Caffeic acid Structure grease powder.
Embodiment 2：The preparation of water soluble coffee acid Structure grease
The caffeic acetate of 3mmol with 30mmol glycerol in reactor, 80 DEG C, 300rpm magnetic agitation 30min, afterwards
After 55 DEG C of magnetic agitation 20min, 12% (w/w) immobilized-lipase Lipozyme 435 is added to react 36h at 75 DEG C.Use 90mL
Distillation water dissolution reactant mixture, decompression sucking filtration obtain filtrate, every time with about 90mL dichloromethane extraction filtrate, extraction 3～5
It is secondary, upper layer of extraction liquid is collected, 55 DEG C of vacuum concentrated by rotary evaporations obtain about 15mL concentrated solutions crystallisation by cooling in 4 DEG C of refrigerators, afterwards
Decompression sucking filtration obtains white crystal, and is washed with cold distilled water more than 3 times, is dried, obtains water solublity under room temperature low moisture environments
Caffeic acid Structure grease powder.
Embodiment 3：The preparation of water soluble coffee acid Structure grease
The caffeic acetate of 3mmol with 90mmol glycerol in reactor, 80 DEG C, 300rpm magnetic agitation 30min, afterwards
After 65 DEG C of magnetic agitation 20min, 10% (w/w) immobilized-lipase Lipozyme RM IM are added to react 48h at 65 DEG C.With
100mL distills water dissolution reactant mixture, and decompression sucking filtration obtains filtrate, every time with about 100mL dichloromethane extraction filtrate, extraction
Take 3～5 times, collect upper layer of extraction liquid, 45 DEG C of vacuum concentrated by rotary evaporations obtain about 20mL concentrated solutions and knot is cooled down in 4 DEG C of refrigerators
Crystalline substance, the sucking filtration that reduces pressure afterwards obtain white crystal, and are washed with cold distilled water more than 3 times, be dried, obtain under room temperature low moisture environments
Water soluble coffee acid Structure grease powder.
Embodiment 4：The Structural Identification of water soluble coffee acid Structure grease
(1) the water soluble coffee acid Structure grease for preparing embodiment 1 is dissolved in methanol (1mg/ml), and using efficient
Liquid chromatograph is detected to which.Sample configures concrete operations：The first of the mono- coffee acid glycerides of 1- of 1mg/ml is prepared accurately
Alcoholic solution, and with 0.22 μm of organic membrane filtration, be stored in the sample bottle of 2mL, it is to be measured.
Wherein high performance liquid chromatography is 2998 types of Waters, and testing conditions are：Chromatographic column：Symmetry C18 are anti-phase
Chromatographic column, 4.6 × 250mm, 5 μm；Column temperature：25℃；Detection wavelength：325nm；Mobile phase：Mobile phase A：Pure methanol, Mobile phase B：
0.75% acetic acid；Flow velocity：1mL/min；Gradient elution program：0～15min, 35%A～75%A (65%～25%B), 15～
19min, 75%A～35%A (25%～65%B), 19～24min, 35%A (65%B).As a result see Fig. 1.
(2) using Agilent 1290/maXis impact high-resolution mass spectrometers to water soluble coffee acid Structure grease
Molecular weight and molecular formula are identified.Testing conditions are：Ion source：ESI+And ESI-；Capillary voltage：3.5kv；Taper hole voltage：
2kv；Mass-to-charge ratio sweep limitss：200～1000m/z.
Method according to above high performance liquid chromatography prepares sample and mobile phase, after sample introduction, the anti-phase colors of sample Jing SB-C18
After spectrum post (1.8 μm of 2.1 × 50mm of RRHD) is separated, in sample material according to polarity from sequentially entering high score to weak order by force
Distinguish in mass spectrograph, ionize Jing after electro-spray ionization, charged particle enters ion hydrazine mass analyzer, detects charged particle, root
According to the liquid phase appearance time of single coffee acid glyceride, so that it is determined that the molecular ion peak and fragment peak of single coffee acid glyceride, from
And determine its molecular weight and molecular formula.As a result see Fig. 2.Product formula is C as seen from the figure12H14O6, molal weight is 254g/
mol。
(3) using 600 nuclear magnetic resonance analyser identification and analysis coffee acyls of Bruker Avance III HD on the glycerol backbone
Position, be dissolved in deuterated dimethyl sulfoxide, use respectively1H/13C-NMR is detected and is mutually authenticated that testing result is shown in figure
3。
Concrete operations are that the 1- of at least 20mg purity more than 95% mono- coffee acid glyceride sample is dissolved in 0.75mL deuteriums
For dimethyl sulfoxide, being sufficiently mixed is completely dissolved sample, draws the solvent of all sample dissolutions in nuclear-magnetism with 1mL liquid-transfering guns
In detection pipe, nuclear magnetic tube is placed on Bruker Avance III HD 600M instrument sample platforms, instrument automatically into magnetic field simultaneously
Start detection.
Determine that product is the sweet ester of the mono- caffeic acids of 1-, wherein the result of detailed nuclear-magnetism is as follows：
1H-NMR (600M) result is：3.38 (bimodal, 12a), 3.41 (bimodal, 12b), 3.70 (multiplet, 11), 4.00
(bimodal, 10a), 4.15 (bimodal, 10b), 4.65 (unimodal, O-3), 4.91 (unimodal, O-4), 6.26 (bimodal, 8), 6.77 is (double
Peak, 1), 7.00 (it is bimodal, 6), 7.04 (unimodal, 4), 7.49 is bimodal, 7), 9.57 (unimodal, O-1), 9.14 (unimodal, O-2)；
13C-NMR (600M) result is：63.13(12),66.09(10),66.91(11),114.51(4),115.25(8),
116.24(1),121.78(6),126.01(5),145.33(3),146.05(7),148.85(2),167.07(9)。
Structural formula is as follows：
Embodiment 5：The antioxidant activity research of water soluble coffee acid Structure grease
DPPH, ABTS method is respectively adopted to be detected, it is specific as follows：
(1) DPPH methods：Prepare DPPH free radical methanol solutions so as to 517nm locate absorbance be 0.5 ± 0.02 work
Liquid.In reaction system, the 1- that the embodiment 1 of 0.3mL variable concentrations (0.02,0.05,0.1 and 0.2mM) is prepared is mono-
The DPPH free radical working solutions of coffee acid glyceride solution and 2.7mL are added in brown reagent bottle, after being sufficiently mixed with it is ultraviolet-
Visible spectrophotometer is surveyed once per 5min, determines change of its absorbance in 30min in 517nm mensuration absorbances
Situation, investigates Scavenging activities of the 1-MCG of variable concentrations to DPPH free radicals, as a result sees Fig. 4.
(2) ABTS methods：ABTS solution is prepared with the phosphate buffer of pH 7.4 so as to be 0.5 in the absorbance of 734nm
± 0.02, obtain ABTS+Working solution.In reaction system, by 0.3mL variable concentrations (0.02,0.05,0.1 and 0.2mM)
The mono- coffee acid glyceride solution of 1- and the ABTS of 2.7mL that embodiment 1 is prepared+Working solution is added to brown reagent bottle
In, after being sufficiently mixed, its absorbance is determined in 734nm ultraviolet-visible spectrophotometers, survey once per 5min, determine which
Situation of change of the absorbance in 30min, investigates the 1-MCG of variable concentrations to ABTS+Scavenging activity, as a result see Fig. 5.
As a result show, the water soluble coffee acid Structure grease of the present invention has stronger removing DPPH, ABTS free radical
Ability, and antioxidation can be played in aqueous solution systems and alcohol-soluble system.
Above-described embodiment is the present invention preferably embodiment, but embodiments of the present invention not by above-described embodiment
Limit, other any spirit without departing from the present invention and the change, modification, replacement made under principle, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Claims (10)
1. a kind of water soluble coffee acid Structure grease, it is characterised in that chemical name is the mono- coffee acid glycerides of 1-, chemical constitution
As shown in following formula ()：
2. the enzymatic synthesis method of the sour Structure grease of water soluble coffee described in a kind of claim 1, it is characterised in that concrete to wrap
Include following steps：
Caffeic acetate is mixed with glycerol, heated and stirred, lowered the temperature, add immobilized-lipase, isothermal reaction to separate, obtain
Water soluble coffee acid Structure grease.
3. the enzymatic synthesis method of water soluble coffee according to claim 2 acid Structure grease, it is characterised in that：Coffee used
Coffee acetoacetic ester is 1 with the mol ratio of glycerol:3～1:30.
4. the enzymatic synthesis method of water soluble coffee according to claim 2 acid Structure grease, it is characterised in that：It is described solid
The consumption of immobilized lipase is the 2～12% of caffeic acetate and glycerol gross mass.
5. the enzymatic synthesis method of water soluble coffee according to claim 2 acid Structure grease, it is characterised in that：Described
Immobilized-lipase be Novozym435, Lipozyme 435, Lipozyme TL IM and Lipozyme RM IM at least
It is a kind of.
6. the enzymatic synthesis method of water soluble coffee according to claim 2 acid Structure grease, it is characterised in that：It is described to add
The temperature of thermal agitation is 75～85 DEG C.
7. the enzymatic synthesis method of water soluble coffee according to claim 2 acid Structure grease, it is characterised in that：The drop
Temperature refers to and is reduced to 55～75 DEG C.
8. the enzymatic synthesis method of water soluble coffee according to claim 2 acid Structure grease, it is characterised in that：The perseverance
The time of temperature reaction is 0.5～48h of stirring reaction.
9. the enzymatic synthesis method of water soluble coffee according to claim 2 acid Structure grease, it is characterised in that：Described point
From comprising the following steps：Water dissolution reactant mixture, sucking filtration is added upper layer of extraction liquid to be obtained by extraction to filtrate with dichloromethane,
Crystallisation by cooling after vacuum concentrated by rotary evaporation.
10. water soluble coffee acid application of the Structure grease in food and cosmetic field described in claim 1.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107831130A (en) * | 2017-10-26 | 2018-03-23 | 清华大学 | A kind of portable unit and method for determining water body water soluble organic substance and electronic capability being provided |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101153866A (en) * | 2007-10-19 | 2008-04-02 | 中国药科大学 | Novel analysis method for complex composition of traditional Chinese medicine injection based on LC/MS-IT-TOF |
| CN101398417A (en) * | 2008-10-29 | 2009-04-01 | 中国药科大学 | Universal method for rapid detection and structural identification for Chinese traditional medicine complex component |
| CN103088078A (en) * | 2013-02-04 | 2013-05-08 | 江苏科技大学 | Lipase synthesis-promoting method of propyl caffeate |
| CN103173502A (en) * | 2013-03-04 | 2013-06-26 | 江苏科技大学 | Biosynthesis method of caffeic acid phenethylester by means of transesterification |
| KR20160070891A (en) * | 2014-12-10 | 2016-06-21 | 강원대학교산학협력단 | Composition comprising caffeoylglycolic acid methyl ester or 1-O-caffeoylglycerol for preventing or treating inflammatory diseases |
| CN106086096A (en) * | 2016-08-26 | 2016-11-09 | 中国农业科学院油料作物研究所 | A kind of method efficiently preparing phenolic acid glyceride |
-
2016
- 2016-11-17 CN CN201611010656.7A patent/CN106588664A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101153866A (en) * | 2007-10-19 | 2008-04-02 | 中国药科大学 | Novel analysis method for complex composition of traditional Chinese medicine injection based on LC/MS-IT-TOF |
| CN101398417A (en) * | 2008-10-29 | 2009-04-01 | 中国药科大学 | Universal method for rapid detection and structural identification for Chinese traditional medicine complex component |
| CN103088078A (en) * | 2013-02-04 | 2013-05-08 | 江苏科技大学 | Lipase synthesis-promoting method of propyl caffeate |
| CN103088078B (en) * | 2013-02-04 | 2015-07-01 | 江苏科技大学 | Lipase synthesis-promoting method of propyl caffeate |
| CN103173502A (en) * | 2013-03-04 | 2013-06-26 | 江苏科技大学 | Biosynthesis method of caffeic acid phenethylester by means of transesterification |
| KR20160070891A (en) * | 2014-12-10 | 2016-06-21 | 강원대학교산학협력단 | Composition comprising caffeoylglycolic acid methyl ester or 1-O-caffeoylglycerol for preventing or treating inflammatory diseases |
| CN106086096A (en) * | 2016-08-26 | 2016-11-09 | 中国农业科学院油料作物研究所 | A kind of method efficiently preparing phenolic acid glyceride |
Non-Patent Citations (1)
| Title |
|---|
| HU, JUN ET AL: "Developmental toxicity of orally administered pineapple leaf extract in rats", 《 FOOD AND CHEMICAL TOXICOLOGY》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107831130A (en) * | 2017-10-26 | 2018-03-23 | 清华大学 | A kind of portable unit and method for determining water body water soluble organic substance and electronic capability being provided |
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