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Method for extracting flavonoid components in prescription for nourishing yin, tonifying qi and activating blood by using natural deep-eutectic solvent and process optimization method thereof

A technology of deep eutectic solvent, nourishing qi and promoting blood circulation, applied in material separation, analysis of materials, organic chemistry, etc., can solve problems that have not been reported

Active Publication Date: 2021-09-07
ZHEJIANG CHINESE MEDICAL UNIVERSITY
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  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there have been related reports on the extraction of Yangyin Yiqi Huoxue prescription and deep eutectic solvent, but there is no report on the application of deep eutectic solvent to the extraction of Yangyin Yiqi Huoxue prescription

Method used

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  • Method for extracting flavonoid components in prescription for nourishing yin, tonifying qi and activating blood by using natural deep-eutectic solvent and process optimization method thereof
  • Method for extracting flavonoid components in prescription for nourishing yin, tonifying qi and activating blood by using natural deep-eutectic solvent and process optimization method thereof
  • Method for extracting flavonoid components in prescription for nourishing yin, tonifying qi and activating blood by using natural deep-eutectic solvent and process optimization method thereof

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0059] Experimental example 1: Preparation and screening of NADESs

[0060] Accurately weigh a certain mass of hydrogen bond acceptor and hydrogen bond donor reagents, mix them in molar ratio, and stir continuously under heating in an oil bath at 80-90°C until a clear and transparent liquid is formed. The NADESs prepared in this experiment are shown in Table 1.

[0061] Table 1 Different types of NADESs prepared in the laboratory

[0062]

[0063] This experiment investigates the effect of the type of NADES on the extraction rate of the three target components in Yangyin Yiqi Huoxue prescription to find the best type of NADES. Precisely weigh 100mg of Yangyin Yiqi Huoxue prescription into a 1.5mL centrifuge tube, add Pro-Ur, ChCl-Ur, Bet-Gly, Pro-Mu, ChCl-La and Bet-Lac at a ratio of 150:1mg / mL to liquid , the mixture was centrifuged at 4500rpm for 10min with a centrifuge, and the supernatant was taken. HPLC analyzed and calculated the extraction rates of the three effect...

experiment example 2

[0067] Experimental Example 2: Single Factor Experiment

[0068] Effect of water content on extraction rate: Make NADES into solvents with water content of 30, 40, 50, 60, 70%, accurately weigh 100mg of formula for nourishing yin, benefiting qi and activating blood, put it in a 1.5mL centrifuge tube, and follow the 100% Ratio of solid to liquid: Add natural deep eutectic solvent, plug tightly, shake well, and extract at 50°C for 30min. After the extraction, the mixture was centrifuged at 4500rpm for 10min with a centrifuge, and the supernatant was diluted with 1:1 methanol water, then suction filtered, and used for HPLC detection of puerarin, calycosin, and formononetin at different water contents. rate as figure 2 shown.

[0069] The water content (water ratio) of NADES is a key factor affecting the physical and chemical properties of the solvent, which can change the polarity and viscosity of NADES. Adding water to NADES can increase its fluidity, while too much water ma...

experiment example 3

[0076] Experimental Example 3: Methodological Investigation

[0077] Specificity test: Accurately draw the appropriate amount of the mixed reference solution and the formula for nourishing yin, nourishing qi and activating blood, and measure according to the aforementioned chromatographic conditions. The peak shapes of puerarin, calycosin and formononetin in the sample and the mixed standard Well, resolution greater than 1.5, see Figure 6 and Figure 7 . It shows that the specificity of the method is good.

[0078] Investigation of the linear relationship: Accurately draw an appropriate amount of No. 1 mixed solution, and perform multiple dilution to obtain No. 2-7 mixed solution. That is, the concentration of puerarin is 0.5000, 0.2500, 0.125, 0.0625, 0.0313, 0.0156 mg / mL, the concentration of calycosin is 0.200, 0.100, 0.050, 0.025, 0.0125, 0.006 mg / mL, and the concentration of formononetin is 0.100, 0.050 , 0.025, 0.013, 0.006, 0.003mg / mL, perform HPLC analysis accordi...

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Abstract

The invention provides a method for extracting the flavonoid components in a prescription for nourishing yin, tonifying qi and activating blood by using a natural eutectic solvent and a process optimization method thereof. According to the method, the total extraction rate of puerarin, calycosin and formononetin in a prescription for nourishing yin, tonifying qi and activating blood is taken as an evaluation index, a test scheme is determined by adopting Box-Benhnken Design on the basis of a single-factor test, high performance liquid chromatography is used for measuring the three flavonoid components, the total extraction rate is calculated, extraction process parameter optimization is performed in combination with the response surface model, and test verification is performed. Compared with a traditional organic solvent, the natural eutectic solvent adopted by the method is more efficient, economical and environmentally friendly, can be used for extracting puerarin, calycosin and formononetin in the prescription for nourishing yin, tonifying qi and activating blood, is a sustainable extraction medium, and provides a new reference for the extraction process of other prescriptions at the same time.

Description

technical field [0001] The invention belongs to the technical field of chemical component extraction, and in particular relates to a method for extracting flavonoid components in a formula for nourishing yin, benefiting qi and activating blood with a natural deep eutectic solvent and a process optimization method thereof. Background technique [0002] The prescription for nourishing yin, benefiting qi and activating blood is made from rehmannia glutinosa, astragalus root, kudzu root and chuanxiong in a ratio of 1:1:1.2:0.67, decocted in water and extracted with alcohol, rotary evaporated to a certain volume, and excipients are added to make granules. The prescription of nourishing yin, replenishing qi and activating blood has the functions of preventing thrombus formation, preventing tissue cell damage, improving cell antioxidant capacity, etc., and has obvious curative effect in the treatment of cardiovascular and cerebrovascular diseases. [0003] Puerarin has the effects ...

Claims

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Application Information

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IPC IPC(8): C07D311/40C07D311/36C07D407/04G01N30/02
CPCC07D311/40C07D311/36C07D407/04G01N30/02
Inventor 万海同虞立李畅何昱万浩芳杨洁红
Owner ZHEJIANG CHINESE MEDICAL UNIVERSITY
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