Dendrobium officinale extract with effect of resistance to gastric cancer and preparation method and application of extract
A technology of Dendrobium officinale and its extract is applied in the field of Dendrobium officinale extract and its preparation, and can solve the problems of less research on gastric cancer and the like
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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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