CN109699999A - A kind of method that amylose embedding improves burst size in p-Coumaric Acid stability and enteron aisle - Google Patents
A kind of method that amylose embedding improves burst size in p-Coumaric Acid stability and enteron aisle Download PDFInfo
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- CN109699999A CN109699999A CN201811603188.3A CN201811603188A CN109699999A CN 109699999 A CN109699999 A CN 109699999A CN 201811603188 A CN201811603188 A CN 201811603188A CN 109699999 A CN109699999 A CN 109699999A
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- coumaric acid
- amylose
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Abstract
Method the invention discloses a kind of amylose embedding p-Coumaric Acid to improve burst size in its stability and enteron aisle.P-Coumaric Acid has different physiological roles, but unstable property limits its extensive use.P-Coumaric Acid itself can not be embedded by amylose, and the present invention connect p-Coumaric Acid to form p-Coumaric Acid hexadecyl ester with hexadecanol by esterification, to realize amylose embedding p-Coumaric Acid.For the embedded object that the method for the present invention is formed after ultraviolet light direct irradiation 72h, retention rate is up to 89%；Without embedding p-Coumaric Acid after same condition is handled retention rate only 43%.Simulation digestion experiment shows that burst size of the p-Coumaric Acid in enteron aisle in embedded object is significantly higher than the burst size for the p-Coumaric Acid not embedded.Embedding method of the invention significantly improves the photostability of p-Coumaric Acid, can play the role of effective protection in food processing, storage and sales process, and can improve its release in enteron aisle, is conducive to digestion and absorption and utilization in human body.
Description
Technical field
The invention belongs to food processing technology field, in particular to a kind of amylose embedding improves p-Coumaric Acid stability
With the method for burst size in enteron aisle.
Technical background
P-Coumaric Acid, i.e. p-Coumaric Acid are a kind of cinnamic acids as derived from ArAA, are widely present in standing grain
In graminaceous plant stem, it is the active constituent in the herbal medicine such as oldenlandia diffusa, Prunella vulgaris, is usually used in Cholagogue drug in pharmaceutical sector
Preparation.Recent study shows that p-Coumaric Acid has the extensive physiological activity such as preferable antibacterial, anti-inflammatory, anti-apoptotic
Function all has biggish potentiality to be exploited in the prevention and treatment of a variety of diseases.However due to active high, the easy oxygen of p-Coumaric Acid
Change, is easily oxidized the feature for losing original activity at photaesthesia in alimentary canal, in the processing, storage, sale of food and health care product
Degradation is easy Deng during, so that original active function, greatly limits its addition and application in food and health care product.
A kind of linear polysaccharide formed by α-Isosorbide-5-Nitrae glucosides key connection of amylose, is widely present in various staple foods.In spy
Amylose can form left hand single coil configuration under fixed condition, spiraled cavity internal drainage and outer relative hydropathic, be easily obtained and make
It is standby, it is a kind of good wall material.Research shows that material stability can be improved in amylose embedding means.Since amylose can not
P-Coumaric Acid is directly embedded, therefore, develops a kind of amylose embedding p-Coumaric Acid to improve its stability and in alimentary canal
The method of burst size is very necessary.
Summary of the invention
In view of the above problems, the object of the present invention is to provide a kind of amylose embedding p-Coumaric Acid method,
The burst size in the stability and enteron aisle of p-Coumaric Acid can be significantly improved.
The purpose of the present invention is what is be achieved through the following technical solutions:
The method that a kind of embedding of amylose improves burst size in p-Coumaric Acid stability and enteron aisle, the method includes with
Lower step:
(1) hexadecanol, p-Coumaric Acid and triphenylphosphine are dissolved in anhydrous tetrahydro furan under condition of ice bath, stirring is mixed
It closes uniformly, obtains mixed solution；
(2) diethyl azodiformate with p-Coumaric Acid equimolar amounts is added dropwise in Xiang Shangshu mixed solution, in ice
Continue after being stirred 15~20min in bath, restores to terminate after to room temperature the reaction was continued 4~5h, obtain reaction solution；
(3) above-mentioned reaction solution is placed in 35~40 DEG C of rotary evaporations, gained crude product is using silica gel column chromatography with just
Hexane/ethyl acetate is isolated and purified as eluent, obtains p-Coumaric Acid hexadecyl ester；
(4) in 85~95 DEG C of water-baths, amylose is dissolved in dimethyl sulphoxide aqueous solution, amylose concentration is made
40~50mg/mL is stirring evenly and then adding into the p-Coumaric Acid hexadecyl ester of amylose weight 10~15%, is then added 2~3 times
Volume of water stirs and evenly mixs；
(5) standing makes temperature be down to room temperature, and the centrifugation of gained turbid takes precipitating, and it is extra to perfume (or spice) to be washed away with ethanol water
Beans acid hexadecyl ester, vacuum drying remove solvent and obtain product.
Further, the molar ratio of hexadecanol, p-Coumaric Acid and triphenylphosphine described in step (1) is 1:1:2~1:
2:2.
Further, in eluent described in step (3) ethyl acetate volume fraction 5~10%.
Further, in dimethyl sulphoxide aqueous solution described in step (4) dimethyl sulfoxide volumetric concentration be 90~
95%.
Further, the mixing time is 15~20min after embedding in step (4).
Further, the volumetric concentration of ethanol water described in step (5) is 45~55%, and washing times are 3~4
It is secondary.
Further, vacuum drying condition described in step (5) is 45~55 DEG C of dry 24~48h.
Further, the condition of ice bath temperature is 0~4 DEG C；The ambient temperature is 20~25 DEG C.
It is another object of the present invention to what is be achieved through the following technical solutions:
A kind of p-Coumaric Acid embedded object, the embedded object are prepared by above-mentioned method.
A kind of food contains the p-Coumaric Acid embedded object in the food.
A kind of health care product contains the p-Coumaric Acid embedded object in the health care product.
The present invention having the beneficial effect that compared with prior art
1, the present invention considers the structure feature of amylose, in conjunction with the method for esterification process and molecule embedding, using dredging
Water active force will be esterified p-Coumaric Acid and be loaded into amylose spiraled cavity, and it is non-helical between, improve the steady of embedded object
It is qualitative.
2, the present invention realizes amylose embedding p-Coumaric Acid, and significantly improves its photostability, provides a kind of new
Embedding techniques: ultraviolet light the experimental results showed that, after treatment with ultraviolet light 72h, the p-Coumaric Acid retention rate that does not embed <
50%, and p-Coumaric Acid retention rate > 85% in embedded object.
3, the embedded object prepared by the present invention has good releasing effect, and has significantly under the conditions of digestive juice
Protective effect can be improved release of the p-Coumaric Acid in enteron aisle, promote its digestion and absorption and utilization in human body.
4, the preparation method in the present invention has edible, easy acquisition, blood glucose increasing effect using amylose as wall material
Low advantage has certain applying value in the production of functional food and drug.
Detailed description of the invention
Fig. 1 is that amylose embeds p-Coumaric Acid schematic diagram.
Fig. 2 is the X ray diffracting spectrum of embedded object and control group.
Fig. 3 is Photostability experiments result.
Fig. 4 is the experimental result for simulating p-Coumaric Acid burst size under digestion condition.
Specific embodiment
To be best understood from the present invention, it is further described below with reference to specific implementation method, embodiment and attached drawing explanation, but this
Invent the range that claimed range is not limited only to embodiment statement.
Amylose embedding p-Coumaric Acid schematic diagram is shown in Fig. 1, synthesizes p-Coumaric Acid hexadecyl ester by esterification, separates pure
After change, p-Coumaric Acid hexadecyl ester is loaded into the spiraled cavity of amylose, is wrapped through centrifugation, washing and drying steps
Bury object powder.When forming embedded object, amylose is dissolved in dimethyl sulphoxide solution at high temperature, forms the left hand of internal drainage
Single coil configuration, under the promotion of intermolecular hydrophobic active force, hydrophobic 16 carbon teminal of p-Coumaric Acid hexadecyl ester enters spiral sky
In chamber, relatively stable complex compound is formed, after subsequent processing is dry, completes embedding.
It is identified with structure of the X-ray diffraction method to embedded object, as a result sees Fig. 2.Wherein a, b, c, d respectively correspond for
P-Coumaric Acid, p-Coumaric Acid hexadecyl ester, p-Coumaric Acid amylose embedded object and p-Coumaric Acid hexadecyl ester amylose embedded object
X ray diffracting spectrum.The diffraction pattern of p-Coumaric Acid amylose embedded object is similar with the Type B starch diffraction pattern presentation not embedded
Diffraction maximum, illustrate that p-Coumaric Acid itself cannot be embedded by amylose.And p-Coumaric Acid hexadecyl ester amylose embedded object is then
Apparent diffraction maximum is showed 2 θ=13 ° and 20 ° respectively, there is V after forming embedding6hThe feature of type starch, illustrates to tonka-bean
Sour hexadecyl ester by successfully embedding enter amylose formed spiraled cavity in, and it is non-helical between.
Specific assay method is as follows for p-Coumaric Acid or p-Coumaric Acid ester in sample:
Using high effective liquid chromatography for measuring content: selecting reverse phase C18 chromatographic column to exist using Shimadzu LC-20AT chromatograph
P-Coumaric Acid is detected under 280nm wavelength；P-Coumaric Acid hexadecyl ester is measured at 300 nm wavelength.Configuration concentration gradient is 0~150 μ
M p-Coumaric Acid/p-Coumaric Acid hexadecyl ester titer.Sample uses dilution in acetonitrile after being dissolved in dimethyl sulfoxide, and 0.22 μm excessively organic
Loading after film.Applied sample amount is 10 μ L, and mobile phase is the elution of 100% acetonitrile, flow velocity 1.0mL/min.
The specific method is as follows for Photostability experiments:
It takes 50mg or so sample to pulverize to pave and be placed in glass dish, is 256nm ultraviolet light (16W) in black box with wavelength
Middle progress direct ultraviolet light irradiation experiment, light source and sample distance are 5cm, are taken within 6,12,24,48 and 72 hours after being put into
Sample is dissolved in dimethyl sulfoxide, the p-Coumaric Acid content being measured in sample after being 100 μM with dilution in acetonitrile to concentration.
Simulating digestion experiment, the specific method is as follows:
(1) simulate the gastric juice: take the sample of 5mg p-Coumaric Acid or the p-Coumaric Acid containing equivalent in (the 1L distillation of 50mL simulate the gastric juice
In water: 2.0g NaCl；7.0mL 36%HCl；1.0mL Tween 80；0.6g pepsin, pH=1.2) in digestion.
(2) simulated intestinal fluid: peptic digest liquid 10mL is mixed with intestinal digestion liquid (in 1L distilled water :) with 1:1 volume ratio, adjusts pH
It is 7.5, adds the configured pancreatic juice of 5mL, makes pancreatin containing 1g/L in system, 3g/L Pig cholate.
Simulation digestion 150rpm in 37 DEG C of water-baths shakes, and is measured by sampling respectively at 0,15,30,60,120min.
Embodiment 1
The method that a kind of embedding of amylose improves burst size in p-Coumaric Acid stability and enteron aisle, the method includes with
Lower step:
(1) hexadecanol, p-Coumaric Acid and triphenylphosphine are dissolved in molar ratio for 1:2:2 under 0 DEG C of condition of ice bath
It in anhydrous tetrahydro furan, is uniformly mixed, obtains mixed solution.
(2) diethyl azodiformate with p-Coumaric Acid equimolar amounts is added dropwise in Xiang Shangshu mixed solution, in ice
Continue after being stirred 15min in bath, restores to terminate after to 25 DEG C of room temperatures the reaction was continued 5h, obtain reaction solution.
(3) above-mentioned reaction solution is placed in 40 DEG C of rotary evaporations, gained crude product using silica gel column chromatography with n-hexane/
Ethyl acetate (ethyl acetate volume fraction 10%) is isolated and purified as eluent, obtains p-Coumaric Acid hexadecyl ester.
(4) in 90 DEG C of water-baths, amylose is dissolved in 95% dimethyl sulphoxide aqueous solution, amylose concentration is made
50mg/mL is stirring evenly and then adding into the p-Coumaric Acid hexadecyl ester of amylose weight 10%, is stirred 30min, is added 2.5
The stirring of times volume of water, dilution 15min.
(5) 12 are stood~so that temperature is down to 25 DEG C of room temperatures for 24 hours, 3000 × g of gained turbid centrifugation takes precipitating, with 50% second
Alcohol solution washs 3-4 times, and to wash away extra p-Coumaric Acid hexadecyl ester, vacuum drying removes solvent and obtains product.Described
Vacuum drying condition is 50 DEG C of dry 48h.
The hexadecanol, p-Coumaric Acid are purchased from Sigma Co., USA；The triphenylphosphine, azoformic acid two
Ethyl ester is purchased from terraced wish and falls in love with Hai company；The dimethyl sulfoxide, anhydrous tetrahydro furan, ethyl alcohol are purchased from Tianjin, and big reagent is public forever
Department.
Each raw material can be gram quantity grade in the present embodiment, or kilogram.
In the present embodiment each temperature, time parameter can in the range of providing flexible combination.
Photostability experiments and simulation digestion experiment are carried out respectively.By in above-mentioned high performance liquid chromatography test sample
P-Coumaric Acid or p-Coumaric Acid hexadecyl ester content.
Photostability result such as Fig. 3: the present embodiment in embedded object the retention rate of p-Coumaric Acid be up to 89%, without
The p-Coumaric Acid retention rate of embedding only has 43%.
Simulation digestion experiment result such as Fig. 4: gained embedded object can stablize release p-Coumaric Acid in enteron aisle in the present embodiment,
And it measures burst size and is significantly higher than the p-Coumaric Acid not embedded and other control groups.
Certain embodiment of the invention above described embodiment only expresses, the description thereof is more specific and detailed, but simultaneously
Limitations on the scope of the patent of the present invention therefore cannot be interpreted as.It should be pointed out that for those of ordinary skill in the art
For, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to guarantor of the invention
Protect range.Therefore, the scope of protection of the patent of the present invention should be with appended claims.
Claims (10)
1. a kind of method that amylose embedding improves burst size in p-Coumaric Acid stability and enteron aisle, the method includes following
Step:
(1) hexadecanol, p-Coumaric Acid and triphenylphosphine are dissolved in anhydrous tetrahydro furan under condition of ice bath, are stirred
It is even, obtain mixed solution；
(2) diethyl azodiformate with p-Coumaric Acid equimolar amounts is added dropwise in Xiang Shangshu mixed solution, in ice bath
Continue after being stirred 15~20min, restores to terminate after to room temperature the reaction was continued 4~5h, obtain reaction solution；
(3) above-mentioned reaction solution is placed in 35~40 DEG C of rotary evaporations, gained crude product using silica gel column chromatography with n-hexane/
Ethyl acetate as eluent isolates and purifies, and obtains p-Coumaric Acid hexadecyl ester；
(4) in 85~95 DEG C of water-baths, amylose is dissolved in dimethyl sulphoxide aqueous solution, make amylose concentration 40~
50mg/mL is stirring evenly and then adding into the p-Coumaric Acid hexadecyl ester of amylose weight 10~15%, and 2~3 times of volumes are then added
Water stirs and evenly mixs；
(5) standing makes temperature be down to room temperature, and the centrifugation of gained turbid takes precipitating, and extra p-Coumaric Acid is washed away with ethanol water
Hexadecyl ester, vacuum drying remove solvent and obtain product.
2. the method that amylose embedding according to claim 1 improves burst size in p-Coumaric Acid stability and enteron aisle,
It is characterized in that, the molar ratio of hexadecanol, p-Coumaric Acid and triphenylphosphine described in step (1) is 1:1:2~1:2:2.
3. the method that amylose embedding according to claim 1 improves burst size in p-Coumaric Acid stability and enteron aisle,
It is characterized in that, in eluent described in step (3) ethyl acetate volume fraction 5~10%.
4. the method that amylose embedding according to claim 1 improves burst size in p-Coumaric Acid stability and enteron aisle,
It is characterized in that, dimethyl sulfoxide volumetric concentration is 90~95% in dimethyl sulphoxide aqueous solution described in step (4).
5. the method that amylose embedding according to claim 1 improves burst size in p-Coumaric Acid stability and enteron aisle,
It is characterized in that, the mixing time is 15~20min after embedding in step (4).
6. the method that amylose embedding according to claim 1 improves burst size in p-Coumaric Acid stability and enteron aisle,
It is characterized in that, the volumetric concentration of ethanol water described in step (5) is 45~55%, washing times are 3~4 times.
7. the method that amylose embedding according to claim 1 improves burst size in p-Coumaric Acid stability and enteron aisle,
It is characterized in that, vacuum drying condition described in step (5) is 45~55 DEG C of dry 24~48h.
8. a kind of amylose embedded object of p-Coumaric Acid, which is characterized in that the embedded object is by any one of claim 1-7 institute
The method stated is prepared.
9. a kind of food, which is characterized in that contain p-Coumaric Acid embedded object as claimed in claim 8 in the food.
10. a kind of health care product, which is characterized in that contain p-Coumaric Acid embedded object as claimed in claim 8 in the health care product.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114522635A (en) * | 2022-01-24 | 2022-05-24 | 华南理工大学 | Antibacterial microcapsule capable of controllably releasing cinnamaldehyde and preparation method thereof |
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| US20040043078A1 (en) * | 2000-12-27 | 2004-03-04 | David Herault | Encapsulation of emulsions |
| US20070155695A1 (en) * | 2004-01-19 | 2007-07-05 | Corinna Wirth | Flavonoid complexes with cyclodextrins |
| WO2013054063A1 (en) * | 2011-10-14 | 2013-04-18 | Institut National De La Recherche Agronomique - Inra | Method for preparing complexes comprising amylose assembled with at least one organic molecule of interest |
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2018
- 2018-12-26 CN CN201811603188.3A patent/CN109699999A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040043078A1 (en) * | 2000-12-27 | 2004-03-04 | David Herault | Encapsulation of emulsions |
| US20070155695A1 (en) * | 2004-01-19 | 2007-07-05 | Corinna Wirth | Flavonoid complexes with cyclodextrins |
| WO2013054063A1 (en) * | 2011-10-14 | 2013-04-18 | Institut National De La Recherche Agronomique - Inra | Method for preparing complexes comprising amylose assembled with at least one organic molecule of interest |
Non-Patent Citations (3)
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| WANG, SIQI 等: "Lipophilization and molecular encapsulation of p-coumaric acid by amylose inclusion complex", 《FOOD HYDROCOLLOIDS》 * |
| 刘鹰翔: "《药物合成反应 新世纪第2版》", 31 August 2017, 中国中医药出版社 * |
| 陆昌琪等: "儿茶素及其衍生物合成研究进展", 《食品科学》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114522635A (en) * | 2022-01-24 | 2022-05-24 | 华南理工大学 | Antibacterial microcapsule capable of controllably releasing cinnamaldehyde and preparation method thereof |
| CN114522635B (en) * | 2022-01-24 | 2023-08-25 | 华南理工大学 | Antibacterial microcapsule capable of controllably releasing cinnamaldehyde and preparation method thereof |
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