CN106520858B - extraction method of ginkgo leaf water-soluble dietary fiber - Google Patents
extraction method of ginkgo leaf water-soluble dietary fiber Download PDFInfo
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Abstract
The invention belongs to the technical field of natural product extraction, and particularly relates to an extraction method of ginkgo leaf water-soluble dietary fibers. The extraction method of the ginkgo leaf water-soluble dietary fiber adopts cellulase and pectinase as a complex enzyme preparation, and uses ginkgo leaves as a raw material and water as an extractant to perform enzymolysis to extract the water-soluble dietary fiber, so that the extraction method is simple and convenient to operate, low in cost and non-toxic, and the extracted water-soluble dietary fiber can be used as a new natural green food additive to be added into foods such as dairy products and beverages, so that the health-care function of the product is increased, and a wide market prospect is provided for comprehensive development and utilization of the ginkgo leaves. The transfer rate of the water-soluble dietary fiber extracted by the method is more than 90%, the purity of the water-soluble dietary fiber in the obtained extraction product is more than 85%, and the extraction reproducibility is good.
Description
Technical Field
the invention belongs to the technical field of natural product extraction, and particularly relates to an extraction method of ginkgo leaf water-soluble dietary fibers.
Background
A plant of deciduous tree of Ginkgoaceae, belonging to gymnosperm, is a subtropical and temperate tree species, which is widely cultivated in China, and is one of the main economic tree species in China. Researches show that the ginkgo preparation has the effects of preventing and treating cardiovascular and cerebrovascular diseases, senile dementia and resisting platelet activating factors, and a plurality of products such as ginkgo botanical drug preparations, functional foods, health-care products, cosmetics and the like are researched and developed at present, so that the ginkgo preparation has obvious economic advantages. The current ginkgo preparation products mostly use ginkgo as an active substance, while the utilization of other parts is less.
In fact, studies have shown that ginkgo leaves also have high medicinal values, such as blood circulation promoting, pain relieving, diarrhea relieving with astringents, lung astringing, asthma relieving, and the like. The ginkgo leaves are rich in nutrition, mainly contain flavonoids, terpenoids, phenols, alkaloids and the like, contain more water-soluble dietary fibers, and are a substance which is homologous to medicine and food. Dietary fibers are classified into water-Soluble Dietary Fibers (SDF) and water-insoluble Dietary Fibers (DF) according to their solubility. The SDF has a physiological function greater than that of DF, has a wide nutrition and health care effect, can prevent diseases such as obesity, heart disease, diabetes and the like, can be thoroughly decomposed by microorganisms in intestinal tracts to generate various short-chain fatty acids, has a certain effect on preventing and treating colorectal cancer and colon cancer, is beneficial to the growth and the propagation of beneficial bacteria such as bifidobacterium and the like, and can improve the gastrointestinal function. With the improvement of living standard of people, various civilization diseases continuously appear, so that the importance of the dietary fiber is gradually known by people, and how to safely and efficiently prepare soluble dietary fiber products is more and more valued by scholars at home and abroad. Therefore, how to further and effectively develop the nutritive value of the ginkgo biloba, develop more ginkgo biloba preparation products and further expand the economic value of the ginkgo biloba industry.
disclosure of Invention
Therefore, the technical problem to be solved by the invention is to provide an extraction method of ginkgo leaf water-soluble dietary fibers, so as to solve the problem of poor development of ginkgo leaf extracts in the prior art.
in order to solve the technical problems, the extraction method of the ginkgo leaf water-soluble dietary fiber comprises the following steps:
(1) Pulverizing dried folium Ginkgo, sieving, adding water as extractant, and adding compound enzyme of cellulase and pectinase for enzymatic extraction to obtain extractive solution;
(2) Filtering the obtained extracting solution, concentrating the filtrate under reduced pressure to obtain a concentrated solution, and precipitating the concentrated solution with ethanol to obtain a precipitate, thereby obtaining the ginkgo leaf water-soluble dietary fiber.
In the step (1), the enzymatic extraction step is to adjust the pH value to 4.5-5.5 and control the enzymolysis temperature to 45-55 ℃ for enzymolysis.
In the step (1), the addition amount of the cellulase is 0.6-1.0 wt% of the mass of the ginkgo leaf raw material, and preferably 0.8 wt%; the addition amount of the pectinase is 0.5-0.8 wt% of the weight of the ginkgo leaf raw material, and preferably 0.6 wt%.
In the step (1), the mass ratio of the volume of the extracting agent water to the ginkgo leaf raw material is 15ml/g-30ml/g, and preferably 20 ml/g.
In the step (1), the step of adjusting the pH value is to adjust by using salicylic acid.
In the step (2), the step of filtering the extracting solution is to filter the extracting solution by medium-speed filter paper under the conditions of the vacuum degree of 0.08MPa to 0.10MPa and the temperature of 15 ℃ to 30 ℃ to obtain filtrate.
In the step (2), the vacuum concentration step is to concentrate the obtained filtrate to 1/3 of the volume of the initial filtrate under the conditions of the vacuum degree of 0.08MPa to 0.10MPa and the temperature of 50 ℃ to 60 ℃ to obtain a concentrated solution.
In the step (2), the alcohol precipitation step is to add 90-95 wt% ethanol with 3-5 times volume of the concentrated solution to carry out alcohol precipitation for 36-48 h.
and (2) freeze-drying the obtained precipitate under the conditions that the vacuum degree is less than 20Pa and the cold trap temperature is-50 to-60 ℃.
The invention also discloses the ginkgo leaf water-soluble dietary fiber extracted by the method.
The extraction method of the ginkgo leaf water-soluble dietary fiber adopts cellulase and pectinase as a complex enzyme preparation, and uses ginkgo leaves as a raw material and water as an extractant to perform enzymolysis to extract the water-soluble dietary fiber, so that the extraction method is simple and convenient to operate, low in cost and non-toxic, and the extracted water-soluble dietary fiber can be used as a new natural green food additive to be added into foods such as dairy products and beverages, so that the health-care function of the product is increased, and a wide market prospect is provided for comprehensive development and utilization of the ginkgo leaves. The transfer rate of the water-soluble dietary fiber extracted by the method is more than 90%, the purity of the water-soluble dietary fiber in the obtained extraction product is more than 85%, and the extraction reproducibility is good.
The health care performance of the ginkgo leaf water-Soluble Dietary Fiber (SDF) and VC shown in the attached drawings 1-4 are compared, the ginkgo leaf water-soluble dietary fiber extracted by the method can effectively remove OH and O2-free radicals, has better removal efficiency on DPPH and NO2-, and the health care effect of the ginkgo leaf water-Soluble Dietary Fiber (SDF) prepared by the method is better than that of VC.
Drawings
In order that the present disclosure may be more readily and clearly understood, the following detailed description of the present disclosure is provided in connection with specific embodiments thereof and the accompanying drawings, in which,
FIG. 1 is a comparison of the OH-scavenging effect of Ginkgo biloba leaf water Soluble Dietary Fiber (SDF) and VC of the present invention;
FIG. 2 is a comparison of the scavenging effect of the ginkgo biloba leaf water Soluble Dietary Fiber (SDF) and VC of the present invention on O2-free radical;
FIG. 3 is a comparison of the effect of Ginkgo biloba leaf water Soluble Dietary Fiber (SDF) and VC on DPPH removal;
FIG. 4 is a comparison of the effect of ginkgo biloba leaf water Soluble Dietary Fiber (SDF) and VC in scavenging NO 2-according to the present invention.
Detailed Description
In the following embodiments of the present invention, a phenol-sulfuric acid method is used for detecting water-soluble dietary fibers, and specifically includes:
1. drawing a standard curve: accurately transferring 0.00, 0.10, 0.20, 0.40, 0.60, 0.80, 1.00 and 1.20mL (100 mu g/mL) of glucose standard solution into a test tube with a plug by using a pipette, supplementing 2.00mL of deionized water, adding 1.00mL of 6% phenol solution, then quickly dropwise adding 5.00mL of concentrated sulfuric acid, shaking uniformly, cooling to room temperature, measuring the absorbance at 490nm, drawing by taking the absorbance (Y) as an ordinate and the glucose concentration (X) as an abscissa to obtain a linear regression equation Y of 0.0564X + 0.0167; r2 ═ 0.9987;
2. sample detection: accurately weighing 0.25g of a soluble dietary fiber sample, dissolving with warm water, quantitatively transferring into a 250mL volumetric flask, fixing the volume, shaking up, accurately transferring 25.00mL of the solution into the 250mL volumetric flask by using a pipette, fixing the volume with deionized water, and shaking up. Accurately transferring 1.00mL of the diluted solution into a stoppered test tube, adding ionized water to supplement 2.00mL, adding 1.00mL of 6% phenol solution, shaking up, quickly transferring 5.00mL of concentrated sulfuric acid into the stoppered test tube, shaking up, measuring absorbance, and calculating the purity of the water-soluble dietary fiber product according to a regression equation.
Example 1
The method for extracting the ginkgo leaf water-soluble dietary fiber comprises the following steps:
(1) drying and crushing ginkgo leaves, sieving the dried and crushed ginkgo leaves with a 50-mesh sieve, taking 1kg of the crushed ginkgo leaves, adding 20L of water as an extracting agent, uniformly mixing, then adding 8g of cellulase and 6g of pectinase, uniformly mixing, adjusting the pH value of a solution to be 4.8 by using salicylic acid, controlling the temperature to be 50 ℃, performing compound enzymolysis for 60min to obtain an extracting solution, and testing that the transfer rate of water-soluble dietary fibers in the extracting solution is 95.10%;
(2) filtering the extractive solution with medium speed filter paper at 25 deg.C under 0.08MPa to obtain filtrate; concentrating the filtrate at 60 deg.C under 0.09MPa to 1/3 of the volume of the initial filtrate to obtain concentrated solution; adding 4 times of 95 wt% ethanol solution into the concentrated solution, and precipitating with ethanol for 48 hr; and (3) after alcohol precipitation, removing supernatant, putting the precipitate in a freeze dryer, and drying until the precipitate is completely removed under the conditions that the vacuum degree is less than 20Pa and the temperature of a cold trap is-50 ℃ to obtain the ginkgo leaf water-soluble dietary fiber product.
Through detection, the purity of the dietary fiber of the ginkgo leaf water-soluble dietary fiber product is 88.62%.
Example 2
The method for extracting the ginkgo leaf water-soluble dietary fiber comprises the following steps:
(1) Drying and crushing ginkgo leaves, sieving the dried and crushed ginkgo leaves with a 50-mesh sieve, taking 1kg of the crushed ginkgo leaves, adding 25L of water as an extracting agent, uniformly mixing, then adding 9g of cellulase and 7g of pectinase, uniformly mixing, adjusting the pH value of a solution to be 5.0 by using salicylic acid, controlling the temperature to be 48 ℃, carrying out compound enzymolysis for 55min to obtain an extracting solution, and testing that the transfer rate of water-soluble dietary fibers in the extracting solution is 94.56%;
(2) Filtering the extractive solution with medium speed filter paper at 25 deg.C and vacuum degree of 0.09MPa to obtain filtrate; concentrating the filtrate at 60 deg.C under 0.08MPa to 1/3 of the volume of the initial filtrate to obtain concentrated solution; adding 3 times of 90 wt% ethanol solution into the concentrated solution, and precipitating with ethanol for 44 h; and (3) after alcohol precipitation, removing supernatant, putting the precipitate in a freeze dryer, and drying until the precipitate is completely removed under the conditions that the vacuum degree is less than 20Pa and the cold trap temperature is-55 ℃ to obtain the ginkgo leaf water-soluble dietary fiber product.
Through detection, the purity of the dietary fiber of the ginkgo leaf water-soluble dietary fiber product is 86.60%.
example 3
The method for extracting the ginkgo leaf water-soluble dietary fiber comprises the following steps:
(1) Drying and crushing ginkgo leaves, sieving the dried and crushed ginkgo leaves with a 50-mesh sieve, taking 1kg of the crushed ginkgo leaves, adding 28L of water as an extracting agent, uniformly mixing, then adding 7g of cellulase and 6.5g of pectinase, uniformly mixing, adjusting the pH value of a solution to be 5.2 by using salicylic acid, controlling the temperature to be 52 ℃, performing compound enzymolysis for 65min to obtain an extracting solution, and testing that the transfer rate of water-soluble dietary fibers in the extracting solution is 93.80%;
(2) filtering the extractive solution with medium speed filter paper at 25 deg.C and vacuum degree of 0.10MPa to obtain filtrate; concentrating the filtrate at 50 deg.C under 0.10MPa to 1/3 of the volume of the initial filtrate to obtain concentrated solution; adding 5 times of 95 wt% ethanol solution into the concentrated solution, and precipitating with ethanol for 40 h; and (3) after alcohol precipitation, removing supernatant, putting the precipitate in a freeze dryer, and drying until the precipitate is completely removed under the conditions that the vacuum degree is less than 20Pa and the temperature of a cold trap is-60 ℃ to obtain the ginkgo leaf water-soluble dietary fiber product.
through detection, the purity of the dietary fiber of the ginkgo leaf water-soluble dietary fiber product is 87.26%.
example 4
The method for extracting the ginkgo leaf water-soluble dietary fiber comprises the following steps:
(1) Drying and crushing ginkgo leaves, sieving the dried and crushed ginkgo leaves with a 50-mesh sieve, taking 1kg of the crushed ginkgo leaves, adding 15L of water as an extracting agent, uniformly mixing, then adding 6g of cellulase and 8g of pectinase, uniformly mixing, adjusting the pH value of a solution to be 4.5 by using salicylic acid, controlling the temperature to be 55 ℃, performing compound enzymolysis for 60min to obtain an extracting solution, and testing that the transfer rate of water-soluble dietary fibers in the extracting solution is 91.38%;
(2) Filtering the extractive solution with medium speed filter paper at 15 deg.C under 0.10MPa to obtain filtrate; concentrating the filtrate at 50 deg.C under 0.08MPa to 1/3 of the volume of the initial filtrate to obtain concentrated solution; adding 4 times of 95 wt% ethanol solution into the concentrated solution, and precipitating with ethanol for 36 h; and (3) after alcohol precipitation, removing supernatant, putting the precipitate in a freeze dryer, and drying until the precipitate is completely removed under the conditions that the vacuum degree is less than 20Pa and the cold trap temperature is-55 ℃ to obtain the ginkgo leaf water-soluble dietary fiber product.
through detection, the purity of the dietary fiber of the ginkgo leaf water-soluble dietary fiber product is 83.59%.
Example 5
the method for extracting the ginkgo leaf water-soluble dietary fiber comprises the following steps:
(1) drying and crushing ginkgo leaves, sieving the dried and crushed ginkgo leaves with a 50-mesh sieve, taking 1kg of the crushed ginkgo leaves, adding 30L of water as an extracting agent, uniformly mixing, then adding 10g of cellulase and 5g of pectinase, uniformly mixing, adjusting the pH value of a solution to be 5.5 by using salicylic acid, controlling the temperature to be 45 ℃, carrying out compound enzymolysis for 60min to obtain an extracting solution, and testing that the transfer rate of water-soluble dietary fibers in the extracting solution is 91.43%;
(2) filtering the extractive solution with medium speed filter paper at 30 deg.C and vacuum degree of 0.08MPa to obtain filtrate; concentrating the filtrate at 55 deg.C under vacuum degree of 0.10MPa to 1/3 of the volume of the initial filtrate to obtain concentrated solution; adding 4 times of 95 wt% ethanol solution into the concentrated solution, and precipitating with ethanol for 48 hr; and (3) after alcohol precipitation, removing supernatant, putting the precipitate in a freeze dryer, and drying until the precipitate is completely removed under the conditions that the vacuum degree is less than 20Pa and the cold trap temperature is-55 ℃ to obtain the ginkgo leaf water-soluble dietary fiber product.
through detection, the purity of the dietary fiber of the ginkgo leaf water-soluble dietary fiber product is 83.18%.
Comparative example 1
The extraction method of the ginkgo leaf water-soluble dietary fiber described in the present comparative example is the same as that of example 1, except that HCl is used to adjust the pH in the step (1).
Through detection, the purity of the dietary fiber of the ginkgo leaf water-soluble dietary fiber product is 58.27%. It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.
Claims (1)
1. The extraction method of the ginkgo leaf water-soluble dietary fiber is characterized by comprising the following steps:
(1) Pulverizing dried folium Ginkgo, sieving, adding water as extractant, adding compound enzyme preparation of cellulase and pectinase, adjusting pH to 4.5-5.5 with salicylic acid, and performing enzymatic extraction at 45-55 deg.C to obtain extractive solution; the addition amount of the cellulase is 0.6-1.0 wt% of the mass of the ginkgo leaf raw material, and the addition amount of the pectinase is 0.5-0.8 wt% of the mass of the ginkgo leaf raw material;
(2) filtering the obtained extract, concentrating the filtrate under reduced pressure to obtain concentrated solution, and precipitating the concentrated solution with ethanol to obtain precipitate to obtain the folium Ginkgo water-soluble dietary fiber;
in the step (1), the mass ratio of the volume of the extractant water to the ginkgo leaf raw material is 15ml/g-30 ml/g;
in the step (2), the step of filtering the extracting solution is to filter the extracting solution by medium-speed filter paper under the conditions of the vacuum degree of 0.08MPa to 0.10MPa and the temperature of 15 ℃ to 30 ℃ to obtain filtrate;
In the step (2), the step of concentrating under reduced pressure refers to concentrating the obtained filtrate to 1/3 of the volume of the initial filtrate under the conditions of the vacuum degree of 0.08MPa to 0.10MPa and the temperature of 50 ℃ to 60 ℃ to obtain a concentrated solution;
in the step (2), the alcohol precipitation step is to add 90-95 wt% ethanol with 3-5 times volume of the concentrated solution to carry out alcohol precipitation for 36-48 h;
and (2) freeze-drying the obtained precipitate under the conditions that the vacuum degree is less than 20Pa and the cold trap temperature is-50 to-60 ℃.
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