CN-122010893-A - Method for green extraction of flavonoid compounds based on deep eutectic solvent

Abstract

The invention discloses a deep eutectic solvent-based flavonoid compound green extraction method, which comprises the following steps: pulverizing the qi-tonifying and mass-eliminating prescription, and sieving to obtain compound powder; adding compound powder and Deep Eutectic Solvent (DES) into a grinding instrument for MCE grinding extraction to obtain compound extract; adding the compound extract into pure water, and taking supernatant to obtain compound extract after vortex and centrifugation; the invention provides the method for extracting flavonoid components (procyanidine B1, hesperidin, quercetin, aloe-emodin 8-glucoside and neohesperidin) in the qi-tonifying and mass-eliminating formula by using the DES solvent to assist MCE ball milling for the first time, so that the extraction efficiency is obviously improved, the extraction process is more environment-friendly, and the method is suitable for industrial production.

Inventors

• CHU CHU
• LI JIAJUN
• LIU BIAO
• Tabusam Tufel
• PENG XIN
• ZHONG GUANGHUI

Assignees

• 浙江工业大学

Dates

Publication Date

20260512

Application Date

20260202

Claims (8)

1. 1. A method for green extraction of flavonoid compounds based on deep eutectic solvent is characterized by comprising the following steps: A. pulverizing the qi-tonifying and mass-eliminating prescription, and sieving to obtain compound powder; B. Adding the compound powder obtained in the step A and a deep eutectic solvent into a grinding instrument for MCE grinding extraction to obtain a compound extract; The composition of the deep eutectic solvent is selected from any one of the following: choline chloride/urea molar ratio 1:2; choline chloride/ethylene glycol molar ratio is 1:2; choline chloride/lactic acid molar ratio is 1:2; Choline chloride/ethylene glycol/lactic acid molar ratio is 1:1:2; C. And C, adding the compound extract obtained in the step B into pure water, and taking the supernatant to obtain the compound extract through vortex and centrifugation.
2. 2. The method for extracting flavonoid compounds based on deep eutectic solvents in green according to claim 1, wherein in the step A, the formula for tonifying qi and eliminating mass comprises the following medicinal materials in parts by weight.
3. 3. The method for green extraction of flavonoid compounds based on deep eutectic solvents as claimed in claim 1, wherein in the step B, the feed liquid ratio of the compound powder to the deep eutectic solvents is 10:1, mg/μl.
4. 4. The method of claim 1, wherein in step B, 240 s is ground and extracted with 540 rpm.
5. 5. The method for green extraction of flavonoids based on deep eutectic solvents according to claim 1, wherein in the step B, the composition of the deep eutectic solvents is choline chloride/lactic acid molar ratio of 1:2.
6. 6. The method for green extraction of flavonoids based on deep eutectic solvents according to claim 1, wherein in the step B, the preparation method of the deep eutectic solvents comprises the steps of mixing the components according to the proportion, heating to 80 ℃ in a nitrogen environment, and magnetically stirring with 300 rpm until a uniform transparent liquid is obtained.
7. 7. The method for green extraction of flavonoid compounds based on deep eutectic solvents according to claim 1, wherein in the step C, the liquid-solid ratio of pure water to the compound powder obtained in the step A is 1:50, mL/mg.
8. 8. The method for green extraction of flavonoids based on deep eutectic solvents according to claim 1, wherein in step C, vortex 3 min is centrifuged 3 min at 15000 rpm.

Description

Method for green extraction of flavonoid compounds based on deep eutectic solvent Technical Field The invention relates to extraction of active ingredients of traditional Chinese medicines, in particular to a novel, green and efficient method for extracting flavonoid compounds in a traditional Chinese medicine compound based on deep eutectic solvents. Background Mechanical cell disruption (MCE) is a novel and efficient pretreatment technique that is widely accepted for its environmental protection and pollution-free properties. The technology has wide application prospect in the field of natural product and traditional Chinese medicine extraction. Through the chemical and physical change of the mechanical force-induced material, the MCE can effectively realize the reduction of the particle size under the normal temperature condition, and has the remarkable advantages of high extraction rate, short treatment time, small environmental impact and the like. Therefore, the MCE method is widely used for extracting flavonoids, alkaloids and other bioactive compounds based on the advantages of low cost, high efficiency, normal temperature operation, pure water instead of an organic solvent as an extraction medium, and the like. However, the traditional MCE is mainly used as a simple extraction process, lacks an integrated enrichment step, and limits the detection efficiency of the traditional MCE on low-abundance components. Furthermore, the process generally relies on solid alkaline reagents (including inorganic and organic bases), with potential environmental risks. Therefore, the development of novel environment-friendly reagents for extraction of target compounds has important significance. Such reagents are particularly critical for microextraction applications and detection of low abundance components in complex matrices. With the increasing importance of global environmental protection and sustainable development, traditional organic solvents face stricter and stricter limits due to high toxicity and poor biodegradability. Therefore, the development of green solvents has become a major focus of green chemistry research. Deep Eutectic Solvents (DESs) are mixtures of two or more components, typically a hydrogen bond acceptor HBA and a hydrogen bond donor HBD, which form a eutectic system at a specific molar ratio by strong intermolecular forces. During DES formation, HBA and HBD form hydrogen bonds through specific molar ratios, forming a mixture with a melting point lower than that of the pure components. This unique composition imparts significant physicochemical properties to the DESs, including low volatility, low toxicity, ease of manufacture, biodegradability, and environmental compatibility, all of which have been demonstrated by scientific research. In recent years, DESs are increasingly replacing traditional organic solvents, becoming green substitutes for extracting and enriching active ingredients from various natural products. However, this extraction method is rarely used for extraction and separation of complex mixtures of compounds, since most methods still rely on toxic organic solvents and green solvents, such as DESs, are rarely used. Therefore, binding MCE to des rather than traditional agents is a promising research direction. Qi-tonifying and mass-eliminating prescription (YQXZF) is developed by modifying a prescription for treating Chronic Kidney Disease (CKD) according to a national famous doctor Zhang Peiqiu in the Kidney department of Ningbo market. The prescription is composed of astragalus root, pilose asiabell root, chinese angelica root, ligusticum wallichii, white muscardine silkworm, rhubarb horsetails, zedoary and dried orange peel, and can reduce urine protein level, improve kidney function, inhibit high sugar induced tubular epithelial cell fibrosis, epithelial-mesenchymal transition and inflammatory reaction. Hesperidin, quercetin and neohesperidin are natural flavonoids. These three compounds are designated as quantitative markers in the chinese pharmacopoeia and exhibit a range of pharmacological activities including antioxidant, anti-inflammatory, cardiovascular protection and hypoglycemic effects. In addition, YQXZF also contains various flavonoid compounds including procyanidin B1 and aloe-emodin 8-glucoside, and has remarkable anti-inflammatory and anti-fibrosis activities. Is widely used for treating kidney meridian blood stasis light symptoms caused by spleen and kidney qi deficiency, blood circulation weakness and qi stagnation and blood stasis clinically. The invention provides a green, practical and basic DES auxiliary MCE method for extracting procyanidine B1, hesperidin, quercetin, aloe-emodin 8-glucoside and neohesperidin from YQXZF. Five target compounds were quantitatively analyzed by HPLC. Key parameters including DES type, DES usage, milling time, solid to liquid ratio, and vortex time were optimized by a single factor test system. The interaction among thre