CN104558642A - Plant extract and method for cross-linking collagen by using plant extract as biological cross-linker - Google Patents
Plant extract and method for cross-linking collagen by using plant extract as biological cross-linker Download PDFInfo
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- CN104558642A CN104558642A CN201410844886.8A CN201410844886A CN104558642A CN 104558642 A CN104558642 A CN 104558642A CN 201410844886 A CN201410844886 A CN 201410844886A CN 104558642 A CN104558642 A CN 104558642A
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Abstract
The invention discloses a plant extract and a method for cross-linking collagen by using the plant extract as a biological cross-linker. The plant extract is an amomum villosum extract, wherein the amomum villosum extract contains polyphenol effective components. The plant extract is prepared through the following steps: firstly, cleaning, drying and crushing amomum villosum; secondly, extracting crushed amomum villosum powder in an extracting solution; thirdly, separating the extracting solution; fourthly, concentrating the extracting solution; fifthly, obtaining a concentrated plant extract solution or dehydrated plant extract powder. The plant extract has the benefits that by using the amomum villosum plant extract as the biological cross-linker, molecular chains of the collagen are cross-linked, the structure of the collagen can be stabilized, and the degradation resistance of the collagen is improved, so that the processing characteristics of a collagen product are improved, the physical properties and the texture characteristics are good, and the application prospect is wide.
Description
Technical field
The present invention relates to biochemical field, particularly a Plant Extracts and the method as Biological cross-linker cross-linked collagen thereof.
Background technology
Collagen protein is present in the reticular tissue of animal and human body, the important feature matrix forming the organs such as tissue, cartilage, skin, animal collagen has abundant nutritive value, often be extracted and utilize as functional food ingredient, but due to the triple helix constructional feature of collagen protein self, be easy to degraded, be difficult to maintain collagen-stabilized triple helix structure, thus have impact on collagen protein texture characteristic and the thermostability of the food being composition.Therefore, for improving the stability of collagen protein and anti-degradation capability, now needing by collagen protein through crosslinked action, with the collagen protein that generating structure is stable, thus improving its anti-degradation capability and thermostability, improve its processing characteristics.
At present to collagen-modified method mainly physical method and chemical process, physical method mainly passes through physical means, as the methods such as uviolizing, severe dehydration, gammairradiation and heat cross-linking carry out modification to collagen protein, its shortcoming is that the method crosslinking degree is low and uneven, only uses as supplementary means at present.And the chemical cross-linking agent that Chemical Crosslinking Methods mainly adopts at present has: aldehydes, carbodiimide (EDC), epoxide (comprising single function and multi-functional two large classes), genipin, hydrazine etc.These compounds can effective cross-linked collagen, but has certain chemical toxicity due to it, should not apply in food processing process.
The object of the invention is for the deficiencies in the prior art, provide a kind of natural plant extracts as the method for linking agent and cross-linked collagen thereof, thus overcome the deficiency of traditional cross-linking method, thus provide multiple choices for collagen protein processing mode.
Summary of the invention
For solving above-mentioned the deficiencies in the prior art, the invention provides a Plant Extracts and as the method for Biological cross-linker cross-linked collagen to improve the stability of its collagen structure and anti-degradation capability.
For solving the problems of the technologies described above, the invention provides a Plant Extracts, it comprises: Fructus Amomi extract.
Wherein, described plant milk extract is Fructus Amomi extract, includes polyphenol effective constituent in described Fructus Amomi extract, and described plant milk extract is prepared according to the following steps, the first step, is cleaned by fructus amomi, dries, and pulverizes; Second step, the lixiviate in vat liquor of the Villous Amomum Fruit after pulverizing; 3rd step, vat liquor is separated; 4th step, vat liquor concentrates; 5th step, obtains the plant extract powders of the plant extract solution after concentrating or dehydration.
For solving the problems of the technologies described above, the invention provides the preparation method of a Plant Extracts, it comprises the following steps:
The first step, cleans fructus amomi, dries, and pulverizes;
Second step, the lixiviate in vat liquor of the Villous Amomum Fruit after pulverizing;
3rd step, vat liquor is separated;
4th step, vat liquor concentrates;
5th step, obtains the plant extract powders of the plant extract solution after concentrating or dehydration.
Wherein, in the described the first step, fructus amomi water for cleaning is deionized water, and bake out temperature is 30 ~ 80 DEG C, is crushed to 40 ~ 120 orders;
Wherein, in described second step, digestion agent is the ethanolic soln of 30 ~ 95%, and solid-liquid ratio is 1:5 ~ 50, and extraction temperature is 30 ~ 95 DEG C, and extraction time is 1 ~ 5 hour, needs continuous stirring in leaching process;
Wherein, the method that in described 3rd step, vat liquor is separated is centrifugation or filtration: the rotating speed of centrifugation is 4000 ~ 10000 revs/min, and the time is 5 ~ 10 minutes; The aperture of filtering membrane is 0.25 ~ 0.60 μm;
Wherein, the method that in described 4th step, vat liquor is concentrated is concentrating under reduced pressure, and concentrating under reduced pressure temperature is 30 ~ 80 DEG C, and vacuum tightness is 0.01 ~ 0.08MPa;
Wherein, in described 5th step, vat liquor is prepared as the method for dehydrated plant extract powder is lyophilize, and cryodesiccated temperature is-10 ~-20 DEG C, and the time is 36 ~ 60 hours.
The present invention also provides the preparation method of a Plant Extracts, further preferably, comprises the following steps:
The first step, by fructus amomi washed with de-ionized water, dries, with pulverizer, amomum powder is broken to 80 ~ 120 orders under 50 ~ 70 DEG C of conditions;
Second step, the lixiviate in the ethanolic soln of 75 ~ 90% of the Villous Amomum Fruit after pulverizing, input solid-liquid ratio is 1:10 ~ 25, and extraction temperature is 50 ~ 80 DEG C, and extraction time is 2 ~ 4 hours, needs continuous stirring in leaching process;
3rd step, vat liquor and residue adopt the film of 0.4 ~ 0.5 μm to carry out filtering separation;
4th step, by vat liquor concentrating under reduced pressure, temperature is 50 ~ 70 DEG C, and vacuum tightness is 0.04 ~ 0.08MPa;
5th step, obtains the plant extract solution after concentrating and adopts cryodesiccated method to obtain the plant extract powders of dewatering further.
For solving the problems of the technologies described above, the invention provides more than one and state the method that plant milk extract is biological cross-linking agents collagen protein, it comprises the following steps:
The first step, the preparation of collagen solution;
Second step, the preparation of Fructus Amomi extract solution;
3rd step, by collagen solution and the mixing of Fructus Amomi extract solution;
4th step, by concentrated for mixed solution also lyophilize.
Wherein, the collagen protein in the described the first step can be aquatic animal collagen protein, as: holothurian collagen, fish scale collagen, collagen of fish skin etc.; Also can be mammalian collagen, as: cow leather collagen, pigskin collagen etc.; Described collagen protein is acid-soluble collagen protein, and the described acid-soluble collagen protein concentration be dissolved in the acetic acid solution of 0.1 ~ 1Mol/L is 5 ~ 10%;
Wherein, in described second step, fructus amomi extract solution is 5 ~ 20% by the concentration of polyphenol effective constituent;
Wherein, in described 3rd step, crosslinking temperature is 50 ~ 90 DEG C, and crosslinking time is 1 ~ 6 hour, and collagen solution and Fructus Amomi extract liquor capacity are than being 1:0.01 ~ 1:0.1;
Wherein, concentrated employing concentrating under reduced pressure method in described 4th step, concentrating under reduced pressure temperature is 30 ~ 50 DEG C, and vacuum tightness is 0.04 ~ 0.08MPa, dry employing lyophilize.
Present invention also offers more than one and state the method that plant milk extract is biological cross-linking agents collagen protein, further preferably, comprise the following steps:
The first step, acid-soluble holothurian collagen is dissolved in the acetic acid solution of 0.1 ~ 0.5Mol/L, and the concentration of collagen solution is 5 ~ 8%;
Second step, dilutes Fructus Amomi extract concentrated solution, and maybe by the Fructus Amomi extract powder dissolution constant volume of dehydration, final Fructus Amomi extract solution is 5 ~ 10% by the concentration of polyphenol effective constituent;
3rd step, by collagen solution and the mixing of Fructus Amomi extract solution, crosslinking temperature is 70 ~ 90 DEG C, and crosslinking time is 1 ~ 3 hour, and collagen solution and Fructus Amomi extract liquor capacity are than being 1:0.01 ~ 1:0.05;
4th step, adopts concentrating under reduced pressure method to concentrate by mixed solution, thickening temperature is 40 ~ 50 DEG C, and vacuum tightness is 0.06 ~ 0.08MPa, and dry employing lyophilize, obtains the cross-linked collagen dewatered further.
The invention has the beneficial effects as follows: collagen protein and plant polyphenol can be made to occur crosslinked, the collagen product that generating structure is stable, and not there is toxicity, in can adding in functional foodstuff or being applied to other modified collagen protein product, the processing characteristics of collagen protein is improve while improving the anti-degradation capability of collagen protein, providing new method for improving collagen-stabilized property in food, having broad application prospects.
Accompanying drawing explanation
Fig. 1 is the DSC thermostability Comparative map in the embodiment of the present invention one before and after collagen cross-linking.
embodiment
The invention discloses a Plant Extracts and the method as Biological cross-linker cross-linked collagen thereof, for making object of the present invention, technical scheme and effect clearly, clearly, the present invention is described in more detail below.Should be appreciated that specific embodiment described herein is only in order to explain the present invention, is not intended to limit the present invention.
Embodiment 1:
One Plant Extracts, Fructus Amomi extract, it includes polyphenol effective constituent, and Fructus Amomi extract is dry powder form.Concrete preparation method's step of Fructus Amomi extract is as follows:
The first step, by fructus amomi washed with de-ionized water, dries, with pulverizer, amomum powder is broken to 80 orders under 60 DEG C of conditions;
Second step, the lixiviate in the ethanolic soln of 50% of the Villous Amomum Fruit after pulverizing, input solid-liquid ratio is 1:15, and extraction temperature is 60 DEG C, and extraction time is 2 hours, needs continuous stirring in leaching process;
3rd step, vat liquor and residue adopt the film of 0.5 μm to carry out filtering separation;
4th step, by vat liquor concentrating under reduced pressure, temperature is 60 DEG C, and vacuum tightness is 0.07MPa;
5th step, obtains the plant extract solution quick freezing under-30 DEG C of conditions after concentrating, and freezes the plant extract powders that the cryodesiccated method of rear employing obtains dewatering.
More than one state the method that Fructus Amomi extract is the cross-linked collagen of Biological cross-linker, and concrete steps are as follows:
The first step, is dissolved in the acetic acid solution of 0.2Mol/L by the acid-soluble holothurian collagen extracted from wall of sea cucumber Stichopus japonicus, is mixed with the holothurian collagen solution that concentration is 5%;
Second step, dilutes Fructus Amomi extract concentrated solution, and maybe by the Fructus Amomi extract powder dissolution constant volume of dehydration, final Fructus Amomi extract solution is 5% by the concentration of polyphenol effective constituent;
3rd step, by the Fructus Amomi extract solution mixing obtained in the collagen solution obtained in the first step and second step, cross-linking reaction, crosslinking temperature is 70 DEG C, and crosslinking time is 3 hours, and collagen solution and Fructus Amomi extract liquor capacity are than being 1:0.01;
4th step, adopts concentrating under reduced pressure method to concentrate by mixed solution, thickening temperature is 50 DEG C, and vacuum tightness is 0.08MPa, and by the solution quick freezing under-30 DEG C of conditions obtained, the sample after freezing adopts freeze-drying method dry, obtains the cross-linked collagen dewatered.
The invention has the beneficial effects as follows: the thermally-stabilised collection of illustrative plates as shown in Figure 1 before and after collagen cross-linking, this is DSC(differential heating scan instrument) the collagen protein thermostability collection of illustrative plates that draws, wherein peak value is the thermal denaturation temperature of collagen protein, the thermal denaturation temperature of collagen protein is higher, then collagen structure is more stable, as can be seen from figure obviously, after crosslinked, the thermostability of the collagen protein of sea cucumber obviously raises, and illustrates that linking agent can improve the thermostability of holothurian collagen.
Embodiment 2
One Plant Extracts, Fructus Amomi extract, it includes polyphenol effective constituent, and Fructus Amomi extract is dry powder form.Concrete preparation method's step of this Fructus Amomi extract is as follows:
The first step, by fructus amomi washed with de-ionized water, dries, with pulverizer, amomum powder is broken to 120 orders under 70 DEG C of conditions;
Second step, the lixiviate in the ethanolic soln of 75% of the Villous Amomum Fruit after pulverizing, input solid-liquid ratio is 1:20, and extraction temperature is 50 DEG C, and extraction time is 4 hours, needs continuous stirring in leaching process;
3rd step, vat liquor and residue use the rotating speed centrifugation of 5000r/min, get supernatant liquor;
4th step, by vat liquor concentrating under reduced pressure, temperature is 50 DEG C, and vacuum tightness is 0.08MPa;
5th step, obtains the plant extract solution after concentrating freezing under-18 DEG C of conditions, freezes the plant extract powders that the cryodesiccated method of rear employing obtains dewatering.
More than one state the method that Fructus Amomi extract is the crosslinked collagen of fish skin of Biological cross-linker, and concrete steps are as follows:
The first step, is dissolved in the acetic acid solution of 0.3Mol/L by the acid soluble fish skin collagen extracted from fish-skin, is mixed with the collagen of fish skin solution that concentration is 8%;
Second step, dilutes Fructus Amomi extract concentrated solution, and maybe by the Fructus Amomi extract powder dissolution constant volume of dehydration, final Fructus Amomi extract solution is 10% by the concentration of polyphenol effective constituent;
3rd step, by the Fructus Amomi extract solution mixing obtained in the collagen solution obtained in the first step and second step, cross-linking reaction, crosslinking temperature is 80 DEG C, and crosslinking time is 2 hours, and collagen solution and Fructus Amomi extract liquor capacity are than being 1:0.02;
4th step, adopts concentrating under reduced pressure method to concentrate by mixed solution, thickening temperature is 50 DEG C, and vacuum tightness is 0.08MPa, and by the solution quick freezing under-30 DEG C of conditions obtained, the sample after freezing adopts freeze-drying method dry, obtains the cross-linked collagen dewatered.
It is crosslinked that the method can make collagen protein and plant polyphenol occur, the collagen product that generating structure is stable, and not there is toxicity, in can adding in functional foodstuff or being applied to other modified collagen protein product, the processing characteristics of collagen protein is improve while improving the anti-degradation capability of collagen protein, providing new method for improving collagen-stabilized property in food, having broad application prospects.
Below the present invention be described in detail, the above, be only the preferred embodiment of the present invention, when not limiting the scope of the present invention, namely allly does impartial change according to the application's scope and modify, all should still belong in covering scope of the present invention.
Claims (9)
1. a Plant Extracts, is characterized in that: described plant milk extract is Fructus Amomi extract, includes polyphenol effective constituent in described Fructus Amomi extract, and described plant milk extract is prepared according to the following steps, the first step, is cleaned by fructus amomi, dries, and pulverizes; Second step, the lixiviate in vat liquor of the Villous Amomum Fruit after pulverizing; 3rd step, vat liquor is separated; 4th step, vat liquor concentrates; 5th step, obtains the plant extract powders of the plant extract solution after concentrating or dehydration.
2. a preparation method for plant milk extract as claimed in claim 1, is characterized in that being made up of following steps:
The first step, cleans fructus amomi, dries, and pulverizes;
Second step, the lixiviate in vat liquor of the Villous Amomum Fruit after pulverizing;
3rd step, vat liquor is separated;
4th step, vat liquor concentrates;
5th step, obtains the plant extract powders of the plant extract solution after concentrating or dehydration.
3. the preparation method of a Plant Extracts according to claim 2, is characterized in that: in the described the first step, fructus amomi water for cleaning is deionized water, and bake out temperature is 30 ~ 80 DEG C, is crushed to 40 ~ 120 orders.
4. the preparation method of the Plant Extracts according to Claims 2 or 3, it is characterized in that: in described second step, digestion agent is the ethanolic soln of 30 ~ 95%, solid-liquid ratio is 1:5 ~ 50, and extraction temperature is 30 ~ 95 DEG C, extraction time is 1 ~ 5 hour, needs continuous stirring in leaching process.
5. the preparation method of the Plant Extracts according to Claims 2 or 3, is characterized in that: the method that in described 3rd step, vat liquor is separated is centrifugation or filtration; The rotating speed of described centrifugation is 4000 ~ 10000 revs/min, and the time is 5 ~ 10 minutes; The aperture of described filtration filtering membrane used is 0.25 ~ 0.45 μm.
6. the preparation method of the Plant Extracts according to Claims 2 or 3, is characterized in that: the method that in described 4th step, vat liquor is concentrated is concentrating under reduced pressure, and concentrating under reduced pressure temperature is 30 ~ 80 DEG C, and vacuum tightness is 0.01 ~ 0.08MPa; In described 5th step, vat liquor is prepared as the method for dehydrated plant extract powder is lyophilize, and cryodesiccated temperature is-10 ~-20 DEG C, and the time is 36 ~ 60 hours.
7., by the method for plant milk extract according to claim 1 as Biological cross-linker cross-linked collagen, it is characterized in that being made up of following steps:
The first step, the preparation of collagen solution;
Second step, the preparation of Fructus Amomi extract solution;
3rd step, by collagen solution and the mixing of Fructus Amomi extract solution;
4th step, by concentrated for mixed solution also lyophilize.
8. according to claim 7 by the method for plant milk extract according to claim 1 as Biological cross-linker cross-linked collagen, it is characterized in that: the collagen protein in the described the first step is the acid-soluble collagen protein in aquatic animal collagen protein or mammalian collagen, and the described acid-soluble collagen protein concentration be dissolved in the acetic acid solution of 0.1 ~ 1Mol/L is 5 ~ 10%; In described second step, fructus amomi extract solution is 5 ~ 20% by the concentration of polyphenol effective constituent.
9. according to claim 7 or 8 by the method for plant milk extract according to claim 1 as Biological cross-linker cross-linked collagen, it is characterized in that: in described 3rd step, crosslinking temperature is 50 ~ 90 DEG C, crosslinking time is 1 ~ 6 hour, and collagen solution and Fructus Amomi extract liquor capacity are than being 1:0.01 ~ 1:0.1; Concentrated employing concentrating under reduced pressure method in described 4th step, concentrating under reduced pressure temperature is 30 ~ 50 DEG C, and vacuum tightness is 0.04 ~ 0.08MPa, dry employing lyophilize.
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| CN111001038A (en) * | 2019-12-30 | 2020-04-14 | 中国科学院苏州纳米技术与纳米仿生研究所 | Collagen-based 3D printing biological ink, and preparation method and application thereof |
| CN112841647A (en) * | 2021-01-18 | 2021-05-28 | 南京中医药大学 | Collagen food fixing agent and fixing method thereof |
| CN114041576A (en) * | 2021-11-24 | 2022-02-15 | 汕头大学 | Fish gelatin processing method and application thereof |
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| CN111001038A (en) * | 2019-12-30 | 2020-04-14 | 中国科学院苏州纳米技术与纳米仿生研究所 | Collagen-based 3D printing biological ink, and preparation method and application thereof |
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