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Method for separating and purifying antioxidant substance from wisteria

A technology for separation and purification of antioxidant substances, applied in the field of separation and purification, can solve problems such as unclear targets, large sample loss, and cumbersome steps, and achieve good separation effects, no sample loss, and large preparation volumes

Active Publication Date: 2019-01-25
HUAIYIN TEACHERS COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, the traditional determination of antioxidant activity of natural products basically only detects the overall antioxidant capacity of the mixture, and it is impossible to determine which compound in the complex mixture has antioxidant activity. The single antioxidant active ingredient can only be detected by column chromatography, etc. The method separates and prepares, and the separated components are measured separately. The goal of this process is not clear, the steps are cumbersome, the workload is large, and the sample loss is large.
[0004] In the application separation of high-speed countercurrent chromatography, the successful selection of the solvent system is the key. The necessary condition for sample separation is that the sample has a suitable distribution ratio in the two immiscible phases of the solvent system. The commonly used method for determining the distribution coefficient is high-performance liquid chromatography. , thin-layer chromatography and spectrophotometry, etc., the principle is to measure the response value of each component in the upper and lower phases respectively, and then calculate the distribution coefficient. The ideal distribution coefficient is 0.5~2.0, but these determination methods can only screen out bright The solvent system for absorbing substances cannot screen out specific high-speed countercurrent chromatography separation solvent systems with antioxidant active substances

Method used

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  • Method for separating and purifying antioxidant substance from wisteria
  • Method for separating and purifying antioxidant substance from wisteria
  • Method for separating and purifying antioxidant substance from wisteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Step 1: Take 50.00 g of wisteria flowers that have been dried and crushed to 20 meshes, add 400 mL of ethanol with a volume concentration of 50% as the extraction solvent, and extract in an ultrasonic-microwave extractor for 30 minutes. The ultrasonic power is 400W and the microwave power is 400W. Ultrasonic-microwave extraction After filtration, the filter residue was repeatedly extracted 4 times, and the filtrate was combined and concentrated in vacuo at 60°C to obtain the wisteria flower extract;

[0035]Step 2: Set the conditions of high performance liquid chromatography, the wavelength is 254nm, Megres C18 column (4.6mm×250mm, 5μm), mobile A phase: water, mobile B phase: acetonitrile; column temperature: 30°C, flow rate 0.8mL / min, injection volume 10μL, gradient elution conditions: 0-5min, 20% B; 5-25min, 20%-30% B; 25-35min, 30%-40% B; 35-40min, 40% B; 40-45min, 40%-20% B; 45-50min, 20% B, under this condition, carry out chromatographic analysis on the extract of...

Embodiment 2

[0042] Step 1: Take 10.00 g of wisteria flowers that have been dried and crushed to 20 meshes, add 60 mL of ethanol with a volume concentration of 70% as the extraction solvent, and extract in an ultrasonic-microwave extractor for 10 minutes. The ultrasonic power is 180W, and the microwave power is 30W. After the completion of filtration, the filter residue was repeatedly extracted 3 times, and the filtrate was combined and concentrated in vacuum at 60°C to obtain the wisteria flower extract;

[0043] Step 2: the implementation mode is the same as step 2 of Example 1;

[0044] Step 3: the implementation mode is the same as Step 3 of Example 1;

[0045] Step 4: Use a solvent system of n-hexane-ethyl acetate-ethanol-water with a volume ratio of 3:5:3:5, the upper phase is the stationary phase, the lower phase is the mobile phase, and the flow rate of the mobile phase is 2 mL / min. The column rotation speed was 900 rpm, and the Wisteria flower extract was separated by high-speed ...

Embodiment 3

[0047] Step 1: Take 100.00 g of wisteria flowers that have been dried and crushed to 20 meshes, add 600 mL of ethanol with a volume concentration of 90% as the extraction solvent, and extract in an ultrasonic-microwave extractor for 30 minutes. The ultrasonic power is 900W, and the microwave power is 900W. Ultrasonic-microwave extraction After filtration, the filter residue was repeatedly extracted 3 times, and the filtrates were combined and concentrated in vacuo at 60°C to obtain the wisteria flower extract.

[0048] Step 2: the implementation mode is the same as step 2 of Example 1;

[0049] Step 3: the implementation mode is the same as Step 3 of Example 1;

[0050] Step 4: using a solvent system of n-hexane-ethyl acetate-ethanol-water with a volume ratio of 4:5:3:5, the upper phase is the stationary phase, the lower phase is the mobile phase, and the flow rate of the mobile phase is 1.5 mL / min. The rotation speed of the separation column was 900 rpm, and the Wisteria flo...

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Abstract

The invention discloses a method for separating and purifying an antioxidant substance from wisteria. The method includes the following steps: the first step of weighing dry pulverized wisteria, usingethanol as an extraction solvent, mixing, putting a mixture in an extractor for extraction or soaking for extraction, filtering, repeating the above extraction step for a residue, combining filtratesand concentrating to obtain a wisteria extract; the second step of adopting a high performance liquid chromatography and an iron ion reduction / antioxidation capability method to screen compound peakswith antioxidant activity from the wisteria extract obtained in the first step on line. According to the method, the cumbersome process of first separating and then re-testing the activity of the traditional method is eliminated. The method has the advantages of being fast, intuitive, accurate and stable, and solves the difficulty that a target is unclear in the traditional high-speed countercurrent chromatography solvent screening, measurement can only be conducted after compound separation by the high-speed countercurrent chromatography, and a solvent system is adjusted according to a measurement result.

Description

technical field [0001] The invention relates to a separation and purification method, in particular to a method for separation and purification of antioxidant substances from wisteria flowers. Background technique [0002] Wisteria is the purple flower of the leguminous plant Wisteria (scientific name: Wisteria sinensis, also known as Wisteria, Zhuteng, Huanghuan, etc.). Wisteria flower is mild in nature, sweet in taste, has the effects of diuresis, swelling, dispelling wind and relieving pain, etc. It is mainly used to treat ascites, edema, dysuria, joint swelling and pain, and gout. Wisteria has a strong fragrance, and it is used to be eaten in many places such as Henan, Shandong, and Hebei in my country. People often pick wisteria flowers to steam food, or make special foods such as wisteria flower cake, wisteria flower cake, and wisteria flower porridge. In addition, wisteria flowers can also extract aromatic oils. The research results [Antioxidant Effect of Wisteria ...

Claims

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Application Information

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IPC IPC(8): C07D311/30C07D311/40
CPCC07D311/30C07D311/40
Inventor 鄢贵龙赵利琴汪伟张洁刘扬孙碧雪王晶杨依晶
Owner HUAIYIN TEACHERS COLLEGE
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