Preparation method and content measuring method of macrothelypteris viridifrons total flavone with anti-tumor activity

A kind of anti-tumor activity, the technology of Pinhair fern, which is applied in the field of total flavonoids, achieves the effect of easy control, good reproducibility and high product yield

Inactive Publication Date: 2011-02-09
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Through the study of the chemical composition of the green needle hair fern, the research group found that the plant contains a large amount of flavonoids. Since the flavonoids have a special B-ring structure without a benzene ring, commonly used sodium nitrite-aluminum nitrate, etc. Colorimetric method is not suitable for the content determination of this flavonoid

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The preparation of the total flavonoids of Pinhairia emeralds with anti-tumor activity provided by the present invention

[0028] (1) Preparation of raw materials: the emerald green needle fern (collected in Xijiujiang, Baijiang in October 2009, identified as Macrothelypteris macrothelypteris by the researcher Tan Tse-ming of the Jiujiang Forest Herbarium) The dried whole herb of viridifrons (Tagawa) Ching) is a raw material, which is cleaned, dried, pulverized, and crossed through a 40-mesh sieve to obtain the raw material coarse powder of emerald green needle hair fern for use;

[0029] (2) Solvent extraction: use organic solvent ethanol, methanol, 60-90% hydrous ethanol or 60-90% hydrous methanol to extract the raw material coarse powder of emerald green needle hair fern, heat to 70-90 ℃ for extraction, and the amount of solvent used is the same as that of emerald green The weight ratio of needle hair fern to coarse powder is 6-15:1, and the extraction is repeated 1 ...

Embodiment 2

[0036] Take the dried whole herb of Pinata emerald, crush it, sieve it, add 10 times the volume of 70% ethanol, and extract under reflux at 90°C for 60 minutes. Repeat the extraction twice, combine the extracts, concentrate under reduced pressure to recover the solvent, and dry in vacuum to obtain the total extract. Weigh a certain amount of total extract, add distilled water to dissolve it ultrasonically, extract twice with three times the amount of petroleum ether, combine the water layer, adjust the pH to 6 with 15% sodium hydroxide, filter, and filter out the precipitate. The filtrate is washed with concentrated hydrochloric acid Adjust the pH value to 1, put it in the refrigerator at 4°C overnight, filter the next day, wash the obtained precipitate with distilled water until it is neutral, and dry it in vacuum to obtain the flavonoids extracted from cloverine. After adjusting the pH to 4 with sodium hydroxide, the remaining filtrate was subjected to macroporous resin adso...

Embodiment 3

[0038] Inhibitory effect of total flavonoids of the method of the present invention on human breast cancer cell MCF-7

[0039] After subculture, human breast cancer cells MCF-7 in logarithmic growth phase were blown and dispersed in the medium containing 10% inactivated calf serum, and the concentration was measured with a cell counting plate under an inverted microscope and adjusted to 2×10 5 cells / mL. Add 100 μL of the cell suspension to each well of a 96-well plate and store at 37°C in 5% CO 2 cultured in an incubator for 24 h. Add 100 μL of sterilized sample-containing culture solution to make the final concentrations of the samples 0.1, 1.0, 10.0, and 100.0 μg / mL, respectively. Continue at 37°C 5% CO 2 cultured in an incubator for 48 h. Afterwards, 10 μL of 5 mg / mL MTT solution was added to each well to continue culturing for 4 h. Carefully aspirate the cell solution, add 200 μL of DMSO to each well to dissolve the formed formazan, shake it on a cell plate shaker for...

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Abstract

The invention provides a macrothelypteris viridifrons total flavone with anti-tumor effect and a preparation method thereof, and also provides a content measuring method of the macrothelypteris viridifrons total flavone. The total flavone with anti-tumor activity, which is provided by the invention, is obtained by solvent extraction, alkaline extraction, acid deposition and macroporous resin purification of the dried herbs of the macrothelypteris viridifrons. The macrothelypteris viridifrons total flavone mainly contains flavonoid compounds and has a non-benzene ring special B ring structure, so that the conventional content measuring method of the total flavone is not suitable. The invention establishes a simple and easy content measuring method of the macrothelypteris viridifrons total flavone and optimizes the extraction and purification processes of the macrothelypteris viridifrons total flavone, so that the finally obtained macrothelypteris viridifrons total flavone has obvious anti-tumor activity. The content measuring method of the macrothelypteris viridifrons total flavone, which is provided by the invention, is beneficial to quality control and effective utilization of macrothelypteris viridifrons resources and has wide application prospects.

Description

technical field [0001] The invention relates to total flavonoids with anti-tumor activity extracted from herbal medicines, in particular to total flavonoids from Pinhairia emerald greens, a preparation method thereof, and a content determination method of the total flavonoids. Background technique [0002] Flavonoids widely exist in nature and are an important class of natural organic compounds, generally divided into: flavones, flavonols, dihydroflavones, dihydroflavonols, anthocyanins, chalcones, biflavones, isoflavones, etc. Flavonoids have a variety of biological activities, such as cardiovascular effects, antiviral effects, anti-inflammatory effects, etc. Among them, the antitumor effect of flavonoids has attracted widespread attention. Malignant tumors are common and frequently-occurring diseases that seriously threaten human health. Drug therapy plays an important role in the treatment of malignant tumors. Discovering new substances with anti-tumor activity from Chine...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61P35/00A61K36/11
Inventor 阮金兰魏安华熊朝梅吴光华
Owner HUAZHONG UNIV OF SCI & TECH
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