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CN-122011230-A - A herba Sedi polysaccharide extraction process

CN122011230ACN 122011230 ACN122011230 ACN 122011230ACN-122011230-A

Abstract

The invention discloses a sedum sarmentosum polysaccharide extraction process which comprises the steps of drying sedum sarmentosum, crushing, extracting with water according to a feed liquid ratio of 1:20-1:60, concentrating, precipitating with alcohol to obtain crude polysaccharide, deproteinizing, dialyzing, freeze-drying, gradient eluting, purifying to obtain sedum sarmentosum polysaccharide SSBP, optimizing the extraction process by a response surface method, selecting the optimal condition, obtaining the crude polysaccharide with the yield reaching 19.06%, structurally characterizing that SSBP is acidic polysaccharide with molecular weight 7.484kDa and contains 36.12% of uronic acid, monosaccharide composition including rhamnose, arabinose, galactose, galacturonic acid and glucuronic acid, the molar ratio of 12.85:19.13:18.98:35.73:3.35, and in vitro activity experiment shows that SSBP has good antioxidant activity and remarkable anti-inflammatory activity, can inhibit release of RAW264.7 macrophage NO and IL-6, IL-1 beta and TNF-alpha secretion induced by LPS by dose-dependently, and the inhibition rates reach 74.41%, 92.53%, 95.27% and 95.73% of sedum sarmentosum polysaccharide under the concentration of 1.2 mg/mL.

Inventors

  • ZHANG YAN
  • LONG LIN

Assignees

  • 天津科技大学

Dates

Publication Date
20260512
Application Date
20260327

Claims (10)

  1. 1. The sedum sarmentosum polysaccharide is characterized by being an acidic polysaccharide, and contains uronic acid, wherein the total sugar content is 85% -87%, the uronic acid content is 35% -37%, the protein content is 1.5% -1.8%, and the molecular weight is 7-8 kDa.
  2. 2. The sedum sarmentosum polysaccharide according to claim 1, wherein the monosaccharide composition of the SSBP comprises rhamnose (Rha), arabinose (Ara), galactose (Gal), galacturonic acid (Gal-UA) and glucuronic acid (Glc-UA) in a molar ratio of (12-13): (19-20): (18-19): (35-36): (3-4).
  3. 3. The sedum sarmentosum polysaccharide according to claim 1, wherein the infrared spectrum of SSBP has characteristic absorption peaks at 3415 cm-1, 2936 cm-1, 1743 cm-1, 1611 cm-1, 833 cm-1, wherein the absorption peaks at 1743 cm-1 and 1611 cm-1 correspond to esterified carboxyl and free carboxyl groups, respectively, indicating that uronic acid units are present, and the absorption peak at 833 cm-1 indicates that alpha-type glycosidic linkages are present.
  4. 4. A process for extracting sedum sarmentosum polysaccharide according to any one of claims 1 to 3, characterized by comprising the following steps: 1) Washing whole herb of sedum sarmentosum, oven drying and crushing for standby; 2) Mixing herba Sedi powder with distilled water, heating for extraction, centrifuging, collecting supernatant, and concentrating; 3) Adding absolute ethyl alcohol according to a certain proportion, standing overnight, centrifuging, collecting precipitate, re-dissolving, and freeze-drying to obtain sedum sarmentosum crude polysaccharide CSSBP; 4) Re-dissolving CSSBP, mixing with Sevage reagent to remove impurities, treating with running water of dialysis bag, treating with still water, collecting dialysate, and lyophilizing; 5) Freeze-drying the sample, dissolving in water, centrifuging, collecting supernatant, purifying with column, gradient eluting, collecting, freezing, and drying to obtain herba Sedi polysaccharide SSBP.
  5. 5. The process for extracting sedum sarmentosum polysaccharide according to claim 4, wherein the drying temperature in the step 1) is 37 ℃, and sedum sarmentosum powder is sieved by a 50-mesh sieve.
  6. 6. The process for extracting sedum sarmentosum polysaccharide according to claim 4, wherein in the step 2), the material-liquid ratio of sedum sarmentosum powder and distilled water is 1:20-1:60 (g/mL), the extraction temperature is 60-100 ℃, the extraction time is 1-5h, and the sedum sarmentosum powder is concentrated to 1/10 of the original volume by rotary evaporation at 65 ℃.
  7. 7. The process for extracting sedum sarmentosum polysaccharide according to claim 4, wherein in the step 3), the absolute ethyl alcohol is added in a volume ratio of 1:4, and the standing condition is 4 ℃.
  8. 8. The process for extracting sedum sarmentosum polysaccharide according to claim 4, wherein in the step 4), the Sevage reagent is a mixed system of n-butanol and chloroform, wherein V N-butanol ：V trichloromethane = 1:4, the mixing volume ratio of the solution of cssbp and the Sevage reagent is 1:5, the dialysis bag 3500Da is used for treating the sedum sarmentosum polysaccharide for 72 hours under the condition of 4 ℃ and 24 hours.
  9. 9. The process for extracting sedum sarmentosum polysaccharide according to claim 4, wherein in step 5), the purification column is DEAE SEPLIFE FF, the flow rate of the column is 4ml/min, gradient elution is sequentially carried out by pure water, 0.1M, 0.2M and 0.3M NaCl solution, and all eluents are collected.
  10. 10. The application of sedum sarmentosum polysaccharide SSBP in preparing anti-inflammatory drugs or functional foods is characterized in that the SSBP can inhibit the release of RAW264.7 macrophage NO and the secretion of inflammatory factors IL-6, IL-1 beta and TNF-alpha induced by LPS.

Description

A herba Sedi polysaccharide extraction process Technical Field The invention belongs to the technical field of extraction of plant active ingredients, and particularly relates to a stringy stonecrop herb polysaccharide extraction process. Background The sedum sarmentosum is perennial herb of Crassulaceae, is used as a whole herb, is cool in nature, sweet and slightly sour in taste, enters liver and gall and small intestine channels, has the effects of clearing heat and removing toxicity, and promoting diuresis and removing jaundice, is originally carried in Qing Dynasty 'Ben Cao gang mu Shi Yi', and is one of common medicinal materials used for treating acute and chronic hepatitis in traditional Chinese medicine of China. Modern pharmacological researches show that sedum sarmentosum has various biological activities of protecting liver, reducing enzyme, resisting inflammation, resisting oxidation, regulating immunity, resisting tumor and the like, and has wide medicinal development prospect. At present, medicines developed based on sedum sarmentosum are mainly compound preparations, including compound sedum sarmentosum syrup, yin-nourishing tea syrup, liver-protecting tablet, yiganle granule, sedum sarmentosum granule, compound sedum sarmentosum capsule and the like, and are mainly used for the auxiliary treatment of acute and chronic hepatitis, and the preparations are mainly used for taking sedum sarmentosum crude extract or compound forms as medicines and mainly aiming at small molecular components such as flavonoid, cyanosides and the like. From the study of chemical components, the active ingredients of the sedum sarmentosum mainly comprise flavonoids (such as quercetin, kaempferol, isorhamnetin, luteolin and the like), triterpenes, alkaloids, cyanosides (mainly sedum sarmentosum glycoside) and polysaccharides and the like. Wherein the flavonoid component is the main active substance of herba Sedi, has anti-inflammatory, anti-fibrosis, antioxidant and anti-angiogenesis activities, and the sedum sarmentosum glycoside is one of the effective components for protecting liver and reducing enzyme. In recent years, as pharmacological activities of polysaccharide components in the fields of immunoregulation, anti-tumor, antioxidation and the like are widely confirmed, research on sedum sarmentosum polysaccharide is attracting attention. The prior researches show that the sedum sarmentosum polysaccharide is arabinogalactan glucan composed of monosaccharides such as arabinose, glucose, galactose and the like, crude polysaccharide is obtained by the steps of hot water extraction, ethanol precipitation, deproteinization by a Sevage method and the like conventionally, and then polysaccharide components are obtained by ion exchange chromatography and gel filtration chromatography purification, and preliminary researches show that the sedum sarmentosum polysaccharide has the functions of resisting oxidation, regulating immunity, resisting tumors and the like. In terms of patent technology, related patent is searched and issued. CN111892662B discloses a sedum sarmentosum uniform polysaccharide, its preparation method and use, crude polysaccharide is obtained by hot water extraction and alcohol precipitation, water washing component is obtained by eluting with distilled water through DE-52 anion exchange cellulose column, uniform polysaccharide CPCW is obtained by eluting and purifying with 0.2M NaCl solution through gel chromatographic column, the purity of the polysaccharide is up to 99%, molecular weight is 1-30kDa, monosaccharide composition is arabinose, glucose and galactose, and it is proved that the polysaccharide has anti-tumor effect of inhibiting liver cancer Huh-7 cell proliferation. CN111297935B discloses a sedum sarmentosum extract, a preparation method and application thereof, mainly relates to a sedum sarmentosum ethyl acetate part and an ethanol part, and comprises organic acids, flavonoids, triterpenes and alkaloids, has an anti-influenza virus effect, and does not relate to extraction and application of polysaccharide components. However, the related technology of the traditional sedum sarmentosum polysaccharide still has obvious defects. In terms of the purification method, CN111892662B adopts distilled water to elute an ion exchange column, a neutral polysaccharide component is obtained, an acidic polysaccharide component possibly obtained by salt gradient elution is not involved, and the comprehensive development and utilization of polysaccharides with different charge properties in sedum sarmentosum are limited. In the aspect of process optimization, the prior art does not systematically optimize the extraction process parameters, the yield parameters are not clear, and the process parameter basis is lacked in the industrial conversion. In the aspect of activity research, the prior art mainly focuses on anti-tumor activity, and the anti-inflammatory activity of the sedum sarmentosum polysaccharid