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Method for screening active constituents capable of eliminating free radicals from natural products

A technology for natural products and active ingredients, applied in material separation, analytical materials, measurement devices, etc., can solve the problems of inability to measure, easily subject to substrate interference, and large demand for natural product raw materials, saving time, reducing Screening cost and the effect of less loss of active ingredients

Inactive Publication Date: 2010-02-03
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This is the basic means for studying the active ingredients of natural products, but there are many defects in this method: 1) the raw materials of natural products require a large amount
However, there are some deficiencies in these free radical determination methods: 1) the single component in the extract cannot be measured, and what is measured is the total activity, and the determination of a single substance needs further separation and purification before determination; 2) due to the use of colorimetry Determination, for the determination of some colored substances such as pigments, etc., it is easy to be interfered by the substrate

Method used

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  • Method for screening active constituents capable of eliminating free radicals from natural products
  • Method for screening active constituents capable of eliminating free radicals from natural products
  • Method for screening active constituents capable of eliminating free radicals from natural products

Examples

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Effect test

Embodiment 1

[0034] The fruit of the plant gardenia is a material with the same source of medicine and food, which has significant effects of protecting the liver, promoting gallbladder, lowering blood pressure and reducing inflammation. Cross 20 mesh sieves after the dried fruit of gardenia is crushed, get 5g dried fruit powder, add 50% ethanol aqueous solution 30 ℃ of water bath oscillations of 20 times of volumes wherein to extract 1 hour, 3000 rev / min centrifugal 10 minutes, after removing precipitation, The upper layer solution was concentrated in vacuo to remove ethanol, and the volume was adjusted to 10 mL with pure water. After passing this solution through the membrane (the pore size of the membrane is 0.45 μm), 10 μL was taken as a sample for high-performance liquid chromatography separation. The chromatographic column was a Zorbax SB-C18 column (4.6mm i.d.×250mm, 5 μm particle size, Agilent Company, the United States) The separation conditions are shown in Table 1 (mobile phase:...

Embodiment 2

[0039] Marigold oil-soluble ingredients are rich in lutein and vitamin E ingredients, which have activities such as scavenging free radicals in the body and anti-aging. After the commercially available marigold particles were pulverized, 20 g of the powder was weighed in a Soxhlet extractor, and 200 mL of n-hexane was used as the extraction solvent for Soxhlet extraction. The heating temperature was 80° C., and the extraction time was 8 hours. After the extraction is completed, the residue and the extract are included, and then the solvent n-hexane (40° C.) is removed from the extract with a vacuum rotary evaporator to obtain an oil-soluble component. Take 0.5 g of the oil, dissolve it in n-hexane to a constant volume of 10 mL, pass it through a membrane (the pore size of the membrane is 0.45 μm), and then take 10 μL of the oil for injection by high performance liquid chromatography. The liquid phase system is a normal phase separation system, and the chromatographic column is...

Embodiment 3

[0041] After the oil-soluble components of marigolds are extracted, the remaining residue is rich in active ingredients, which have good antioxidant and free radical scavenging activities. Take by weighing 1 g of the marigold residue after the oil-soluble component was extracted in Example 2, add 30 ml of 70% ethanol aqueous solution, shake and extract in a water bath at 30° C. for 24 hours, centrifuge at 3000 rpm for 10 minutes, remove the precipitate, and use the extract for 70 % ethanol solution to a volume of 50 mL. After the solution passes through the membrane (0.45 μm), 10 μL of sample is taken for HPLC separation, and the chromatographic column is a Zorbax SB-C18 column (4.6mm i.d.×250mm, 5 μm particle size, Agilent Company, the United States). The separation conditions are shown in Table 2 (Mobile phase: 20% methanol: water (0.25% formic acid) and 100% methanol gradient elution), the separated components flow through the post-column reaction system, and then detected ...

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Abstract

The invention discloses a method for screening active constituents capable of eliminating free radicals from natural products, belonging to the technical field of separation and detection of natural products. The method comprises the following steps: 1) obtaining a constituent chromatogram by separating the constituents in natural products through chromatography ; 2) pumping the free radical solution into a post column reaction system; 3) carrying out liquid phase chromatographic separation on the extracted natural products again according to the operational conditions in step 1), and carryingout the reaction between the chromatographic separation constituents and the free radicals in the post column system; 4) sending the reaction product to a detector, and eliminating the free radicalsby active constituents capable of eliminating free radicals to form a negative peak; and 5) comparing the negative peak with the constituent chromatogram to determine the substance capable of eliminating free radicals. In the method of the invention, a fast separation means of high performance liquid chromatography and a measuring method of free radicals are combined to realize fast and efficientscreening of the active constituents capable of eliminating free radicals from natural products, thus reducing the waste of resources and manpower and having little requirement on natural products.

Description

technical field [0001] The invention belongs to the technical field of separation and detection of natural products, and in particular relates to a method for screening active ingredients with the ability to scavenge free radicals from natural products. Background technique [0002] Natural products generally refer to organic compounds of natural origin that are unique to a certain organism, or shared by a few closely related organisms. It is a substance that is produced by organisms through secondary metabolism and has a special structure and special activity. At present, the extraction, separation and structure determination of the chemical components of natural products have been greatly developed, many of which have been used in various aspects such as medicine, health care and environmental protection. In particular, the research on active ingredients in natural products has received a lot of attention, and the development of new drugs for anti-cancer and anti-infectio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/74G01N30/86
Inventor 高彦祥贺文浩刘璇龚英何李
Owner CHINA AGRI UNIV
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