Zushi daphneol extract, its preparation method and use
A technology of alcohol extract and extract, which is applied in the direction of drug combination, pharmaceutical formula, plant raw materials, etc., can solve the problems of high price and large side effects
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Embodiment 1
[0040] The preparation of embodiment 1 Zuma alcohol extract
[0041] 1. Experimental equipment and reagents:
[0042] Pulverizer (Beijing Guangming Instrument Factory FZ-102) Isotope bottle
[0043] Rotary evaporator (Shanghai Shensheng Instrument Co., Ltd.) Four-hole water bath (Beijing Instrument Factory)
[0044] 95% ethanol (analytical grade, Shanghai Reagent No. 1 Factory) large beaker
[0045] Electronic balance (accurate 0.1g, Beijing Medical Balance Factory DT-500) experimental filter paper
[0046] TP300 Ultrasonic Cleaner (Beijing Tianpeng Electronic Technology Co., Ltd.)
[0047] (The relevant instrument models mentioned below are no longer indicated)
[0048] 2. Source of experimental medicinal materials:
[0049] It was purchased in Anguo, Anhui in November 2001. It was identified by Professor Yan Yuning of Beijing University of Traditional Chinese Medicine and put into storage with a serial number.
[0050] Identification results: stem bark and root bark of D...
Embodiment 2
[0058] The liquid phase separation of embodiment 2 Zuma alcohol extract
[0059] 1. Experimental equipment
[0060] HPLC preparation system (Shimadazu Class_vp HPLC, Shimadazu Instrument Analysis Institute, Japan)
[0061] Methanol (HPLC grade, 4L, Fisher)
[0062] h 2 O (Millipore 0.22um filter)
[0063] Chromatographic column (YWG C18 10u 250×10, Beijing shuntu biotech.co., LTD)
[0064] 2. Experimental steps:
[0065] (1) After shaking the ethanol extract of Zumima (approximately liquid) obtained in Example 1, take 200ul respectively in four eppendorfs (1.5ml, domestic). And take 800ul of pure methanol in these four eppendorf tubes. Shake well to dissolve it fully. Centrifuge and filter. Take the supernatant in a 4ml injection bottle. Take out 1ml of the supernatant, centrifuge and concentrate, and calculate the concentration of the supernatant after drying (to ensure reproducibility, this configuration method is almost the same as the first preparation, and the fi...
Embodiment 3
[0074] Example 3 Zumima alcohol extract reduces density lipoprotein activity test
[0075] 1. Materials and Methods
[0076] Drugs to be tested: the ethanol extract of Zumima obtained in Example 1, and the F15 and F16 component samples obtained in Example 2.
[0077] Plasmid: pLDLR270-luc was constructed by Shanghai Huada Tianyuan Biotechnology Co., Ltd., and the LDLR promoter region cloned from the genome was inserted into the pGL3-basic vector (Promega, USA).
[0078] Cells: pLDLR270-luc stably transfected human embryonic kidney cells (HEK 293) (CRL-1573), human liver cancer cells HepG2 (ATCC HB-8065).
[0079] Reagents: KpnI, BglII (NewEngland Biolab), pGL3-basic vector (Promega, USA), T4 ligase (Promega, USA), Lipofeetamine2000 (Invitrogen, USA), DMEM medium (GIBCO), streptomycin, L-Glutamine (Invitrogen, USA), Hygromycin (Roche, Germany), Fetal Bovine Serum (Hyclone, USA), Heparin (Hepes), Luciferase Assay System and Bright-Glo Luciferase Assay System (Promega, USA), R...
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