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Lignan compound as well as method for extracting and separating lignan compound from litsea coreana and application

A technology of lignans and separation method, applied in the field of lignans and their extraction and separation from eagle tea, to achieve the effects of good controllability and reproducibility, reduced sample loss and simple operation

Active Publication Date: 2018-10-26
ZUNYI MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the existing technology, the compounds in eagle tea extracted and separated from Litsea coreana Lévl.var.lanuginosa (Migo) Yang et P.H.Huang are mostly flavonoids, and no active ingredients of lignans have been reported.

Method used

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  • Lignan compound as well as method for extracting and separating lignan compound from litsea coreana and application
  • Lignan compound as well as method for extracting and separating lignan compound from litsea coreana and application
  • Lignan compound as well as method for extracting and separating lignan compound from litsea coreana and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The method for extracting and separating the above-mentioned lignans from eagle tea comprises the following:

[0028] 1) Take 15kg of the leaves of the eagle tea tree that Leopard skin camphora is derived from, after drying and crushing, extract 3 times with methanol at a concentration of 90% under reflux, each extraction for 2-5 hours, remove the chlorophyll by MCI column chromatography after the extracts are combined, and then Removed the extract of chlorophyll and concentrated under reduced pressure to obtain 843g of methanol extract;

[0029] 2) After the methanol extract in 1) is dispersed in water, it is sequentially extracted three times with petroleum ether, ethyl acetate and n-butanol, and each extracted part is concentrated under reduced pressure to obtain extracts of various parts;

[0030] 3) Separate the extract from the ethyl acetate part in 2) by silica gel column chromatography, and use petroleum ether-ethyl acetate at a volume ratio of 10:1, 10:1, 5:1, ...

Embodiment 2

[0034] Above-mentioned 1) the aqueous methanol concentration 95% that reflux extraction is used, reflux extraction 3 times; Beneficial effects described in the present invention.

[0035] The present invention carries out the condition of TLC identification: developer is sherwood oil-ethyl acetate system and dichloromethane-methanol system, chromogenic agent a: observe fluorescence under ultraviolet lamp (254nm); chromogenic agent b: 5% ethanol sulfate Spray, bake at 105°C until color develops; color developer c: iodine jar for color development.

[0036] Structural identification: The spectroscopic techniques used mainly include nuclear magnetic resonance spectroscopy ( 1 H-NMR, 13 C-NMR, NOESY, HSQC, HMBC) and mass spectrometry (HR-ESI-MS) identified the structures of the compounds.

[0037] The compound is a yellow oil, HR-ESI-MS [M–H]–m / z 373.1287 (calculated value 373.1293), combined with NMR spectrum to determine the molecular formula of the compound is C 20 h 22 o ...

Embodiment 3

[0042] The inhibitory effect of the lignan component on human liver cancer cells HepG2 was determined.

[0043] Using the conventional MTT method, select the above-mentioned cells in the logarithmic growth phase, adjust the cell concentration to 10×105 cells / mL with culture medium containing 10% fetal bovine serum, inoculate 100 μL per well into a 96-well flat-bottomed cell culture plate, place in After culturing in a 37° C., 5% CO2 incubator for 24 hours, the samples were added at concentrations of 0, 10, 20, 40, 80, and 160 μg / mL, and three replicate wells were set up. Continue culturing in a 37° C., 5% CO2 incubator for 48 hours, then discard the culture solution with the sample, and add 100 μL of culture solution. Then add 20 μL of MTT solution (5 mg / mL) to each well to continue culturing for 4 hours, discard the supernatant after centrifugation, wash with PBS for 2 to 3 times, and then add MTT-containing culture solution. The culture was terminated, and the culture mediu...

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Abstract

The invention discloses a method for extracting and separating a lignan compound from litsea coreana and application of the compounds in the chemistry field of separation and extraction technologies.The method disclosed by the invention is characterized in that the lignan compound is separated and purified from the litsea coreana by combining the traditional extraction method and a modern chromatographic separation technology, and the method comprises the following steps: performing reflux extraction, solvent extraction, column chromatography separation and semi-preparative high performance liquid chromatographic separation. According to the separating preparation method provided by the invention, the lignan component is successfully separated and prepared from the litsea coreana, and activity tests show that the lignan component can be used for preparing anticancer drugs.

Description

technical field [0001] The invention relates to separation and extraction technology in the field of chemistry, in particular to a lignan compound and its extraction and separation method and application from eagle tea. Background technique [0002] Eagle tea is originally produced in Dalou Mountain, and it is still in wild and semi-wild state. It is a plant substitute tea that has been drunk by various ethnic groups in southern my country for a long time, especially in Sichuan, Chongqing and Zunyi, Guizhou. Studies have shown that eagle tea has anti-oxidation, hypoglycemic, lipid-lowering, anti-inflammatory, antibacterial, antiviral and other effects, and has great development and application prospects, which is in line with the theme of general health research advocated by the state. [0003] In the existing technology, the compounds in eagle tea extracted and separated from Litsea coreana Lévl.var.lanuginosa (Migo) Yang et P.H.Huang are mostly flavonoids, and no active in...

Claims

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Application Information

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IPC IPC(8): C07D317/54A61P35/00
CPCA61P35/00C07D317/54
Inventor 肖世基成蕾张茂生何芋歧
Owner ZUNYI MEDICAL UNIVERSITY
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