A method for screening antioxidant active ingredients

A technology of anti-oxidation activity and thin-layer plate, which is applied in the direction of analyzing materials through chemical reactions, material analysis through observing the influence of chemical indicators, and preparation of test samples, etc., to achieve easy identification, memory, and operation Simple, low-cost experimental results

Inactive Publication Date: 2017-05-31
SHANGHAI UNIV OF T C M +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, TLC-biological autoradiography, which is applied to the screening of antioxidant activity, mostly uses the DPPH method. Because DPPH is a stable free radical, the difference between the active spot (yellow) and the background color (purple) is significant after color development, so it is a good method. It is a good method for screening antioxidant activity; but DPPH free radical is an inorganic free radical, not a free radical that actually exists in organisms; Screening of Antioxidative Active Components for Oxidative Lipid Reaction

Method used

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  • A method for screening antioxidant active ingredients
  • A method for screening antioxidant active ingredients
  • A method for screening antioxidant active ingredients

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] TLC plate A: Take 1g of perilla seed powder, weigh it accurately, add 5mL of methanol, and after ultrasonication for 25 minutes, take the supernatant and filter, and the filtrate is used as the test solution; take rosmarinic acid and luteolin reference substances separately , add methanol to make a solution containing 1mg per 1mL, as the reference solution; draw 1 μL of the reference solution and 10 μL of the test solution respectively, and spot the samples on the same silica gel GF254 thin-layer plate. After the methanol is evaporated, use n-hexane- Dichloromethane-ethyl acetate-formic acid (volume ratio: 2:5:2.5:0.5) is used as the developer for development; take it out and dry it for later use.

[0037] TLC plate B: Take 0.5g of Cistanche deserticola powder, weigh it accurately, add 10mL of methanol, take the supernatant after ultrasonication for 25 minutes, filter, and use the filtrate as the test solution; take another ergosteroside reference substance, add methanol...

Embodiment 2

[0043] TLC plate A: Accurately weigh 1 mg of water-soluble vitamin E (Trolox), dissolve it in 1 mL of absolute ethanol, and prepare a solution with a concentration of 1 mg / mL; draw 2 μL of the solution and spot on silica gel GF 254 On the TLC plate A, shake off the ethanol and set aside.

[0044]TLC plate B: Accurately weigh 1 mg β-sitosterol, dissolve it in 1 mL of absolute ethanol, and prepare a solution with a concentration of 1 mg / mL; absorb 2 μL of the solution and apply it on silica gel GF254 thin-layer plate B, shake off the ethanol for later use .

[0045] Thin-layer plate C: Accurately weigh 1mg of nuciferine, dissolve it in 1mL of absolute ethanol, and prepare a solution with a concentration of 1mg / mL; absorb 2μL of the solution and place a sample on silica gel GF254 thin-layer plate C, and shake off the ethanol for later use .

[0046] Preparation of linoleic acid solution: Accurately measure 0.6 mL of linoleic acid, dissolve it in 20 mL of n-hexane, and prepare a...

Embodiment 3

[0051] Preparation of the test solution: Accurately weigh 1 mg of water-soluble vitamin E (Trolox), dissolve it in 1 mL of absolute ethanol, and prepare a solution with a concentration of 1 mg / mL.

[0052] Preparation of linoleic acid solution: Accurately measure 0.2, 0.4, 0.6, 0.8 and 1.0mL of linoleic acid, dissolve them in 20mL of n-hexane respectively, and prepare linoleic acid n-hexane solutions of different concentrations (recorded as linoleic acid in sequence acid solutions 1, 2, 3, 4, 5).

[0053] Preparation of developer solution: Add 0.87g of thiobarbituric acid and 2.5g of trichloroacetic acid into 100mL of 50% ethanol aqueous solution, stir to dissolve and mix evenly.

[0054] Take the test solution and place samples in parallel on 5 silica gel GF254 thin-layer plates.

[0055] Immerse the above five thin-layer boards in linoleic acid solutions 1, 2, 3, 4, and 5 respectively (the corresponding thin-layer boards are sequentially recorded as thin-layer boards 1, 2, ...

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Abstract

The invention discloses a method for screening an antioxidant active component. The method comprises the following steps: a) applying the sample of a test sample solution on a thin-layer plate, evaporating a sample solvent for further use or further developing the sample solvent with an appropriate developing solvent for further use; b) dipping the obtained product in a linoleic acid solution, taking the product out from an evaporating solvent, and placing the obtained product under an ultraviolet lamp of 254nm or 366nm for illumination; c) dipping the obtained product in a developer solution, taking the product out and heating the product for color development; d) inspecting the obtained product, wherein a blue spot in a green background under the ultraviolet lamp of 366nm and a white spot in a pink background under visible light are the antioxidant active component. According to the method, complex instrument and equipment are eliminated, the antioxidant active component can be screened out quickly in a high throughput, the screening result is vivid, and easy to identify and remember, and the sensitivity and specificity are higher than those obtained by an ordinary screening method, so that a convenient and practical way is provided for screening the antioxidant active component.

Description

technical field [0001] The invention relates to a method for screening antioxidant active ingredients, in particular to a method for screening antioxidant active ingredients with anti-linoleic acid peroxidation lipid reaction by using thin-layer chromatography-bioautographic technology. Background technique [0002] Oxygen free radical reaction and lipid peroxidation reaction play an important role in the metabolic process of the body. Under normal circumstances, the two are in a state of coordination and dynamic balance, maintaining many physiological and biochemical reactions and immune reactions in the body. Once this coordination and Disturbance and imbalance of dynamic balance will cause a series of metabolic disorders and immune function reduction, forming a chain reaction of oxygen free radicals, damaging biofilm and its functions, resulting in the formation of cell hyaline lesions and fibrosis, and large-scale cell damage causing further damage. Damage to nerves, tis...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/78G01N1/28
Inventor 谷丽华秦志鹏廖立平侴桂新王峥涛
Owner SHANGHAI UNIV OF T C M
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