CN113876754B - Preparation method of lycium ruthenicum anthocyanin extract and freeze-dried powder and application of lycium ruthenicum anthocyanin extract and freeze-dried powder in diabetic cataract prevention and treatment products - Google Patents
Preparation method of lycium ruthenicum anthocyanin extract and freeze-dried powder and application of lycium ruthenicum anthocyanin extract and freeze-dried powder in diabetic cataract prevention and treatment products Download PDFInfo
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- CN113876754B CN113876754B CN202111270287.6A CN202111270287A CN113876754B CN 113876754 B CN113876754 B CN 113876754B CN 202111270287 A CN202111270287 A CN 202111270287A CN 113876754 B CN113876754 B CN 113876754B
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- lycium ruthenicum
- anthocyanin
- freeze
- resin
- dried powder
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Abstract
The invention relates to the technical field of lycium ruthenicum anthocyanin extracts, products and application thereof, in particular to a preparation method of lycium ruthenicum anthocyanin extracts and freeze-dried powder and application of the lycium ruthenicum anthocyanin extracts and freeze-dried powder in products for preventing and treating diabetic cataract, and the lycium ruthenicum anthocyanin extracts and freeze-dried powder are prepared through the steps of extraction, high-speed centrifugation and ceramic membrane filtration, organic membrane degumming and concentration, simulated moving bed chromatographic separation, low-temperature concentration, freeze drying and freeze-dried powder preparation. The preparation method of the lycium ruthenicum anthocyanin extract and the freeze-dried powder provided by the invention are suitable for industrial production, and have the advantages of high anthocyanin content, high recovery rate, good product color and no exogenous acidic substance residue. The lycium ruthenicum anthocyanin extract and the freeze-dried powder provided by the invention can inhibit the generation of ROS and AGEs in a crystalline lens under a high-sugar environment, inhibit the reduction of enzyme activity related to oxidative stress, improve an antioxidant defense system, have a certain prevention and treatment effect on diabetic cataract, and can be used for functional foods, health-care foods and medicines for preventing and treating the diabetic cataract.
Description
Technical Field
The invention relates to the technical field of lycium ruthenicum anthocyanin extracts, products and application thereof, in particular to a preparation method of lycium ruthenicum anthocyanin extracts and freeze-dried powder and application thereof in products for preventing and treating diabetic cataract.
Background
Diabetic cataract is one of the main complications of diabetes, is the first blinding eye disease and is closely related to the change of the osmotic pressure of the crystalline lens of the diabetes, the accumulation of glycosylation end products, oxidative stress and the like. According to statistics, the probability of diabetic cataract in diabetic patients is 63%, and the disease course is fast in development. Unlike age-related cataracts, juvenile diabetic patients can develop the disease and the incidence increases significantly as the course of diabetes increases. At present, no medicine can cure cataract, and the treatment is mainly performed in an operation mode. However, the epidemiological survey research center for diabetic retinopathy reports that diabetic patients are at high risk of cataract extraction, and the operation may lead to rapid acceleration of retinopathy, and induce or cause iris and macular changes such as iris adhesion or cystoid macular edema, and the recurrence rate is more than 24.8% 10 years after the operation. Studies have shown that diabetic cataract formation is reversible before lenticular opacity. Not only reducing the high sugar environment, normal metabolism of crystalline lens, but also gradually clearing the cortical opacity. After the lenticular nucleus develops opacity, it becomes an irreversible diabetic cataract. Therefore, in view of the problems of risk and economic stress existing in the operation and easy recurrence after the operation, the focus of the treatment of diabetic cataract in the future should be on the aspects of preventing and delaying cataract formation, controlling blood sugar and the like by using medicaments, and the method has important significance for improving the survival rate of the diabetic patients and improving the life quality of the diabetic patients.
Lycium ruthenicum Murr, also called as "lateral Ma" in Tibetan medicine, is a perennial shrub of Lycium of Solanaceae, has strong vitality, can resist salt, cold and drought, and is mainly distributed in Qinghai, xinjiang, ningxia, tibet, northern Shaanxi, gansu and other places in China. Lycium ruthenicum Murr is collected in Tibetan medicine classic works such as Jingzhu Bencao (four medical classics), has sweet taste, mild nature and heart heat clearing effect, and is a plant used as both medicine and food. The lycium ruthenicum murr is rich in anthocyanin and alkaloid components, the anthocyanin component has multiple physiological functions of anti-inflammation, antioxidation, anti-aging, anti-tumor, vision protection and the like, is a natural antioxidant and free radical scavenger, and has wide development and application prospects in the industries of medicine, food, health care products and cosmetics. The research on alkaloid components is very few, and except a few analysis and determination researches, the researches on chemical components, biological activity and the like are basically blank.
Zl201010542518.X "preparation method of antioxidant lycium ruthenicum extract", discloses a method for preparing lycium ruthenicum extract by water extraction, colloid mill grinding, high pressure homogenization, microfiltration ceramic membrane impurity removal, macroporous resin separation and spray drying or freeze drying of fresh lycium ruthenicum; ZL 201610548250.8' A method for extracting anthocyanin from fructus Lycii, and discloses a method for preparing anthocyanin from fructus Lycii powder by mixing with ethanol solution, extracting, membrane treating, purifying with macroporous resin, concentrating under reduced pressure, and drying; ZL201711220791.9 'A composition with eyesight improving effect', discloses a composition with eyesight improving effect and its preparation, namely a composition with eyesight improving effect comprising Lycium ruthenicum Murr, lycium barbarum or their extracts as effective components, and lutein is also added into all the compositions. The composition provided by the invention has the functions of relieving visual fatigue, preventing and assisting in treating visual problems such as visual deterioration, nyctalopia, muscae volitantes, glaucoma, cataract and the like caused by macular degeneration, retinitis pigmentosa and diabetic retinopathy, and can improve the vision to the greatest extent.
According to the existing scheme, water or ethanol extraction and macroporous resin adsorption purification are mostly adopted for extracting, separating and purifying lycium ruthenicum anthocyanin, and acidic ethanol solution extraction and elution are mostly adopted in the processing treatment process, so that the added exogenous acidic substances cannot be removed in the whole extraction, separation and purification, concentration and drying processes, the obtained product contains a large amount of acid, the anthocyanin is dark red (the anthocyanin is bluish purple in a neutral state), the anthocyanin activity is reduced, and meanwhile, the product is increased in heavy metal chromium pollution risk due to acid corrosion in the production process. Colloid mill grinding, high pressure homogenization, superfine grinding, ultrasonic-microwave reaction extraction, bionic extraction, ultrasonic-assisted extraction and other pretreatment and extraction equipment are expensive, and the implementation of industrialization is difficult. In addition, the high temperature of the spray drying process causes the anthocyanin to be decomposed or inactivated, and the process is not suitable for producing the lycium ruthenicum anthocyanin. The medicine or health-care product composition has the effects of improving vision, relieving eye fatigue and improving eyesight, and preventing and treating macular degeneration, cataract, glaucoma, presbyopia, eye fatigue, diabetic retinopathy, amblyopia, nyctalopia, hypopsia, blurred vision and the like, is a mixture of various extracts containing lycium ruthenicum extract, is added with lutein, carotenoid, vitamin and the like, and has unclear medicinal components and action mechanism. The research on the pure lycium ruthenicum anthocyanin extract for preventing and treating the diabetic cataract is not reported.
Disclosure of Invention
In order to solve the technical problems, the invention aims to provide a preparation method of an anthocyanin extract and freeze-dried powder of lycium ruthenicum and application of the anthocyanin extract and freeze-dried powder in products for preventing and treating diabetic cataract.
The invention provides a preparation method of an anthocyanin extract of lycium ruthenicum, which specifically comprises the following steps:
and 4, simulated moving bed chromatographic separation: adding the membrane concentrated solution obtained in the step 3 into a simulated moving bed chromatography for separation to obtain a lycium ruthenicum fine anthocyanin separation solution; the simulated moving bed separation method comprises the following steps: the adsorbent filled by the simulated moving bed chromatography adopts any one or more of nonpolar macroporous adsorption resin, weak-polarity macroporous resin and polar macroporous resin, and the flow rate of an adsorption area is 1-2 BV/h; the water washing area is purified water, the dosage of the purified water is 1 to 2 times of the volume of the resin, and the flow rate is 1 to 3BV/h; the desorbent is 5-90% ethanol solution, the dosage is 2-4 times of the resin volume, the flow rate is 1-3 BV/h; the resin regeneration solvent is 95% ethanol solution, and the dosage of the resin regeneration solvent is 1 time of that of the resin; the flow rate is 2-3 BV/h; wherein the nonpolar macroporous adsorbent resin is HPD-100 and HPD-750; the weak-polarity macroporous resin comprises AB-8, DM130, D860021, DS-401 and CAD-40; the polar macroporous resin is ADS-21, ADS-7, BS-75, BS-45, NKA-2 and NKA-9;
and step 5, low-temperature concentration: recovering ethanol from the fine anthocyanin separation liquid of the lycium ruthenicum murr obtained in the step 4, and carrying out vacuum low-temperature concentration at the temperature of 10-50 ℃ and the vacuum degree of-0.06 MPa to obtain an anthocyanin concentration liquid of the lycium ruthenicum murr;
The invention also provides a preparation method of the lycium ruthenicum anthocyanin freeze-dried powder, which specifically comprises the following steps:
and 4, simulated moving bed chromatographic separation: adding the membrane concentrated solution obtained in the step 3 into a simulated moving bed chromatography for separation to obtain a lycium ruthenicum fine anthocyanin separation solution; the simulated moving bed separation method comprises the following steps: the adsorbent filled by the simulated moving bed chromatography adopts any one or more of nonpolar macroporous adsorption resin, weak-polarity macroporous resin and polar macroporous resin, and the flow rate of an adsorption area is 1-2 BV/h; the water washing area is purified water, the dosage of the purified water is 1 to 2 times of the volume of the resin, and the flow rate is 1 to 3BV/h; the desorbent is 5-90% ethanol solution, the dosage is 2-4 times of the resin volume, the flow rate is 1-3 BV/h; the resin regeneration solvent is 95% ethanol solution, and the dosage of the resin regeneration solvent is 1 time of that of the resin; the flow rate is 2-3 BV/h; wherein, the nonpolar macroporous adsorption resin is HPD-100 and HPD-750; the weak-polarity macroporous resin is AB-8, DM130, D860021, DS-401 and CAD-40; the polar macroporous resin is ADS-21, ADS-7, BS-75, BS-45, NKA-2 and NKA-9;
and step 5, low-temperature concentration: recovering ethanol from the fine anthocyanin separation liquid of the lycium ruthenicum mill obtained in the step 4, and carrying out vacuum low-temperature concentration at the temperature of 10-50 ℃ and the vacuum degree of-0.06 MPa to obtain an anthocyanin concentration liquid of the lycium ruthenicum mill;
Further, the auxiliary materials added in the step 6 are one of starch, maltodextrin, fructo-oligosaccharide, galacto-oligosaccharide, lactose, inulin, beta-cyclodextrin, anhydrous glucose, polydextrose, milk powder, calcium citrate, xylitol, sorbitol and D-mannitol, and then a freeze dryer is adopted to freeze and dry the mixture at the temperature of between 50 ℃ below zero and 70 ℃ below zero to obtain the lycium ruthenicum anthocyanin freeze-dried powder.
The invention also provides the lycium ruthenicum anthocyanin extract and lycium ruthenicum anthocyanin freeze-dried powder prepared by the method, wherein the lycium ruthenicum anthocyanin extract and the lycium ruthenicum anthocyanin freeze-dried powder contain one or more or all of the following chemical components: n-trans-silapropylhydrazine, (E) -N- (4-Acetamidobutyl) -2- [4,5-dihydroxy-2- [3- [2- (4-hydroxyphenyl) ethyllamino ] -3-oxoprophyl ] -phenyl ] -3- (4-hydroxy-3-methoxyphenyl) prop-2-enamine (1S, 2R) -N3- (4-Acetamidobutyl) -1- (3, 4-dihydroxy-phenyl) -7-hydroxy-N2- (4-hydroxyphentyl) -6, 8-dihydroxy-1, 2-dihydroxy-naphthalene-2, 3-dicarboamide, petuniadin 3-O- [6-O- (4-O-trans- (beta-D-glucopyranoside) -p-coumaroyl) -alpha-L-hydroxyamnopyranosyl) -beta-D-glucopyranoside ] -5-O- [ beta-D-glucopyranoside ], (N), n-dihydrocaffeoylspersmidine, N-hydroxyyl-N-hydroxycarboylspersmidine, N-hydroxycarboyl-N-hydroxyospermimidine, petuninidin-3-O- (6-O-p-coumaryl) -rutinoside-5-O-glucoside, malvidin-3-O-rutinoside (cis-p-coumaryl) -5-O-lucinoside, petuninidin-3-O-rutinoside (caffeine) -5-O-lucinoside (capsular-p-coumaryl) -5-O-lucinoside, and Malvidin-3-O-rutinoside (cis-p-coumaryl) -5-O-gcosinide;
the structural formula of the chemical components is as follows in sequence:
the anthocyanin content in the lycium ruthenicum anthocyanin extract and the lycium ruthenicum anthocyanin freeze-dried powder is 0.1-90%, and the total alkaloid content is 0.1-90%.
The invention also protects the application of the lycium ruthenicum anthocyanin extract and the lycium ruthenicum anthocyanin freeze-dried powder prepared by the method; the lycium ruthenicum anthocyanin extract and the lycium ruthenicum anthocyanin freeze-dried powder are applied to preparation of products for preventing and treating diabetic cataract, the products for preventing and treating diabetic cataract are food, health-care food or medicine, and the lycium ruthenicum anthocyanin extract and/or the freeze-dried powder are/is used as effective components and are prepared into various dosage form medicines according to any pharmaceutically acceptable carrier or various food or health-care food according to any carrier which is acceptable in food science.
Furthermore, the food, the health food or the medicine is in the form of powder, tablets or capsules.
Further, the food is lycium ruthenicum anthocyanin solid beverage, and the health-care food is lycium ruthenicum anthocyanin tablet candy; the medicine is a lycium ruthenicum anthocyanin capsule medicine.
Compared with the prior art, the invention has the following beneficial effects:
the preparation methods of the lycium ruthenicum anthocyanin extract and the lycium ruthenicum anthocyanin freeze-dried powder are suitable for industrial production, the preparation process is green and pollution-free, the anthocyanin content of the prepared extract and freeze-dried powder is high, no exogenous acid is required to be added, the natural attribute of the lycium ruthenicum anthocyanin is completely maintained, and harmful substances such as heavy metal, pesticide residue and the like in raw materials are removed. The prepared lycium ruthenicum anthocyanin extract is researched by Streptozotocin (STZ) -induced diabetes rat experimental models, and the result shows that the lycium ruthenicum anthocyanin can improve weight loss of STZ-induced diabetes rats, reduce blood sugar concentration, improve SOD activity in retinas and reduce MDA content, so that oxidative damage to the retinas is reduced, and the prevention and treatment effects on diabetic retinopathy are exerted; can inhibit STZ-induced retinal cell apoptosis of diabetic rats, and has certain protective effect on diabetic retinopathy; can inhibit the generation of ROS and AGEs in crystalline lens under high sugar environment, inhibit the decrease of enzyme activity related to oxidative stress, and improve antioxidant defense system, thereby having certain preventing and treating effect on diabetic cataract. Can be used as functional food, health food and medicine for preventing and treating diabetic cataract.
Drawings
FIG. 1 is a diagram of the chemical composition of 11 Lycium ruthenicum anthocyanin extracts prepared by the invention;
FIG. 2 is a HPLC chart of the anthocyanin extract of Lycium ruthenicum Murr obtained in example 1 of the present invention;
FIG. 3 is a schematic diagram of an HPLC/MS-MS analysis ion flow chart of the anthocyanin extract of Lycium ruthenicum Murr obtained in example 1 of the present invention;
FIG. 4 is a diagram of simulated moving bed separation of anthocyanin extract of Lycium ruthenicum Murr obtained in example 1 of the invention;
FIG. 5 is a graph of the effect of the present invention of anthocyanidin extract of Lycium ruthenicum on the STZ-induced expression of Bax in retinas of diabetic rats;
FIG. 6 is a graph of the effect of the anthocyanidin extract of Lycium ruthenicum Murr of the present invention on STZ-induced Bcl-2 expression in diabetic rat retinas;
FIG. 7 is a graph of the effect of the anthocyanidin extract of Lycium ruthenicum Murr of the present invention on the induction of ROS in lens tissues of STZ-diabetic rats;
FIG. 8 shows the effect of the anthocyanidin extract of Lycium ruthenicum Murr of the present invention on the induction of AGEs in the lens tissue of STZ-induced diabetic rats.
In FIGS. 5 and 6, normal blank Control group, control galactose aging model Control group, A Lycium ruthenicum anthocyanin 1g crude drug/kg dose group, B Lycium ruthenicum anthocyanin 2g crude drug/kg dose group, C Lycium ruthenicum anthocyanin 5g crude drug/kg dose group; Δ P <0.0001, <0.05, <0.001.
In FIGS. 7 and 8, a Normal blank Control group, a Control galactose aging model Control group, a Lycium ruthenicum anthocyanin 1g crude drug/kg dose group, B Lycium ruthenicum anthocyanin 2g crude drug/kg dose group, and C Lycium ruthenicum anthocyanin 5g crude drug/kg dose group; Δ P <0.0001, # P <0.01, # P <0.001.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be obtained by a person skilled in the art without making any creative effort based on the embodiments in the present invention, belong to the protection scope of the present invention.
Example 1
The lycium ruthenicum raw material used in the embodiment of the invention is provided by Qinghai Jinmaiqi Biotech limited and purchased from gelmu City of Qinghai province.
Test equipment: CSJ-20 coarse crusher (Jiangyin Square and round machines manufacturing Co., ltd.), ZD-LZ-1.5 juicer (Zhongde food machinery Jingjiang Co., ltd.), extraction equipment (Zhejiang warm brother machinery valve industry Co., ltd.), GQ105B tubular centrifuge (Shanghai Pudongtian centrifugal machinery Co., ltd.), PTSX40 butterfly centrifuge (Yixing Huading machinery Co., ltd.), SMB-H-08-A simulated moving bed (Hanbang science Co., ltd.), GLZ-1B freeze dryer (Shanghai Pudong freeze-drying equipment Co., ltd.), SCIENTZ-10 freeze dryer (Ningbo New Zhi Biotechnology Co., ltd.), ZPSX tablet press (Shanghai Xiangshun pharmaceutical machinery Co., ltd.), NJP-300 full-automatic capsule filling machine (Reynaud Tian pharmaceutical machinery Co., ltd.).
An experimental instrument: roche Diagnostics GmbH model rapid glucometer and dipstick (Roche, germany), spectraMax190 enzyme-linked immunosorbent assay (MD, usa), 5424R desk-top high-speed refrigerated centrifuge (Eppendorf); MY-20 hand-held tissue grinder (Shanghai Jingxin), 9700PCR amplificator (ABI, USA), rad-IQ5 real-time fluorescence quantitative PCR instrument (Bio-Rad, USA), and QubitTM nucleic acid protein analyzer (Invitrogen).
The experimental reagent: lycium ruthenicum anthocyanin extract (supplied by Qinghai Jinmaiqi Biotech Co., ltd., lot No. 20201001); DPPH (Sigma, USA), ascorbic acid (VC) and Phenazine Methosulfate (PMS) are all purchased from Allantin; reduced coenzyme I (NADH) and Nitrotetrazolium Blue (NBT) were purchased from Biyunyan; 30% of H 2 O 2 Iron sulfate and salicylic acid are all purchased from Tianjin Damao reagent company; physiological saline (batch No. C17010902, a product of Hebei Tian Chengyou GmbH); streptozotocin (sigma, usa); high-sugar high-fat diet (Ready bit D12492); pentobarbital (Shanghai chemical reagent company, pharmaceutical group, china); superoxide dismutase (SOD) and glutathionePeptide peroxidase (GSH-Px), glutathione (GSH), oxidized Glutathione (GSSH), glutathione Reductase (GR), catalase (CAT), advanced glycosylation end products (AGEs) and Malondialdehyde (MDA) kits are all products of Nanjing institute of engineering; an intracellular Reactive Oxygen Species (ROS) detection kit (pecan corporation), an RNA extraction kit (Promega corporation, usa); a reverse transcription kit and a fluorescent quantitative PCR kit (Takara).
Animals: SD rat, SPF grade, supplied by beijing weitonglihua laboratory animal technology ltd, license number: SCXK (Jing) 2016-0006.
Example 1
The preparation method of the lycium ruthenicum anthocyanin extract specifically comprises the following steps:
and step 5, low-temperature concentration: recovering ethanol from the fine anthocyanin separation liquid of the lycium ruthenicum mill obtained in the step 4, and carrying out vacuum low-temperature concentration at 10 ℃ and under the vacuum degree of-0.06 MPa to obtain about 25Kg of anthocyanin concentration liquid of the lycium ruthenicum mill;
The anthocyanin content of the lycium ruthenicum anthocyanin extract is measured by a pH differential method to be 56.0%, and the total alkaloid content is measured by a spectrophotometry method to be 5.5%.
The fructus Lycii anthocyanin extract has scavenging capacity of 92.9% for organic free radical (DPPH. Cndot.) and superoxide anion (. O. Cndot.) 2 - ) The radical scavenging ability was 86.6%, and the hydroxyl radical (. OH) scavenging ability was 96.3%.
By HPLC-MS/MS analysis, the fructus Lycii anthocyanin extract contains chemical components 1,2, 3,4, 5, 6, 7, 8, 9, 10 and 11, and the structural formula is shown in figure 1.
Example 2
The preparation method of the lycium ruthenicum anthocyanin extract specifically comprises the following steps:
and 4, simulated moving bed chromatographic separation: adding the membrane concentrated solution obtained in the step 3 into a simulated moving bed chromatography for separation to obtain 260L of lycium ruthenicum fine anthocyanin separation solution; the simulated moving bed separation method comprises the following steps: the adsorbent filled by the simulated moving bed chromatography adopts AB-8 macroporous resin, and the flow rate of an adsorption zone is 1-2 BV/h; the water washing area is purified water, the dosage of the purified water is 1.5 times of the volume of the resin, and the flow rate is 2BV/h; the desorbent is 30-60% ethanol solution, the dosage of which is 3 times of the volume of the resin, and the flow rate is 1-2 BV/h; the resin regeneration solvent is 95% ethanol solution, and the dosage is 1 time of the resin; the flow rate is 2-3 BV/h;
and 5, low-temperature concentration: recovering ethanol from the fine anthocyanin separation liquid of the lycium ruthenicum mill obtained in the step 4, and carrying out vacuum low-temperature concentration at 50 ℃ and under the vacuum degree of-0.06 MPa to obtain about 22Kg of anthocyanin concentration liquid of the lycium ruthenicum mill;
The anthocyanin content in the lycium ruthenicum anthocyanin extract is 88.6% through pH differential method determination, and the total alkaloid content is 0.12% through spectrophotometry determination.
The fructus Lycii anthocyanin extract has scavenging capacity of 92.9% for organic free radical (DPPH. Cndot.) and superoxide anion (. O. Cndot.) 2 - ) The radical scavenging ability was 86.6%, and the hydroxyl radical (. OH) scavenging ability was 96.3%.
By HPLC-MS/MS analysis, the anthocyanidin extract of Lycium ruthenicum Murr contains chemical components 4, 8, 9, 10 and 11, and its structural formula is shown in figure 1.
Example 3
The preparation method of the lycium ruthenicum anthocyanin extract specifically comprises the following steps:
and 5, low-temperature concentration: recovering ethanol from the fine anthocyanin separation liquid of the lycium ruthenicum murr obtained in the step 4, and carrying out vacuum low-temperature concentration at 20 ℃ and under the vacuum degree of-0.06 MPa to obtain about 26Kg of anthocyanin concentration liquid of the lycium ruthenicum murr;
The anthocyanin content of the lycium ruthenicum anthocyanin extract is 32.6% by a pH differential method, and the total alkaloid content is 18.2% by a spectrophotometry method.
The anthocyanin extract of Lycium ruthenicum Murr has scavenging ability of 82.3% for organic free radical (DPPH.), and scavenging ability for superoxide anion (. O.) 2 - ) Radical scavenging ability of 75.6%, forThe hydroxyl radical (. OH) scavenging ability was 82.9%.
By HPLC-MS/MS analysis, the anthocyanidin extract of Lycium ruthenicum Murr contains chemical components 1,2, 3,4, 5, 6, 7, 8, 9, 10 and 11, and its structural formula is shown in figure 1.
Example 4
The preparation method of the lycium ruthenicum anthocyanin extract specifically comprises the following steps:
and 5, low-temperature concentration: recovering ethanol from the fine anthocyanin separation liquid of the lycium ruthenicum mill obtained in the step 4, and carrying out vacuum low-temperature concentration at 25 ℃ and under the vacuum degree of-0.06 MPa to obtain about 15Kg of anthocyanin concentration liquid of the lycium ruthenicum mill;
The anthocyanin content of the lycium ruthenicum anthocyanin extract is 15.2% by a pH differential method, and the total alkaloid content is 75.6% by a spectrophotometry method.
The anthocyanin extract of Lycium ruthenicum Murr has scavenging ability of 77.8% for organic free radical (DPPH.), and can scavenge superoxide anion (. O.) 2 - ) The radical scavenging ability was 66.5%, and the hydroxyl radical (. OH) scavenging ability was 82.3%.
By HPLC-MS/MS analysis, the fructus Lycii anthocyanin extract contains chemical components 1,2, 3,4, 5, 6, 7, 8, 9, 10 and 11, and the structural formula is shown in figure 1.
Example 5
The preparation method of the lycium ruthenicum anthocyanin extract specifically comprises the following steps:
and 4, simulated moving bed chromatographic separation: adding the membrane concentrated solution obtained in the step 3 into a simulated moving bed chromatography for separation to obtain 80L of the lycium ruthenicum fine anthocyanin separation solution; the simulated moving bed separation method comprises the following steps: the adsorbent filled by the simulated moving bed chromatography adopts NKA-2 resin, and the flow rate of an adsorption zone is 1-2 BV/h; the water washing area is purified water, the dosage of the purified water is 1.5 times of the volume of the resin, and the flow rate is 2BV/h; the desorbent is 5-25% ethanol solution, the dosage is 3 times of the resin volume, and the flow rate is 1-2 BV/h; the resin regeneration solvent is 95% ethanol solution, and the dosage of the resin regeneration solvent is 1 time of that of the resin; the flow rate is 2-3 BV/h;
and 5, low-temperature concentration: recovering ethanol from the fine anthocyanin separation liquid of the lycium ruthenicum murr obtained in the step 4, and carrying out vacuum low-temperature concentration at 30 ℃ and under the vacuum degree of-0.06 MPa to obtain about 8Kg of anthocyanin concentration liquid of the lycium ruthenicum murr;
The anthocyanin content in the lycium ruthenicum anthocyanin extract is determined to be 3.2% by a pH differential method, and the total alkaloid content is determined to be 89.2% by a spectrophotometry method.
The anthocyanin extract of Lycium ruthenicum Murr has scavenging ability of 62.8% for organic free radical (DPPH.), and scavenging ability for superoxide anion (. O.) 2 - ) The radical scavenging ability was 56.5%, and the hydroxyl radical (. OH) scavenging ability was 72.6%.
By HPLC-MS/MS analysis, the anthocyanidin extract of Lycium ruthenicum Murr contains chemical components 1,2, 3,4, 5, 6, 7, 8 and 10, and its structural formula is shown in figure 1.
Example 6
The preparation method of the lycium ruthenicum anthocyanin freeze-dried powder specifically comprises the following steps:
and step 5, low-temperature concentration: recovering ethanol from the fine anthocyanin separation liquid of the lycium ruthenicum murr obtained in the step 4, and carrying out vacuum low-temperature concentration at 35 ℃ and under the vacuum degree of-0.06 MPa to obtain about 28Kg of anthocyanin concentration liquid of the lycium ruthenicum murr;
The anthocyanin content in the lycium ruthenicum anthocyanin freeze-dried powder is measured to be 8.6% by a pH differential method, and the content of total alkaloids is measured to be 1.8% by a spectrophotometry method.
The fructus lycii anthocyanin freeze-dried powder has the capacity of removing organic free radicals (DPPH. Of 75.4 percent) and superoxide anion (O) 2 - ) The radical scavenging ability was 65.6%, and the hydroxyl radical (. OH) scavenging ability was 68.5%.
Through HPLC-MS/MS analysis, the lycium ruthenicum anthocyanin freeze-dried powder contains chemical components 1,2, 3,4, 5, 6, 7, 8, 9 and 10, and the structural formula is shown in figure 1 in detail.
Example 7
The preparation method of the lycium ruthenicum anthocyanin freeze-dried powder specifically comprises the following steps:
and 4, simulated moving bed chromatographic separation: adding the membrane concentrated solution obtained in the step 3 into a simulated moving bed chromatography for separation to obtain 120L of lycium ruthenicum fine anthocyanin separation solution; the simulated moving bed separation method comprises the following steps: the adsorbent filled by the simulated moving bed chromatography adopts CAD-40: BS-45= 1; the water washing area is purified water, the dosage of the purified water is 1.5 times of the volume of the resin, and the flow rate is 2BV/h; the desorbent is 15-40% ethanol solution, the dosage of which is 3 times of the volume of the resin, and the flow rate is 1-2 BV/h; the resin regeneration solvent is 95% ethanol solution, and the dosage of the resin regeneration solvent is 1 time of that of the resin; the flow rate is 2-3 BV/h;
and step 5, low-temperature concentration: recovering ethanol from the fine anthocyanin separation liquid of the lycium ruthenicum mill obtained in the step 4, and carrying out vacuum low-temperature concentration at 40 ℃ and under the vacuum degree of-0.06 MPa to obtain about 22Kg of anthocyanin concentration liquid of the lycium ruthenicum mill;
The anthocyanin content in the lycium ruthenicum anthocyanin freeze-dried powder is measured to be 5.5% by a pH differential method, and the content of total alkaloids is measured to be 3.2% by a spectrophotometry method.
The fructus lycii anthocyanin freeze-dried powder has 70.6% of capability of removing organic free radicals (DPPH), and has an effect of removing superoxide anion (O) 2 - ) The radical scavenging ability was 61.2%, and the hydroxyl radical (. OH) scavenging ability was 65.8%.
By HPLC-MS/MS analysis, the fructus Lycii anthocyanidin lyophilized powder contains chemical components 1,2, 3,4, 5, 6, 7, 8, 9 and 10, and the structural formula is shown in figure 1.
Example 8
The effect of lycium ruthenicum anthocyanin extract on the level of oxidative stress induced by STZ on diabetic rat retina.
Rat experimental model for inducing diabetes by Streptozotocin (STZ)
50 healthy male SD rats with the weight of 180 +/-10 g are selected and randomly divided into 5 groups according to the weight, wherein the groups are respectively a normal control group, a model control group, and a Lycium ruthenicum anthocyanin 1,2 and 5g crude drugs/kg dose group, and each group comprises 10 animals. After the normal control group rats are fed with common feed, and the other groups are fed with high-sugar high-fat feed for 2 weeks, the model control group rats and the lycium ruthenicum anthocyanin 1,2 and 5g crude drug/kg dose group rats are fasted for 24 hours, water is freely drunk in the period, STZ50mg/kg is injected into the abdominal cavity at one time, normal diet is given, and meanwhile, normal control group rats are injected with normal saline with the same volume into the abdominal cavity at one time. After 72h, the tail vein is blood-taken to measure the fasting blood sugar of the rat, and the rat with the blood sugar more than 16.7mmol/L is a diabetes model rat after continuous measurement for 3 days. The stomach filling is started the next day after the model building is successful, the raw medicine/kg dose groups of the anthocyanin of the lycium ruthenicum 1,2 and 5g are subjected to stomach filling administration for 10 weeks according to the dose, and the normal saline of the blank control group and the model control group is subjected to stomach filling for 10 weeks to eat food freely. After the completion of the gavage, 5 rats were randomly selected from each group, were anesthetized by intraperitoneal injection of 2.5% pentobarbital, and their eyeballs were quickly removed, ice-surface-treated, and then the retinas were peeled off from a dissecting microscope, manually homogenized, centrifuged at 2500r/min for 15min, and the supernatants were removed, and the kit was used to detect the MDA content and SOD activity.
Data are all provided with
Statistical analysis of data was performed using the SPSS16.0 software and comparisons between groups were performed using the t-test.
Results of the experiment
TABLE 1 Effect of Lycium ruthenicum anthocyanin extract on STZ-induced body weight and blood glucose in diabetic rats
Note: comparison with normal control group: Δ P<0.001， ΔΔ P<0.0001; comparison with model control group: * P is<0.05，**P<0.0005.
As a result, compared with the blank control group, the STZ-induced diabetic rats showed a significant increase in body weight after 2 weeks of high-sugar and high-fat feeding (P < 0.001), and the fasting blood glucose after the STZ-induction (P < 0.0001), and the modeling was successful. After 6 weeks of intragastric administration, STZ induced significant decrease in body weight (P < 0.0001) and increase in fasting plasma glucose (P < 0.0001) in the diabetic rats compared to the placebo group; compared with the model group of the STZ-induced diabetes group, the weights of rats in the Lycium ruthenicum Murr anthocyanin group with 1g, 2g and 5g crude drugs/kg dose groups are increased, but are still lower than those of the blank control group, the fasting blood glucose is reduced to different degrees, and is still higher than that of the blank control group, and the effect of the Lycium ruthenicum Murr anthocyanin group with 5g crude drugs/kg dose group is most obvious (P < 0.0005).
TABLE 2 influence of Lycium ruthenicum anthocyanin extract on MDA content and SOD activity in retinas of STZ-induced diabetic rats
Note: comparison with normal control group: Δ P<0.0001; comparison with model control group: * P<0.05， ** P<0.005， *** P<0.0001.
As a result, compared with a blank control group, the SOD activity in the retina of the rat in the STZ diabetes model group is obviously reduced (P < 0.0001), and the MDA content is obviously increased (P < 0.0001); compared with the model group of the STZ-induced diabetes mellitus group, the SOD activity of the rat with the dose of 1,2 and 5g of lycium ruthenicum anthocyanin is obviously improved, the MDA content is obviously reduced, and the rat with the dose of 1g of crude drug/kg has significance (P is less than 0.05).
The retina is the initial site for sensing and transmitting visual information, and is metabolized vigorously in the body. The glucose metabolism disorder is easy to cause retina tissue metabolism disorder, lipid peroxidation is increased, SOD activity is reduced, and oxidative stress damage is caused. The results show that the lycium ruthenicum anthocyanin can improve the weight loss of STZ-induced diabetic rats, reduce the blood sugar concentration, improve the SOD activity in retinas and reduce the MDA content, thereby reducing the oxidative damage of the retinas and playing a role in preventing and treating diabetic retinopathy.
Example 9
Influence of lycium ruthenicum anthocyanin extract on STZ-induced apoptosis of diabetic rat retinal cells
Streptozotocin (STZ) was used to induce a diabetic rat model. After the completion of the gavage, 5 rats were randomly selected from each group, 2.5% pentobarbital was intraperitoneally injected for anesthesia, the eyeballs were rapidly taken out, ice surface operation was performed, the retinas were stripped off in a dissecting microscope, homogenate was manually performed, total RNA of retinal cells was extracted with an RNA kit, total RNA of retinal cells was reverse-transcribed into cDNA with a reverse transcription kit, and the cDNA was stored at 4 ℃ for detection of Bax and Bcl-2 expression with a fluorescence quantitative PCR kit.
TABLE 3 primers for RT-PCR experiments designed by pubmed and Primerpromierv 5.0, synthesized by Huada Gene
Δ Ct (calibration sample) = average Ct value for control group-GAPDH average Ct value for control group; Δ Ct (processing sample) = average Ct value of processing sample-GAPDH average Ct value of processing sample; the Δ Ct =: (calibration sample) the processing sample Δ Ct.
Results of the experiment
Effect of Lycium ruthenicum anthocyanin on STZ-induced expression of Bax and Bcl-2 in retinas of diabetic rats (see FIGS. 5 and 6 for details). A Normal blank Control group, a Control galactose aging model Control group, an A lycium ruthenicum anthocyanin 1g crude drug/kg dose group, a B lycium ruthenicum anthocyanin 2g crude drug/kg dose group, and a C lycium ruthenicum anthocyanin 5g crude drug/kg dose group; Δ P<0.0001，*P<0.05，***P<0.001。
bax and Bcl-2 are apoptosis-related genes, bcl-2 can prevent apoptosis and prolong the survival period of cells, bax can antagonize the apoptosis inhibition effect of Bcl-2 and promote apoptosis, and the balance of the Bax and the Bcl-2 ensures the steady state of the cells. The Bcl-2/Bax ratio is increased to inhibit apoptosis, so that the cells are protected. As a result, bax expression was significantly increased (P < 0.0001) and Bcl-2 expression was significantly decreased (P < 0.0001) in retinas of rats in the STZ diabetes model group, compared to the blank control group; compared with the model group of the STZ-induced diabetes group, the Bax expression of the lycium ruthenicum anthocyanin group with 1g crude drug/kg dose group rat has no obvious change, and the Bcl-2 expression is increased (P is less than 0.05); the Bax expression of rats in the lycium ruthenicum anthocyanin 2g crude drug/kg dose group is reduced (P < 0.05), and the Bcl-2 expression is increased (P < 0.05); the Bax expression of rats in the lycium ruthenicum anthocyanin 5g crude drug/kg dose group is remarkably reduced (P < 0.0001), and the Bcl-2 expression is remarkably increased (P < 0.0001). The lycium ruthenicum anthocyanin can inhibit retinal cell apoptosis of diabetic rats induced by STZ and has a certain protective effect on diabetic retinopathy.
Example 10
Prevention and treatment effect of lycium ruthenicum anthocyanin on diabetic cataract
Rat experimental model for inducing diabetes by Streptozotocin (STZ)
Selecting 30 healthy male SD rats with weight of 180 +/-10 g, randomly grouping according to weight, and respectively comprising a normal control group, a model control group, and 5 crude drug/kg dose groups of 1,2 and 5g of lycium ruthenicum anthocyanin, wherein each group comprises 6 animals. After the rats in the normal control group are fed with common feed, and the rest groups are fed with high-sugar high-fat feed for 2 weeks, the rats in the model control group and the lycium ruthenicum anthocyanin 1,2 and 5g crude drug/kg dose group are fasted for 24 hours, water is freely drunk in the process, STZ45mg/kg is injected into the abdominal cavity at one time, normal diet is given, and meanwhile, normal control group is injected into the abdominal cavity at one time with physiological saline with the same volume. After 72h, the tail vein is blood-taken to measure the fasting blood sugar of the rat, and the rat with the blood sugar more than 16.7mmol/L after continuous measurement for 3 days is a diabetes model rat. The stomach filling is started the next day after the model building is successful, the crude drug/kg dose groups of the anthocyanin of the lycium ruthenicum 1,2 and 5g are filled with the normal saline according to the dose for 6 weeks, and the blank control group and the model control group are filled with the normal saline for 6 weeks and eat freely. Killing the rat by a decapitation method after the completion of the intragastric perfusion, removing an eyeball, completely separating out a crystalline lens, grinding the crystalline lens by using physiological saline in an ice bath to prepare ice-cold homogenate with the mass concentration of 10%, centrifuging the homogenate at the temperature of 4 ℃ for 15min at 3000g, taking supernatant, and measuring CAT, SOD, MDA, GSH-Px, GSH, GR and GST values in tissues and organs by using a kit; grinding with normal saline in ice bath to obtain ice-cold homogenate with mass concentration of 10%, centrifuging at 4 deg.C for 30min at 3000g, collecting supernatant, and determining AGEs value in tissue and organ with kit.
Data are all provided with
Statistical data analysis was performed using SPSS16.0 software and comparisons between groups were performed using the t-test.
Results of the experiment
The effect of lycium ruthenicum anthocyanin on the induction of ROS and AGEs in the lens tissues of STZ-diabetic rats (see fig. 7 and 8 for details). A Normal blank Control group, a Control galactose aging model Control group, an A lycium ruthenicum anthocyanin 1g crude drug/kg dose group, a B lycium ruthenicum anthocyanin 2g crude drug/kg dose group, and a C lycium ruthenicum anthocyanin 5g crude drug/kg dose group; Δ P<0.0001，**P<0.01，***P<0.001。
as a result, the content of ROS and AGEs in the lenses of the rats in the STZ diabetes model group is remarkably increased compared with that of the blank control group (P < 0.0001). Compared with the model group of the STZ-induced diabetes mellitus group, the content of ROS and AGEs in the crystalline lens of the rat with the dose of 1g crude drug/kg anthocyanin of lycium ruthenicum Murr is slightly reduced, but the statistical significance is avoided; the content of ROS and AGEs in rat lens of the lycium ruthenicum anthocyanin group with 2g crude drug/kg dose is obviously reduced, and the effect is enhanced along with the increase of the dose.
TABLE 4 influence of Lycium ruthenicum anthocyanin on the activity of SOD, CAT and MDA in STZ-induced diabetic rat lenses
Note: comparison with normal control group: Δ P <0.0001; comparison with model control group: * P <0.05, P <0.005, P <0.0005.
As a result, compared with a blank control group, the SOD and CAT activities in the lens of the rat in the STZ diabetes model group are obviously reduced (P < 0.0001), and the MDA content is obviously increased (P < 0.0001); compared with the model group of the STZ-induced diabetes mellitus group, the lycium ruthenicum anthocyanin group rats with 1,2 and 5g crude drugs/kg dose can obviously improve the SOD and CAT activities and reduce the MDA content, and the effect is enhanced along with the increase of the dose.
TABLE 5 Effect of Lycium ruthenicum anthocyanin on STZ-induced GSH-Px, GR, GSSH and GSH in diabetic rat lenses
Note: comparison with normal control group: Δ P <0.0005, Δ Δ P <0.0005; comparison with model control group: * P <0.05, P <0.01, P <0.0005.
The lens has a GSH synthesis and regeneration system that is capable of maintaining high levels of GSH, protecting it from oxidative stress damage. As a result, the GSH-Px and GR activities in the lenses of the rats of the STZ diabetic model group were significantly reduced (P < 0.0001), the GSH content limitation was reduced (P < 0.0001), and the GSSH content was significantly increased (P < 0.0005) compared with the blank control group; compared with an STZ-induced diabetes group model group, the Lycium ruthenicum Murr anthocyanin 1g crude drug/kg dose group rat can obviously improve GR activity (P < 0.05) and GSH content (P < 0.01), and has no obvious influence on GSH-Px activity and GSSH content change. The lycium ruthenicum anthocyanin 2 and 5g crude drug/kg dose group rats can obviously improve the GSH content and the GSH-Px and GR activities, reduce the GSSH content, and the effect is enhanced along with the increase of the dose.
Oxidative damage and protein glycosylation can both cause opacification of the lens leading to the formation of diabetic cataracts. The results show that the lycium ruthenicum anthocyanin can inhibit the generation of ROS and AGEs in a lens under a high-sugar environment, inhibit the reduction of enzyme activity related to oxidative stress, and improve an antioxidant defense system, so that the lycium ruthenicum anthocyanin has a certain prevention and treatment effect on diabetic cataract.
Example 11
Lycium ruthenicum anthocyanin solid beverage food
The lycium ruthenicum anthocyanin freeze-dried powder prepared in the embodiment 6 is added with 18% of maltodextrin, 0.25% of sodium carboxymethylcellulose and 4% of sucrose or xylitol, mixed uniformly by a CH-200 trough type mixing machine, and granulated by a conventional method to obtain the lycium ruthenicum anthocyanin solid beverage.
Example 12
Lycium ruthenicum anthocyanin tablet candy health food
The lycium ruthenicum anthocyanin extract prepared in example 3 was added with 18% of maltitol or xylitol or aspartame or stevioside, 16% of microcrystalline cellulose and 6% of magnesium stearate, mixed by a CH-200 tank mixer, dry granulated, and tabletted by a ten-punch tablet press (zsx) (pressure 40-60 KN) to obtain tabletted candies.
Example 13
Lycium ruthenicum anthocyanin capsule medicine
The lycium ruthenicum anthocyanin extract obtained in example 1 or the lycium ruthenicum anthocyanin freeze-dried powder prepared in example 6 is added with 40-50% of lactose or pregelatinized starch, 1.8% of magnesium stearate and 4.5% of talcum powder or superfine silica gel powder, mixed uniformly by a CH-200 groove type mixing machine, granulated by a dry method and filled by an NJP-300 capsule filling machine to obtain capsules.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (3)
1. The preparation method of the lycium ruthenicum anthocyanin extract is characterized by comprising the following steps: the method specifically comprises the following steps:
step 1, extraction process: crushing dry lycium ruthenicum mill, adding deionized water with 5-10 times of volume to soak, stirring and extracting for 3-5 times to obtain coarse lycium ruthenicum mill juice; or crushing fresh Lycium ruthenicum Murr, juicing, extracting the residue with water for 3 times, and mixing filtrates to obtain coarse Lycium ruthenicum Murr juice;
step 2, high-speed centrifugation and ceramic membrane filtration: centrifuging the coarse lycium ruthenicum juice obtained in the step 1 by using a centrifuge with the rotating speed of 15000-20000 r/min to remove impurities, and clarifying by using a ceramic membrane of 50-200 nm to obtain clarified lycium ruthenicum juice;
step 3, degumming and concentrating the organic membrane: removing pectin, protein and polysaccharide macromolecular substances from the clarified lycium ruthenicum juice obtained in the step 2 by an ultrafiltration organic membrane with the molecular weight cut-off of 50000-100000, adding pure water for cleaning, collecting effluent liquid, and dehydrating and concentrating by the ultrafiltration organic membrane with the molecular weight cut-off of 300-5000 to obtain membrane concentrated solution;
and 4, simulated moving bed chromatographic separation: adding the membrane concentrated solution obtained in the step 3 into a simulated moving bed chromatography for separation to obtain a lycium ruthenicum fine anthocyanin separation solution; the simulated moving bed separation method comprises the following steps: the adsorbent filled by the simulated moving bed chromatography adopts any one or more of nonpolar macroporous adsorption resin, weak-polarity macroporous resin and polar macroporous resin, and the flow rate of an adsorption area is 1-2 BV/h; the water washing area is purified water, the dosage of the purified water is 1 to 2 times of the volume of the resin, and the flow rate is 1 to 3BV/h; the desorbent is 5-90% ethanol solution, the dosage is 2-4 times of the resin volume, the flow rate is 1-3 BV/h; the resin regeneration solvent is 95% ethanol solution, and the dosage of the resin regeneration solvent is 1 time of that of the resin; the flow rate is 2-3 BV/h; wherein the nonpolar macroporous adsorbent resin is HPD-100 and HPD-750; the weak-polarity macroporous resin is AB-8, DM130, D860021, DS-401 and CAD-40; the polar macroporous resin is ADS-21, ADS-7, BS-75, BS-45, NKA-2 and NKA-9;
and 5, low-temperature concentration: recovering ethanol from the fine anthocyanin separation liquid of the lycium ruthenicum mill obtained in the step 4, and carrying out vacuum low-temperature concentration at the temperature of 10-50 ℃ and the vacuum degree of-0.06 MPa to obtain an anthocyanin concentration liquid of the lycium ruthenicum mill;
step 6, freeze drying: directly freeze-drying the lycium ruthenicum anthocyanin concentrated solution obtained in the step 5 to obtain a lycium ruthenicum anthocyanin extract; wherein, the lycium ruthenicum anthocyanin extract is obtained by freeze drying at-50 to-70 ℃ by a freeze dryer.
2. The preparation method of the lycium ruthenicum anthocyanin freeze-dried powder is characterized by comprising the following steps of: the method specifically comprises the following steps:
step 1, extraction process: crushing dry lycium ruthenicum mill, adding deionized water with 5-10 times of volume to soak, stirring and extracting for 3-5 times to obtain coarse lycium ruthenicum mill juice; or crushing fresh Lycium ruthenicum Murr, juicing, extracting the residue with water for 3 times, and mixing the filtrates to obtain coarse Lycium ruthenicum Murr juice;
step 2, high-speed centrifugation and ceramic membrane filtration: centrifuging the coarse lycium ruthenicum juice obtained in the step 1 by using a centrifuge with the rotating speed of 15000-20000 r/min to remove impurities, and clarifying by using a 50-200 nm ceramic membrane to obtain clarified lycium ruthenicum juice;
step 3, degumming and concentrating the organic membrane: removing pectin, protein and polysaccharide macromolecular substances from the clarified lycium ruthenicum juice obtained in the step 2 by an ultrafiltration organic membrane with the molecular weight cut-off of 50000-100000, adding pure water for cleaning, collecting effluent liquid, and dehydrating and concentrating by the ultrafiltration organic membrane with the molecular weight cut-off of 300-5000 to obtain membrane concentrated solution;
step 4, simulated moving bed chromatographic separation: adding the membrane concentrated solution obtained in the step 3 into a simulated moving bed chromatography for separation to obtain a lycium ruthenicum fine anthocyanin separation solution; the simulated moving bed separation method comprises the following steps: the adsorbent filled by the simulated moving bed chromatography adopts any one or more of nonpolar macroporous adsorption resin, weak-polarity macroporous resin and polar macroporous resin, and the flow rate of an adsorption area is 1-2 BV/h; the water washing area is purified water, the dosage of the purified water is 1 to 2 times of the volume of the resin, and the flow rate is 1 to 3BV/h; the desorbent is 5-90% ethanol solution, the dosage is 2-4 times of the resin volume, the flow rate is 1-3 BV/h; the resin regeneration solvent is 95% ethanol solution, and the dosage is 1 time of the resin; the flow rate is 2-3 BV/h; wherein the nonpolar macroporous adsorbent resin is HPD-100 and HPD-750; the weak-polarity macroporous resin comprises AB-8, DM130, D860021, DS-401 and CAD-40; the polar macroporous resin is ADS-21, ADS-7, BS-75, BS-45, NKA-2 and NKA-9;
and 5, low-temperature concentration: recovering ethanol from the fine anthocyanin separation liquid of the lycium ruthenicum murr obtained in the step 4, and carrying out vacuum low-temperature concentration at the temperature of 10-50 ℃ and the vacuum degree of-0.06 MPa to obtain an anthocyanin concentration liquid of the lycium ruthenicum murr;
step 6, freeze-dried powder preparation: and (5) adding auxiliary materials into the lycium ruthenicum anthocyanin concentrated solution obtained in the step 5, uniformly stirring, and freeze-drying to obtain lycium ruthenicum anthocyanin freeze-dried powder.
3. The preparation method of lycium ruthenicum anthocyanin freeze-dried powder according to claim 2, characterized by comprising the following steps: and (3) adding an auxiliary material selected from starch, maltodextrin, fructo-oligosaccharide, galacto-oligosaccharide, lactose, inulin, beta-cyclodextrin, anhydrous glucose, polydextrose, milk powder, calcium citrate, xylitol, sorbitol and D-mannitol in the step 6, and freeze-drying at-50 to-70 ℃ by using a freeze dryer to obtain the lycium ruthenicum anthocyanin freeze-dried powder.
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