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CN-121987727-A - Method for extracting gastrodia elata polyphenol by ultrasonic-assisted eutectic solvent, product and application

CN121987727ACN 121987727 ACN121987727 ACN 121987727ACN-121987727-A

Abstract

The invention discloses a method for extracting gastrodia elata polyphenol by using ultrasonic-assisted eutectic solvent, a product and application thereof. The invention takes betaine as HBA and glycerol as HBD, synthesizes DES solvent to extract gastrodin, screens out optimal extraction process conditions of betaine-glycerol molar ratio of 1:2, water content of 26%, extraction temperature of 50 ℃, extraction time of 50min and liquid-material ratio of 1:20 (g/mL), and finally uses AB-8 macroporous adsorption resin for purification. The extraction effect of the invention is higher than that of the traditional solvent, and the invention has high safety, economy and durability, has feasibility of becoming a solvent for extracting the gastrodia elata polyphenol, and provides theoretical basis for efficient and green extraction and deep development and utilization of the gastrodia elata polyphenol.

Inventors

  • GAO MING
  • YANG XIAOSHENG
  • ZHANG JUNMIN
  • PAN XIONG
  • Yang Lishou
  • LI QIJI
  • CHEN FAJU

Assignees

  • 贵州省天然产物研究中心

Dates

Publication Date
20260508
Application Date
20260126

Claims (10)

  1. 1. A method for extracting gastrodia elata polyphenol by using ultrasonic-assisted eutectic solvent is characterized by comprising the following steps of: (1) Pretreating rhizoma Gastrodiae by steaming fresh rhizoma Gastrodiae, slicing, drying, pulverizing, sieving with 60-100 mesh sieve to obtain rhizoma Gastrodiae powder, and sealing for use; (2) The preparation of the DES solvent comprises the steps of heating, stirring and mixing hydrogen bond acceptor betaine and hydrogen bond donor glycerol with the molar ratio of 0.5-1.5:1.5-2.5 until uniform and transparent liquid is formed, standing, uniformly and transparent, not layering, and diluting with water until the water content of DES is 10% -30%, thus obtaining the DES solvent; (3) Extracting rhizoma Gastrodiae polyphenols by weighing rhizoma Gastrodiae powder, adding DES solvent, performing ultrasonic extraction for 40-60min at ultrasonic temperature of 30-60deg.C at ratio of rhizoma Gastrodiae powder to DES solvent of 1g:15-25mL, centrifuging the extractive solution, and collecting supernatant to obtain rhizoma Gastrodiae polyphenols crude extract; (4) And (3) macroporous resin purification, namely concentrating the crude gastrodia elata polyphenol extract, then carrying out enrichment purification on the crude gastrodia elata polyphenol extract by using an AB-8 macroporous adsorption resin column, adjusting the pH value of the sample-loading concentrated solution to be 5-6, eluting impurities by using distilled water after the sample loading is finished, carrying out polyphenol analysis by using 70% ethanol, collecting an analysis solution, carrying out rotary evaporation concentration, and drying to obtain the purified gastrodia elata polyphenol extract.
  2. 2. The method for extracting gastrodia elata polyphenol by using the ultrasound-assisted eutectic solvent according to claim 1, wherein the gastrodia elata pretreatment in the step (1) is characterized in that fresh gastrodia elata is taken, steamed and sliced, dried at 45-55 ℃ for 22-26 hours in a drying box, crushed by using a high-speed traditional Chinese medicine crusher, and sample powder is sieved by a 60-100-mesh sieve to obtain gastrodia elata powder, and the gastrodia elata powder is stored in a dryer in a sealing manner for standby.
  3. 3. The method for extracting gastrodia elata polyphenol by using the ultrasound-assisted eutectic solvent according to claim 1, wherein the preparation of the DES solvent in the step (2) is characterized in that hydrogen bond acceptor betaine and hydrogen bond donor glycerol are mixed for 0.5-2.5h under stirring, the molar ratio of the betaine to the glycerol is 0.5-1.5:1.5-2.5, the heating temperature is 50-90 ℃ until uniform transparent liquid is formed, the liquid is still uniform and transparent and is not layered after being placed, and the DES solvent is obtained after the liquid is diluted by adding water until the water content of the DES is 10-30%.
  4. 4. The method for extracting gastrodia elata polyphenol from the ultrasound-assisted eutectic solvent according to claim 3, wherein the preparation of the DES solvent in the step (2) is characterized in that a magnetic heating stirrer is used for stirring and mixing hydrogen bond acceptor betaine and hydrogen bond donor glycerol for 1.5h, the molar ratio of betaine to glycerol is 1:2, the heating temperature is 70 ℃ until uniform transparent liquid is formed, the liquid is still uniform and transparent and is not layered after being placed for 24h, and water is added for dilution until the water content of the DES is 26%, so that the DES solvent is obtained.
  5. 5. The method for extracting gastrodia elata polyphenol by using the ultrasonic-assisted eutectic solvent according to claim 1, wherein the step (3) is characterized in that 1.5-2.5g of gastrodia elata powder is weighed and placed in a centrifuge tube, DES solvent is added for ultrasonic extraction for 40-60min, the ratio of the gastrodia elata powder to the DES solvent is 1g:15-25mL, the ultrasonic power is 200-400W, the ultrasonic temperature is 30-60 ℃, and supernatant fluid is obtained after the extract is centrifuged for 5-15 min at 10000 r/min, so that the gastrodia elata polyphenol crude extract is obtained.
  6. 6. The method for extracting gastrodia elata polyphenol by using the ultrasonic-assisted eutectic solvent according to claim 5, wherein the step (3) is characterized in that 2g of gastrodia elata powder is weighed and placed in a 50mL centrifuge tube, DES solvent is added for ultrasonic extraction for 50min, the ratio of the gastrodia elata powder to the DES solvent is 1g to 20mL, the ultrasonic power is 300W, the ultrasonic temperature is 50 ℃, and the supernatant is obtained after the extracting solution is centrifuged at 10000r/min for 10min, so that the gastrodia elata polyphenol crude extracting solution is obtained.
  7. 7. The method for extracting gastrodia elata polyphenol by using the ultrasonic-assisted eutectic solvent, which is characterized in that the macroporous resin purification in the step (4) is characterized in that crude gastrodia elata polyphenol extract is concentrated to a concentration of 0.8-1.2mg/mL, then the concentrated gastrodia elata polyphenol extract is subjected to enrichment purification by using an AB-8 macroporous adsorption resin column, the pH of the loaded concentrated solution is adjusted to be 5-6, the adsorption speed is 0.9-1.1mL/min, distilled water is used for impurity elution after loading, polyphenol analysis is performed by using 3.5-4.5BV of 70% ethanol, the analysis flow rate is 1.8-2.2mL/min, the analysis liquid is collected, and the purified gastrodia elata polyphenol extract is obtained after spin evaporation concentration and drying.
  8. 8. The method for extracting gastrodia elata polyphenol with the ultrasonic-assisted eutectic solvent, as claimed in claim 7, characterized in that the macroporous resin purification in the step (4) is characterized in that a rotary evaporator is used for concentrating the crude gastrodia elata polyphenol extract to a concentration of 1.0mg/mL, an AB-8 macroporous adsorption resin column is used for enrichment purification, 1% dilute hydrochloric acid is used for adjusting the pH value of the loaded concentrated solution to be 5.5, the adsorption speed is 1mL/min, distilled water is used for impurity elution after loading, 4BV of 70% ethanol is used for polyphenol analysis, the analysis flow rate is 2mL/min, the analysis liquid is collected, and the purified gastrodia elata polyphenol extract is obtained after rotary evaporation concentration and drying.
  9. 9. A gastrodia elata polyphenol extract obtained by the method of any one of claims 1 to 8.
  10. 10. The use of the gastrodia elata polyphenol extract according to claim 9 as an antioxidant in food additives, cosmetic raw materials and medicines.

Description

Method for extracting gastrodia elata polyphenol by ultrasonic-assisted eutectic solvent, product and application Technical Field The invention belongs to the technical field of natural product extraction, and particularly relates to a method for extracting gastrodia elata polyphenol by using an ultrasonic-assisted eutectic solvent, a product and application. Background The eutectic solvent (Deep eutectic solvents, DES) has the advantages of simple preparation, good stability, high chemical purity, low cost, good degradability and the like, and the polarity of the solvent can be regulated by adjusting the material ratio of the eutectic solvent components, so that the eutectic solvent has been widely applied to extracting the effective components of the traditional Chinese medicine. Gastrodia elata (Gastrodia elata Bl., GE) is a dry tuber of Gastrodia elata (Gastrodia elata Blume) which is an annual parasitic herb of Gastrodia elata Blume of Gastrodia of Orchidaceae, also called as Caesalpinia crista, he Ji Cao and Ding Feng Cao, etc. In modern pharmacological research, gastrodia elata has antioxidant, neuroprotective, antidepressant, anxiolytic and sedative effects. Researches show that the gastrodia elata is rich in active substances such as gastrodin, polysaccharide, polyphenol, protein and the like, and the content of the polyphenol active substances is relatively high. The existing extraction method of gastrodia elata polyphenol is many, but various problems exist, such as solvent extraction method has the problems of solvent residue, poor selectivity, high energy consumption, environmental pollution and the like, extraction rate of water extraction method is low, mildewing is easy, impurities are more, oxidation is possibly promoted and the like, supercritical CO 2 extraction equipment is extremely expensive, effect on polar polyphenol is poor, operation is complex, and environmental protection eutectic solvent is adopted for extracting gastrodia elata polyphenol, so that no report is yet available. The invention aims to obtain a green and efficient method for extracting the gastrodia elata polyphenol by comparing the extraction rate and stability of various gastrodia elata polyphenol extraction methods, and provides technical reference for development and utilization of the gastrodia elata polyphenol. Disclosure of Invention In order to solve the technical problems, the invention provides a method for extracting gastrodia elata polyphenol by using ultrasonic-assisted eutectic solvent, which is used for extracting gastrodia elata by using betaine as HBA and glycerol as HBD and synthesizing DES solvent, so that high-efficiency and rapid extraction of gastrodia elata polyphenol substances is realized. The technical scheme of the invention is as follows: A method for extracting gastrodia elata polyphenol by using ultrasonic-assisted eutectic solvent comprises the following steps: (1) Pretreating rhizoma Gastrodiae by steaming fresh rhizoma Gastrodiae, slicing, drying, pulverizing, sieving with 60-100 mesh sieve to obtain rhizoma Gastrodiae powder, and sealing for use; (2) The preparation of the DES solvent comprises the steps of heating, stirring and mixing hydrogen bond acceptor betaine and hydrogen bond donor glycerol with the molar ratio of 0.5-1.5:1.5-2.5 until uniform and transparent liquid is formed, standing, uniformly and transparent, not layering, and diluting with water until the water content of DES is 10% -30%, thus obtaining the DES solvent; (3) Extracting rhizoma Gastrodiae polyphenols by weighing rhizoma Gastrodiae powder, adding DES solvent, performing ultrasonic extraction for 40-60min at ultrasonic temperature of 30-60deg.C at ratio of rhizoma Gastrodiae powder to DES solvent of 1g:15-25mL, centrifuging the extractive solution, and collecting supernatant to obtain rhizoma Gastrodiae polyphenols crude extract; (4) And (3) macroporous resin purification, namely concentrating the crude gastrodia elata polyphenol extract, then carrying out enrichment purification on the crude gastrodia elata polyphenol extract by using an AB-8 macroporous adsorption resin column, adjusting the pH value of the sample-loading concentrated solution to be 5-6, eluting impurities by using distilled water after the sample loading is finished, carrying out polyphenol analysis by using 70% ethanol, collecting an analysis solution, carrying out rotary evaporation concentration, and drying to obtain the purified gastrodia elata polyphenol extract. In the method, the gastrodia elata pretreatment in the step (1) is that fresh gastrodia elata is taken, steamed and sliced, dried in a drying oven at 45-55 ℃ for 22-26 hours (preferably at 50 ℃ for 24 h), crushed by a high-speed traditional Chinese medicine crusher, and the sample powder is sieved by a 60-100-mesh sieve, so that gastrodia elata powder is obtained, and the gastrodia elata powder is stored in a dryer in a sealing manner for standby. In the method, the preparat