Rhizoma anemarrhenae polysaccharide AAP1-2 as well as preparation method, identification method and application thereof

Abstract

The invention relates to the technical field of medicines and health foods, in particular to rhizoma anemarrhenae polysaccharide AAP1-2 as well as a preparation method, an identification method and application thereof. According to the specific technical scheme, the rhizoma anemarrhenae polysaccharide AAP1-2 is a homogeneous polysaccharide, and the molecular weight range of the rhizoma anemarrhenae polysaccharide AAP1-2 is 1000-100000Da. The preparation method is simple, reaction conditions are mild, and large-scale production can be achieved; the chemical structure of the obtained high-purity rhizoma anemarrhenae polysaccharide is preliminarily identified, and a structural basis is provided for exploring the pharmacological activity mechanism of the rhizoma anemarrhenae polysaccharide. Meanwhile, the obtained rhizoma anemarrhenae polysaccharide AAP1-2 lays a foundation for rhizoma anemarrhenae polysaccharide anti-aging active drugs, health-care foods and functional foods, quality control of the rhizoma anemarrhenae polysaccharide anti-aging active drugs, health-care foods and functional foods, and deep research of structure-function relationships and action mechanisms of the rhizoma anemarrhenae polysaccharide

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of medicine and health food, in particular to a polygonatum sibiricum polysaccharide AAP1-2, a preparation method, an identification method and an application thereof. BACKGROUND

[0002] The global aging process continues to intensify, leading to an increase in global morbidity, mortality and health care costs. Aging is usually accompanied by the decline of cell and tissue function, leading to chronic diseases such as cardiovascular disease, diabetes and neurodegenerative disease. However, traditional drugs for treating aging and age-related diseases face some challenges, including low bioavailability, high cost and serious side effects. Therefore, finding low-toxic and high-efficiency drugs is a major problem to be solved now.

[0003] Polysaccharide, also known as polyose, is a kind of biological information macromolecule widely existing in animals, plants and microorganisms. Natural polysaccharide plays an important role in a variety of diseases due to its outstanding antioxidant and immunomodulatory activity, including tumor, anti-inflammatory and cardiovascular disease. More and more evidence shows that plant polysaccharide has become a hot spot in anti-aging research due to its natural source, low toxicity and multiple pharmacological activities. Therefore, finding potential therapeutic drugs from natural polysaccharide is expected to become an effective strategy for anti-aging.

[0004] Rhodomyrtus tomentosa (Ait.) Hassk is the rhizome of Rhodomyrtus tomentosa (Ait.) Hassk of Liliaceae. It is harvested in spring and autumn, and the dry or baked product is obtained after removing dry leaves and roots and shaking off the soil. Rhodomyrtus tomentosa (Ait.) Hassk has the effects of nourishing yin, reducing fire, moistening dryness and lubricating intestines. It is used for treating heat consumption, bone heat, lung heat, dry constipation and difficult urination. The chemical components of Rhodomyrtus tomentosa (Ait.) Hassk mainly include glycosides, flavones and saponins. It has the effects of reducing blood sugar, reducing blood lipids, improving senile dementia, relieving fever and anti-inflammatory. As one of the main active ingredients of Rhodomyrtus tomentosa (Ait.) Hassk, the pharmacological effects of polysaccharide have not been widely studied. At present, there are few reports on the homogenate polysaccharide and anti-aging effect of Rhodomyrtus tomentosa (Ait.) Hassk. Therefore, it is necessary to provide a method for extracting and purifying Rhodomyrtus tomentosa (Ait.) Hassk polysaccharide, so as to lay a foundation for quality control and in-depth study of anti-aging activity of Rhodomyrtus tomentosa (Ait.) Hassk polysaccharide. SUMMARY

[0005] In view of the deficiencies of the prior art, the present application provides a polygonatum sibiricum polysaccharide AAP1-2, a preparation method, an identification method and an application thereof, which has a remarkable effect on anti-aging. The present application combines water extraction and alcohol precipitation, and high-concentration ethanol can separate polysaccharides with large polarity and good water solubility from polysaccharides with small polarity and poor water solubility, thereby solving the problem of complicated and difficult separation caused by traditional water boiling method for extracting polysaccharides.

[0006] To achieve the above object, the present application is implemented by the following technical solutions:

[0007] The application discloses an Anemarrhena asphodeloides polysaccharide AAP1-2, which is a homogeneous polysaccharide with a molecular weight ranging from 1000 to 100000 Da.

[0008] Correspondingly, a preparation method of the Anemarrhena asphodeloides polysaccharide AAP1-2 comprises the following steps.

[0009] (1) crushing dried rhizome of Anemarrhena asphodeloides to obtain Anemarrhena asphodeloides rhizome powder, adding the Anemarrhena asphodeloides rhizome powder into water, heating extraction, filtering to obtain an extract;

[0010] (2) performing multiple alcohol precipitation on the extract obtained in step (1), and performing alcohol precipitation again on supernatant obtained after each alcohol precipitation; and obtaining crude polysaccharide AA1, crude polysaccharide AA2 and crude polysaccharide AA3 respectively after each alcohol precipitation;

[0011] (3) removing protein from the crude polysaccharide AA1, dialyzing, freeze-drying to obtain Anemarrhena asphodeloides polysaccharide AA1; performing ion exchange column chromatography on the Anemarrhena asphodeloides polysaccharide AA1, gradient elution is performed on the Anemarrhena asphodeloides polysaccharide AA1 with 0-2M NaCl solution, and according to an elution curve, the sugar part is collected and concentrated, freeze-dried to obtain Anemarrhena asphodeloides polysaccharide AAP1;

[0012] (4) dissolving the Anemarrhena asphodeloides polysaccharide AAP1 in water, centrifuging, taking supernatant to perform molecular sieve gel column chromatography, eluting with water, according to an elution curve, the sugar part is collected and concentrated, freeze-dried to obtain Anemarrhena asphodeloides polysaccharide AAP1-2.

[0013] Preferably, in step (1), the water is added in an amount of 6-10 times the weight of the Anemarrhena asphodeloides powder.

[0014] Preferably, in step (1), the heating extraction is performed at a temperature of 60-80 DEG C, the heating extraction is performed for 1-3h, and the heating extraction is performed for 2-4 times.

[0015] Preferably, in step (2), the multiple alcohol precipitation is performed as follows: the extract obtained in step (1) is reduced pressure concentrated to obtain concentrated solution 1; ethanol is added into the concentrated solution 1 until the volume concentration of the ethanol is a%, and then standing, collecting the precipitate and supernatant to obtain crude polysaccharide AA1 and supernatant 1; the supernatant 1 is reduced pressure concentrated to obtain concentrated solution 2; ethanol is added into the concentrated solution 2 until the volume concentration of the ethanol is b%, and then standing, collecting the precipitate and supernatant to obtain crude polysaccharide AA2 and supernatant 2; the supernatant 2 is reduced pressure concentrated to obtain concentrated solution 3; ethanol is added into the concentrated solution 3 until the volume concentration of the ethanol is c%, and then standing, collecting the precipitate to obtain crude polysaccharide AA3; wherein, 10≤a

[0016] Preferably, the temperature of the reduced pressure concentration is 40-70 DEG C, and the standing time is 10-15h.

[0017] Correspondingly, an identification method of the Anemarrhena asphodeloides polysaccharide AAP1-2 comprises the following steps.

[0018] (1) Take the Anemarrhena asphodeloides polysaccharide AAP1-2, and detect homogeneity and molecular weight by high performance liquid chromatography;

[0019] (2) Take the Anemarrhena asphodeloides polysaccharide AAP1-2, dry, tablet, and detect by infrared spectroscopy.

[0020] Correspondingly, the Anemarrhena asphodeloides polysaccharide AAP1-2 is applied to preparation of anti-aging drugs or health products.

[0021] The present application has the following beneficial effects:

[0022] 1. The present application adopts water extraction and alcohol precipitation to preliminarily separate the Anemarrhena asphodeloides polysaccharide, and the effect is remarkable, and the preparation method is simple, the reaction condition is mild, and large-scale production can be realized.

[0023] 2. The present application performs secondary separation and purification on the Anemarrhena asphodeloides crude polysaccharide by column chromatography, and the effect is remarkable, and the Anemarrhena asphodeloides polysaccharide AAP1-2 is prepared for the first time.

[0024] 3. The structure of the Anemarrhena asphodeloides polysaccharide AAP1-2 is identified, and the physicochemical properties of each polysaccharide component are clear, which provides structural basis for exploring the pharmacological activity mechanism.

[0025] 4. The Anemarrhena asphodeloides polysaccharide AAP1-2 obtained in the present application is well preserved, the quality is controllable, can restore the movement ability and swallowing ability of the aging nematodes, enhance the resistance to heat stress and oxidative stress of the aging nematodes, and inhibit the levels of lipofuscin and ROS in the aging nematodes to play the anti-aging activity, which provides basis for the application of the Anemarrhena asphodeloides polysaccharide in the fields of medicine, health products and the like.

[0026] 5. The present application lays a foundation for the Anemarrhena asphodeloides polysaccharide drugs, quality control, and further research on the structure-activity relationship and action mechanism. BRIEF DESCRIPTION OF DRAWINGS

[0027] Figure 1 It is a high performance liquid chromatogram of the Anemarrhena asphodeloides polysaccharide AAP1-2;

[0028] Figure 2 It is an infrared spectrum of the Anemarrhena asphodeloides polysaccharide AAP1-2;

[0029] Figure 3 It is the influence of the Anemarrhena asphodeloides polysaccharide AAP1-2 on the movement ability and swallowing ability of the aging nematodes;

[0030] Figure 4 It is the influence of the Anemarrhena asphodeloides polysaccharide AAP1-2 on the heat stress and oxidative stress of the aging nematodes;

[0031] Figure 5 Effects of Anemarrhena asphodeloides Bunge polysaccharide AAP1-2 on lipofuscin in aging nematodes;

[0032] Figure 6 Effects of Anemarrhena asphodeloides Bunge polysaccharide AAP1-2 on ROS in aging nematodes. DETAILED DESCRIPTION

[0033] The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work fall within the protection scope of the present application.

[0034] If not specifically indicated, the technical means used in the examples is the conventional means well known to those skilled in the art.

[0035] The present application discloses Anemarrhena asphodeloides Bunge polysaccharide AAP1-2, which is a uniform polysaccharide with a molecular weight ranging from 1000 to 100000 Da, and has a remarkable anti-aging effect.

[0036] The present application discloses a preparation method of the Anemarrhena asphodeloides Bunge polysaccharide AAP1-2, comprising the following steps:

[0037] (1) The dried rhizome of Anemarrhena asphodeloides Bunge is crushed, cleaned with water, and dried to obtain Anemarrhena asphodeloides Bunge rhizome powder, which is added into water and heated for extraction, and then filtered to obtain an extract; the amount of water added is 6-10 times the weight of the Anemarrhena asphodeloides Bunge powder; the heating extraction temperature is 60-80 DEG C; the heating extraction time is 1-3 h; and the heating extraction times is 2-4 times.

[0038] (2) The extract obtained in step (1) is subjected to multiple alcohol precipitation, and after each alcohol precipitation, the supernatant obtained after alcohol precipitation is subjected to re-alcohol precipitation; and after each alcohol precipitation, crude polysaccharide AA1, crude polysaccharide AA2 and crude polysaccharide AA3 are obtained, respectively.

[0039] The process of multiple alcohol precipitation is as follows: the extract obtained in step (1) is concentrated under reduced pressure (vacuum degree-0.1 MPa) to obtain concentrated solution 1; ethanol is added to the concentrated solution 1 to a volume concentration of a%, and then the precipitate and supernatant are collected to obtain crude polysaccharide AA1 and supernatant 1; the supernatant 1 is concentrated under reduced pressure to obtain concentrated solution 2; ethanol is added to the concentrated solution 2 to a volume concentration of b%, and then the precipitate and supernatant are collected to obtain crude polysaccharide AA2 and supernatant 2; the supernatant 2 is concentrated under reduced pressure to obtain concentrated solution 3; ethanol is added to the concentrated solution 3 to a volume concentration of c%, and then the precipitate is collected to obtain crude polysaccharide AA3; wherein 10≤a<b<c<100. The temperature for concentration under reduced pressure is 40-70 DEG C, and the standing time is 10-15 h.

[0040] (3) The crude polysaccharide AA1 is subjected to protein removal, dialysis and freeze-drying to obtain the crude anemarrhena rhizome polysaccharide AA1; the crude anemarrhena rhizome polysaccharide AA1 is subjected to ion exchange column chromatography, gradient elution is performed using a NaCl solution with a concentration of 0-2 M, and the phenol-sulfuric acid method is used to track the elution curve; according to the elution curve, the sugar part is collected and concentrated, and freeze-drying is performed to obtain the crude anemarrhena rhizome polysaccharide AAP1;

[0041] (4) The crude anemarrhena rhizome polysaccharide AAP1 is dissolved in water, centrifuged, and the supernatant is subjected to gel column chromatography using a molecular sieve; elution is performed using water, the phenol-sulfuric acid method is used to detect the elution curve, and according to the elution curve, the sugar part is collected, concentrated and freeze-dried to obtain the crude anemarrhena rhizome polysaccharide AAP1-2.

[0042] The present application adopts the combination of water extraction and alcohol precipitation, and high-concentration ethanol can separate polysaccharides with large polarity and good water solubility from polysaccharides with small polarity and poor water solubility, thereby solving the problem of complicated and difficult separation in the later stage caused by the traditional water boiling method for extracting polysaccharides.

[0043] The present application discloses a kind of identification method of the anemarrhena rhizome polysaccharide AAP1-2, and the basic characterization information of anemarrhena rhizome polysaccharide AAP1-2 is successfully obtained, including the following steps:

[0044] (1) take anemarrhena rhizome polysaccharide AAP1-2, and use high performance liquid chromatography to detect uniformity and molecular weight;

[0045] (2) take anemarrhena rhizome polysaccharide AAP1-2, dry, tablet, and detect by infrared spectroscopy.

[0046] The present application uses anemarrhena rhizoma dry powder as raw material, and separates crude polysaccharide by water extraction and alcohol precipitation; the extracted crude polysaccharide is subjected to deproteinization; then ion exchange chromatography and gel molecular sieve column chromatography are used to purify the crude anemarrhena rhizome polysaccharide; a kind of anemarrhena rhizome polysaccharide pure product is prepared for the first time; the physicochemical properties, molecular weight, infrared spectrum and the like of the polysaccharide pure product are systematically analyzed and identified; the basic characterization information of the anemarrhena rhizome polysaccharide is successfully obtained; and the anemarrhena rhizome polysaccharide AAP1-2 is a homogeneous polysaccharide.

[0047] The application discloses application of the Anemarrhena asphodeloides polysaccharide AAP1-2 in preparation of anti-aging drugs or health products.

[0048] The application will be further described in combination with specific examples.

[0049] Example 1

[0050] A preparation method of the Anemarrhena asphodeloides polysaccharide AAP1-2 comprises the following steps:

[0051] S1, crushing

[0052] 1 kg of dried Anemarrhena asphodeloides fruits is crushed, washed quickly with cold water, and dried to obtain Anemarrhena asphodeloides powder;

[0053] S2, water extraction

[0054] 8 times water by weight of the Anemarrhena asphodeloides powder obtained in step S1 is added, heated to 70 DEG C for extraction, and the extraction is carried out for 1 h; the extraction liquid is collected, and the residue is dried to obtain the extraction liquid and the residue;

[0055] S3, fractional alcohol precipitation

[0056] The extraction liquid obtained in step S2 is concentrated under reduced pressure (vacuum degree -0.1 MPa) at 60 DEG C to obtain concentrated liquid 1; ethanol is added to the concentrated liquid 1 until the volume concentration of the ethanol is 50%, and the mixture is left overnight; the precipitate and the supernatant are collected to obtain crude polysaccharide AA1 and supernatant 1;

[0057] The supernatant 1 is concentrated under reduced pressure to obtain concentrated liquid 2; ethanol is added to the concentrated liquid 2 until the volume concentration of the ethanol is 70%, and the mixture is left overnight; the precipitate and the supernatant are collected to obtain crude polysaccharide AA2 and supernatant 2;

[0058] The supernatant 2 is concentrated under reduced pressure to obtain concentrated liquid 3; ethanol is added to the concentrated liquid 3 until the volume concentration of the ethanol is 90%, and the mixture is left overnight; the precipitate is collected to obtain crude polysaccharide AA3;

[0059] Wherein, 10≤a

[0060] S4, purification

[0061] S4-01, primary purification

[0062] The crude polysaccharide AA1 obtained in step S3 is subjected to protein removal by using the Sevage method; after the protein removal, the crude polysaccharide is subjected to dialysis by using a dialysis bag (molecular weight cut-off 1000 Da) and freeze-drying to obtain the Anemarrhena asphodeloides polysaccharide AA1;

[0063] S4-02, secondary purification

[0064] Take 150 mg of Anemarrhena asphodeloides Bunge polysaccharide AA1, dissolve in 5 mL of deionized water, and load on a DEAE-FF column, and use 0-2 M NaCl solution to perform gradient elution. One elution peak appears, in which the elution peak is 0.15 M NaCl elution part (phenol-sulfuric acid method is used to track the elution curve during the elution process, and the sugar part is collected according to the elution curve). After the obtained elution liquid is concentrated and freeze-dried, an Anemarrhena asphodeloides Bunge polysaccharide AAP1 is obtained.

[0065] The freeze-dried Anemarrhena asphodeloides Bunge polysaccharide AAP1 is dissolved with water, centrifuged, and the supernatant is taken and loaded on a Sephadex G-100 column and eluted with water. The phenol-sulfuric acid method is used to track the elution curve, and one single symmetrical peak appears. The main peak is collected, concentrated, freeze-dried, and Anemarrhena asphodeloides Bunge polysaccharide AAP1-2 is obtained.

[0066] Example 2

[0067] Structure analysis of Anemarrhena asphodeloides Bunge polysaccharide AAP1-2

[0068] (I) Test material: Anemarrhena asphodeloides Bunge polysaccharide AAP1-2 in Example 1.

[0069] (II) Test method:

[0070] 1. Uniformity and molecular weight analysis

[0071] Sample treatment:

[0072] Precisely take 5 mg of Anemarrhena asphodeloides Bunge polysaccharide AAP1-2 and dissolve in 1 mL of deionized water. After filtration with a 0.22 μm filter membrane, it is used for HPLC sample analysis.

[0073] Chromatographic column:

[0074] TSKgel G3000PWXL, 7.8 x 300 mm, 7 μm; mobile phase: 0.02 M phosphate buffer; detector: RID; flow rate: 0.5 mL / min; sample volume: 10 μL.

[0075] 2. Infrared spectrum detection

[0076] Grind 2.0 mg of dried Anemarrhena asphodeloides Bunge polysaccharide AAP1-2 test material with KBr, press into a tablet, and scan in the range of 4000-400 cm -1 using IR Affinity-1.

[0077] (III) Test results:

[0078] 1. Structure characterization of Anemarrhena asphodeloides Bunge polysaccharide AAP1-2

[0079] (1) Monosaccharide composition analysis

[0080] AsFigure 1 As shown in the HPLC spectrum, the Anemarrhena asphodeloides polysaccharide AAP1-2 presents a single symmetrical peak, indicating that the AAP1-2 is a homogeneous polysaccharide.

[0081] (2) Infrared spectrum analysis

[0082] As shown in the infrared spectrum of the Anemarrhena asphodeloides polysaccharide AAP1-2, the Anemarrhena asphodeloides AAP1-2 contains the infrared characteristic absorption peaks of polysaccharides. Figure 2

[0083] Example 3

[0084] Study on the anti-aging effect of the Anemarrhena asphodeloides polysaccharide AAP1-2 on nematodes

[0085] (I) Test material: the Anemarrhena asphodeloides polysaccharide AAP1-2 in Example 1.

[0086] (II) Test object: nematodes (purchased from Caenorhabditis Genetics Center).

[0087] (III) Test method:

[0088] 1. Nematode culture:

[0089] OP50 bacterial liquid was sucked and added to the NGM plate and evenly coated. The plate coated with the bacterial liquid was cultured in a 37°C incubator overnight and then stored at room temperature. The nematodes were inoculated on the NGM containing OP50 and cultured at 20°C. When the food on the NGM plate was consumed, the Caenorhabditis elegans was washed off using M9, and after sedimentation, it was transferred to a new NGM plate.

[0090] 2. Synchronization

[0091] The oogenesis period of the Caenorhabditis elegans was lysed into a 1.5 mL centrifuge tube using a Caenorhabditis elegans lysis solution (NaClO4:2M NaOH:H2O=1:1:1), centrifuged, and washed 3 times to obtain worm eggs. The worm eggs were transferred to a centrifuge tube containing 9 mL S Medium and cultured at 20°C for 24 h.

[0092] 2. Drug administration

[0093] After 24 h of synchronization, the nematodes were transferred to the NGM medium containing OP50 and cultured at 20°C for 44 h. Then the Caenorhabditis elegans was washed off using M9, washed 3 times for drug administration, and cultured at 20°C for 7 days after drug administration.

[0094] Blank group: S Medium with a final concentration of 5-fluorouracil (75 μg / mL) + NA22 (OD value of 0.4).

[0095] ​Drug administration group: S Medium medium containing 5-fluorouracil (75 μg / mL) + NA22 (OD value 0.4) + different concentrations of polysaccharides (50, 100, 200 μg / mL).

[0096] 3. Experiments on nematode motility and swallowing ability

[0097] After administration, the nematodes were washed three times with M9 solution and transferred to blank NMG medium. Their body bending movements were recorded for 20 seconds. The remaining nematodes were transferred to NMG medium containing OP50, and their pharyngeal pump swallowing was recorded for 20 seconds.

[0098] 4. Nematode heat stress and oxidative stress experiments

[0099] After administration, the nematodes were washed three times with M9 solution. For the heat stress experiment, the nematodes were transferred to 96-well plates and incubated at 37°C. Observations and records were recorded every 2 hours until the last nematode died. For the oxidative stress experiment, the nematodes were transferred to 96-well plates, 500 μM juglone was added to each well, and the plates were incubated at 20°C. Observations and records were recorded every 2 hours until the last nematode died.

[0100] 5. Detection experiment of lipofuscin in nematodes

[0101] After administration, the sample was washed three times with M9 solution, fixed with paraformaldehyde, and the fluorescence intensity was observed under a fluorescence microscope and quantified using Image J.

[0102] 6. Detection experiment of lipofuscin in nematodes

[0103] After administration, the probe was washed three times with M9 solution, and then incubated with 200 μM H2DCFDA at 20°C in the dark for 2 hours. After incubation, the probe was washed away with M9 solution, and the fluorescence intensity was observed under a fluorescence microscope and quantified using ImageJ.

[0104] (IV) Experimental Results

[0105] like Figures 3-6 As shown, Anemarrhena asphodeloides polysaccharide AAP1-2 can restore the motility and swallowing ability of senescent nematodes, enhance their resistance to heat stress and oxidative stress, and inhibit the content of lipofuscin and ROS in senescent nematodes to exert anti-aging activity.

[0106] In summary, the pure polysaccharide AAP1-2 prepared by this invention is a homogeneous polysaccharide that can restore the motility and swallowing ability of senescent nematodes, enhance their resistance to heat stress and oxidative stress, and inhibit the content of lipofuscin and ROS in senescent nematodes to exert anti-aging activity.

[0107] The above described embodiments are only to illustrate the preferred modes of the present application, and are not intended to limit the scope of the present application. Any modification and improvement made by those skilled in the art to the technical solutions of the present application without departing from the design spirit of the present application shall fall within the protection scope of the present application as defined by the claims.

Claims

1. An Anemarrhena asphodeloides polysaccharide AAP1-2, characterized in that: The Anemarrhena polysaccharide AAP1-2 is a homogeneous polysaccharide with a molecular weight range of 1000-100000 Da.

2. A method for preparing the Anemarrhena asphodeloides polysaccharide AAP1-2 according to claim 1, characterized in that: Includes the following steps: (1) The dried rhizome of Anemarrhena asphodeloides is crushed to obtain rhizome powder, which is added to water, heated and extracted, filtered to obtain the extract; (2) The extract obtained in step (1) is subjected to multiple alcohol precipitations. After each alcohol precipitation, the supernatant obtained after alcohol precipitation is subjected to alcohol precipitation again. After each alcohol precipitation, crude polysaccharide AA1, crude polysaccharide AA2 and crude polysaccharide AA3 are obtained respectively. (3) The crude polysaccharide AA1 was deproteinized, dialyzed, and freeze-dried to obtain the mother polysaccharide AA1; the mother polysaccharide AA1 was subjected to ion exchange column chromatography, and gradient elution was performed with 0-2M NaCl solution. The sugar fraction was collected according to the elution curve and concentrated and freeze-dried to obtain the mother polysaccharide AAP1. (4) Dissolve Anemarrhena polysaccharide AAP1 in water, centrifuge, take the supernatant for molecular sieve gel column chromatography, elute with water, collect the sugar fraction according to the elution curve, concentrate and freeze dry to obtain Anemarrhena polysaccharide AAP1-2.

3. The preparation method according to claim 2, characterized in that: In step (1), the amount of water added is 6-10 times the weight of Anemarrhena asphodeloides powder.

4. The preparation method according to claim 2, characterized in that: In step (1), the temperature for heating and extraction is 60-80℃, the heating and extraction time is 1-3h, and the number of heating and extraction times is 2-4.

5. The preparation method according to claim 2, characterized in that: In step (2), the multiple alcohol precipitation process is as follows: the extract obtained in step (1) is concentrated under reduced pressure to obtain concentrate 1; ethanol is added to concentrate 1 until the ethanol volume concentration is a%, and the mixture is allowed to stand. The precipitate and supernatant are collected to obtain crude polysaccharide AA1 and supernatant 1; supernatant 1 is concentrated under reduced pressure to obtain concentrate 2; ethanol is added to concentrate 2 until the ethanol volume concentration is b%, and the mixture is allowed to stand. The precipitate and supernatant are collected to obtain crude polysaccharide AA2 and supernatant 2; supernatant 2 is concentrated under reduced pressure to obtain concentrate 3; ethanol is added to concentrate 3 until the ethanol volume concentration is c%, and the mixture is allowed to stand. The precipitate is collected to obtain crude polysaccharide AA3; wherein, 10≤a<b<c<100.

6. The preparation method according to claim 5, characterized in that: The concentration under reduced pressure is carried out at a temperature of 40-70℃ and a settling time of 10-15 hours.

7. A method for identifying Anemarrhena asphodeloides polysaccharide AAP1-2 as described in claim 1, characterized in that: Includes the following steps: (1) Take Anemarrhena polysaccharide AAP1-2 and use high performance liquid chromatography to detect its homogeneity and molecular weight; (2) Take Anemarrhena polysaccharide AAP1-2, dry it, compress it into tablets, and detect it by infrared spectroscopy.

8. The use of the Anemarrhena asphodeloides polysaccharide AAP1-2 as described in claim 1 in the preparation of anti-aging drugs or health products.

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