CN113004433A

CN113004433A - Rhizoma polygonati fructan and preparation method and application thereof

Info

Publication number: CN113004433A
Application number: CN202110255156.4A
Authority: CN
Inventors: 彭腾; 吴明一; 张军银; 罗兰; 周志鹏; 杨莲; 陈胡兰
Original assignee: Chengdu University of Traditional Chinese Medicine
Current assignee: Chengdu University of Traditional Chinese Medicine
Priority date: 2021-03-09
Filing date: 2021-03-09
Publication date: 2021-06-22
Also published as: CN116253806A

Abstract

The invention relates to polygonatum fructan and a preparation method and application thereof, wherein the polygonatum fructan has a structure represented by the following general formula I: in the formula (I), m, n and p represent the number of sugar repeating units, m, n and p are natural numbers including 0, and m + n > p. The polygonatum fructan has a structure represented by the following general formula I, wherein beta (2 → 6) fructose glycosidic bond is taken as a main chain, and side chain fructose is connected to the main chain by the beta (2 → 1) glycosidic bond. The polygonatum fructan and the pharmaceutical composition containing the polygonatum fructan both have certain activity of promoting the proliferation of probiotics, so the polygonatum fructan and the pharmaceutical composition containing the polygonatum fructan have medical value in the preparation of the beneficial bacteria drugs.

Description

Rhizoma polygonati fructan and preparation method and application thereof

Technical Field

The invention relates to the technical field of food and medicine, in particular to polygonatum sibiricum fructan and a preparation method and application thereof.

Background

Rhizoma Polygonati is a plant of genus rhizoma Polygonati of family Liliaceae, and is named because of its tonic effect. The genus plant mainly includes Polygonatum sibiricum (Polygonatum sibiricum), Polygonatum kingianum (Polygonatum kingianum), Polygonatum cyrtonema (Polygonatum cyrtonema), Polygonatum conatum (Polygonatum cathayensis), Polygonatum sibiricum (Polygonatum cathayensis), Polygonatum odoratum (Polygonatum odoratum), Polygonatum minor (Polygonatum humile), Polygonatum odoratum (Polygonatum inflatum), Polygonatum falcatum (Polygonatum flagatum), Polygonatum sibiricum curvatum (Polygonatum curvatum), Polygonatum cyrtonema (Polygonatum desoxyphyllum), Polygonatum longum (Polygonatum sibiricum), Polygonatum crudella (Polygonatum hirtum), Polygonatum giganteum (Polygonatum sibiricum), Polygonatum giganteum (Polygonatum giganteum), Polygonatum sibiricum (Polygonatum sibiricum), Polygonatum sibiricum minor rhizome (Polygonatum), Polygonatum sibiricum uncitum rhizome minor rhizome (Polygonatum sibiricum grandiflorum), Polygonatum sibiricum (Polygonatum sibiricum grandiflorum medicine (Polygonatum), Polygonatum sibiricum grandiflorum crispum), Polygonatum (Polygonatum rhizome), Polygonatum sibiricum grandiflorum), Polygonatum sibiricum (Polygonatum sibiricum), Polygonatum sibiricum) blend root of Polygonatum sibiricum, Polygonatum sibiricum (Polygonatum sibiricum), Polygonatum sibiricum grandiflorum paniculatum medicine (Polygonatum), Polygonatum sibiricum grandiflorum paniculatum), Polygonatum sibiricum (Polygonatum sibiricum), Polygonatum sibiricum grandiflorum paniculatum rhizome (Polygonatum sibiricum), Polygonatum sibiricum grandiflorum paniculatum), Polygonatum (Polygonatum sibiricum) blend root of, Rhizoma Polygonati (Polygonatum macrocarpium), rhizoma Polygonati Sinapis (Polygonatum sibiricum), rhizoma Polygonati officinalis (Polygonatum verticillium), and rhizoma Polygonati stenophyllum (Polygonatum stenophyllum). The medicinal part of the sealwort is rhizome, sweet and mild in taste. Tonify qi and yin, invigorate spleen, moisten lung and tonify kidney. Can be used for treating spleen deficiency, stomach weakness, asthenia, xerostomia, dry cough due to lung deficiency, essence and blood deficiency, internal heat, and diabetes. Rhizome of Polygonatum plant can be used as medicine, and the rhizome mainly comprises the following chemical components: rhizoma Polygonati polysaccharide, mucus, starch, steroid saponin, anthraquinone compounds, alkaloid, cardiac glycoside, lignan, vitamins, fat, protein and various amino acids useful for human body, wherein rhizoma Polygonati polysaccharide is the most effective component in rhizoma Polygonati.

However, the chemical structure of plant polysaccharide is very complex, the same plant often contains a plurality of different types of polysaccharide, and the research on the biological efficacy of polysaccharide is less developed, so the research and development of the application of plant polysaccharide in the fields of biological medicine and food are slow.

Disclosure of Invention

The invention aims to: aiming at the problems of insufficient research depth of rhizoma polygonati, complex components of plant polysaccharide in rhizoma polygonati and slow development of research on plant polysaccharide in rhizoma polygonati in the prior art, the invention provides rhizoma polygonati fructan and a preparation method and application thereof.

In order to achieve the purpose, the invention adopts the technical scheme that:

a polygonatum fructan having a structure represented by the following general formula I:

in the formula (I), m, n and p represent the number of sugar repeating units, m, n and p are natural numbers including 0, and m + n > p.

Pharmacological research shows that the polygonatum fructan with a novel structure can promote the proliferation of probiotics including lactobacillus and bifidobacterium, and has an obvious effect within 2-12 hours; with the increase of the drug concentration, the proliferation promoting capability of the polygonatum fructan on the probiotics is obviously enhanced, which shows that the polygonatum fructan can promote the growth of the probiotics.

In a preferred embodiment of the invention, the weight-average molecular weight of the polygonatum sibiricum fructan is 2000 Da-8000 Da, and the polydispersity is 1-3.

In order to better obtain a high-purity product of the polygonatum sibiricum fructan, the invention also provides a preparation method, so that the separation effect in the polygonatum sibiricum extraction process is better ensured, and the purity of the obtained product meets the application requirements.

A preparation method of the polygonatum sibiricum fructan comprises the following steps:

extracting crude rhizome polysaccharide of the polygonatum plants;

purifying the components with the weight average molecular weight of 2000 Da-8000 Da in the crude polygonatum rhizome polysaccharide to obtain the polygonatum rhizome levan.

Further, the step of extracting the polygonatum fructan comprises the following steps: extracting rhizome of Polygonatum plant with hot water, precipitating Polygonatum polysaccharide with 95% ethanol to alcohol concentrations of 40 v%, 60 v% and 80 v%, centrifuging, and collecting the precipitate with 80 v% ethanol concentration. According to the mode of fractional precipitation, the concentration of the ethanol is gradually increased, and the purposes of precipitation, separation and purification are achieved.

Further, the polygonatum plants refer to each polygonatum of polygonatum. Can be one or more than one of rhizoma Polygonati in rhizoma Polygonati or their mixture. The rhizoma Polygonati is at least one of the existing rhizoma Polygonati.

Further, the preparation method comprises the following steps: s1, extracting crude polysaccharide of the sealwort under the concentration of 80 v% ethanol; s2, purifying components with the weight-average molecular weight of 2000 Da-8000 Da in the polygonatum polysaccharide to obtain the polygonatum fructan.

Further, the crude polysaccharide with 80 v% ethanol concentration is obtained by extracting rhizome of Polygonatum plant with hot water, and precipitating with ethanol to obtain 80 v% ethanol precipitate.

In one embodiment, the step of extracting the polygonatum fructan comprises the following steps: firstly, precipitating the sealwort by 95 percent ethanol until the final ethanol concentration reaches 80 percent, centrifuging and collecting alcohol precipitation precipitate.

In one embodiment, the method for purifying the components with the weight-average molecular weight of 2000 Da-8000 Da in the polygonatum polysaccharide is selected from one or more of gel exclusion chromatography column method, dialysis method and ultrafiltration method.

In one embodiment, the polygonatum plant comprises one or more of polygonatum, polygonatum kingianum, polygonatum cyrtonema, polygonatum clavatum, polygonatum odoratum, kadin polygonatum, polygonatum minor, polygonatum pubescens, polygonatum pendula, polygonatum longum, polygonatum hirsutum, polygonatum macrospermanum, polygonatum stipulatum, polygonatum brevifolium, polygonatum muticum, polygonatum stigmata, polygonatum solomorpha, polygonatum bicolor, polygonatum nodosum, polygonatum paratyphyllae, polygonatum laevigatum, polygonatum griffithii, polygonatum falcatum, polygonatum tenuifolium polygonati, polygonatum hotriver polygonatum, polygonatum Xinjiang, polygonatum.

A pharmaceutical composition comprising an effective amount of polygonatum fructan.

An application of the polygonatum sibiricum fructan in preparation of food and/or medicines for regulating gastrointestinal flora, promoting intestinal peristalsis and/or promoting digestion.

An application of the rhizoma polygonati fructan in promoting the growth of probiotics is provided.

In one embodiment, the dosage form of the pharmaceutical composition is one of lyophilized powder, oral liquid, capsule and the like.

Application of the above medicinal composition in preparing food and medicine for promoting digestion and intestinal peristalsis is provided.

The rhizoma Polygonati fructan can be used in facial mask, cosmetic, daily chemical product, etc.

In summary, due to the adoption of the technical scheme, the invention has the beneficial effects that:

1. the polygonatum fructan is obtained by first separation, has the effect of promoting the proliferation of probiotics, shows obvious concentration dependence characteristic, and can be used as an active ingredient for promoting various preparations of the probiotics.

2. The polygonatum fructan disclosed by the invention can promote the proliferation of probiotics and regulate the proliferation of intestinal probiotics, so that the polygonatum fructan has a dietary therapy and health care application value. Can be used as main or auxiliary active ingredients of food, health care products or medicines in the aspects of prevention, treatment or conditioning.

Description of the drawings:

FIG. 1 is an HPLC chart of polygonatum sibiricum fructan;

FIG. 2 is a MALDI-TOF MS diagram of rhizoma Polygonati fructan;

FIG. 3 is a HPLC chart of monosaccharide composition analysis of rhizoma Polygonati fructan, wherein chart a is an HPLC chart of monosaccharide standard, chart b is an HPLC chart of rhizoma Polygonati fructan, Fru represents fructose, and Glc represents glucose;

FIG. 4 is an infrared spectrum of fructosan from Polygonatum sibiricum;

FIG. 5 is a nuclear magnetic resonance spectrum of rhizoma Polygonati fructan, wherein FIG. a is a 1H NMR spectrum, and FIG. b is a 13C NMR spectrum;

FIG. 6 is a nuclear magnetic resonance spectrum of rhizoma Polygonati fructan, wherein panel a is HSQC-TOCSY NMR chart, and panel b is HMBC NMR chart;

figure 7 is a graph of the effect of polygonatum fructan on the pH of the medium inoculated with probiotics including lactobacillus and bifidobacterium at different times (n-3);

fig. 8 is a graph of the effect of polygonatum fructan on the OD values of media inoculated with probiotics including lactobacillus and bifidobacterium at different times (n-3);

fig. 9 shows the effect of polygonatum fructan on the number of bacteria in a medium inoculated with probiotics including lactobacillus and bifidobacterium at different times (n-3).

Detailed Description

In order that the invention may be more fully understood, a more particular description of the invention will now be rendered by reference to specific embodiments thereof that are illustrated in the appended drawings. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

The ethanol solution used in the present invention is an aqueous ethanol solution in volume percentage, and is expressed in percentage by%, which should be understood as vol%.

The polygonatum fructan of one embodiment of the invention has a structure represented by the following general formula I:

The polygonatum fructan takes a beta (2 → 6) glycosidic bond as a main chain, and fructose as a side chain is connected to the main chain by the beta (2 → 1) glycosidic bond.

The inventor of the invention has conducted intensive research on polygonatum, and surprisingly found that polygonatum fructan with a novel structure can be obtained by extracting and purifying polygonatum plants such as polygonatum cyrtonema and the like by an innovative method. Through further pharmacological activity research, the newly discovered polygonatum fructan can obviously promote the proliferation of probiotics, so that the polygonatum fructan has the application of preparing medicines and/or foods for intestinal regulation and digestion assistance.

Therefore, the polygonatum fructan with a novel structure, the preparation method thereof, the medicinal composition and the application thereof in preparing the intestinal tract regulating and digestion promoting medicines are disclosed and reported for the first time.

In a specific example, the weight average molecular weight of the polygonatum sibiricum fructan is 2000Da to 8000Da, and the polydispersity number is 1 to 2. Preferably, the weight average molecular weight of the polygonatum sibiricum fructan is 3000 Da-5000 Da.

In one specific example, when p is 0 in formula I, and m + n > p is satisfied, then m and n are minimum values, m and n, 1, and the polygonatum fructan has the structure represented by formula II below:

the preparation method of the polygonatum sibiricum fructan of one embodiment of the invention comprises the following steps of S1-S2:

s1: extracting crude rhizoma Polygonati fructan of rhizoma Polygonati.

S2: purifying the components with the weight average molecular weight of 2000 Da-8000 Da in the crude polygonatum sibiricum fructan to obtain the polygonatum sibiricum fructan.

In one specific example, the polygonatum genus plant includes, but is not limited to, polygonatum kingianum, polygonatum cyrtonema, polygonatum clavatum, polygonatum, prandium, polygonatum minor, polygonatum odoratum, polygonatum prolapseum, polygonatum longum, polygonatum glabrum, polygonatum macrosperma, polygonatum stipulatum, polygonatum brevifolium, polygonatum muticum, polygonatum stipulatum, polygonatum solomorpha, polygonatum bicolor, polygonatum nodosum, polygonatum folicum, polygonatum laeberry, polygonatum kingianum, polygonatum pentaphyllum, polygonatum convolvulus, polygonatum tenuifolium, polygonatum tenuipilatum, polygonatum hotriver, polygonatum Xinjiang, polygonatum sibiricum, polygonatum cyrto. In view of the planting resource condition of polygonatum plants, preferably, the polygonatum plants are one and/or more of polygonatum, polygonatum kingianum, polygonatum cyrtonema and polygonatum.

As a preferred scheme of the invention, the crude polysaccharide of the sealwort under the concentration of 80 v% ethanol is extracted by the following method:

weighing rhizoma Polygonati (preferably Polygonatum sibiricum Red.), adding 5-10 times of water, reflux-extracting for 1-4 times (each for 1-5 hr), filtering, mixing extractive solutions, centrifuging to obtain supernatant, and concentrating to 0.5-3 times of original rhizoma Polygonati. Then, the ethanol concentration is adjusted to 40 v%, and centrifugation is carried out to obtain 40% ethanol precipitate and first supernatant. And (4) continuously adjusting the ethanol concentration to 60 v% in the first supernatant, and centrifuging to obtain a 60% ethanol precipitate and a second supernatant. Adjusting ethanol concentration to 80 v% under stirring, centrifuging to obtain 80% ethanol precipitate and 80% ethanol supernatant, washing 80% ethanol precipitate with pure ethanol or 95% ethanol for 2-4 times, and volatilizing to remove ethanol to obtain crude rhizoma Polygonati fructan.

Further, the concentrations of 40 v%, 60 v% and 80 v% ethanol were adjusted and controlled so as to be up to and down to 5 v%.

In a specific example, the step of extracting polygonatum fructan comprises the following steps: weighing 200g of polygonatum cyrtonema crude product, carrying out reflux extraction for 2 times by 10 times of water, carrying out 4 hours each time, filtering, combining extracting solutions, centrifuging to obtain a supernatant, and concentrating to 680 mL. Adding 95% ethanol until the alcohol concentration is 40%, and centrifuging to obtain 40% ethanol precipitate and 1370mL supernatant. 783mL of 95% ethanol is added into the supernatant after 40% ethanol precipitation under stirring, and centrifugation is carried out to obtain 60% ethanol precipitate and 2000mL of 60% ethanol precipitation supernatant. Adding 2667mL of 95% ethanol into the supernatant under stirring, centrifuging to obtain 80% ethanol precipitate and 80% ethanol precipitate supernatant, washing the 80% ethanol precipitate with 95% ethanol for 3 times, and volatilizing the alcohol to obtain crude rhizoma Polygonati fructan.

In a specific example, the method for purifying the components with the weight-average molecular weight of 2000 Da-8000 Da in the polygonatum polysaccharide is selected from one or more of anion exchange column chromatography, gel exclusion column chromatography, dialysis and ultrafiltration.

Specifically, the crude polygonatum sibiricum fructan component is redissolved by adding deionized water, a small amount of insoluble substances are removed by centrifugation, the crude polygonatum sibiricum fructan component is purified by an optional alcohol precipitation fractionation method, a gel exclusion chromatography method, a dialysis method or an ultrafiltration method and other methods, the fraction containing the fructan or the trapped fluid or the permeate is collected, and after desalination, direct vacuum freeze drying or reduced pressure concentration, alcohol precipitation and reduced pressure drying are carried out to obtain the purified polygonatum sibiricum fructan.

As will be readily understood by those skilled in the art, for Gel exclusion chromatography, Gel materials such as Sephadex series, Sepharose series, polyacrylamide Bio-Gel P series, and Gel fillers formed by cross-linking them with each other are appropriately selected according to the molecular weight of the polysaccharide substance in the polysaccharide fraction of Polygonatum sibiricum, and then the Gel fillers are subjected to column packing, sample loading, and sequential elution with a salt-containing or salt-free elution solution and fractions are collected according to the actual properties of each filler. Because the saccharide has no characteristic ultraviolet absorption, the flow can be detected by color development such as a sulphuric acid phenol method, etc., an outflow curve is drawn, each outflow peak is merged, the outflow liquid can be concentrated or does not need to be concentrated, the outflow liquid is put into a dialysis bag for dialysis or an ultrafiltration membrane for ultrafiltration desalination, the trapped fluid after desalination is collected, and the purified polygonatum fructan is obtained by vacuum freeze drying or vacuum reduced pressure drying. Or purifying rhizoma Polygonati polysaccharide component by dialysis or ultrafiltration, and selecting ultrafiltration membrane with appropriate molecular weight for tangential flow ultrafiltration interception treatment. For example, the aqueous solution of rhizoma Polygonati polysaccharide component is first dialyzed or ultrafiltered fully with a dialysis bag or an ultrafiltration membrane having a molecular weight greater than that of rhizoma Polygonati fructan, the permeate or effluent is collected to remove the substances having a large molecular weight, then dialyzed or ultrafiltered fully with a dialysis bag or an ultrafiltration membrane having a molecular weight less than that of rhizoma Polygonati fructan, the trapped fluid is collected, concentrated and then vacuum freeze-dried or vacuum reduced pressure dried to obtain purified rhizoma Polygonati fructan.

The invention also provides a structure analysis method of the polygonatum sibiricum fructan, which comprises the following steps:

(1) and (3) measuring the molecular weight: taking a purified rhizoma Polygonati fructan sample, and optionally adopting high performance gel exclusion chromatography-differential detector detection method (HPGPC-RI) and matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) to analyze the molecular weight and distribution of rhizoma Polygonati fructan.

(2) Analysis of monosaccharide composition: taking a purified rhizoma Polygonati fructan sample, partially hydrolyzing with low-concentration acid to obtain monosaccharide, directly introducing into a chromatographic column such as sugar-D, and analyzing monosaccharide composition of rhizoma Polygonati fructan by high performance liquid chromatography.

(3) Methylation analysis: taking a purified rhizoma polygonati fructan sample, carrying out partial methylation of polysaccharide hydroxyl groups by using a halogenated methane reagent such as methyl iodide and the like under an alkaline condition, hydrolyzing under an alkaline condition, reducing by using sodium borohydride, carrying out acetylation on methylated sugar alcohol by using acid anhydride, carrying out extraction and volume fixing, and then carrying out gas chromatograph combined mass spectrometer (GC-MS) analysis to judge the glycosidic bond connection mode of the rhizoma polygonati fructan.

(4) Infrared spectrum analysis: taking a purified rhizoma polygonati fructan sample, fully drying, and measuring the infrared spectrum of the sample on an infrared spectrometer by adopting a solid potassium bromide tabletting method.

(5) Nuclear magnetic resonance analysis: dissolving purified rhizoma Polygonati fructan sample in deuterium-substituted heavy water, freeze vacuum drying, repeating three times of heavy water exchange, dissolving in deuterium-substituted heavy water, and detecting nuclear magnetic resonance spectrum of sample including one dimension¹H and¹³c spectrum and¹H-¹H COSY、¹H-¹H TOCSY、¹H-¹H ROESY、¹H-¹³C HSQC、¹H-¹³c HMBC two-dimensional correlation spectrum.

(6) Analyzing comprehensive data: comprehensively analyzing the analysis data of the steps to clarify the chemical structure of the polygonatum fructan.

According to the structure analysis method, the structure analysis is carried out on the polygonatum fructan extracted and purified from polygonatum cyrtonema by the preparation method. The result shows that the polygonatum fructan only shows one chromatographic peak with good symmetry on the chromatographic peak of HPGPC gel, the weight-average molecular weight of the polygonatum fructan is 3kDa, and the polydispersity index of the polygonatum fructan is 1.20; the monosaccharide composition shows that the polygonatum fructan consists of fructose and glucose; methylation analysis shows that the polygonatum fructan has beta (2 → 6) glycosidic bonds and beta (2 → 1) glycosidic bonds; infrared spectrum analysis shows that absorption peaks of hydroxyl groups on the sugar ring and the ring and absorption signal peaks of furanose in a fingerprint spectrum region all accord with absorption peaks of fructan; can clearly belong to rhizoma Polygonati fructan according to nuclear magnetic resonance spectrum data¹H and¹³NMR signals of C.

Combining the above data, the polygonatum fructan has the structural characteristics shown in the general formula I: the main chain is a beta (2 → 6) glycosidic bond, and the side chain fructose is connected to the main chain by a beta (2 → 1) glycosidic bond.

According to the preparation method and the structure analysis method, the polygonatum plants contain polygonatum fructan with the structure shown in the chemical formula II.

Obviously, the polygonatum fructan with the structure shown in chemical formula II is the polygonatum fructan shown in general formula I, when p is 0 and m + n > p is satisfied, the minimum value of m and n is 1.

According to the invention, pharmacological research shows that the polygonatum fructan can promote the proliferation of probiotics and has a remarkable effect within 2-12 hours; with the increase of the drug concentration, the ability of the polygonatum fructan for promoting the probiotics is obviously enhanced, which shows that the polygonatum fructan can promote the proliferation of the probiotics. Therefore, the polygonatum fructan with a novel structure has the effect of promoting the proliferation of probiotics, and has the practical application value of regulating the intestinal flora.

Therefore, the invention further provides application of the polygonatum fructan in preparing medicines and/or foods for regulating intestinal flora, promoting digestion and enhancing intestinal peristalsis.

The pharmaceutical composition provided by the embodiment of the invention comprises the polygonatum fructan, the pharmaceutically acceptable salt or the pharmaceutically acceptable solvent thereof and pharmaceutically and/or food acceptable auxiliary materials. Optionally, the adjuvant comprises a pharmaceutical excipient, a carrier and/or a diluent and/or a flavoring agent, etc.

Furthermore, the pharmaceutical composition also contains medicinal and/or food auxiliary materials. For example, at least one of an excipient or a flavoring agent.

Further, the pharmaceutical composition comprises an effective amount of polygonatum fructan.

In a specific example, the dosage form of the pharmaceutical composition is an injection or a solid preparation, such as a water injection, a freeze-dried powder injection for injection, a capsule, an effervescent agent and the like, and the auxiliary material comprises an excipient and/or a flavoring agent and the like.

The invention also provides application of the pharmaceutical composition in preparing medicines and/or functional foods for regulating intestinal flora, promoting digestion and enhancing intestinal peristalsis.

An "effective amount" as described herein refers to an amount sufficient to achieve a desired result or to have an effect on an undesired condition. For example, a "therapeutically effective amount" refers to an amount sufficient to achieve a desired therapeutic result or effect on an undesirable symptom, but generally insufficient to cause an adverse side effect.

The application of the invention in the directions of regulating intestinal flora, promoting digestion, enhancing intestinal peristalsis and the like is to be applied to a patient aiming at improving and stabilizing pathological conditions or symptoms related to the digestive tract. Both active treatment, direct improvement of pathological conditions or disorders; prophylactic treatment, i.e. reduction or partial or complete inhibition of the occurrence of a pathological condition or disorder, is also included. Where prevention refers to the exclusion, avoidance, elimination, prevention, deterrence, or hindrance of something happening, especially through prior action.

Generally, the above-mentioned medicine or the composition containing the above-mentioned medicine of the present invention is used for preparing edible food or health-care product for mammal (for example human). Of course, it is also contemplated in some instances to apply it to other mammals, such as feeding pet cats, dogs, etc.; or livestock, such as cow, horse, pig, sheep, goat, etc., and others such as mouse, rabbit, rat, guinea pig, etc.

When the polygonatum polysaccharide is prepared into various production profiles, necessary auxiliary materials and/or auxiliary agents can be added. For example, a pharmaceutically acceptable carrier, broadly refers to an aqueous or non-aqueous solution, dispersion, suspension, emulsion, or the like. Non-aqueous solutions may be simply enumerated as including one or more of such materials as ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, and the like), carboxymethyl cellulose, vegetable oils, and the like. If used for preparing oral dosage forms, such as tablet and capsule, other fillers, disintegrants, sustained release agents, preservatives, dispersants, etc. can also be included. By way of example, the preservative may be paraben, chlorobutanol, sorbic acid, and the like.

The present invention will be described in detail with reference to specific embodiments in the following drawings, which should not be construed as limiting the scope of the claims of the present invention.

The starting materials, compounds, compositions or components used in the following examples are obtained primarily commercially or by preparation using techniques generally known to those skilled in the art. Next, experimental procedures of the preparation processes in the following examples are conducted in accordance with general experimental guidelines, and the experimental sections in the examples are illustrative of forms clear to those skilled in the art and are not limited to the methods in the following examples. The person skilled in the art can carry out the necessary derivations to obtain experimental procedures suitable for the particular raw materials, according to routine knowledge.

Example 1 extraction and purification of Polygonatum sibiricum Fructrs

1. Extraction of polygonatum polysaccharide

Weighing 200g of polygonatum cyrtonema crude product, carrying out reflux extraction for 2 times with 10 times of water for 4 hours each time, filtering, combining extracting solutions, centrifuging to obtain supernatant, and concentrating to obtain 680mL of concentrated solution. 495mL of 95 v% ethanol was added to an alcohol concentration of 40 v%, and centrifugation (4000 rpm. times.15 min) was carried out to obtain a 40% ethanol precipitate and a supernatant. 783mL of 95 v% ethanol was added to the 40 v% ethanol precipitated supernatant with stirring until the alcohol concentration reached 60 v%, and the mixture was centrifuged (4000 rpm. times.15 min) to obtain 60 v% ethanol precipitate and supernatant. Under stirring, 2667mL of 95 v% ethanol is added into the 60% ethanol precipitation supernatant until the ethanol concentration reaches 80 v%, centrifugation (4000rpm is multiplied by 15min) is carried out to obtain 80 v% ethanol precipitate and supernatant, the 80% ethanol precipitate is washed for 3 times by 95% ethanol, and the crude polygonatum fructan is obtained after ethanol volatilization. Wherein, the ethanol can be removed by reduced pressure evaporation and volatilization.

2. Purification of rhizoma polygonati fructan

Dissolving the crude rhizoma Polygonati fructan in deionized water, loading onto DEAE-52(3.2cm × 30cm) weak anion exchange column, eluting with deionized water and 0.1, 0.5 and 1.0M NaCl solution, and collecting eluate about 10mL per tube. Detecting with phenol-sulfuric acid method, drawing outflow curve, collecting the final fraction containing no or little polysaccharide, concentrating under reduced pressure, and freeze vacuum drying to obtain purified rhizoma Polygonati fructan 22.83 g.

Example 2 Structure analysis of Polygonatum sibiricum Fructo-glycan

1. Procedure of experiment

1.1. Molecular weight and distribution thereof

The molecular weight and distribution of the polygonatum fructan prepared in example 1 were analyzed by high performance gel exclusion chromatography-differential detector assay (HPGPC-RI).

Chromatography apparatus: agilent technologies 1260series high performance liquid chromatograph;

chromatographic conditions are as follows: shodex Ohpak SB-804HQ (7.8 mm. times.300 mm) column; the column temperature was 35 ℃; a difference detector; the mobile phase is NaCl with the concentration of 0.1M, and the flow rate is 0.5 mL/min;

the determination process comprises the following steps: adding mobile phase into 5mg/mL solution of 5mg rhizoma Polygonati fructan sample or dextran reference substance with known molecular weight, filtering with 0.22 μm microporous membrane, analyzing 50 μ L filtrate with high performance liquid chromatograph, and recording chromatogram. And processing the data by adopting GPC software, drawing a standard curve, substituting the data into an equation, and calculating the molecular weight.

Analyzing the molecular weight and distribution of rhizoma Polygonati fructan by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS)

The instrument comprises the following steps: bruker UltrafleXtreme matrix-assisted laser desorption ionization time-of-flight mass spectrometer;

the instrument conditions were as follows: a linear positive ion mode; an N2 laser source, and a substrate is 2, 5-dihydroxy benzoic acid.

The determination process comprises the following steps: weighing a certain amount of rhizoma polygonati fructan sample, adding deionized water to prepare a solution, mixing 1 mu L of sample solution with 1 mu L of matrix solution, spotting on a sample target, naturally drying, measuring the molecular weight of the rhizoma polygonati fructan, and recording a mass spectrogram. The data were processed using flexAnalysis software and the molecular weight was calculated.

1.2 monosaccharide composition analysis

Taking a rhizoma polygonati fructan sample of about 20mg, placing the sample in 2 COD test tubes, adding 2mL of 0.05M sulfuric acid solution into the test tubes for dissolving, carrying out hydrolysis reaction for 40min in an oven at 80 ℃, cooling to room temperature after the reaction is finished, taking out the sample, placing the sample in a 5mL EP tube (centrifugal tube), adding barium carbonate solid powder for neutralizing sulfuric acid, centrifuging (15000rpm multiplied by 15min), taking supernatant, and passing through a 0.22 mu M microporous filter membrane for chromatographic analysis.

Chromatographic conditions are as follows: the instrument comprises the following steps: agilent technologies 1260series high performance liquid chromatograph; a chromatographic column: a Cosmosil Sugar-D column (2.0 mm. times.250 mm, 5 μm); the column temperature is 30 ℃; the mobile phase is acetonitrile, water (85: 15); flow rate: 0.2 mL/min; the injection volume is 20 mu L; and a RID detector.

1.3 methylation analysis

Methylation reaction: weighing 10mg of a polygonatum sibiricum fructan sample, placing the sample in a 5mL reaction bottle, dissolving the sample in 4mL DMSO, sequentially adding 100mg of sodium hydroxide and 600 mu L of methyl iodide, filling nitrogen, and carrying out ultrasonic reaction at room temperature for 1 h. After the reaction is finished, 2mL of pure water is added, the pH value of the solution is adjusted to be neutral by using 1M hydrochloric acid solution, 6mL of trichloromethane is added for extraction, and the organic phase is dried under reduced pressure at 40 ℃ and then dried in vacuum for 12 hours.

Acid hydrolysis of methylated polysaccharides: the dried sample was dissolved in 2mL of 0.5M TFA solution, and the tube was sealed and hydrolyzed at 120 ℃ for 5 hours.

Reduction of methylated monosaccharides: and (3) adjusting the pH value of the hydrolysate to 10-12 by using 1M NaOH, adding 50mg of sodium borohydride, stirring in a water bath at 50 ℃ for 2 hours, adding 250 mu L of glacial acetic acid to stop the reaction, and freeze-drying the reaction solution.

Acetylation of methylated sugar alcohols: 1mL of pyridine and 1mL of acetic anhydride were added to the above lyophilized sample, and the mixture was reacted in a test tube at 100 ℃ for 1 hour with tube sealing. After cooling, 1mL of water are added, 2mL portions of dichloromethane are extracted 3 times, and the organic phase is extracted with N₂After blowing, 100. mu.L of methylene chloride was added to dissolve the residue, and GC-MS analysis was performed.

GC conditions were as follows: HP-5MS quartz capillary column (50 mm. times.0.25 mm, 0.25 μm); column temperature: the initial temperature is 80 ℃, the temperature is kept for 11min, the temperature is programmed to be 5 ℃/min to 250 ℃, and the temperature is kept for 5 min; the carrier gas is high-purity helium; the carrier gas flow is 1.5 mL/min; the temperature of a sample inlet is 270 ℃; column front pressure 100 KPa; no flow diversion; sample introduction amount: 2 μ L.

MS conditions: an ionization mode EI; electron energy 70; the transmission line temperature is 290 ℃; the ion source temperature is 230 ℃; the temperature of the quadrupole rods is 150 ℃; the mass range is 50-600; and (4) adopting a CCRC standard spectrum library computer to search and determine the quality.

1.4 Infrared Spectroscopy

According to the fourth part of 2020 edition Chinese pharmacopoeia, a solid tabletting method is adopted: taking 2mg of a sample, vacuum-drying the sample for 24 hours at 40 ℃, adopting a KBr tabletting method, and carrying out 4000-400 cm on the sample by using a Tensor 27 Fourier transform mid-infrared spectrometer^-1Scanning and recording a spectrogram.

1.5 nuclear magnetic resonance analysis

Dissolving 10mg of rhizoma Polygonati fructan sample in 0.5mL of D₂In O, freeze-drying, D₂O exchange IIIAfter this time, the freeze-dried sample was dissolved in 0.5mL of D₂O (99.9 atom% D, containing 0.05 wt.% 3- (trimethylsilyl) propionic-2,2,3,3-D as an internal standard₄acid, sodium salt). Measurement with Bruker 800MHz NMR spectrometer¹H/¹³C NMR spectrum and two-dimensional spectrum (¹H-¹H COSY、¹H-¹H TOCSY、¹H-¹H ROESY、¹H-¹³C HMBC、¹H-¹³C HSQC). And analyzing and processing the detection result by adopting MestReNova software.

2. Results of the experiment

The properties of the polygonatum fructan are as follows: white-like solid, tasteless, easily soluble in water, slightly soluble in organic solvents such as ethanol, etc., and has hygroscopicity.

The high performance gel exclusion chromatography (HPGPC-RI) showed that (FIG. 1) the rhizoma Polygonati fructan has only one symmetrical peak, its weight average molecular weight (Mw) is 3803Da and its polydispersity index is 1.21. The matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) result shows (figure 2) that the molecular weight range of rhizoma Polygonati fructan is 2000.9-7027.5Da, and the corresponding molecular weight of rhizoma Polygonati fructan with highest intensity is 2973.7 Da.

The analysis result of monosaccharide composition shows (figure 3), an HPLC (high performance liquid chromatography) chart of the polygonatum fructan after being directly hydrolyzed by acid and then being subjected to a Sugar-D column shows that only one peak of fructose and glucose exists, wherein the peak area of the fructose is far higher than that of the glucose, which indicates that the polygonatum fructan contains only fructose and glucose, and the glucose content is far lower than that of the fructose, and accords with the structural characteristics of the fructan.

Methylation analysis results show that the polygonatum fructan has beta (2 → 6) fructosyl glycosidic bond, beta (2 → 1,6) fructosyl glycosidic bond, beta (→ 2) fructosyl glycosidic bond, beta (2 → 1) fructosyl glycosidic bond and alpha (1 → 6) glucosyl glycosidic bond.

As shown in FIG. 4, infrared spectrum (cm)^-1) The data are as follows: 3385cm^-1The characteristic absorption of the O-H stretching vibration of the hydroxyl on the sugar ring is realized; 2935cm^-1Stretching and vibrating methylene C-H on a sugar ring; 1129-1028 cm^-1C-O-C stretching vibration on the sugar ring; 933cm^-1And 878cm^-1Is the configurational characteristic absorption of beta-D-fructose.

The results of NMR detection and analysis are shown in FIGS. 5 and 6, in detail¹H and¹³the C NMR signals are assigned in tables 1 and 2. Chemical shift 5.43ppm is the anomeric proton signal of 1,6-O- α -D-glucose (1,6-O- α -D-Glcp, abbreviated as G), the 6-position (H-6 and H-6') proton signals of 1,6-O- α -D-Glcp is 4.08(3.77) ppm and its corresponding chemical shift of C-1 is 95.0 ppm. The chemical shifts are shifted significantly to a low field relative to the corresponding chemical shifts for unsubstituted glucose, indicating that the hydroxyl groups at the 1-and 6-positions of the glucose group G are substituted. The coupling constant of anomeric C-H of glucosyl group G is 180Hz, indicating that its anomeric configuration should be alpha type. Fructose is furanose, which has no anomeric hydrogen at position 2, and only anomeric carbon signal in NMR spectrum, which has 4 anomeric carbon signals in the anomeric carbon signal region, and chemical shift values of 106.9, 106.7, 106.5 and 106.1ppm from high to low, corresponding to 2, 6-O-. alpha.0-D-fructosyl group (2, 6-O-. alpha.1-D-Fruf, abbreviated as A), 1,2, 6-O-. beta.D-fructosyl group (1,2, 6-O-. beta. -D-Fruf, abbreviated as B), 2-O-. beta. -D-fructosyl group (2-O-. beta. -D-Fruf, abbreviated as D) and 2, 1-O-. beta. -D-fructosyl group (2, 1-O-. beta. -D-Fruf, c-2 signal abbreviated E)

¹H and¹³the C remote correlation HMBC spectrum clearly shows that H-6(G6) of glucosyl G and C-2(D2) of D have obvious correlation signals; h-1(B1) of the fructosyl group B and C-2(E2) of the fructosyl group E have related signals, H-6(B6) of the fructosyl group B and C-2(A2) of the fructosyl group A have related signals, and H-6(A6) of the fructosyl group A and C-2(B2) of the fructosyl group B have related signals, H-1(E1) of the fructosyl group E and C-2(E2) of the fructosyl group E have strong related signals. Thus, the way fructan is linked is: the fructosyl A forms a main chain with a beta (2 → 6) glycosidic bond, while the fructosyl E is linked to the backbone of the fructosyl A twice with a beta (2 → 1) glycosidic bond.

The data are combined to show that the chemical structural formula of the polygonatum fructan with a novel structure is shown as a general formula I.

The structure is characterized in that: (1) the rhizoma Polygonati fructan is polysaccharide extracted from rhizoma Polygonati rhizome and is plant fructan; (2) in the monosaccharide composition of the polygonatum sibiricum fructan, fructose is the main component and a small amount of glucose is contained; (3) the monosaccharide connection mode of the polygonatum sibiricum fructan is as follows: the main chain is beta (2 → 6) glycosidic bond, and the side chain fructose is connected to the main chain by beta (2 → 1) glycosidic bond. Through the search of published documents at home and abroad, the arrangement mode of the polygonatum sibiricum fructan is not reported in a public way and is discovered by the inventor for the first time.

TABLE 1 preparation of rhizoma Polygonati fructan¹Assignment of H NMR

TABLE 2 preparation of rhizoma Polygonati fructan¹³Assignment of C NMR

Wherein the content of the first and second substances,

2,6-O- β -D-Fruf denotes → 6) - β -D-Fruf- (2 →;

1,2,6-O- β -D-Fruf denotes → 1,6) - β -D-Fruf- (2 →;

2-O- β -D-Fruf means β -D-Fruf- (2 →;

2,1-O- β -D-Fruf denotes → 1) - β -D-Fruf- (2 →;

1,6- α -D-Glcp means → 1) - α -D-Glcp- (6 →.

Example 3 experiment of promoting growth of probiotic bacteria with rhizoma Polygonati fructan

1. Test article, reagent and strain

Polygonatum fructan, abbreviated DP1, prepared as in example 1; sugar-free Man-Rogosa-Sharpe (MRS) medium, Inc., Coolaber, Beijing; the 4 strains are respectively Bifidobacterium animalis subsp.

2. Experimental methods

2.1 preparation of the solution

600mg of Fructooligosaccharide (FOS), 600mg of polygonatum fructosan (DP1) and 700mg of Glucose (Glucose) were weighed out, and each solution was prepared into 10mg/mL solution with sterile water, and after filtering with a 0.22 μm sterile filter, each solution was added to a sugar-free MRS medium at a concentration of 5 mg/mL. 56mL of sterilized MRS medium was taken, and 56mL of sterile water was added to dilute the filtered sample, followed by mixing to prepare 5mg/mL of MRS medium. Bifidum and L.plantarum on the corresponding medium 1X 10⁷Amounts of CFU/mL, B.animalis subsp.lactis and L.acidophilus were inoculated at 1X 10 onto the respective media⁸Amount of CFU/mL.

2.2 bacterial culture

Bacteria culture medium: MRS (L-cysteine) medium contains 0.05% of L-cysteine;

and (3) recovering bacteria: prepare 1 relative liquid medium and 2 plates, scribe the tube body 1/2 with a small wheel in a clean bench, wipe the tube wall with 75 vol% alcohol, and break with tweezers. Injecting 0.3-0.5mL of liquid culture medium into a freeze-drying tube, gently blowing and uniformly mixing, fully dissolving into bacterial suspension, sucking 1/3, adding into the liquid culture medium, averagely adding the rest onto 2 plates, and culturing under corresponding conditions until bacterial colonies grow out.

And (3) expanding bacteria: individual colonies were picked from the plates and cultured on a shaker at 37 ℃ in liquid medium.

Seed preservation: centrifuging 25mL of bacterial solution at 4 deg.C (4100 rpm. times.10 min), removing supernatant, adding 2.5mL of liquid culture medium, suspending the precipitate, adding glycerol of the same volume, mixing, and freezing at-80 deg.C.

2.3 cultivation of the respective bacterial culture Medium

Each bacterial strain is placed in the anaerobic chamberOxygen environment (85% N)₂,10％CO₂And 5% of H₂) All in percent by volume%, and samples were taken at 0h, 2h, 4h, 6h, 8h, 12h, 24h, 36h and 48h time points.

2.4 detection of pH

The culture broth was removed at each sampling time point, in triplicate, and after centrifugation (4000 rpm. times.10 min), the pH was determined 3 times per time point.

2.5 detection of OD values

The culture broth was taken out at each sampling time point, in parallel, in triplicate, shaken up and down for 15s, poured into a cuvette, and the OD was determined with an ultraviolet spectrophotometer at a wavelength of 600nm, 3 times per time point.

2.6 detection of the number of bacteria

The culture medium was taken out at each sampling time point, in triplicate, shaken up and down for 15s, from which a certain amount of culture medium was taken out, diluted by a certain multiple and counted on a hemocytometer for 3 times per time point.

3. Results of the experiment

3.1 Effect on the pH value in the culture Medium of the respective bacteria

As shown in FIG. 7, the pH of the treated group was decreased to a different extent than that of the blank group, indicating that the pH of the medium was decreased by the increased amounts of short-chain fatty acid metabolites such as lactic acid and acetic acid in the presence of the fructosan from Polygonatum sibiricum Red (DP1), Fructooligosaccharide (FOS) and Glucose (Glucose) from each bacterial strain.

3.2 Effect on OD value in culture Medium for respective bacteria

As shown in FIG. 8, the OD values of the culture solutions obtained by culturing in different media increased rapidly from 0 to 12 hours, and after 12 hours, the OD values of the culture solutions remained substantially unchanged; the OD values of the polygonatum fructan (DP1), fructo-oligosaccharide (FOS) and Glucose (Glucose) groups were all significantly higher than the blank group at the corresponding times. The polygonatum fructan, fructo-oligosaccharide and glucose can promote the proliferation of each bacterial strain, and the proliferation amount of bacteria treated by the polygonatum fructan is obviously different from that of a blank group.

3.3 Effect on the number of bacterial strains in the respective bacterial media

As shown in figure 9, in the culture medium containing rhizoma Polygonati fructan, 4 bacterial strains enter exponential phase within 2-12 hours and rapidly proliferate; the comparison of the maximum bacterial strain number of 24h or 48h bacteria entering the platform stage shows that the rhizoma polygonati fructan group is equivalent to the oligofructan group and the glucose group and is obviously higher than the blank group; the proliferation conditions of 4 different strains are different with the strains, and the proliferation promoting activity of the lactobacillus bifidus (B.animalis subsp. lactis) on animal bifidobacteria is obviously higher than that of lactobacillus acidophilus (L.acidophilus).

The results prove that the polygonatum fructan has remarkable activity of promoting the growth of probiotics, and the polygonatum fructan with a novel structure has practical application values of improving intestinal flora, promoting digestion and enhancing intestinal peristalsis.

Example 4 preparation of lyophilized powder injection of rhizoma Polygonati fructan

1. Material

The polygonatum fructan obtained by the method in the example 1 and the pharmaceutical grade sodium chloride are used.

2. Prescription

Name of raw and auxiliary materials	Dosage of
		Rhizoma Polygonati fructan	50g
Sodium chloride	4.5g
		Water for injection	500mL
Are co-produced into	1000 pieces

3. Preparation process

Weighing rhizoma Polygonati fructan and sodium chloride according to prescription amount, adding water for injection to full amount, stirring to dissolve completely, and sterilizing by intermittent hot pressing method. Adding 0.3% medicinal active carbon, and stirring for 20 min; the heat source was removed by decarburizing filtration using a Buchner funnel and a 3.0 μm microporous membrane. Filtering with 0.22 μm microporous membrane after the content is qualified; and (3) subpackaging into tube-type penicillin bottles, wherein each bottle is 0.5mL, half plugging, placing into a freezing and drying box, performing freeze drying according to a set freeze drying curve, plugging, taking out of the box, rolling a cover, performing visual inspection to obtain a qualified product, and packaging to obtain a finished product.

And (3) freeze-drying: putting the sample into a box, cooling the temperature of a partition plate to-40 ℃, and keeping for 4 hours; the cold trap was lowered to-50 ℃ and vacuum was started to 250 μ bar. Starting sublimation: heating to-20 deg.C at constant speed for 1h, and maintaining for 3 h; heating to-10 ℃ at constant speed for 3h, keeping for 8h, and keeping in vacuum for 100-250 μ bar; and then drying: heating to-5 ℃ for 2h, keeping for 2h, and keeping the vacuum state at 150-200 mu bar; heating to 10 ℃ within 0.5h, keeping for 2h, and keeping vacuum at 80-100 μ bar; heating to 40 deg.C for 0.5h, maintaining for 4h, and vacuum pumping to minimum.

Example 5 preparation of Polygonatum sibiricum Fructo-glycan capsules

1. Material

The polygonatum fructan obtained by the method of example 1, food or pharmaceutical grade starch.

2. Prescription

Name of raw and auxiliary materials	Dosage of
		Rhizoma Polygonati fructan	300g
Starch	500g
		Are co-produced into	5000 granules

3. Preparation process

Weighing the polygonatum fructan and the starch according to the prescription amount, and stirring to completely mix. Adding appropriate amount of pulvis Talci, granulating with ethanol by wet method, sieving, drying, and packaging into number 2 capsule shell, and filling 60mg of rhizoma Polygonati fructan into each capsule body to obtain rhizoma Polygonati fructan capsule.

Example 6 preparation of Polygonatum sibiricum fructan oral liquid

1. Material

The polygonatum fructan obtained by the method in the example 1, the food or medicine grade flavoring agent is aspartame and apple essence.

2. Prescription

Name of raw and auxiliary materials	Dosage of
		Rhizoma Polygonati fructan	50g
Asparasaminidophenylalanine methyl ester	0.3g
		Apple essence	0.05g
Purified water	1000mL
		Are co-produced into	500 pieces

3. Preparation process

Weighing rhizoma Polygonati fructan, aspartame and apple essence according to prescription amount, adding purified water to dissolve completely, filtering with 0.22 μm microporous membrane, bottling the filtrate according to 2mL per bottle by oral liquid canning machine, sealing, and sterilizing.

Example 7 preparation of Polygonatum sibiricum fructan facial cleanser

1. Material

The polygonatum fructan obtained by the method in the example 1 adopts cosmetic grade or food grade raw materials.

2. Prescription

7g of polygonatum fructan, 2.0g of glycerol, 4.0g of butanediol, 0.5g of propylene glycol, 0.1g of EDTA sodium, 0.3g of guar gum, 2.0g of zinc dioxide, 4.0g of C12-15 alcohol benzoate, 3.0g of C12-20 alkyl glucoside, 0.5g of C14-22 alcohol, 1.2g of cetearyl alcohol, 0.2g of nipagin ester, 0.3g of sodium stearate, 0.5g of dimethiconol, 0.2g of polysorbate and 55g of deionized water.

3. Preparation process

Weighing rhizoma Polygonati fructan according to the prescription amount, and adding purified water to completely dissolve; dissolving glycerol, butanediol, propylene glycol, sodium EDTA, guar gum, zinc dioxide, sodium stearate, C12-15 alcohol benzoate, C12-20 alkyl glucoside, C14-22 alcohol, and cetearyl alcohol in purified water; taking the nipagin ester, the dimethiconol and the polysorbate, and adding purified water to completely dissolve. Mixing the above solutions, stirring, filtering with 0.22 μm microporous membrane, bottling the filtrate 30mL per bottle, sealing, and sterilizing.

Example 8 preparation of Polygonatum sibiricum fructan facial mask

1. Material

2. Prescription

12g of polygonatum fructan, 3.5g of transparent xanthan gum, 2.0g of hydrolyzed hyaluronic acid, 2.0g of sodium hyaluronate, 0.3g of propylene glycol, 0.5g of carbomer, 0.1g of nipagin ester, 0.25g of PPG-10 methyl glucose ether, 1.0g of glycerol glucoside and 45g of deionized water.

3. Preparation process of facial mask

Step 1, adding deionized water, transparent xanthan gum, hydrolyzed hyaluronic acid, sodium hyaluronate, propylene glycol, carbomer and nipagin ester into an emulsifying pot, heating to 75 ℃ while stirring, keeping the temperature until the solution is transparent and uniform, keeping the temperature for 20 minutes, and cooling to 42 ℃.

And 2, adjusting the pH value to 7.6 +/-0.2.

And 3, continuously cooling to 35 ℃, adding PPG-10 methyl glucose ether, rhizoma polygonati fructan and glycerol glucoside, continuously stirring uniformly, and discharging after the inspection is qualified. The coating was applied to a nonwoven substrate at a weight of 2.1g per square centimeter.

The descriptions of each patent, patent application, and publication cited in this application are incorporated herein by reference in their entirety. Citation of any reference shall not be construed as an admission that such reference is available as "prior art" to the present application.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims

1. A polygonatum fructan, characterized in that, the polygonatum fructan has a structure represented by the following general formula I:

2. The polygonatum fructan of claim 1, wherein the polygonatum fructan has a weight average molecular weight of 2000-8000 Da and a polydispersity of 1-3.

3. A method for preparing polygonatum fructan according to any one of claims 1-2, comprising the following steps:

extracting crude rhizome polysaccharide of the polygonatum plants;

4. The method of claim 3, wherein the step of extracting the polygonatum fructan comprises the steps of: extracting rhizome of Polygonatum plant with hot water, precipitating Polygonatum polysaccharide with 95% ethanol to alcohol concentrations of 40 v%, 60 v% and 80 v%, centrifuging, and collecting the precipitate with 80 v% ethanol concentration.

5. The method of claim 4, wherein the rhizoma Polygonati fructan is a fructan obtained by precipitation from a solution with an ethanol concentration of 80 v%.

6. The method for preparing polygonatum fructan according to claim 4, wherein the method for purifying the components of polygonatum polysaccharides with a weight average molecular weight of 2000 Da-8000 Da is selected from one or more of alcohol precipitation fractionation, gel exclusion chromatography column, dialysis and ultrafiltration.

7. The method of claim 4, wherein the Polygonatum cyrtonema Hua, Polygonatum sibiricum Red, Polygonatum Canadensis, Polygonatum Canaliculatum, Polygonatum sibiricum Red, Polygonatum Cantonese, Polygonatum Canaliculatum, Polygonatum Canaliculat.

8. A pharmaceutical composition comprising an effective amount of the polygonatum fructan of claim 1 or 2.

9. The pharmaceutical composition of claim 8, wherein the pharmaceutical composition is in the form of lyophilized powder, oral liquid or capsule.

10. Use of the polygonatum fructan according to claim 1 or 2 in the preparation of a food and/or a medicament for regulating the gastrointestinal flora, promoting intestinal motility and/or promoting digestion;

or, the use of the polygonatum fructan according to claim 1 or 2 for the preparation of a facial mask, a cosmetic or a daily chemical.