Dendrobium candidum extract with anti-gastric cancer effect and its preparation method and application
A technology of dendrobium candidum and its extract, applied in the field of dendrobium candidum extract and its preparation, can solve the problems of less research on gastric cancer
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[0028] The invention provides a preparation method of Dendrobium officinale extract with anti-gastric cancer effect, comprising the following steps:
[0029] (1) complete the dendrobium candidum, obtain the dendrobium candidum after completing;
[0030] (2) the dendrobium candidum after described finishing is pulverized to make dendrobium powder;
[0031] (3) mixing the dendrobium powder with absolute ethanol, and separating the solid and liquid after leaching to obtain the extract;
[0032] (4) Concentrating the extract in vacuo, then blowing and drying with nitrogen to obtain anhydrous ethanol extract;
[0033] (5) mixing the absolute ethanol extract with water to obtain an aqueous extract;
[0034] (6) Mix the aqueous extract solution with petroleum ether, leave to separate phases after extraction, and collect the first lower phase;
[0035] (7) Mix the first lower phase with n-butanol, leave to separate the phases after extraction, and collect the second lower phase;
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Embodiment 1
[0050] Preparation of Dendrobium candidum anti-gastric cancer extract
[0051] 1) Take the whole plant of Dendrobium officinale, heat it at 100-110°C for 15-25 minutes, make it into a dry powder of Dendrobium, and pass it through a 40-mesh sieve.
[0052] 2) According to the solid-liquid ratio of 1:20, add absolute ethanol to the dry powder of Dendrobium officinale, soak for 48 hours, and ultrasonically treat for 45 minutes (conditions are 120W, 60Hz, 55°C). Then place it in a centrifuge and centrifuge at 6000r / min for 10min. The supernatant was filtered with a 220nm filter membrane, concentrated under vacuum conditions at 55°C, and then dried by nitrogen blowing to obtain anhydrous ethanol extract.
[0053] 3) Redissolve the absolute ethanol extract with pure water according to the ratio of solid to liquid 1:12, extract three times with one-third volume of petroleum ether, each time for 10 min, let stand to separate the phases, and collect the lower phase.
[0054] 4) Add o...
Embodiment 2
[0057] Gastric cancer cell proliferation inhibition experiment
[0058] Take the logarithmic phase cells and inoculate them in 96-well plates. After culturing for 24 hours, add different concentrations of Dendrobium candidum extracts and act for 48 hours respectively. Then discard the original medium, add 180 μL serum-free medium, and add 20 μL MTT to each well. (Concentration: 5 mg / mL), then placed in a carbon dioxide incubator for 4 hours, took out the 96-well plate, carefully sucked off the supernatant, added 150 μL DMSO to each well to dissolve the blue-purple crystal formazan, detected the absorbance with a microplate reader, and set The parameters of the microplate reader were to vibrate for 3 minutes after entering the plate, and then detect the ultraviolet absorption at a wavelength of 490 nm. Processed by Soft Max Pro6.5.1 data processing software, using the 4-Parameter method to calculate the half-inhibition rate (IC 50 value).
[0059] The formula for calculating ...
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