[go: up one dir, main page]

CN107789376B - A kind of two-phase deep eutectic solvent for extracting active components of Ginkgo biloba, and preparation method and extraction method thereof - Google Patents

A kind of two-phase deep eutectic solvent for extracting active components of Ginkgo biloba, and preparation method and extraction method thereof Download PDF

Info

Publication number
CN107789376B
CN107789376B CN201711064250.1A CN201711064250A CN107789376B CN 107789376 B CN107789376 B CN 107789376B CN 201711064250 A CN201711064250 A CN 201711064250A CN 107789376 B CN107789376 B CN 107789376B
Authority
CN
China
Prior art keywords
phase
substance
deep eutectic
eutectic solvent
extraction
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201711064250.1A
Other languages
Chinese (zh)
Other versions
CN107789376A (en
Inventor
苏二正
曹君
李默涵
曹福亮
陈璐瑶
赵林果
王耀松
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nanjing Forestry University
Original Assignee
Nanjing Forestry University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Nanjing Forestry University filed Critical Nanjing Forestry University
Priority to CN201711064250.1A priority Critical patent/CN107789376B/en
Publication of CN107789376A publication Critical patent/CN107789376A/en
Application granted granted Critical
Publication of CN107789376B publication Critical patent/CN107789376B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/16—Ginkgophyta, e.g. Ginkgoaceae (Ginkgo family)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization

Landscapes

  • Health & Medical Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Biotechnology (AREA)
  • Botany (AREA)
  • Medical Informatics (AREA)
  • Medicinal Chemistry (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

The invention discloses a two-phase deep eutectic solvent for extracting active ingredients of ginkgo leaves, a preparation method and an extraction method thereof, wherein the two-phase deep eutectic solvent consists of a hydrophilic phase and a hydrophobic phase which are equal in volume, the hydrophilic phase consists of a substance A and a substance B, the substance A consists of DES1 and water, and DES1 is prepared from choline chloride and levulinic acid; the substance B consists of DES2 and water, and DES2 is prepared from choline chloride and malonic acid; the hydrophobic phase is prepared from methyl trioctyl ammonium chloride, octanol and octanoic acid. The two-phase deep eutectic solvent prepared by the invention is a green solvent, is safe and pollution-free, has small viscosity, stable hydrogen bond and good fluidity; can simultaneously extract hydrophobic active ingredients of polyprenol acetate and hydrophilic active ingredients of ginkgetin, terpene lactone and procyanidine in the ginkgo leaves, has high extraction rate and safe use, and does not form residues. The preparation method of the deep eutectic solvent is simple and convenient, low in cost and high in biodegradability.

Description

Two-phase deep eutectic solvent for extracting active ingredients of ginkgo leaves and preparation method and extraction method thereof
Technical Field
The invention relates to the utilization of agricultural and forestry biological resources, in particular to a two-phase deep eutectic solvent for extracting active components of ginkgo leaves, a preparation method and an extraction method thereof.
Background
Ginkgo biloba (known as Ginkgo biloba L.), Gongsun tree and Ginkgo biloba belong to gymnospermum, Ginkgoaceae Ginkgo, and are individually a single species. Ginkgo leaves, fruits and the like have medicinal development value and are known as activated stones which are precious. In China, the gingko resource possession accounts for more than 70% of the total world amount.
The active components of folium Ginkgo mainly comprise ginkgetin, terpene lactone, procyanidine, and polyprenol. The content of ginkgo flavonoids is in the first place, the total content is about 2.5-5.9%, and more than 40 flavonoids are separated from ginkgo leaves. The ginkgo leaf flavonoid compound has the effects of inhibiting platelet aggregation, reducing blood viscosity, improving brain circulation, resisting atherosclerosis, resisting oxidation, promoting lipolysis of fat cells, resisting tumors and the like, and can be effectively applied to treating diseases such as coronary heart disease, angina pectoris, cerebral arteriosclerosis, senile dementia, hypertension and the like.
Bilobalide is another important active ingredient in folium Ginkgo, belongs to terpenoids, is also called ginkgo terpene lactones, and mainly comprises sesquiterpene lactone and diterpene lactone. Bilobalide is the only sesquiterpene lactone compound isolated from ginkgo leaves. Bilobalide A, B, C, M, J is diterpene lactone compound. Ginkgolide B is a potent antagonist of specific Platelet Activating Factor (PAF). The pharmacological action research of various scientists on the ginkgo leaf terpene lactone compounds discovers that the ginkgo leaf terpene lactone is the most promising drug in the current natural PAF receptor antagonists, and has good clinical application prospect in the aspects of protecting ischemic brain injury, promoting the differentiation of neural stem cells to neurons, inhibiting the apoptosis of nerve cells, treating the degenerative diseases of the central nervous system, the diseases of the cardiovascular system and the like.
The polyprenol is a lipoid compound, and the structure of the polyprenol mainly comprises a series of isopentenyl units and terminal isopentenol units. The polyprenol contained in folium Ginkgo belongs to birch polyprenol, and exists in leaves mainly in the form of acetate, and exists in leaves in the form of free alcohol at least. The number of the isoamylene units in the ginkgo leaf polyprenol molecule is 14-24, the main component is 17-19 isopentene units, and the content of the total polyprenol compounds is about 80%. The polyprenols compounds have immunological activity, can resist HIV virus, can be used for treating immunodeficiency diseases such as septicemia, can induce tumor cell apoptosis, has obvious biological activity in resisting hepatitis virus and assisting chemotherapy leukemia, and has therapeutic effect on immunologic function diseases such as hypertension, hyperlipemia, diabetes, ventilation, lupus erythematosus, etc.
Procyanidin is a plant polyphenol compound with flavan-3-ol structure, and the content of procyanidin in the ginkgo leaf extract is about 4-12%. Besides the well-known antioxidant activity, the ginkgo leaf procyanidine has the effects of resisting hypertension, endothelium-dependent vasodilation activity, ischemia-reperfusion injury, atherosclerosis, platelet aggregation and the like in a cardiovascular system, and also has a remarkable protective effect on acute renal failure. In addition, procyanidin has anticancer, immunity regulating, capillary permeability reducing, antibacterial and anti-inflammatory effects, and can be used for treating peripheral venous insufficiency, lymphedema, etc.
At present, water or an organic solvent is generally used as an extraction solvent for extracting active ingredients of ginkgo leaves, the extraction effect of the water as the extraction solvent is poor, and the traditional organic solvent is volatile and easy to remain, causes harm to the environment and human bodies, and has high cost. In addition, since ginkgo flavone, terpene lactone and procyanidin have hydrophilicity and poly-isopropenyl alcohol acetate has hydrophobicity, the previous extraction method of ginkgo leaf active ingredients extracts the ingredients separately or extracts the hydrophilic ingredients and the hydrophobic ingredients separately, which not only wastes bioactive ingredients, but also increases extraction steps and increases extraction cost. Therefore, it is necessary to find a green, safe, efficient and synchronous method for extracting active components from ginkgo biloba.
Disclosure of Invention
The purpose of the invention is as follows: in order to solve the problems in the prior art, the invention provides a two-phase deep eutectic solvent (TPDES). The two-phase deep eutectic solvent provided by the invention is used as a green solvent, is safe, pollution-free, recyclable, easily biodegradable, simple in preparation process and low in cost, and has the properties of low steam pressure, incombustibility, excellent solubility and conductivity, stable electrochemical window and the like; can synchronously extract a plurality of active ingredients in the ginkgo leaves, including flavone, terpene lactone, procyanidine and polyprenol acetate, in a green, safe and high-efficiency way.
The invention also provides a preparation method of the two-phase deep eutectic solvent and a method for extracting ginkgo leaf active ingredients by using the two-phase deep eutectic solvent.
The technical scheme is as follows: in order to achieve the purpose, the two-phase deep eutectic solvent for extracting the active ingredients of the ginkgo biloba leaves consists of a hydrophilic phase and a hydrophobic phase which have equal volumes, wherein the hydrophilic phase consists of a substance A and a substance B according to a volume ratio of 10-0:0-10, the substance A consists of DES1 and water according to a mass ratio of 6:3.5-4.5, and DES1 is prepared from choline chloride and levulinic acid according to a molar ratio of 1: 1.5-2.5; the substance B consists of DES2 and water according to the mass ratio of 4.5:5-6, and DES2 is prepared from choline chloride and malonic acid according to the molar ratio of 1: 1.5-2.5; the hydrophobic phase is a deep eutectic solvent DES3 prepared from methyl trioctyl ammonium chloride, octanol and octanoic acid according to the molar ratio of 1:2: 2.5-3.5.
Preferably, the hydrophilic phase consists of substance A and substance B in a volume ratio of 7.75-8.75: 2.25-1.25.
Most preferably, the hydrophilic phase in the deep eutectic solvent consists of substance a and substance B in a volume ratio of 8.75: 1.25.
Preferably, the substance A consists of DES1 and water in a mass ratio of 6:4-4.5, and the DES1 is prepared from choline chloride and levulinic acid in a molar ratio of 1: 2-2.5; the substance B consists of DES2 and water according to the mass ratio of 4.5:5.5-6, and DES2 is prepared from choline chloride and malonic acid according to the molar ratio of 1: 1.5-2; the hydrophobic phase is prepared from methyl trioctyl ammonium chloride, octanol and octanoic acid according to a molar ratio of 1:2:2.5-3.
Most preferably, the substance A consists of DES1 and water in a mass ratio of 6:4, and the DES1 is prepared from choline chloride and levulinic acid in a molar ratio of 1: 2; the substance B is composed of DES2 and water according to the mass ratio of 4.5:5.5, and DES2 is prepared from choline chloride and malonic acid according to the molar ratio of 1: 2; the hydrophobic phase DES3 is prepared from methyl trioctyl ammonium chloride, octanol and octanoic acid according to a molar ratio of 1:2:3.
The preparation method of the two-phase deep eutectic solvent comprises the following steps:
(1) weighing choline chloride and levulinic acid according to a molar ratio, uniformly mixing, heating and stirring at 80-110 ℃ until a transparent liquid is formed, namely a deep eutectic solvent DES1, and mixing DES1 and water according to a mass ratio to obtain a substance A;
(2) weighing choline chloride and malonic acid according to a molar ratio, uniformly mixing, heating and stirring at 80-110 ℃ until a transparent liquid is formed, namely a deep eutectic solvent DES2, and mixing DES2 and water according to a mass ratio to obtain a substance B;
(3) mixing the substance A and the substance B according to the volume ratio to obtain a hydrophilic phase of the two-phase deep cosolvent;
(4) weighing methyl trioctyl ammonium chloride, octanol and octanoic acid according to a molar ratio, uniformly mixing, and heating at 80-110 ℃ until a transparent liquid is formed, namely a deep co-dissolving solvent DES3, serving as a hydrophobic phase of a two-phase deep co-dissolving solvent;
(5) and mixing the hydrophilic phase and the hydrophobic phase in equal volume, standing and layering to obtain the two-phase deep eutectic solvent.
The method for extracting the active components of the ginkgo leaves by the two-phase deep eutectic solvent is characterized by comprising the following steps:
(1) preparing a two-phase deep eutectic solvent according to a material-liquid ratio by a preparation method of the two-phase deep eutectic solvent, placing the two-phase deep eutectic solvent in an extraction container, accurately weighing ginkgo leaf powder, adding the ginkgo leaf powder into the two-phase deep eutectic solvent in the extraction container, and uniformly mixing;
(2) mixing, heating for extraction, centrifuging after extraction, and layering two-phase system, wherein the upper phase is hydrophobic phase and the lower phase is hydrophilic phase. And (3) detecting the contents of flavone, terpene lactone and procyanidine in the hydrophilic phase, and detecting the content of polyprenol acetate in the hydrophobic phase.
Wherein the mass-volume ratio of the ginkgo leaf powder in the step (1) to the diluted deep eutectic solvent is 1:15-25 g/mL. The mass volume ratio of the ginkgo leaf powder to the two-phase deep eutectic solvent is preferably 1:20-25 g/mL. The optimal mass-to-volume ratio is 1:20 g/mL.
Wherein the heating extraction in the step (2) is oscillation heating extraction or stirring heating extraction.
The rotation speed of the oscillation or stirring is 100-300rpm, the temperature is 50-70 ℃, and the time is 30-60 min.
The preferred rotation speed of oscillation or stirring is 150-160rpm, the temperature is 60-65 ℃ and the time is 40-45 min.
The optimum speed of oscillation or stirring is 150rpm, the temperature is 60 ℃ and the time is 40 min.
The properties of the two-phase deep eutectic solvents (TPDESs) have an important relationship with the components, the composition molar ratio and the water content of each phase of Deep Eutectic Solvents (DESs), the properties of the DESs can be greatly changed due to the slight difference of the components, and the properties of the DESs can be adjusted to meet different application requirements by utilizing the characteristics. The TPDESs can efficiently extract a plurality of active ingredients from ginkgo leaves at the same time, and because polypentenol compounds have larger hydrophobicity and large polarity difference with flavone, terpene lactone and other substances, the feasibility of the combined efficient extraction of a single solvent is not large. The two phases can also be separated rapidly after extraction with stirring, as shown in FIG. 1.
Has the advantages that: compared with the prior art, the invention has the following advantages:
(1) the two-phase deep eutectic solvent prepared by the invention is a green solvent, is safe and pollution-free, can be repeatedly used, is easy to biodegrade, and has the properties of low steam pressure, incombustibility, excellent solubility and conductivity, stable electrochemical window and the like; meanwhile, the TPDESs have small viscosity, stable hydrogen bonds and good fluidity; the TPDESs can be used for simultaneously extracting a plurality of active ingredients in the ginkgo leaves, and no residue is formed, so that no harm is caused to the environment and human bodies.
(2) The preparation method of the deep eutectic solvent is simple and convenient, and has wide raw material sources, low cost and high biodegradability.
(3) The method for extracting the active components of the ginkgo leaves by using the two-phase deep eutectic solvent has the advantages of simple and convenient operation, high extraction efficiency, safe use and no resource waste.
Drawings
FIG. 1 is a schematic diagram of a two-phase system before and after extraction of DESS;
FIG. 2 is a graph showing the effect of different extraction temperatures on the extraction rate of active components from ginkgo leaves by DESS;
FIG. 3 is a graph showing the effect of different extraction times on the extraction rate of active components from ginkgo leaves by DESS.
Detailed Description
The invention is further illustrated by the following figures and examples.
Experiment raw materials:
the experimental raw material folium ginkgo is from the production base of iron-enriched folium ginkgo in the State of Bizhou province, and the picking time of the folium ginkgo is 5 months.
The standard rutin is purchased from Shanghai leaf Biotechnology limited, the standard ginkgolide A is from China pharmaceutical and biological product institute, isopropanol, methanol and n-hexane are chromatographically pure, and other used drugs are analytically pure.
P1201 high performance liquid chromatograph and UV1201 detector, macrogol analysis instruments ltd;
UV-1200 type ultraviolet visible spectrophotometer, Shanghai Mei Puda instruments, Inc.;
JY-2 constant temperature stirring oil bath, gold jar city Tian Bing laboratory instrument factory;
constant temperature culture shaker ZWYR-2112B, Shanghai Zhicheng Analyzer manufacture, Inc.;
TG16A-WS desk-top high speed centrifuge, Shanghai Luxiang apparatus centrifuge, Inc.
Other raw materials are commercially available, and the raw materials of choline chloride, levulinic acid, malonic acid, methyltrioctylammonium chloride, octanol and octanoic acid are dried in a vacuum drying oven to remove water.
Example 1
(1) Weighing choline chloride and levulinic acid according to a molar ratio of 1:2, uniformly mixing, heating and stirring at 80-110 ℃ until a transparent liquid is formed, namely a deep eutectic solvent DES1, and mixing DES1 and water according to a mass ratio of 6:4 to obtain a substance A;
(2) weighing choline chloride and malonic acid according to a molar ratio of 1:2, uniformly mixing, heating and stirring at 80-110 ℃ until a transparent liquid is formed, namely a deep eutectic solvent DES2, and mixing DES2 and water according to a mass ratio of 4.5:5.5 to obtain a substance B;
(3) mixing the substance A and the substance B according to the volume ratio of 8.75:1.25 to obtain a hydrophilic phase of the two-phase deep cosolvent;
(4) weighing methyl trioctyl ammonium chloride, octanol and octanoic acid according to the molar ratio of 1:2:3, uniformly mixing, and heating at 80-110 ℃ until a transparent liquid is formed, namely a deep eutectic solvent DES3, serving as a hydrophobic phase of a two-phase deep eutectic solvent;
(5) mixing the hydrophilic phase and the hydrophobic phase according to the volume ratio of 1:1, standing and layering to obtain the two-phase deep eutectic solvent.
Example 2
(1) Weighing choline chloride and levulinic acid according to the molar ratio of 1:2.5, uniformly mixing, heating and stirring at 80-110 ℃ until a transparent liquid is formed, namely a deep eutectic solvent DES1, and mixing DES1 and water according to the mass ratio of 6:4.5 to obtain a substance A;
(2) weighing choline chloride and malonic acid according to a molar ratio of 1:2.5, uniformly mixing, heating and stirring at 80-110 ℃ until a transparent liquid is formed, namely a deep eutectic solvent DES2, and mixing DES2 and water according to a mass ratio of 4.5:6 to obtain a substance B;
(3) mixing the substance A and the substance B according to the volume ratio of 7.75:2.25 to obtain a hydrophilic phase of the two-phase deep cosolvent;
(4) weighing methyl trioctyl ammonium chloride, octanol and octanoic acid according to the molar ratio of 1:2:3.5, uniformly mixing, and heating at 80-110 ℃ until a transparent liquid is formed, namely a deep eutectic solvent DES3, serving as a hydrophobic phase of a two-phase deep eutectic solvent;
(5) mixing the hydrophilic phase and the hydrophobic phase according to the volume ratio of 1:1, standing and layering to obtain the two-phase deep eutectic solvent.
Example 3
(1) Weighing choline chloride and levulinic acid according to the molar ratio of 1:1.5, uniformly mixing, heating and stirring at 80-110 ℃ until a transparent liquid is formed, namely a deep eutectic solvent DES1, and mixing DES1 and water according to the mass ratio of 6:3.5 to obtain a substance A;
(2) weighing choline chloride and malonic acid according to a molar ratio of 1:1.5, uniformly mixing, heating and stirring at 80-110 ℃ until a transparent liquid is formed, namely a deep eutectic solvent DES2, and mixing DES2 and water according to a mass ratio of 4.5:5 to obtain a substance B;
(3) mixing the substance A and the substance B according to the volume ratio of 8.75:1.25 to obtain a hydrophilic phase of the two-phase deep cosolvent;
(4) weighing methyl trioctyl ammonium chloride, octanol and octanoic acid according to the molar ratio of 1:2:2.5, uniformly mixing, and heating at 80-110 ℃ until a transparent liquid is formed, namely a deep eutectic solvent DES3, serving as a hydrophobic phase of a two-phase deep eutectic solvent;
(5) mixing the hydrophilic phase and the hydrophobic phase according to the volume ratio of 1:1, standing and layering to obtain the two-phase deep eutectic solvent.
Example 4
Example 4 the same preparation as in example 1 was carried out except that: and mixing the hydrophilic phase and the hydrophobic phase only containing the substance A according to the volume ratio of 1:1, standing and layering to obtain the two-phase deep eutectic solvent.
Example 5
Example 5 the same preparation as in example 1 was carried out except that: and mixing the hydrophilic phase and the hydrophobic phase only containing the substance B according to the volume ratio of 1:1, standing and layering to obtain the two-phase deep eutectic solvent.
Example 6
(1) Adding 4mL of any one of the two-phase deep eutectic solvents prepared in examples 1-5 into a 20mL glass bottle with a plug, accurately weighing 0.2g of 40-mesh ginkgo leaf powder, adding the ginkgo leaf powder into the two-phase deep eutectic solvent in the glass bottle with the plug, and uniformly mixing by vortex;
(2) mixing, extracting at 60 deg.C under magnetic stirring at 150rpm for 40min, centrifuging at 10000rpm for 10min, collecting hydrophilic phase, detecting flavone, terpene lactone and procyanidin content, and collecting hydrophobic phase to detect polyprenol acetate content.
Example 7
(1) Adding 3mL of any one of the two-phase deep eutectic solvents prepared in examples 1-5 into a 20mL glass bottle with a plug, accurately weighing 0.2g of 40-mesh ginkgo leaf powder, adding the ginkgo leaf powder into the two-phase deep eutectic solvent in the glass bottle with the plug, and uniformly mixing by vortex;
(2) mixing, magnetically stirring at 50 deg.C and 160rpm for 45min, centrifuging at 10000rpm for 10min, collecting hydrophilic phase, detecting flavone, terpene lactone and procyanidin content, and collecting hydrophobic phase to detect polyprenol acetate content.
Example 8
(1) Adding 5mL of any one of the two-phase deep eutectic solvents prepared in examples 1-5 into a 20mL glass bottle with a plug, accurately weighing 0.2g of 40-mesh ginkgo leaf powder, adding the ginkgo leaf powder into the two-phase deep eutectic solvent in the glass bottle with the plug, and uniformly mixing by vortex;
(2) mixing, extracting at 70 deg.C and 100rpm under magnetic stirring for 60min, centrifuging at 10000rpm for 10min, collecting hydrophilic phase, detecting flavone, terpene lactone and procyanidin content, and collecting hydrophobic phase to detect polyprenol acetate content.
Example 9
(1) Adding 4mL of any one of the two-phase deep eutectic solvents prepared in examples 1-5 into a 20mL glass bottle with a plug, accurately weighing 0.2g of 40-mesh ginkgo leaf powder, adding the ginkgo leaf powder into the two-phase deep eutectic solvent in the glass bottle with the plug, and uniformly mixing by vortex;
(2) mixing, extracting at 65 deg.C and 300rpm under shaking and stirring for 30min, centrifuging at 10000rpm for 10min, collecting hydrophilic phase, detecting flavone, terpene lactone and procyanidin content, and collecting hydrophobic phase to detect polyprenol acetate content.
Example 10
(1) Adding 4mL of any one of the two-phase deep eutectic solvents prepared in examples 1-5 into a 20mL glass bottle with a plug, accurately weighing 0.2g of 40-mesh ginkgo leaf powder, adding the ginkgo leaf powder into the two-phase deep eutectic solvent in the glass bottle with the plug, and uniformly mixing by vortex;
(2) mixing, magnetically stirring at 50 deg.C and 300rpm for 30min, centrifuging at 10000rpm for 10min, collecting hydrophilic phase, detecting flavone, terpene lactone and procyanidin content, and collecting hydrophobic phase to detect polyprenol acetate content.
EXAMPLE 11 method for measuring active ingredient
(1) Detection of ginkgo leaf flavone content
The flavone detection adopts a rutin method of an ultraviolet spectrophotometer. Taking 0.5mL of a sample to be detected, adding 0.3mL of 5% sodium nitrite solution, standing for 6min, adding 0.3mL of 10% aluminum nitrate solution, standing for 6min, adding 4mL of 4% sodium hydroxide solution and 4.5mL of 70% ethanol, uniformly mixing, standing for 20min, and detecting at 510 nm.
Preparing standard solution with rutin standard substance, diluting to corresponding times, with concentration gradient of 0.008, 0.016, 0.024, 0.032, 0.04, 0.048, 0.056, 0.064, and 0.072mg/mL, detecting after developing, and drawing standard curve.
The standard curve of flavone detected by rutin method is y-6.1786 x-0.0689, R2The detection range is 0.9989, and the detection range is 0-0.072 mg/mL.
(2) Detection of terpene lactone content in ginkgo leaf
Terpene lactones can undergo a ring-opening reaction under alkaline conditions (pH > 7.5) to form hydroxamic fatty acids in the presence of hydroxylamine, which are then reacted with FeCl3The colored complex is generated by reaction, based on the reaction mechanism, ginkgolide A can be used as a standard sample, and the total terpene lactones content in the ginkgo leaves can be measured by adopting a spectrophotometry.
The process of making the standard curve is as follows: accurately preparing 10mL of 1.42mg/mL ginkgolide A standard solution, sucking 0.0, 0.2, 0.4, 0.6, 0.8 and 1.0mL of ginkgolide A standard solution, respectively placing the solutions into 6 colorimetric tubes with 20mL, adding 70% ethanol solution to 1.0mL of the solutions, then adding 0.4mL of alkaline hydroxylamine solution (13.9% hydroxylamine hydrochloride aqueous solution and 3.5mol/L NaOH solution (1:2) to mix in situ), reacting for 5min, adding 0.4mL of 3mol/L HCI solution and 6% FeCl30.2mL of the solution is mixed uniformly, 5mL of 70% ethanol solution is added, the mixture is shaken uniformly and measured at a wavelength of 517mn, and a standard curve is drawn.
The standard curve of terpene lactone is y-1.2243 x-0.00154, R20.9992, detection range 0-1 mg/mL.
(3) Content determination of ginkgo leaf procyanidin
4-Dimethylcinnamaldehyde (DMAC) spectrophotometry is selected for determination of procyanidine content, and the specific detection method comprises the following steps: accurately measuring 12.5mL of concentrated hydrochloric acid and 12.5mL of water, and diluting to 100mL with absolute ethyl alcohol to obtain acidic ethyl alcohol which needs to be prepared for use. 50mg of DMAC was made up to 50mL with acidic ethanol to prepare a developer. Adding 3mL of color developing agent into 1mL of sample solution, standing at room temperature for 10min for color development reaction, and detecting at 644nm after reaction.
Preparing standard solution with standard substance of procyanidin from folium Ginkgo, diluting to corresponding times, with concentration gradient of 0.0129, 0.0258, 0.0387, 0.0516, 0.0645, 0.0774, 0.0903, 0.1032, 0.1161, and 0.129mg/mL, detecting after developing color, and drawing standard curve.
Drawing a standard curve in an Excel chart tool by using the light absorption value and the procyanidine concentration obtained by detection according to 10 groups of diluted standard solutions, drawing a corresponding linear relation equation, and obtaining the standard curve equation of the procyanidine as follows:
y＝8.8157x+0.0114(0<x<0.129)
R2when the concentration is 0.9994, the linear type of the procyanidin is good in the concentration range of 0-0.129 mg/mL, and the method can be used for quantitative analysis of the procyanidin.
(4) Method for detecting content of polyprenol acetate in ginkgo leaves
Determination of analytical chromatographic conditions
The detection adopts HPLC high performance liquid chromatography to analyze the polyprenol acetate compounds, and sample injection is carried out under the conditions of different mobile phases, column temperatures and detection wavelengths to determine the optimal detection conditions.
Establishment of a Standard Curve for polyprenol acetate
Accurately weighing 14.7mg of polyprenol acetate standard substance, and dissolving in 6mL of n-hexane to prepare polyprenol acetate standard solution. Diluting the standard solution into 11 groups of solutions with different concentrations, which are respectively as follows: 0.0613, 0.0245, 0.49, 0.735, 0.98, 1.225, 1.47, 1.715, 1.96, 2.205 and 2.45 (unit: mg/mL), detecting peak areas at each group of concentrations by HPLC, and drawing a standard curve according to the relationship between the peak areas and the concentrations.
Drawing a coordinate graph in an Excel graph tool by using the peak area obtained by detection and the concentration of the polyprenol acetate according to the diluted 11 groups of standard solutions, drawing a corresponding linear relation equation, and obtaining a standard curve of the polyprenol acetate: 1179.4x +12.746 (0)<x<1.96)，R2Under 0.9991, polyprenols are illustratedThe acetate is good in linear type within the concentration range of 0-1.96 mg/mL, and the method can be used for quantitative analysis of the polyprenol acetate.
Test example 1
The extraction rate of active ingredients from ginkgo leaves was investigated by TPDESS, and the results are shown in Table 1.
Wherein TPDESs of examples 1-3 and the extraction method of example 6 were used to examine the extraction rate of active ingredients from ginkgo biloba leaves by the TPDESs and extraction method of the present invention;
comparative example 1 is where the extraction solvent was water and the extraction method of example 4 was used.
Comparative example 2 was 70% ethanol as an extraction solvent, and the extraction method of example 4 was used.
TABLE 1 Effect of TPDESS on the extraction yield of active ingredients from Ginkgo biloba leaves
Procyanidins (mg/g) Flavone (mg/g) Terpene lactones (mg/g) Polyprenol acetate (mg/g)
Example 1 21.284±0.9169 2.2165±0.0458 22.9552±0.0578 74.2829±1.9386
Example 2 21.145±0.9124 2.2127±0.0567 22.8436±0.0479 74.1857±1.9367
Example 3 21.168±0.9135 2.2089±0.0538 22.8735±0.0597 74.1753±1.8769
Comparative example 1 11.2574±0.2537 1.0537±0.0236 12.3582±0.0247 Not measured out
Comparative example 2 18.4372±0.2742 1.6832±0.5218 19.3527±0.6831 Not detected out
As can be seen from the results in Table 1, the extraction rate of the two-phase deep eutectic solvent and the extraction method of the invention is significantly higher than the extraction rate of the traditional extraction solvent water and 70% ethanol, and the highest extraction rate of the two-phase deep eutectic solvent and the extraction method of the invention can reach 22.0956 + -0.7129 mg/g. The TPDESs can be applied to synchronous extraction of active ingredients of ginkgo leaves, and the extraction efficiency is superior to that of a traditional solvent, wherein the extraction rate of procyanidin is 21.284 +/-0.9169 mg/g, the extraction rate of flavone is 2.2165 +/-0.0458 mg/g, the extraction rate of terpene lactone is 22.9552 +/-0.0578 mg/g, the extraction rate of polyprenol acetate is 74.2829 +/-1.9386 mg/g, and the primary extraction efficiencies of procyanidin, flavone, terpene lactone and polyprenol acetate are 86.07%, 77.72%, 93.29% and 94.63% respectively. Therefore, the TPDESs can efficiently and synchronously extract a plurality of active ingredients in the ginkgo leaves. And the TPDESs are efficient green solvents, do not cause any pollution compared with the traditional organic solvents, and are safe to use.
Test example 2
Influence of the proportion of the DESs components on the extraction rate of active ingredients in the ginkgo leaves.
The TPDESs of examples 1-3 were used, and the extraction procedure of example 6 was used; examining the extraction rate of the deep eutectic solvent of the embodiment 1 of the invention on the active ingredients in the ginkgo leaves; the results are shown in Table 2.
Wherein comparative example 3 is the same as the deep eutectic solvent raw material and preparation method of example 1, except that the molar ratio of choline chloride and malonic acid is 1:1, the molar ratio of choline chloride and levulinic acid is 1:1, the molar ratio of methyltrioctylammonium chloride, octanol and octanoic acid is 1:1:4, and the same extraction method is adopted.
Wherein comparative example 4 is the same as the deep eutectic solvent raw material and preparation method of example 1, except that the molar ratio of choline chloride and malonic acid is 1:3, the molar ratio of choline chloride and levulinic acid is 1:3, and the molar ratio of methyltrioctylammonium chloride, octanol and octanoic acid is 1:3:2, and the same extraction method is adopted.
TABLE 2 influence of different molar ratios of TPDESs components on the extraction rate of active components from ginkgo leaves
Figure BDA0001455409310000101
As can be seen from the results in Table 2, the two-phase deep eutectic solvent obtained by using the molar ratio of the examples of the present invention has a significantly higher extraction rate of active ingredients from ginkgo biloba leaves than the comparative examples. Wherein the molar ratio of comparative example 4 failed to form a stable two-phase system and thus failed to perform extraction. The component molar ratios have a large influence on the formation and properties of the DESs. The component molar ratio of the DESs influences the properties of the DESs, such as viscosity, surface tension, etc., and also influences intermolecular forces, such as hydrogen bonding, van der Waals force, hydrophobic force, etc., between the DESs components, between the DESs and the extract, and further influences the extraction rate.
Example 3
The influence of TPDESs prepared from different hydrophilic phases and hydrophobic phases on the extraction rate of active ingredients in ginkgo leaves.
The two-phase deep eutectic solvents (TPDES) of example 1, example 4 and example 5 are adopted, the mass-to-volume ratio of the ginkgo leaves to the TPDES is 1:20(g/ml, feed-to-liquid ratio), the magnetic stirring is carried out at 150rpm, the extraction temperature is 50 ℃, and the extraction time is 30 min.
TPDESs composed of a substance A and a substance B in a volume ratio of 8.75:1.25 are selected as an extraction solvent for combined extraction, the extraction rate of procyanidine is 17.5974 +/-0.8514 mg/g, the extraction rate is 10.71% higher than that of a single substance A/hydrophobic phase two-phase system in example 4, and the extraction rate is 9.02% higher than that of a single substance B/hydrophobic phase two-phase system in example 5; the extraction rate of flavone is 2.1815 +/-0.0168 mg/g, is 6.98% higher than that of a single substance A/hydrophobic phase two-phase system, and is 15.91% higher than that of a single substance B/hydrophobic phase two-phase system; the extraction rate of terpene lactone is 17.7957 + -0.8657 mg/g, which is 24.83% higher than that of single substance A/hydrophobic phase two-phase system, but 2.47% higher than that of single substance B/hydrophobic phase two-phase system; the extraction rate of the polyprenol acetate is 75.7373 +/-2.0148 mg/g, is the same as that of a single substance A/hydrophobic phase two-phase system, is 78.53% higher than that of a single substance B/hydrophobic phase two-phase system, and has an obvious effect.
Test example 4
Influence of TPDESs extraction method on extraction rate of folium Ginkgo active ingredients.
The TPDESs of example 1 were used while substituting the ratio of substance A to substance B in example 1 to 8.25: 1.75; respectively taking magnetic stirring (150rpm), ultrasonic extraction (150w) and shaking table oscillation (150rpm) as extraction methods to be selected under extraction conditions, wherein the rest conditions are as follows: the mass-volume ratio of the ginkgo leaves to the TPDESs is 1:20(g/mL), the equal volume of the hydrophilic phase and the hydrophobic phase is mixed, the extraction temperature is 50 ℃, and the extraction time is 30 min. The extraction results are shown in Table 3.
TABLE 3 Effect of different extraction methods on the extraction of active ingredients from Ginkgo biloba leaves by TPDESS
Procyanidins (mg/g) Flavone (mg/g) Terpene lactones (mg/g) Polyprenol acetate (mg/g)
8.75:1.25 stirring 17.5974±0.8514 2.1815±0.0168 17.7957±0.8657 75.7373±2.0148
8.75:1.25 ultrasound 12.1337±0.9780 1.8766±0.0531 13.0311±0.5659 47.8669±1.1068
8.75:1.25 oscillation 14.1188±0.9098 1.7998±0.0515 12.5002±0.0578 57.8056±1.7012
8.25:1.75 stirring 16.3307±1.1017 2.0493±0.0910 18.3675±0.3683 74.4370±1.5761
8.25:1.75 ultrasound 10.6401±0.9625 1.6743±0.0343 15.5223±0.0578 46.6246±1.8307
8.25:1.75 oscillations 14.4401±0.4813 1.7957±0.0584 14.4469±0.6238 51.1173±1.7426
7.75:2.25 stirring 17.0113±0.9169 2.0196±0.0168 20.0828±0.4991 74.8909±1.5994
7.75:2.25 ultrasound 8.7495±0.8589 1.3210±0.0474 15.9171±0.3401 43.6770±0.6890
7.75:2.25 oscillation 14.7048±0.4723 1.6689±0.0327 15.0731±0.231 44.3698±1.0139
From the results in table 3, it can be seen that the extraction rates of the four active ingredients for the same extraction solvent are given by the three extraction methods: magnetic stirring > shaking table extraction > ultrasonic extraction. In the magnetic stirring method, as the proportion of the substance B is increased, the extraction rates of polyprenol acetate and procyanidine are basically similar and are within an error range, the extraction rate of flavone is reduced, and the extraction rate of terpene lactone is increased; in the ultrasonic extraction method, as the proportion of the substance B is increased, the extraction rate of terpene lactones is increased, and the extraction rates of the other three indexes are reduced; in the shaking extraction method, procyanidin extraction rate is kept unchanged, flavone and polyprenol acetate extraction rate is slightly reduced, and terpene lactone extraction rate is increased. By integrating the extraction capacities of the two-phase system in the three extraction methods, it can be concluded that as the content of DES1 increases, the capacities of the TPDESs two-phase system for extracting procyanidins are basically similar, the capacities for extracting flavone and polypentenol acetate decrease, and the capacity for extracting terpene lactones increases. Therefore, the best extraction efficiency was achieved when the magnetic stirring was carried out at 150rpm and the volume ratio of substance A to substance B was 8.75: 1.25.
Test example 5
The mass-to-volume ratio (g/mL) of ginkgo leaves to TPDESs (namely the feed-to-liquid ratio) has influence on the extraction rate of active components in ginkgo leaves.
The extraction conditions are as follows: magnetically stirring at 150rpm, extracting at 50 deg.C for 30 min; examining the influence of different mass-to-volume ratios (g/mL) of the ginkgo leaves and TPDESs in example 1 of the invention on the extraction rate of active components in ginkgo leaves, the results are shown in Table 4 when the material-liquid ratios are 1:5, 1:10, 1:15, 1:20, 1:25 and 1:30 (g/mL);
TABLE 4 Effect of feed liquid ratio on extraction of active ingredients from ginkgo leaves
Figure BDA0001455409310000111
Figure BDA0001455409310000121
As is clear from the results in Table 4, the extraction yield of the active ingredient was better at an extract-to-solution ratio of 1: 15-25. The extraction rate gradually increased with increasing TPDESs until 1:20 equilibrium was reached, at which time the extraction rate no longer increased by increasing the volume of TPDESs, and decreased significantly by 1: 30. According to analysis, when the feed-liquid ratio is 1:20, the extraction rate is higher. For TPDESs, the ratio of hydrophilic phase to hydrophobic phase is 1:1(v/v), and when the ratio of hydrophilic phase is decreased, the ratio of hydrophilic phase to liquid decreases, the extraction rate of hydrophilic components decreases, and the ratio of hydrophobic phase to liquid increases, but the extraction rate of hydrophobic substances does not increase, so that equal volumes of hydrophilic DESs and hydrophobic DESs are mixed in an optimum mixing ratio.
Test example 6
Influence of different temperatures on the extraction rate of the active ingredients of the ginkgo leaves extracted by TPDESs.
The extraction conditions are as follows: magnetically stirring at 150rpm, wherein the mass-to-volume ratio of the ginkgo leaves to the TPDESs is 1:20(g/mL), and the extraction time is 30 min; examining the extraction rate of the TPDESs in the embodiment 1 of the invention on the active components of ginkgo leaves at different temperatures, wherein the selected temperatures are 45, 50, 55, 60, 65 and 70 ℃ for extracting the active components; the results are shown in FIG. 2.
As can be seen from FIG. 2, the equilibrium value of polyprenol acetate is reached at 50 deg.C, the extraction equilibrium of flavone is reached at 55 deg.C, and the extraction equilibrium of procyanidin and terpene lactone is reached at 60 deg.C. In order to comprehensively consider the extraction rates of the four substances, the extraction temperature is selected to be 50-70 ℃; preferably 60-65 deg.C, and the most preferable temperature is 60 deg.C.
Test example 7
Influence of different extraction time on the extraction rate of the active components in the ginkgo leaves extracted by TPDESs.
The extraction conditions are as follows: magnetically stirring at 150rpm, wherein the mass-to-volume ratio of the ginkgo leaves to the TPDESs is 1:20(g/mL), the extraction time is 30min, and the extraction temperature is 60 ℃; examining the extraction rate of the TPDESs in the embodiment 1 of the invention on the active components of ginkgo leaves under different extraction time, and extracting the active components with the extraction time of 25 min, 30min, 35min, 40min, 45min, 50 min, 55 min, 60min and 65 min; the results are shown in FIG. 3.
As shown by the results in FIG. 3, polyprenol acetate reached extraction equilibrium at 30min, flavone and terpene lactone reached extraction equilibrium at 35min, and procyanidin equilibrium time was 40 min. So far, the extraction time is selected to be 30-60min, preferably 40-45min, and most preferably 40 min.
In conclusion, the deep eutectic solvent prepared by the invention is stirred at the rotating speed of 150rpm, the extraction temperature is 60 ℃, the material-liquid ratio is 1:20, and the time is 40 min. The extraction rate of procyanidin is 21.284 +/-0.9169 mg/g, the extraction rate of flavone is 2.2165 +/-0.0458 mg/g, the extraction rate of terpene lactone is 22.9552 +/-0.0578 mg/g, the extraction rate of polyprenol acetate is 74.2829 +/-1.9386 mg/g, and the primary extraction efficiency of procyanidin, flavone, terpene lactone and polyprenol acetate is 86.07%, 77.72%, 93.29% and 94.63% respectively. Therefore, the two-phase system deep eutectic solvent prepared by the invention can simultaneously and efficiently extract a plurality of active ingredients in the ginkgo leaves.

Claims (10)

1. A two-phase deep eutectic solvent for extracting active ingredients of ginkgo leaves is characterized by consisting of a hydrophilic phase and a hydrophobic phase which have the same volume, wherein the hydrophilic phase consists of a substance A and a substance B according to the volume ratio of 10-7.75:1.25-10, the substance A consists of DES1 and water according to the mass ratio of 6:3.5-4.5, and DES1 is prepared from choline chloride and levulinic acid according to the molar ratio of 1: 1.5-2.5; the substance B consists of DES2 and water according to the mass ratio of 4.5:5-6, and DES2 is prepared from choline chloride and malonic acid according to the molar ratio of 1: 1.5-2.5; the hydrophobic phase DES3 is prepared from methyl trioctyl ammonium chloride, octanol and octanoic acid according to a molar ratio of 1:2: 2.5-3.5.
2. The two-phase deep eutectic solvent of claim 1, wherein the hydrophilic phase consists of substance a and substance B in a volume ratio of 7.75-8.75: 2.25-1.25.
3. The two-phase deep eutectic solvent of claim 1, wherein the substance A is composed of DES1 and water in a mass ratio of 6:4-4.5, and the DES1 is prepared from choline chloride and levulinic acid in a molar ratio of 1: 2-2.5; the substance B consists of DES2 and water according to the mass ratio of 4.5:5.5-6, and DES2 is prepared from choline chloride and malonic acid according to the molar ratio of 1: 1.5-2.5; the hydrophobic phase DES3 is prepared from methyl trioctyl ammonium chloride, octanol and octanoic acid according to a molar ratio of 1:2:2.5-3.
4. A method for preparing the two-phase deep eutectic solvent according to claim 1, comprising the steps of:
(1) weighing choline chloride and levulinic acid according to a molar ratio, uniformly mixing, heating and stirring at 80-110 ℃ until a transparent liquid is formed, namely a deep eutectic solvent DES1, and mixing DES1 and water according to a mass ratio to obtain a substance A;
(2) weighing choline chloride and malonic acid according to a molar ratio, uniformly mixing, heating and stirring at 80-110 ℃ until a transparent liquid is formed, namely a deep eutectic solvent DES2, and mixing DES2 and water according to a mass ratio to obtain a substance B;
(3) mixing the substance A and the substance B according to the volume ratio to obtain a hydrophilic phase of the two-phase deep cosolvent;
(4) weighing methyl trioctyl ammonium chloride, octanol and octanoic acid according to a molar ratio, uniformly mixing, and heating at 80-110 ℃ until a transparent liquid is formed, namely a deep co-dissolving solvent DES3, serving as a hydrophobic phase of a two-phase deep co-dissolving solvent;
(5) and mixing the hydrophilic phase and the hydrophobic phase in equal volume, standing and layering to obtain the two-phase deep eutectic solvent.
5. A method for extracting active components from ginkgo biloba leaves by using the two-phase deep eutectic solvent as claimed in claim 1, comprising the steps of:
(1) placing the two-phase deep eutectic solvent in an extraction container, accurately weighing folium Ginkgo powder, adding into the two-phase deep eutectic solvent in the extraction container, and mixing well;
(2) mixing, heating for extraction, centrifuging after extraction, and layering a two-phase system, wherein the upper phase is a hydrophobic phase and the lower phase is a hydrophilic phase; and (3) detecting the contents of flavone, terpene lactone and procyanidine in the hydrophilic phase, and detecting the content of poly (isopentenol acetate) in the hydrophobic phase.
6. The method for extracting active components from ginkgo biloba leaves as claimed in claim 5, wherein the mass volume ratio of the ginkgo biloba leaf powder to the two-phase deep eutectic solvent in the step (1) is 1:15-25 g/mL.
7. The method for extracting active components from ginkgo biloba leaves as claimed in claim 6, wherein the mass volume ratio of the ginkgo biloba leaf powder in the step (1) to the two-phase deep eutectic solvent is 1:20-25 g/mL.
8. The method of extracting ginkgo leaf active ingredients according to claim 5, wherein the heating extraction in the step (2) is a shaking heating extraction or a stirring heating extraction.
9. The method of claim 5, wherein the rotation speed of the shaking or stirring is 100-300rpm, the temperature is 50-70 ℃, and the time is 30-60 min.
10. The method as claimed in claim 9, wherein the rotation speed of the oscillation or stirring is 150-160rpm, the temperature is 60-65 ℃, and the time is 40-45 min.
CN201711064250.1A 2017-11-02 2017-11-02 A kind of two-phase deep eutectic solvent for extracting active components of Ginkgo biloba, and preparation method and extraction method thereof Active CN107789376B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201711064250.1A CN107789376B (en) 2017-11-02 2017-11-02 A kind of two-phase deep eutectic solvent for extracting active components of Ginkgo biloba, and preparation method and extraction method thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201711064250.1A CN107789376B (en) 2017-11-02 2017-11-02 A kind of two-phase deep eutectic solvent for extracting active components of Ginkgo biloba, and preparation method and extraction method thereof

Publications (2)

Publication Number Publication Date
CN107789376A CN107789376A (en) 2018-03-13
CN107789376B true CN107789376B (en) 2021-05-18

Family

ID=61548809

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201711064250.1A Active CN107789376B (en) 2017-11-02 2017-11-02 A kind of two-phase deep eutectic solvent for extracting active components of Ginkgo biloba, and preparation method and extraction method thereof

Country Status (1)

Country Link
CN (1) CN107789376B (en)

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108614059B (en) * 2018-06-26 2020-09-22 河南师范大学 Method for detecting nitrite by using hydrophobic eutectic solvent vortex-assisted dispersion liquid microextraction-high performance liquid chromatography
CN110742913B (en) * 2019-09-20 2022-06-21 南京林业大学 A kind of deep eutectic solvent for solubilization of Ginkgo biloba extract and its preparation method and application
WO2021113568A1 (en) * 2019-12-05 2021-06-10 The Procter & Gamble Company Method of making a cleaning composition
EP4069810A1 (en) * 2019-12-05 2022-10-12 The Procter & Gamble Company Cleaning composition
CN111171095B (en) * 2020-01-09 2023-03-14 广西壮族自治区中国科学院广西植物研究所 Deep eutectic solvent and method for extracting flavone-diglucoside from war bone
CN111359260B (en) * 2020-03-30 2021-12-28 南京林业大学 Hydrophobic deep eutectic solvent for removing ginkgolic acid in crude ginkgo leaf extract by liquid-liquid micro extraction, and preparation method and removal method thereof
CN114053754B (en) * 2021-11-30 2023-07-21 南京林业大学 Two-phase deep eutectic solvent and extraction method for simultaneous separation and extraction of multiple active ingredients from waste ginkgo fruit outer testa
CN114656386A (en) * 2022-04-19 2022-06-24 中国科学院兰州化学物理研究所 A kind of method for extracting lycium barbarum components from wolfberry fruit

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104887723A (en) * 2015-06-18 2015-09-09 天津理工大学 Method for extracting flavonoids from ginkgo biloba leaves with deep eutectic solvent
CN105777696A (en) * 2016-04-06 2016-07-20 华南理工大学 Method for extracting anthocyanin by utilizing deep eutectic solvent (DES)

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2650345B1 (en) * 2012-04-13 2015-01-21 King Saud University Method for removal of sulfur containing compounds from hydrocarbon mixtures

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104887723A (en) * 2015-06-18 2015-09-09 天津理工大学 Method for extracting flavonoids from ginkgo biloba leaves with deep eutectic solvent
CN105777696A (en) * 2016-04-06 2016-07-20 华南理工大学 Method for extracting anthocyanin by utilizing deep eutectic solvent (DES)

Also Published As

Publication number Publication date
CN107789376A (en) 2018-03-13

Similar Documents

Publication Publication Date Title
CN107789376B (en) A kind of two-phase deep eutectic solvent for extracting active components of Ginkgo biloba, and preparation method and extraction method thereof
CN110618211B (en) Method for extracting scutellaria chemical components by using eutectic solvent
CN101392000B (en) High efficiency separation purification method of ginkgolide A, B, C, J and bilobalide monomer
CN104147054B (en) A kind of ginkgo biloba p.e and its preparation method and application
CN103123344A (en) Method for measuring fingerprints of medicinal composition
CN105669782A (en) Method for solid-phase extraction of flavonoid compounds in radix puerariae
CN101816694B (en) Echinacea purpurea oral liquid and preparation method and application thereof
CN1899323A (en) Ginkgo leaf extract, injection containing said extract and its preparing method
CN106588838A (en) Hydroxydihydrobovolide in portulaca oleracea, and extraction and separation method of hydroxydihydrobovolide
CN101029889A (en) Method for inspecting Chinese medicinal preparation quality in treatment of old man eyes dieases
CN111228354A (en) Anti-virus active site in cortex dictamni, preparation method, content determination method and application thereof
CN103822888A (en) Quality inspection method of chasing yellow grass
CN114053754B (en) Two-phase deep eutectic solvent and extraction method for simultaneous separation and extraction of multiple active ingredients from waste ginkgo fruit outer testa
CN111595972A (en) Preparation method and quality detection method of lithocarpus polystachyus rehd total flavone extract
CN102670670B (en) Preparation method of ginkgo dipyridolum injection with high content of ginkgo terpene lactones
CN103923138B (en) A kind of preparation method of Nicotifiorin and application thereof
CN114890974A (en) A method for extracting osthole by response surface design method
CN103316087B (en) Semen Myristicae five tastes effective site and preparation method thereof, quality determining method and application
CN1305870C (en) Novel flavane derivative and its preparation method and uses
CN101596274A (en) The method of quality control of Fructus Schisandrae Chinensis in the YIXINSHU Chinese medicine preparation
CN105037466B (en) A kind of method of the extraction separation Quercetin-7-BETA-D-rhamnose from hypericum japonicum
CN1528758A (en) Separation method of main monomer components of soybean isoflavones
CN104546727B (en) A kind of quinocetone soluble powder and preparation method
CN110954618A (en) Method for detecting Cannabigerol (CBG) in industrial cannabis sativa by using HPLC
CN101721531A (en) Shendeng 4-decoction target field and preparation method thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant