Coumarins compound and extracting method thereof
A technology of coumarins and extraction methods, applied in the direction of organic chemistry, etc., can solve the problems of unfavorable marketization of tea leaf extracts, unclear components, and many impurities, and achieve wide market application value, high purity, and extraction The effect of mild operating conditions
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Embodiment 1
[0035] The extraction method of coumarin compounds is as follows:
[0036] 1) Test equipment and reagent preparation:
[0037] Fresh Camellia oleifera: Collected from Tianjiling National Forest Park in Hunan Province, identified as Camellia oleifera Abel. by Professor Tan Xiaofeng of Central South University of Forestry and Technology;
[0038] Methanol, acetonitrile: American Fluka Company;
[0039] Ethanol: Sinopharm;
[0040] Dichloromethane: Sinopharm;
[0041] Dimethyl sulfoxide (DMSO): Sinopharm;
[0042] Ultrasonic cleaner, AS series, Tianjin Autosines Instrument Co., Ltd.;
[0043] Rotary evaporator, R-205B, Shanghai Shensheng Instrument Company;
[0044] D-101 macroporous resin: Tianjin Fubang Chemical Technology Co., Ltd.;
[0045] Column chromatography silica gel: produced by Qingdao Ocean Chemical Factory;
[0046] Thin-layer chromatography silica gel plate: produced by Qingdao Ocean Chemical Factory;
[0047] ZFB type three-purpose ultraviolet analyzer: Sh...
Embodiment 2
[0067] The extraction method of coumarin compounds is as follows:
[0068] 1) used test instrument and reagent preparation are basically the same as embodiment 1;
[0069] 2) Specific extraction method: crush the dried leaves of Camellia oleifera in the shade into a powder of more than 70 meshes, take 20 kg and divide it into two batches, heat and reflux with 60% ethanol at a material-to-liquid ratio of 1:3, and repeat the extraction twice, each extraction for 2 hours, heating The temperature of the extraction is controlled at 80°C, the obtained supernatants are combined, filtered to obtain the supernatants, concentrated under reduced pressure until there is no alcohol smell, and half of the original volume of the crude extract sample is obtained, and set aside;
[0070] 3) The 50% alcohol extract sample of camellia oleifera was analyzed by mass spectrometry, and 18 components were identified, including anthocyanins, and a large amount of hydrophilic components such as polysac...
experiment example 1
[0080] The target compound extracted in Example 1 of the present invention was subjected to the following experiment: MTS method was used to detect the effect of the compound on the proliferation activity of RAW264.7 cells. The cells were inoculated in 96-well plates, and the blank control group and the test group (three different concentration groups of 10 μg / mL, 30 μg / mL and 100 μg / mL) were respectively set up, and three replicate holes were set up for each group. Add 20 μL of MTS solution to each well, incubate for 1-2 hours, and use a microplate reader to measure its absorbance at a wavelength of 490 nm. It was found that the test group (three groups) and the control group had no significant difference in absorbance, indicating that the compound would not affect the proliferative activity of RAW264.7 cells. From this experiment, it can be seen that the target compound extracted by the present invention has no side effects on the body;
[0081] Then the target compound of E...
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