Extraction method and application of antitumor active components of Cordyceps militaris
An anti-tumor activity and extraction method technology, applied in the field of biological product processing, can solve problems such as hindering the progress of Cordyceps militaris, loss of active components, etc., and achieve anti-tumor active components with high content, high consistency, and large amount of separation, purification and preparation. Effect
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Embodiment 1
[0042] Example 1: Extraction of Cordyceps militaris anti-tumor active components
[0043] 1. Raw materials and materials:
[0044] The fruiting bodies of Cordyceps militaris were produced in this laboratory.
[0045] 2. Reagents:
[0046] Ultrapure water was produced by Millipore Water Purifier; acetone was purchased from Tianjin North Tianyi Chemical Reagent Factory; chromatography-grade ethyl acetate, n-hexane, and methylene chloride were purchased from Honeywell Company.
[0047] 3. Instruments and equipment:
[0048] The ultrafine pulverizer was purchased from Shandong Sanqing Stainless Steel Equipment Co., Ltd.; the desktop refrigerated centrifuge was purchased from Thermo Company; the rotary evaporator was purchased from Shanghai Yarong Biochemical Instrument Factory;
[0049] The extraction steps of the anti-tumor active components of the fruiting body of Cordyceps militaris are as follows:
[0050] (1) Ultra-low temperature crushing:
[0051] Take the dried fruiti...
Embodiment 2
[0060] Embodiment 2: Inhibition of human lung cancer A549 cell growth test
[0061] Main materials and reagents:
[0062] Human lung cancer A549 cells were donated by Aisen Bio (Hangzhou) Co., Ltd.; F-12K medium was donated by Gibco; cyclophosphamide was purchased from Tianjin Jinshi Pharmaceutical Co., Ltd.
[0063] Blank control well: no drug added;
[0064] Positive control well: add cyclophosphamide;
[0065] Experimental well: Add the anti-tumor active component C310-6-2 obtained in Example 1.
[0066] Cell culture:
[0067] at 5%C0 2 , 37°C, under saturated humidity, culture human lung cancer A549 cells in F-12K (10%FBS+1%PS) medium, and select the cells growing in the logarithmic phase as experimental cells. After cell counting, dilute with culture medium to a cell suspension of about 57,000 / mL.
[0068] Cell growth monitoring:
[0069] Put the cell real-time monitor into 5%C0 2 , 37°C saturated humidity incubator. Take an 8-well plate, add 150 μL of F-12K medi...
Embodiment 3
[0070] Embodiment 3: Inhibition of human liver cancer Hep-G2 cell growth test
[0071] Main materials and reagents:
[0072] Human liver cancer Hep-G2 cells were donated by Tianda Science and Technology Park; MEM medium was donated by Gibco; cyclophosphamide was purchased from Tianjin Jinshi Pharmaceutical Co., Ltd.
[0073] Blank control well: no drug added;
[0074] Positive control well: add cyclophosphamide;
[0075] Experimental well: Add the anti-tumor active component C310-6-2 obtained in Example 1.
[0076] Cell culture:
[0077] at 5%C0 2 , 37°C, under saturated humidity, human lung cancer Hep-G2 cells were cultured with MEM (10% FBS, 1% PS), and cells with good growth status were selected as experimental cells. After cell counting, dilute with culture medium to a cell suspension of about 57,000 / mL.
[0078] Cell growth monitoring:
[0079] Put the cell real-time monitor into 5%C0 2 , 37°C saturated humidity incubator. Take an 8-well plate, add 150 μL of MEM ...
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