CN108635408B

CN108635408B - Mulberry leaf extract, preparation method and application thereof, and health care wine

Info

Publication number: CN108635408B
Application number: CN201810901351.8A
Authority: CN
Inventors: 江丹; 石万银; 陈志元
Original assignee: Jingpai Zhengtang Pharmaceutical Co ltd
Current assignee: Jingpai Zhengtang Pharmaceutical Co ltd
Priority date: 2018-08-09
Filing date: 2018-08-09
Publication date: 2022-02-18
Anticipated expiration: 2038-08-09
Also published as: CN108635408A

Abstract

A mulberry leaf extract, a preparation method and application thereof and health care wine relate to the field of extraction of plant components. The preparation method of the mulberry leaf extract provided by the embodiment of the invention is that the mulberry leaf water crude extract is filtered to obtain mulberry leaf extract; coating the mulberry leaf extracting solution with cation exchange resin, and standing and adsorbing for 1-3 hours after finishing sample coating; eluting with purified water, and eluting with ammonia water to obtain ammonia water eluate; the mulberry leaf extract is obtained by concentrating and drying the ammonia water eluent, and the preparation method has the advantages of simple process, low production cost, high DNJ purity and high yield, and is suitable for industrial mass production; the DNJ in the mulberry leaf extract of the embodiment of the invention has high purity; the mulberry leaf extract provided by the embodiment of the invention is used for preparing health care wine; the health wine provided by the embodiment of the invention contains a large amount of effective functional components DNJ.

Description

Mulberry leaf extract, preparation method and application thereof, and health care wine

Technical Field

The invention relates to the field of extraction of plant components, and particularly relates to a mulberry leaf extract, a preparation method and application thereof, and health care wine.

Background

Mulberry leaves (mulberry leaves), also known as indisputable fans, are leaves of Morus alba l, a family of moraceae, native to china and korea, distributed in tropical asia, tropical africa, temperate north and american regions. The mulberry leaves are rich in nutrition, contain various active ingredients, have the characteristics of mild drug properties, no toxic or side effect and the like, are classified as medicine and food by the Ministry of health of China, and have very high medicinal and edible values. The mulberry leaves contain various effective functional components, such as amino acids, proteins, polysaccharides, flavones, alkaloids, phytosterols, isoquercitrin, various vitamins, trace elements and the like. The mulberry leaves in China have abundant resources, various varieties and wide distribution area, but the utilization rate is low. Except for silkworm breeding, most of the silkworm droppings and decays, which causes great economic resource waste. Therefore, the method has important significance in reasonably and effectively utilizing the waste mulberry leaf resources and developing products with high added values.

1-Deoxynojirimycin (DNJ) is a unique polyhydroxy alkaloid in folium Mori, has a chemical name of (2R,3R,4R) -2- (hydroxymethyl) -3,4, 5-trihydroxypiperidine and a molecular formula of C₆H₁₃NO₄The molecular mass is 163.17, and the lone electron pair on the nitrogen atom in the molecular structure can accept proton to show weak alkalinity. The Morus plant is the highest content of all DNJ-containing plants discovered at present, wherein the DNJ content in mulberry leaves is 0.11%, and the DNJ content in mulberry roots is 0.14%. DNJ is an azasugar which is not metabolized by a human body, has a structure similar to that of monosaccharide, can simulate enzymatic hydrolysis reaction of a plurality of saccharides and inhibit glycosidase activity, so that the DNJ is a good glycosidase inhibitor, can effectively and competitively inhibit the activity of alpha-glycosidase and reduce the decomposition and digestion of the saccharides, thereby reducing the blood sugar concentration in the human body. DNJ and the derivative thereof have multiple functional activities, can be used for developing new medicines and researching certain biochemical approaches, or preparing medicines and health-care foods for treating diabetes, cancer, obesity and AIDS, and have important application values in the aspects of clinical treatment of diabetes, molecular biology and the like.

DNJ is the only internationally recognized zero-injury biological agent in the current diabetes treatment medicines, has obvious effect on treating type II diabetes, but has a plurality of difficulties in preparation and acquisition. Most DNJ sold in the market is obtained by chemical synthesis, but the existing DNJ synthesis method has the defects of harsh conditions, complex process, high cost, unknown safety and the like, and the medicinal activity of the DNJ is far lower than that of natural DNJ, so that the method for obtaining high-purity DNJ from natural products is the main direction of the current and future research. The extraction method of DNJ in mulberry leaves mainly comprises a decoction method, an immersion method, a microwave-assisted method and the like, which are all available, but have the defects of long extraction time and low extraction rate, and are not beneficial to large-scale production. The methods for separating and purifying alkaloids include: two-phase solvent extraction, crystallization, silica gel column chromatography, resin method, membrane separation, etc. The DNJ is prepared by a specific extraction method, namely heating reflux extraction is carried out to obtain a coarse mulberry leaf water extract with more impurities, then adsorption is carried out through macroporous resin to remove impurities such as flavone and polysaccharide, and enrichment and purification are carried out through ion exchange resin.

Disclosure of Invention

The invention aims to provide a preparation method of a mulberry leaf extract, which has the advantages of simple process, low production cost, high DNJ purity and high yield and is suitable for industrial mass production.

Another object of the present invention is to provide a mulberry leaf extract in which DNJ has high purity.

The invention also aims to provide application of the mulberry leaf extract in preparing health care wine.

Another object of the present invention is to provide a health wine containing a large amount of DNJ as an effective functional ingredient.

The technical problem to be solved by the invention is realized by adopting the following technical scheme.

The invention provides a preparation method of a mulberry leaf extract, which comprises the following steps:

filtering the folium mori water crude extract to obtain a folium mori extract;

coating the mulberry leaf extracting solution with cation exchange resin, and standing and adsorbing for 1-3 hours after finishing sample coating;

eluting with purified water, and eluting with ammonia water to obtain ammonia water eluate;

concentrating and drying the ammonia water eluate to obtain folium Mori extract.

Further, in the preferred embodiment of the present invention, the preparation method of the aqueous folium mori crude extract comprises the following steps: taking mulberry leaf medicinal materials and deionized water, wherein the dosage ratio of the mulberry leaf medicinal materials to the deionized water is 1 g: 15-20 mL, heating and refluxing for extraction for 2-3 times at 80-90 ℃, each time for 1-2 hours, and filtering by a 100-mesh filter screen to obtain a mulberry leaf water crude extract.

Further, in the preferred embodiment of the present invention, the sample loading speed is 1-4 BV/h.

Further, in a preferred embodiment of the present invention, the elution method is: eluting with 3-5 volume times of resin of purified water, and then eluting with 3-5 volume times of resin of ammonia water with the concentration of 0.25-0.50 mol/L.

Further, in a preferred embodiment of the present invention, the elution speed is 2-4 BV/h.

The invention provides a mulberry leaf extract which is prepared by adopting the preparation method of the mulberry leaf extract.

The invention provides application of a mulberry leaf extract, and the mulberry leaf extract is used for preparing health care wine.

The invention provides health wine which comprises white spirit and the mulberry leaf extract, wherein the addition amount of the mulberry leaf extract is 1-8% of the mass of the white spirit.

Further, in the preferred embodiment of the invention, the alcohol content of the white spirit is 40-60% vol.

Further, in a preferred embodiment of the present invention, the wine further comprises phytic acid enzyme and pancreatic juice substitute enzyme, wherein the addition amount of the phytic acid enzyme is 0.5% -1% of the mass of the white wine, and the addition amount of the pancreatic juice substitute enzyme is 0.5% -1% of the mass of the white wine.

The mulberry leaf extract, the preparation method and the application thereof of the embodiment of the invention and the health care wine have the beneficial effects that: the preparation method of the mulberry leaf extract provided by the embodiment of the invention is that the mulberry leaf water crude extract is filtered to obtain mulberry leaf extract; coating the mulberry leaf extracting solution with cation exchange resin, and standing and adsorbing for 1-3 hours after finishing sample coating; eluting with purified water, and eluting with ammonia water to obtain ammonia water eluate; the mulberry leaf extract is obtained by concentrating and drying the ammonia water eluent, and the preparation method has the advantages of simple process, low production cost, high DNJ purity and high yield, and is suitable for industrial mass production; the DNJ in the mulberry leaf extract of the embodiment of the invention has high purity; the mulberry leaf extract provided by the embodiment of the invention is used for preparing health care wine; the health wine provided by the embodiment of the invention contains a large amount of effective functional components DNJ.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.

FIG. 1 is a process flow chart of a method for preparing mulberry leaf extract according to an embodiment of the present invention.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.

The mulberry leaf extract, the preparation method and the application thereof and the health care wine of the embodiment of the invention are specifically explained below.

Referring to fig. 1, an embodiment of the present invention provides a method for preparing a mulberry leaf extract, which includes the following steps:

(1) extraction: taking mulberry leaf medicinal materials and deionized water, wherein the dosage ratio of the mulberry leaf medicinal materials to the deionized water is 1 g: 15-20 mL, heating and refluxing for extraction for 2-3 times at 80-90 ℃ for 1-2 hours each time, coarsely filtering with a 100-mesh filter screen, and collecting and combining filtrates to obtain the mulberry leaf water coarse extract.

(2) And (3) filtering: filtering the folium Mori water crude extract, generally by disc centrifuge, to obtain clear folium Mori extract.

(3) Resin adsorption: and (3) applying the mulberry leaf extracting solution to cation exchange resin, wherein the sample application speed is generally 1-4 BV/h, and standing and adsorbing for 1-3 hours after sample application is finished. As the molecular structure of DNJ contains-NH, the DNJ exists in a positive ion form under neutral and acidic conditions, the DNJ can be enriched and separated from the mulberry leaf extract by using cation exchange resin.

(4) Resin elution: firstly adding purified water for elution, and then adding ammonia water for elution, wherein the general elution method comprises the following steps: firstly, eluting with purified water with the volume of 3-5 times of that of the resin, and then eluting with ammonia water with the volume of 3-5 times of that of the resin, wherein the concentration of the ammonia water is 0.25-0.50 mol/L, and the elution speed is generally 2-4 BV/h, so as to obtain ammonia water eluent.

(5) Concentrating and drying: concentrating and drying the ammonia water eluate to obtain folium Mori extract.

The preparation method of the mulberry leaf extract has the following advantages:

the method has the advantages of simple process, short production period, low cost and easy popularization: the conventional separation and purification process of the mulberry leaf extract requires macroporous resin for adsorption and impurity removal. The embodiment of the invention removes impurities and most of cations from the crude mulberry leaf water extract by filtration, then directly enriches and purifies the mulberry leaf DNJ by cation exchange resin, and then elutes the purified DNJ by using a proper solvent, thereby obtaining the mulberry leaf extract with higher DNJ content, and simultaneously achieving the purposes of simplifying the process, shortening the production period and reducing the production cost.

(II) high transfer rate: the conventional extraction, separation and purification process of the mulberry leaf DNJ requires twice resin, one time is used for removing impurities from the macroporous resin, one time is used for adsorbing and purifying the cationic exchange resin, the process is complex, and the DNJ loss can be caused.

(III) the extraction purity is high: according to the embodiment of the invention, the process method of filtering the mulberry leaf water crude extract and then separating and purifying the mulberry leaf water crude extract by using the cation exchange resin can play a role in removing impurities, enriching and eluting, so that the mulberry leaf extract with higher DNJ content is obtained.

The embodiment of the invention provides a mulberry leaf extract, which is prepared by adopting the preparation method of the mulberry leaf extract.

The embodiment of the invention provides application of a mulberry leaf extract in preparing health care wine.

The embodiment of the invention provides health wine which comprises white spirit and the mulberry leaf extract, wherein the addition amount of the mulberry leaf extract is 1-8% of the mass of the white spirit, and the alcoholic strength of the white spirit is 40-60% vol. Generally, white spirit with the alcoholic strength of 40-60 vol% is taken, folium mori extract is added, the mixture is uniformly stirred, and the health wine can be obtained after standing for 5-30 days and filtering, and is particularly suitable for being drunk by diabetics.

In the embodiment, the wine also comprises phytic acid enzyme and pancreatin substitute enzyme, wherein the addition amount of the phytic acid enzyme is 0.5-1% of the mass of the wine, and the addition amount of the pancreatin substitute enzyme is 0.5-1% of the mass of the wine. Phytase, also known as Phytic acid hydrolase, catalyzes the hydrolysis of Phytic acid (Phytic acid) to release Inositol and orthophosphate. The phytic acid can be easily combined with important minerals in the intestinal tract, so the phytic acid enzyme is a molecule for hindering nutrient absorption, and the phytic acid enzyme is supplemented to decompose the ingested phytic acid, so that the intestinal tract can absorb various important minerals.

The pancreatin substitute enzyme is a mixed fungal enzyme obtained by fermenting aspergillus oryzae and rhizopus, and the mixed fungal enzyme comprises fungal protease, amylase and lipase. The pancreatin substitute enzyme has the function of pancreatic enzyme and is prepared by preparing fungal protease, amylase and lipolytic enzyme obtained by fermenting special microorganisms. The ability of hydrolyzing proteins by proteolytic enzymes (exo-and endo-peptide enzymes) in pancreatic juice substitute enzymes is quite excellent, and most of soluble proteins can be effectively hydrolyzed: exo-peptide peptidase (exoprotease) can hydrolyze peptide at the end of the protein peptide chain and separate amino acids; endopeptidase (endoprotease) can hydrolyze the peptide chain inside the protein and divide the protein into several peptide chains. Pancreatic juice-replacement enzymes randomly break down alpha-1, 4-glucoside bonds (alpha-1,4-glucosidic bonds) of polysaccharides. In addition, the method can help dextrinization of soluble starch and decompose glucose and maltose. Pancreatin-substituting enzymes also have lipase activity and, like pancreatic enzymes, catalyze the hydrolysis of Triglycerides (TG) to produce Monoglycerides (MG), Diglycerides (DG), glycerol and free fatty acids. The phytic acid enzyme and the pancreatic juice enzyme are added to replace the enzyme, so that the harm of alcohol to the body can be reduced, and the health-care effect is achieved.

The features and properties of the present invention are described in further detail below with reference to examples.

Example 1

The embodiment provides a mulberry leaf extract, which is extracted by adopting the following preparation method:

(1) extraction: extracting 100kg folium Mori with 1500L water for 2 times at 80 deg.C for 2 hr, coarse-filtering with 100 mesh filter screen, collecting filtrate, and mixing to obtain folium Mori water crude extract.

(2) And (3) filtering: filtering the folium Mori water coarse extractive solution with disc centrifuge to obtain clear folium Mori extractive solution.

(3) Resin adsorption: and (3) allowing the mulberry leaf extracting solution to stand and adsorb for 2 hours after the loading of a 001X 7 type cation (H type) exchange resin at the flow rate of 2 BV/H.

(4) Resin elution: after adsorption, firstly adding 4 volume times of purified water for elution, and then adding 5 volume times of resin and 0.25mol/L ammonia water for elution, wherein the elution speed is 2BV/h, and obtaining ammonia water eluent.

(5) Concentrating and drying: concentrating and drying the ammonia water eluate to obtain 1.05kg folium Mori extract. The DNJ content of the product is 14.69%, and the yield is 1.05%.

The embodiment also provides health wine, which is prepared by taking 5kg of white spirit with the alcoholic strength of 50% vol, adding 100g of mulberry leaf extract, uniformly stirring, completely dissolving the mulberry leaf extract in the white spirit, standing for 10 days, allowing a small amount of precipitate to exist at the bottom of a wine body, and filtering.

Example 2

(1) extraction: extracting 200kg folium Mori with 4000L water for 3 times at 85 deg.C, coarse-filtering with 100 mesh filter screen, collecting filtrate, and mixing to obtain folium Mori water crude extract.

(3) Resin adsorption: adding the filtered folium Mori extractive solution into R120 type (Na type) cation exchange resin at flow rate of 2BV/h, standing for adsorbing for 2 hr.

(4) Resin elution: after adsorption, firstly adding 4 volume times of purified water for elution, and then adding 4 volume times of resin and 0.30mol/L ammonia water for elution, wherein the elution speed is 2BV/h, and obtaining ammonia water eluent.

(5) Concentrating and drying: concentrating and drying the ammonia water eluate to obtain 2.20kg folium Mori extract. The detection proves that the DNJ content is 13.98%, and the yield is 1.10%.

The embodiment also provides health wine, which is prepared by taking 5kg of white spirit with the alcoholic strength of 50% vol, adding 200g of mulberry leaf extract, uniformly stirring, completely dissolving the mulberry leaf extract in the white spirit, standing for 10 days, allowing a small amount of precipitate to exist at the bottom of a wine body, and filtering.

Example 3

(1) extraction: extracting 300kg folium Mori with water for 2 times (6000L each time), extracting at 90 deg.C for 2 hr, coarse-filtering with 100 mesh filter screen, collecting and mixing filtrates to obtain folium Mori water coarse extractive solution.

(3) Resin adsorption: adding the filtered folium Mori extractive solution into LSI-010 type (H type) cation exchange resin at flow rate of 2BV/H, standing for adsorbing for 2 hr.

(4) Resin elution: after adsorption, firstly adding 4 volume times of purified water for elution, and then adding 3 volume times of resin and 0.50mol/L ammonia water for elution, wherein the elution speed is 2BV/h, and obtaining ammonia water eluent.

(5) Concentrating and drying: concentrating and drying the ammonia water eluate to obtain folium Mori extract 3.45 kg. The DNJ content of the product is 13.88 percent, and the yield is 1.15 percent.

The embodiment also provides health-care wine, which is prepared by taking 5kg of white wine with the alcoholic strength of 50% vol, adding 250g of mulberry leaf extract, 25g of phytic acid enzyme and 25g of pancreatic juice substitute enzyme, uniformly stirring, dissolving all the mulberry leaf extract, the phytic acid enzyme and the pancreatic juice substitute enzyme in the white wine, standing for 10 days, only a small amount of precipitate exists at the bottom of the wine body, and filtering.

In conclusion, the preparation method of the mulberry leaf extract provided by the embodiment of the invention has the advantages of simple process, low production cost, high DNJ purity and high yield, and is suitable for industrial mass production; the DNJ in the mulberry leaf extract of the embodiment of the invention has high purity; the mulberry leaf extract provided by the embodiment of the invention is used for preparing health care wine; the health wine provided by the embodiment of the invention contains a large amount of effective functional components DNJ.

The embodiments described above are some, but not all embodiments of the invention. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Claims

1. A preparation method of a mulberry leaf extract is characterized by being extracted by adopting the following preparation method:

(1) extraction: extracting 300kg folium Mori with water for 2 times (6000L each time), extracting under reflux at 90 deg.C for 2 hr, coarse-filtering with 100 mesh filter screen, collecting and mixing filtrates to obtain folium Mori water coarse extractive solution;

(2) and (3) filtering: filtering the folium Mori water coarse extractive solution with a disc centrifuge to obtain clear folium Mori extractive solution;

(3) resin adsorption: putting the filtered mulberry leaf extracting solution on an LSI-010 type cation exchange resin at the flow rate of 2BV/h, and standing and adsorbing for 2 hours after the sample loading is finished;

(4) resin elution: after adsorption, firstly adding 4 volume times of resin into purified water for elution, and then adding 3 volume times of resin into 0.50mol/L ammonia water for elution, wherein the elution speed is 2BV/h, so as to obtain ammonia water eluent;

(5) concentrating and drying: concentrating and drying the ammonia water eluate to obtain 3.45kg folium Mori extract, wherein DNJ content is 13.88% and yield is 1.15% by detection.