CN105661556A - Herba cistanche dietary fibers and preparation method and application thereof - Google Patents

Abstract

The invention provides a cistanche dietary fiber, its preparation method and application. The preparation method comprises the following steps: (1) mixing the cistanche residue with an extract, extracting, and separating solid and liquid to obtain a supernatant and a precipitate. The above precipitate is dried to obtain insoluble dietary fiber, wherein the extract is any one of acid solution, alkali solution or enzyme solution; (2) supernatant or concentrated supernatant is mixed with absolute ethanol for Alcohol precipitation to obtain flocs, and drying the flocs to obtain soluble dietary fiber. The preparation method of the cistanche dietary fiber is simple, and soluble dietary fiber and insoluble dietary fiber can be obtained from the cistanche residue, which realizes the comprehensive utilization of cistanche and is convenient for industrial production and application, and the cistanche dietary fiber obtained by the preparation method has high purity, It has good color and luster, has excellent laxative effect and inhibits the activity of α-glucosidase, and has wide application in the field of health food.

Description

A kind of cistanche dietary fiber, its preparation method and application

technical field

The invention belongs to the technical field of bioengineering, and relates to cistanche dietary fiber, its preparation method and application, in particular to a method for extracting cistanche dietary fiber from cistanche residue and its application.

Background technique

Dietary fiber (dietary fiber, DF) refers to carbohydrates and lignin that cannot be digested and absorbed by the small intestine of the human body, have health significance for the human body, and have a degree of polymerization ≥ 3, including cellulose, hemicellulose, and pectin. Dietary fiber has physiological effects such as preventing cardiovascular disease, obesity, diabetes, high blood pressure, improving the function of intestinal flora, reducing the occurrence of gastrointestinal cancer, removing harmful substances from exogenous sources, anti-oxidation, and scavenging free radicals. The "seventh type of nutrient" after protein, fat, vitamins, minerals and water.

According to the solubility of dietary fiber, it can be divided into soluble dietary fiber and insoluble dietary fiber. Soluble dietary fiber mainly refers to the storage substances and secretions in the cell wall, such as pectin, some hemicellulose and vegetable gum, etc.; insoluble dietary fiber mainly refers to the components of the cell wall, such as cellulose, some hemicellulose, lignin, and chitosan Sugar and Vegetable Wax etc. At present, the methods for extracting dietary fiber at home and abroad mainly include chemical method, enzymatic method, chemical-enzyme combination method, membrane separation method and fermentation method. The chemical method produces three wastes in the production process, which seriously pollutes the environment. The membrane method is limited by the production conditions, and it is not easy to realize industrial production at present. The advantages of the enzymatic method are that it does not require high temperature and high pressure, saves energy, is convenient to operate, and is conducive to environmental protection.

Cistanche, also known as golden bamboo shoots, goblin, Cistanche or Dayun, is the dry fleshy stem with scales of Cistanche deserticola, which is mainly produced in Inner Mongolia, Gansu, Xinjiang and Qinghai; modern pharmacological studies have shown that Cistanche has the effect of improving Sexual function, anti-aging, improving learning and memory ability, anti-senile dementia, laxative and many other effects, it has the reputation of "desert ginseng". At present, the main active ingredients of Cistanche deserticola are polysaccharides and phenylethanol glycosides, and the utilization of the residues after extraction of polysaccharides and phenylethanol glycosides is very low. The residue accounts for 30-40% of the raw materials of Cistanche deserticola and contains a large amount of dietary fiber; at present, Only CN104957640A reports the preparation method of Cistanche dietary fiber. In this method, fresh Cistanche is taken, washed and sliced, mixed with water according to a weight ratio of 1: (1-2), crushed to obtain Cistanche slurry, and the slurry is filtered to obtain Cistanche residue and Cistanche. Cistanche cistanche slag was enzymolyzed by α-amylase and papain to obtain an enzymolyzed liquid; after the Cistanche deserticola juice was made into a starter, it was inserted into the enzymolyzed liquid for fermentation; Water-insoluble dietary fiber and water-soluble dietary fiber are obtained. Although this method utilizes the whole fruit of Cistanche, the high-value utilization of Cistanche is not carried out, and this method is only applicable to fresh Cistanche, and the time required for the whole technological process is longer, as: the preparation of starter requires 6- 10 hours, 1.5-3 hours for enzymatic hydrolysis, 18-22 hours for culture after inoculation of starter, 1-2 hours for alcohol precipitation, which is not suitable for wide application.

At present, the main products of Herba Cistanche are dried slices of Herba Cistanche, water extract (based on polysaccharides) and alcohol extract (based on phenylethanol glycosides), and the extraction residue in production is not fully utilized. Therefore, the present invention is aimed at the above problems , proposed a new enzymatic method for the preparation of Cistanche dietary fiber, and used it to assist in the treatment of diabetes and the development of laxative-related products.

Contents of the invention

In view of the deficiencies in the prior art, the object of the present invention is to provide Cistanche dietary fiber, its preparation method and application. The preparation method of the Cistanche dietary fiber is simple, and soluble dietary fiber and insoluble dietary fiber can be obtained from the residue of Cistanche. The comprehensive utilization of cistanche is convenient for industrial production and application, and the cistanche dietary fiber obtained by the preparation method has high purity and good color, and has wide application in the field of health food.

For reaching this purpose, the present invention adopts following technical scheme:

One of object of the present invention is to provide a kind of preparation method of cistanche dietary fiber, described preparation method comprises the steps:

(1) Mix the cistanche residue with the extract, extract and separate the solid and liquid to obtain a supernatant and a precipitate, and the precipitate is dried to obtain an insoluble dietary fiber, wherein the extract is an acid solution, an alkali solution or an enzyme solution any of the

(2) The supernatant or the concentrated supernatant is mixed with absolute ethanol for alcohol precipitation, solid-liquid separation to obtain flocs, and the flocs are dried to obtain soluble dietary fiber.

The cistanche dietary fiber provided by the invention has a simple preparation method, can obtain soluble dietary fiber and insoluble dietary fiber, realizes comprehensive utilization of cistanche deserticola, and is convenient for large-scale industrial production and application.

The residue of Cistanche described in step (1) is the residue after water extraction of Cistanche, the residue after alcohol extraction of Cistanche, the residue after water extraction and then alcohol extraction of Cistanche, or the residue after alcohol extraction and then water extraction of Cistanche. Any one or a combination of at least two. A typical but non-limiting combination is: the residue after water extraction of Cistanche and the residue after alcohol extraction of Cistanche, the residue after alcohol extraction of Cistanche and the residue after water extraction and then alcohol extraction of Cistanche, first alcohol extraction of Cistanche The residue after water extraction and Cistanche first carried out water extraction and then the residue after alcohol extraction, the residue after Cistanche alcohol extraction, the residue after Cistanche first water extraction and then alcohol extraction and Cistanche first alcohol extraction and then water extraction Residue, the residue after water extraction of Cistanche, the residue after alcohol extraction of Cistanche, the residue after first water extraction and then alcohol extraction of Cistanche, and the residue after first alcohol extraction and then water extraction of Cistanche.

Preferably, the Cistanche is any one or a combination of at least two of desert Cistanche, Cistanche tuberosa, Cistanche salina, Lanzhou Cistanche or the fleshy stems of Cistanche deserticola. Typical but non-limiting combinations are: the fleshy stems of Cistanche desertica and Cistanche tuberosa, the fleshy stems of Cistanche saline, Cistanche lanzhou and Cistanche cistanche, the fleshy stems of Cistanche tuberosa, Cistanche saline and Cistanche lanceolata, desert cistanche, The fleshy stems of Cistanche tuberosa and Cistanche saline, the fleshy stems of desert Cistanche, Cistanche tuberosa, Cistanche saline, Lanzhou Cistanche and Cistanche deserticola.

The cistanche residue described in step (1) is pulverized and then mixed with the extract. The particle size of the crushed cistanche residue is preferably 100-600 mesh. The pulverization is conducive to better mixing of the Cistanche residue and the extract, and shortens the extraction time.

Preferably, the acid solution in step (1) is any one or a combination of at least two of hydrochloric acid, formic acid, acetic acid, citric acid, malic acid, oxalic acid, sulfuric acid, nitric acid or lactic acid, a typical but non-limiting combination For: hydrochloric acid and formic acid, acetic acid and citric acid, citric acid, malic acid and oxalic acid, acetic acid, citric acid and malic acid, oxalic acid, sulfuric acid, nitric acid and lactic acid. It is preferably a mixed acid solution of hydrochloric acid and formic acid, citric acid and lactic acid. The volume ratio of hydrochloric acid and formic acid is preferably 1-4:1, such as 2:1 or 3:1.

Preferably, the concentration of the acid solution in step (1) is 0.1%-10%, such as 0.2%, 0.5%, 0.8%, 1%, 2%, 3%, 4%, 5%, 6%, 7% , 8% or 9%, etc., preferably 0.5%-5%.

Preferably, the amount of the acid solution in step (1) is 8-30 times the mass of Cistanche residue, such as 9 times, 10 times, 12 times, 15 times, 18 times, 20 times, 22 times, 25 times or 28 times. Times, etc., preferably 10-20 times.

Preferably, the extraction time of the acid solution in step (1) is 1-3 hours, such as 2 hours or 2.5 hours.

The lye in step (1) is NaOH solution and/or KOH solution, preferably a mixed lye of NaOH solution and KOH solution, and the volume ratio of NaOH solution and KOH solution is preferably 1:1.

Preferably, the concentration of the lye is 0.5%-10%, such as 0.8%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 5%, 6%, 7%, 8% or 9%, etc., preferably 0.5%-3%.

Preferably, the amount of lye added in step (1) is 8-30 times the mass of Cistanche residue, such as 9 times, 10 times, 12 times, 15 times, 18 times, 20 times, 22 times, 25 times or 28 times. Times, etc., preferably 10-20 times.

Preferably, the extraction time of the lye in step (1) is 1-3h, such as 2h or 2.5h.

The enzyme solution in step (1) is a buffer containing hydrolase, and the hydrolase is amylase, protease and glucosidase.

Preferably, the Cistanche cistanche residue in step (1) is first mixed with a buffer, and then amylase enzymatic hydrolysis, protease enzymatic hydrolysis and glucosidase enzymatic hydrolysis are performed sequentially. Mixing the extracted raw material with the buffer first can provide a good reaction buffer system for the subsequent enzymatic reaction; secondly, the amylase hydrolyzes the starch and glycogen and other macromolecules in the raw material into oligosaccharides, and the protease hydrolyzes the protein into Small molecule amino acid, glucosidase hydrolyzes the product of amylase, that is, oligosaccharides, into glucose, which is ultimately beneficial to the improvement of the purity and color of dietary fiber.

Preferably, the amylase hydrolysis condition is 50-95°C, such as 52°C, 53°C, 54°C, 55°C, 56°C, 58°C, 60°C, 62°C, 65°C, 68°C, 70°C , 75°C, 80°C, 85°C or 90°C, etc., 20-60min, such as 22min, 25min, 28min, 30min, 32min, 35min, 40min, 50min or 55min, etc.

Preferably, the protease hydrolysis condition is 40-70°C, such as 42°C, 45°C, 48°C, 50°C, 52°C, 55°C, 58°C, 60°C, 62°C, 65°C or 68°C, etc. , 20-60min, such as 22min, 25min, 28min, 30min, 32min, 35min, 40min, 50min or 55min, etc.

Preferably, the enzymatic hydrolysis conditions of the glucosidase are 40-60°C, such as 42°C, 45°C, 48°C, 50°C, 52°C, 55°C or 58°C, etc., 20-60min, such as 22min, 25min, 28min, 30min, 32min, 35min, 40min, 50min or 55min, etc.

Preferably, the addition of the amylase, protease and glucosidase is independently 100-3000U/g, such as 200U/g, 500U/g, 800U/g, 1000U/g, 1200U/g, 1500U/g, 1800U/g, 2000U/g, 2300U/g, 2500U/g or 2800U/g, etc., preferably 2000U/g.

Preferably, the buffer is any one or a combination of at least two of phosphate buffer, Tris-HCl buffer or citrate buffer, a typical but non-limiting combination is: phosphate buffer and Tris - HCl buffer, phosphate buffer and citrate buffer, phosphate buffer, Tris-HCl buffer and citrate buffer, preferably a combination of phosphate buffer and Tris-HCl buffer.

Preferably, the buffer solution has a pH value of 6.5-8.5, such as 6.8, 7.0, 7.2, 7.5, 7.8, 8.0, 8.2 or 8.4.

The solid-liquid separation in step (1) is a conventional operation in the field, and a typical but non-limiting solid-liquid separation method is centrifugation.

The drying described in step (1) is a conventional operation in the art, and those skilled in the art can select the drying temperature according to the actual situation.

The amount of absolute ethanol added in step (2) is 3-5 times the volume of the supernatant or the concentrated supernatant, such as 3.5 times, 4 times or 4.5 times.

The drying described in step (2) is a routine operation in the art, and those skilled in the art can select the drying temperature according to the actual situation.

The solid-liquid separation in step (2) is a conventional operation in the field, and a typical but non-limiting solid-liquid separation method can be centrifugation or filtration.

As preferred technical scheme, the present invention provides a kind of preparation method of cistanche dietary fiber, described preparation method comprises the steps:

(1) Mix the cistanche residue with a buffer solution with a pH value of 6.5-8.5, then add amylase to it, and enzymolyze it at 50-95°C for 20-60min; then add protease, and enzymolyze it at 40-70°C for 20 minutes -60min; then add glucosidase, and enzymolyze at 40-60°C for 20-60min to obtain an enzymatic solution, wherein the addition of the amylase, protease and glucosidase is independently 100-3000U/g; solid-liquid separation of the enzymolysis solution to obtain a supernatant and a precipitate, and the precipitate is dried to obtain insoluble dietary fiber;

(2) The supernatant or the concentrated supernatant is mixed with absolute ethanol to carry out alcohol precipitation to obtain flocs, wherein the addition of absolute ethanol is 3% of the volume of the supernatant or the concentrated supernatant -5 times; the floc is dried to obtain soluble dietary fiber.

The cistanche dietary fiber obtained by the enzymatic hydrolysis method has a good color and is milky white, and has excellent laxative effect and inhibitory activity of α-glucosidase.

Another object of the present invention is to provide a dietary fiber of Cistanche deserticola prepared by the preparation method, and the dietary fiber of Cistanche deserticola includes soluble dietary fiber and insoluble dietary fiber.

One of the objects of the present invention is also to provide an application of the cistanche dietary fiber prepared by the preparation method in the field of food or biomedicine.

The cistanche dietary fiber can be used to prepare foods or health preparations for auxiliary treatment of diabetes, auxiliary treatment of laxative or improvement of constipation, and the health preparations include tablets, granules, pills, granules, tea drinks, capsules, oral liquids or suspensions etc.

Compared with prior art, the beneficial effect of the present invention is:

(1) The preparation method of Cistanche dietary fiber provided by the present invention is simple, can prepare soluble dietary fiber and insoluble dietary fiber from the residue of Cistanche extraction, realizes the comprehensive utilization of Cistanche, and is convenient for large-scale industrial production and application;

(2) The Cistanche dietary fiber prepared by the preparation method of the Cistanche dietary fiber provided by the present invention has better color and luster, is milky white, has excellent bowel laxative and inhibits α-glucosidase activity, and can be prepared according to the technological requirements of modern preparations. It can be made into a functional health food with auxiliary treatment of diabetes and laxative effects, and can also be made into various oral preparations by adding auxiliary materials.

detailed description

The following examples are used to further illustrate the technical solution of the present invention to better understand the present invention, its advantages and effects, but the examples are only used to illustrate the present invention rather than limit the present invention.

Example 1: Preparation of Dietary Fiber in Cistanche Cistanche Water Extraction Residue

Crushed Cistanche deserticola, weighed 500g, added 6000mL distilled water, extracted for 1 hour, filtered, and dried the residue to obtain the water-extracted residue of Cistanche deserticola.

(1) Pulverize the residue of desert cistanche by water extraction, add 0.1% hydrochloric acid solution 8 times its weight, mix well, extract for 3 hours, centrifuge the extract to obtain supernatant and precipitate, dry the precipitate to obtain insoluble dietary fiber ;

(2) After concentrating the supernatant, add 4 times the volume of absolute ethanol, ethanol precipitation for 2 hours to obtain flocs, and dry the flocs to obtain soluble dietary fiber.

Embodiment 2: the preparation of dietary fiber in the cistanche deserticola alcohol extraction residue

Grind Cistanche cistanche, weigh 200g, add 1800mL of 50% ethanol solution, extract for 70min, filter, and dry the residue to obtain the alcohol extraction residue of Cistanche cistanche.

(1) Grind the residue of Cistanche tubulosa alcohol extraction, add 10% lactic acid solution 30 times its weight, mix well, extract for 2 hours, centrifuge the extract to obtain supernatant and precipitate, dry the precipitate to obtain insoluble meal fiber;

(2) After concentrating the supernatant, add 5 times the volume of dehydrated ethanol, ethanol precipitation for 4 hours to obtain flocs, and dry to obtain soluble dietary fiber.

Embodiment 3: Preparation of dietary fiber in Cistanche deserticola water extraction residue

Grind Cistanche saline, weigh 1000g, add 10000mL of distilled water, extract for 5 hours, filter, and dry the residue to obtain the water-extracted residue of Cistanche saline.

(1) Pulverize the water extraction residue of Cistanche saline, add 10 times the weight of 5% mixed acid (1:1 by volume) solution of hydrochloric acid and formic acid, mix well, extract for 1 hour, and centrifuge the extract to obtain Supernatant and precipitation, the precipitation is dried to obtain insoluble dietary fiber;

(2) After concentrating the supernatant, add 4 times the volume of dehydrated ethanol, ethanol precipitation for 5 hours to obtain flocs, and dry to obtain soluble dietary fiber.

Embodiment 4: the preparation of dietary fiber in the cistanche deserticola alcohol extraction residue

Crush Cistanche, weigh 600g, add 9000mL of 60% ethanol solution, extract for 90min, filter, and dry the residue to obtain the alcohol extraction residue of Cistanche.

(1) Grind the residue of desert cistanche alcohol extraction, add 0.5% sulfuric acid solution of 20 times its weight, mix well, extract for 1.5h, centrifuge the extract to obtain supernatant and precipitate, dry the precipitate to obtain insoluble Dietary fiber;

(2) After concentrating the supernatant, add 5 times the volume of dehydrated ethanol, ethanol precipitation for 4 hours to obtain flocs, and dry to obtain soluble dietary fiber.

Example 5: Preparation of Dietary Fiber in Cistanche Cistanche Water Extraction Residue

Crushed Cistanche deserticola, weighed 500g, added 6000mL distilled water, extracted for 1 hour, filtered, and dried the residue to obtain the water-extracted residue of Cistanche deserticola.

(1) Crush the residue of Cistanche cistanche, add 12 times the weight of phosphate buffer (pH8.0), mix well, add 100U/g amylase, 50℃, enzymatic hydrolysis for 60min; then add 500U/g protease at 40°C for 60 minutes; then add 2500U/g glucosidase at 40°C for 60 minutes to obtain an enzymolysis solution; centrifuge the enzymolysis solution to obtain a supernatant and a precipitate, and dry the precipitate. Obtain insoluble dietary fiber;

(2) After concentrating the supernatant, add 4 times the volume of dehydrated ethanol, ethanol precipitation for 2 hours to obtain flocs, and dry to obtain soluble dietary fiber.

Embodiment 6: Preparation of dietary fiber in cistanche water extraction residue

Grind Cistanche cistanche, weigh 300g, add 5000mL of distilled water, extract for 2 hours, centrifuge, and dry the residue to obtain the water-extracted residue of Cistanche cistanche.

(1) Crush the residue of Cistanche tubulosa water extraction, add 16 times its weight of Tris-HCl buffer solution (pH7.5), mix well; add 3000U/g amylase, 95°C, enzymolysis for 20min; after that, add 100U/g protease, 70°C, enzymolysis for 20min; then add 500U/g glucosidase, 60°C, enzymolysis for 20min, to obtain the enzymolysis solution; centrifuge the enzymolysis solution to obtain the supernatant and precipitate, and The precipitation is dried to obtain insoluble dietary fiber;

(2) After concentrating the supernatant, add 3 times the volume of absolute ethanol, ethanol precipitation for 3 hours to obtain flocs, and dry to obtain soluble dietary fiber.

Embodiment 7: Preparation of dietary fiber in cistanche deserticola alcohol extraction residue

Grind Cistanche saline, weigh 200g, add 2000mL of 50% ethanol solution, extract for 50min, filter, and dry the residue to obtain the alcohol extraction residue of Cistanche salina.

(1) Grind the residue of cistanche cistanche ethanol extraction, add 18 times its weight of citric acid buffer (pH6.5), mix well; add 2000U/g amylase, 60°C, enzymatic hydrolysis for 50min; then add 3000U/g g of protease at 55°C for 40 minutes; then add 100 U/g of glucosidase at 50°C for 40 minutes to obtain an enzymatic solution; centrifuge the enzymatic solution to obtain supernatant and precipitate, and dry the precipitate , to obtain insoluble dietary fiber;

(2) After concentrating the supernatant, add 5 times the volume of dehydrated ethanol, ethanol precipitation for 4 hours to obtain flocs, and dry to obtain soluble dietary fiber.

Embodiment 8: Preparation of dietary fiber in cistanche deserticola alcohol extraction residue

Grind Cistanche lanzhou, weigh 1000g, add 20000mL of 40% ethanol solution, extract for 3 hours, centrifuge, and dry the residue to obtain the alcohol extraction residue of Cistanche lanzhou.

(1) Grind the alcohol extraction residue of Cistanche cistanche Lanzhou, add 20 times the weight of the mixture of phosphate buffer and Tris-HCl buffer (pH7.0), mix well; add 1550U/g of amylase, 72.5 ° C, Enzymolysis for 40min; then add 2000U/g protease, 60°C, enzymolysis for 50min; then add 3000U/g glucosidase, 50°C, enzymolysis for 30min, to obtain the enzymolysis solution; centrifuge the enzymolysis solution to obtain the above Supernatant and precipitate, the precipitate is dried to obtain insoluble dietary fiber;

(2) After concentrating the supernatant, add 4 times the volume of dehydrated ethanol, ethanol precipitation for 5 hours to obtain flocs, and dry to obtain soluble dietary fiber.

Embodiment 9: Preparation of dietary fiber in cistanche deserticola alcohol extraction residue

Crush Cistanche, weigh 800g, add 9000mL of 60% ethanol solution, extract for 1 hour, centrifuge, and dry the residue to obtain the alcohol extraction residue of Cistanche.

(1) Grind the residue of desert cistanche alcohol extraction, add 25 times its weight of phosphate buffer (pH8.5), mix well; add 1000U/g amylase, 80°C, enzymolysis for 30min; then add 1550U/g g of protease at 55°C for 40 minutes; then add 2000 U/g of glucosidase at 55°C for 30 minutes to obtain an enzymatic solution; centrifuge the enzymatic solution to obtain supernatant and precipitate, and dry the precipitate , to obtain insoluble dietary fiber;

(2) After concentrating the supernatant, add 4 times the volume of dehydrated ethanol, ethanol precipitation for 2 hours to obtain flocs, and dry to obtain soluble dietary fiber.

Embodiment 10: Preparation of dietary fiber in cistanche residue

Crush desert cistanche, weigh 5000g, add 100000mL of water, extract for 1.5h, centrifuge, and dry the residue to obtain the residue of desert Cistanche water extraction; then add 60000mL of 50% ethanol solution, extract for 1h, filter, and dry the residue to obtain Desert Cistanche alcohol extraction residue.

(1) Crush desert cistanche alcohol extraction residue, add 26 times the weight of phosphate buffer (pH8.2), mix well; add 2500U/g amylase, 75°C, enzymatic hydrolysis for 35min; then add 1000U/g protease at 45°C for 30 minutes; then add 1550 U/g of glucosidase at 50°C for 50 minutes to obtain the enzymolysis solution; centrifuge the enzymolysis solution to obtain the supernatant and precipitate, and dry the precipitate. Obtain insoluble dietary fiber;

(1) After concentrating the supernatant, add 4 times the volume of absolute ethanol, ethanol precipitation for 2 hours to obtain flocs, and dry to obtain soluble dietary fiber.

Example 11: Preparation of Dietary Fiber in Cistanche Residue

Crushed Cistanche deserticola, weighed 5000g, added 10000mL of 52% ethanol solution, extracted for 1h, centrifuged, and dried the residue to obtain the alcohol extraction residue of Cistanche deserticola. Then add 80000mL of water, extract for 1 hour, filter, and dry the residue to obtain the residue of Cistanche deserticola water extraction.

(1) Crush desert cistanche water extraction residue, add 28 times its weight of Tris-HCl buffer (pH8.0), mix well; add 500U/g amylase, 65°C, enzymatic hydrolysis for 55min; then add 2500U/g g of protease at 65°C for 55 minutes; then add 1000 U/g of glucosidase at 60°C for 35 minutes to obtain an enzymatic solution; centrifuge the enzymatic solution to obtain a supernatant and precipitate, then dry the precipitate , to obtain insoluble dietary fiber;

(1) After concentrating the supernatant, add 4.5 times the volume of absolute ethanol, ethanol precipitation for 1.5 hours to obtain flocs, and dry to obtain soluble dietary fiber.

Example 12: Preparation of Dietary Fiber in Cistanche Cistanche Water Extraction Residue

Grind Cistanche saline, weigh 1000g, add 10000mL of distilled water, extract for 5 hours, filter, and dry the residue to obtain the water-extracted residue of Cistanche saline.

(1) Pulverize the water extraction residue of Cistanche saline, add 15 times the weight of 2.5% mixed acid solution of citric acid and lactic acid, mix well, extract for 2 hours, centrifuge the extract to obtain supernatant and precipitate, and The precipitation is dried to obtain insoluble dietary fiber;

(2) After concentrating the supernatant, add 3 times the volume of absolute ethanol, ethanol precipitation for 5 hours to obtain flocs, and dry to obtain soluble dietary fiber.

Example 13: Preparation of Dietary Fiber in Cistanche Cistanche Water Extraction Residue

Grind Cistanche saline, weigh 1000g, add 10000mL of distilled water, extract for 5 hours, filter, and dry the residue to obtain the water-extracted residue of Cistanche saline.

(1) Pulverize the water extraction residue of Cistanche saline, add 15 times the weight of 5% mixed acid (4:1 by volume) solution of hydrochloric acid and formic acid, mix well, extract for 2.5 hours, and centrifuge the extract, Obtain supernatant and precipitate, and dry the precipitate to obtain insoluble dietary fiber;

(2) After concentrating the supernatant, add 3 times the volume of absolute ethanol, ethanol precipitation for 5 hours to obtain flocs, and dry to obtain soluble dietary fiber.

Example 14: Preparation of Dietary Fiber in Cistanche Cistanche Water Extraction Residue

Grind Cistanche cistanche, weigh 1000g, add 10000mL of distilled water, extract for 5h, filter, and dry the residue to obtain the water extraction residue of Cistanche cistanche.

(1) Cistanche tubulosa water extraction residue is pulverized, 3% hydrochloric acid and formic acid mixed acid (volume ratio is 2.5:1) solution of 25 times of its weight are added, mixed, extracted for 2.5h, the extract is centrifuged, Obtain supernatant and precipitate, and dry the precipitate to obtain insoluble dietary fiber;

(2) After concentrating the supernatant, add 5 times the volume of dehydrated ethanol, ethanol precipitation for 5 hours to obtain flocs, and dry to obtain soluble dietary fiber.

Example 15: Preparation of Dietary Fiber in Cistanche Cistanche Water Extraction Residue

Crushed Cistanche deserticola, weighed 500g, added 6000mL distilled water, extracted for 1 hour, filtered, and dried the residue to obtain the water-extracted residue of Cistanche deserticola.

(1) Crush desert cistanche water extraction residue, add 0.5% NaOH solution 8 times its weight, mix well, extract for 1 hour, centrifuge the extract to obtain supernatant and precipitate, dry the precipitate to obtain insoluble dietary fiber ;

(2) After concentrating the supernatant, add 4 times the volume of absolute ethanol, ethanol precipitation for 2 hours to obtain flocs, and dry the flocs to obtain soluble dietary fiber.

Example 16: Preparation of Dietary Fiber in the Alcohol Extraction Residue of Cistanche Deserticola

Grind Cistanche cistanche, weigh 200g, add 1800mL of 50% ethanol solution, extract for 70min, filter, and dry the residue to obtain the alcohol extraction residue of Cistanche cistanche.

(1) Grind the residue of Cistanche tubulosa alcohol extraction, add 10 times the weight of 10% KOH solution, mix well, extract for 3 hours, centrifuge the extract to obtain supernatant and precipitate, dry the precipitate to obtain insoluble diet fiber;

(2) After concentrating the supernatant, add 5 times the volume of dehydrated ethanol, ethanol precipitation for 4 hours to obtain flocs, and dry to obtain soluble dietary fiber.

Example 17: Preparation of Dietary Fiber in Cistanche Cistanche Water Extraction Residue

Grind Cistanche saline, weigh 1000g, add 10000mL of distilled water, extract for 5 hours, filter, and dry the residue to obtain the water-extracted residue of Cistanche saline.

(1) Crush the residue of Cistanche saline, add 20 times the weight of 3% NaOH solution and KOH solution mixed lye (volume ratio is 1:1), mix well, extract for 2 hours, and centrifuge the extract , obtain the supernatant and precipitate, and dry the precipitate to obtain insoluble dietary fiber;

(2) After concentrating the supernatant, add 4 times the volume of dehydrated ethanol, ethanol precipitation for 5 hours to obtain flocs, and dry to obtain soluble dietary fiber.

Example 18: Preparation of Dietary Fiber in Cistanche Alcohol Extraction Residue

Crush Cistanche, weigh 600g, add 9000mL of 60% ethanol solution, extract for 90min, filter, and dry the residue to obtain the alcohol extraction residue of Cistanche.

(1) Grind the residue of desert cistanche alcohol extraction, add 1.75% NaOH solution 30 times its weight, mix well, extract for 1.5h, centrifuge the extract to obtain supernatant and precipitate, dry the precipitate to obtain insoluble Dietary fiber;

(2) After concentrating the supernatant, add 3 times the volume of absolute ethanol, ethanol precipitation for 4 hours to obtain flocs, and dry to obtain soluble dietary fiber.

Example 19: Preparation of Dietary Fiber in the Alcohol Extraction Residue of Cistanche Deserticola

Grind Cistanche cistanche, weigh 200g, add 1800mL of 50% ethanol solution, extract for 70min, filter, and dry the residue to obtain the alcohol extraction residue of Cistanche cistanche.

(1) Crush the residue of Cistanche tubulosa alcohol extraction, add 15 times the weight of 5.25% KOH solution, mix well, extract for 2.5 hours, centrifuge the extract to obtain supernatant and precipitate, dry the precipitate to obtain insoluble Dietary fiber;

(2) After concentrating the supernatant, add 3.5 times the volume of absolute ethanol, ethanol precipitation for 4 hours to obtain flocs, and dry to obtain soluble dietary fiber.

Example 20: Preparation of Dietary Fiber in the Alcohol Extraction Residue of Cistanche Deserticola

Grind Cistanche cistanche, weigh 200g, add 1800mL of 50% ethanol solution, extract for 70min, filter, and dry the residue to obtain the alcohol extraction residue of Cistanche cistanche.

(1) Grind the residue of Cistanche tubulosa alcohol extraction, add 19 times the weight of 6% NaOH solution and KOH solution mixed lye (volume ratio is 2:1), mix well, extract for 2h, and centrifuge the extract , obtain the supernatant and precipitate, and dry the precipitate to obtain insoluble dietary fiber;

(2) After concentrating the supernatant, add 4.5 times the volume of absolute ethanol, ethanol precipitation for 4 hours to obtain flocs, and dry to obtain soluble dietary fiber.

Moistening bowel and laxative experiment of dietary fiber of Cistanche deserticola

The dietary fiber prepared in Example 8 was taken to carry out the small intestine motility experiment.

Animals: ICR male mice, weighing 32-38 g, 10 in each group, purchased from Beijing Weitong Lihua Experimental Animal Co., Ltd.

The experiment set up a blank control group, a model group, a positive control group and three dosage groups. After giving the tested samples for 14 days, the mice in each group were fasted for 16 hours, the model group, the positive control group and the three dosage groups were intragastrically given compound diphenoxylate (10 mg/kg), and the blank control group was given distilled water.

After compound diphenoxylate was given 0.5h, the dose group was given 5% gac (containing 10% gum arabic) containing the corresponding test sample respectively, the blank group and the model group were gavaged with gac, and the positive control group was gavaged with diacetate. Pyridine.

After 25 minutes of gavage with activated charcoal, the mice were killed by dislocation of the cervical spine, the abdominal cavity was opened to separate the mesentery, the upper end of the intestinal tube from the pylorus, the lower end to the ileocecal part was cut, placed on a tray, the small intestine was gently pulled into a straight line, and the length of the intestinal tube was measured as "small intestine total length", from the pylorus to the front of the activated carbon is the "activated carbon propulsion length", and the activated carbon propulsion rate is calculated according to the following formula:

Activated carbon propulsion rate (%) = activated carbon propulsion length (cm) / total length of small intestine (cm) × 100%

The experimental results are shown in Table 1.

Table 1 Effect of cistanche dietary fiber on intestinal motility in mice with constipation

*P<0.05: Compared with the model group, the difference is significant

The above experimental results show that the dietary fiber of Cistanche can promote the propulsion of activated carbon in the small intestine of mice with constipation in a dose-dependent manner, indicating that the dietary fiber of Cistanche can promote small intestinal peristalsis and laxative effect.

Inhibition of α-Glucosidase Activity by Cistanche Dietary Fiber

The dietary fiber prepared in Example 10 was used for the experiment of inhibiting α-glucosidase activity.

Add 50 μL of 50 mmol/L pH6.8 phosphate buffer solution, 2.5 μL of 5 mmol/L reduced glutathione, 2.5 μL of 0.2 U/mL α-glucosidase, and 10 μL of inhibitors of different concentrations in a 96-well plate, and mix After uniformity, keep the temperature in a constant temperature box at 37°C for 10 minutes, add 5 μL of 20 mmol/L pNPG, then place it in a constant temperature box at 37 °C for 10 minutes, add 0.1mol/L Na ₂ CO ₃ 50 μL to stop the reaction, and measure the absorbance at 405 nm with a microplate reader value. Inhibitors were not added to the blank group, pNPG and inhibitors were not added to the blank background group, pNPG was not added to the inhibitor background group, and the volume was made up with distilled water, and other operations were the same as the inhibitor group. The inhibition rate of inhibitor to α-glucosidase activity is calculated according to the following formula:

Inhibition rate (%)=[(A1-A2)-(A3-A4)]/(A1-A2)×100

Among them: A1 is the absorbance value of the blank group, A2 is the absorbance value of the blank background group, A3 is the absorbance value of the inhibitor group, and A4 is the absorbance value of the inhibitor background group.

Table 2 The inhibitory rate of Cistanche dietary fiber to α-glucosidase

Dietary Fiber Concentration Inhibition rate(%) 1 23.06 2 37.27 3 41.02 4 48.26 5 56.20

It can be seen that the dietary fiber of Cistanche deserticola has obvious inhibitory effect on α-glucosidase, and with the increase of dietary fiber concentration, the inhibitory effect on α-glucosidase is gradually enhanced.

The cistanche dietary fiber prepared in other examples also has excellent laxative properties and inhibits α-glucosidase activity.

The applicant declares that the above description is only a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto, and those skilled in the art should understand that any person skilled in the art should be aware of any disclosure in the present invention Within the technical scope, easily conceivable changes or substitutions all fall within the scope of protection and disclosure of the present invention.

Claims (10)

1. the preparation method of a Herba Cistanches dietary fiber, it is characterised in that described preparation method comprises the steps:

(1) Herba Cistanches residue is mixed with extracting solution, extracted, solid-liquid separation, obtain supernatant and precipitation, described precipitation drying obtains insoluble dietary fiber, and wherein, described extracting solution is any one in acid solution, alkali liquor or enzyme liquid;

(2) supernatant after supernatant or concentration is mixed with dehydrated alcohol carry out precipitate with ethanol, solid-liquid separation, obtain floccule, floccule is dried, obtain water soluble dietary fiber.

2. preparation method according to claim 1, it is characterized in that, the described Herba Cistanches residue of step (1) is that the residue after Herba Cistanches water carries, the residue after Herba Cistanches alcohol extraction, Herba Cistanches first carry out water and carry the residue after carrying out alcohol extraction again or Herba Cistanches first carries out alcohol extraction and carries out the combination of any one or at least two in the residue after water proposes again;

Preferably, described Herba Cistanches is the combination of any one or at least two in the fleshy stem of Desert Herba Cistanches, Cistanche Tubulosa, Saline Cistanche Herb, Lanzhou Herba Cistanches or Herba Cistanches sinensis.

3. preparation method according to claim 1 and 2, it is characterised in that the described Herba Cistanches residue of step (1) mixes with extracting solution after smashing again;

Preferably, step (1) described acid solution is the combination of any one or at least two in hydrochloric acid, formic acid, acetic acid, citric acid, malic acid, oxalic acid, sulphuric acid, nitric acid or lactic acid, the volume ratio being preferably the mix acid liquor of hydrochloric acid and formic acid, hydrochloric acid and formic acid is preferably 1-4:1;

Preferably, the concentration of step (1) described acid solution is 0.1%-10%, it is preferred to 0.5%-5%;

Preferably, 8-30 times that addition is Herba Cistanches mass of residue of step (1) described acid solution, it is preferred to 10-20 times;

Preferably, the extraction time of step (1) described acid solution is 1-3h.

4. according to the preparation method one of claim 1-3 Suo Shu, it is characterized in that, step (1) described alkali liquor is NaOH solution and/or KOH solution, it is preferred to the volume ratio of the mixed alkali liquor of NaOH solution and KOH solution, NaOH solution and KOH solution is preferably 1:1;

Preferably, the concentration of described alkali liquor is 0.5%-10%, it is preferred to 0.5%-3%;

Preferably, 8-30 times that addition is Herba Cistanches mass of residue of step (1) described alkali liquor, it is preferred to 10-20 times;

Preferably, the extraction time of step (1) described alkali liquor is 1-3h.

5. according to the preparation method one of claim 1-4 Suo Shu, it is characterised in that the described enzyme liquid of step (1) is the buffer containing hydrolytic enzyme, and described hydrolytic enzyme is amylase, protease and glucosidase;

Preferably, Herba Cistanches residue described in step (1) first mixes with buffer, is sequentially carried out amylase enzymolysis, protease hydrolyzed and glucosidase enzymolysis afterwards;

Preferably, the condition of described amylase enzymolysis is 50-95 DEG C, 20-60min;

Preferably, the condition of described protease hydrolyzed is 40-70 DEG C, 20-60min;

Preferably, the condition of described glucosidase enzymolysis is 40-60 DEG C, 20-60min;

Preferably, the addition of described amylase, protease and glucosidase independently be 100-3000U/g, it is preferred to 2000U/g;

Preferably, described buffer is the combination of any one or at least two in phosphate buffer, Tris-HCl buffer or citrate buffer solution, it is preferred to the combination of phosphate buffer and Tris-HCl buffer;

Preferably, the pH value of described buffer is 6.5-8.5.

6. according to the preparation method one of claim 1-5 Suo Shu, it is characterised in that the addition of step (2) described dehydrated alcohol is 3-5 times of supernatant volume after supernatant volume or concentration.

7. according to the preparation method one of claim 1-6 Suo Shu, it is characterised in that described preparation method comprises the steps:

(1) Herba Cistanches residue is mixed with the buffer that pH value is 6.5-8.5, afterwards, be added thereto to amylase, at 50-95 DEG C of enzymolysis 20-60min;Then, protease is added, at 40-70 DEG C of enzymolysis 20-60min; Adding glucosidase, at 40-60 DEG C of enzymolysis 20-60min, obtain enzymolysis solution, wherein, the addition of described amylase, protease and glucosidase independently be 100-3000U/g; By enzymolysis solution solid-liquid separation, obtaining supernatant and precipitation, described precipitation drying obtains insoluble dietary fiber;

(2) being mixed with dehydrated alcohol by the supernatant after supernatant or concentration and carry out precipitate with ethanol, obtain floccule, wherein, the addition of dehydrated alcohol is 3-5 times of supernatant or the supernatant volume after concentrating; Floccule is dried, obtains water soluble dietary fiber.

8. the Herba Cistanches dietary fiber obtained according to the preparation method one of claim 1-7 Suo Shu, described Herba Cistanches dietary fiber includes water soluble dietary fiber and insoluble dietary fiber.

9. the Herba Cistanches dietary fiber according to claim 8 application in food or biomedicine field.

10. the application of Herba Cistanches dietary fiber according to claim 9, it is characterised in that described Herba Cistanches dietary fiber is for preparing auxiliary treatment diabetes, auxiliary treatment loosening bowel to relieve constipation or improving food or the health preparation of constipation.