A kind of flavonoid compound and its preparation method and application
A technology of flavonoids and compounds, applied in organic chemistry methods, drug combinations, organic chemistry and other directions, can solve problems such as carcinogenesis, achieve high safety and effectiveness, simple and easy separation path, and expand application value.
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Embodiment 1
[0034] Example 1 Separation and purification of flavonoid compound Rhynchone A
[0035] The Luhuo of this embodiment is collected from Zaoyang County, Xiangyang City, Hubei Province, and is identified as the dried root of Rhynchosia volubilis Lour., a plant of the genus Luhuo of the legume family. Grinding with a pulverizer until passing through a 24-mesh sieve, soaking and extracting with 70% acetone water for 5 times, adding 6L of solvent for the first time, completely submerging the traditional Chinese medicine Luhuo, standing at room temperature for 24 hours, filtering out the extract, and then adding 70% Acetone water 4.5L, stand at room temperature for 24h, filter out the extract again, repeat the same method until cold soaking 5 times, finally combine the extracts, concentrate under reduced pressure to obtain the first crude extract with a volume of 1L.
[0036] Fully suspend the first crude extract with distilled water to a volume of 1.5L, extract with 1.5 times the vo...
Embodiment 2
[0044] The identification of embodiment 2 flavonoid compound Rhynchone A
[0045] The compound obtained by extracting the examples is subjected to high-resolution mass spectrometry, NMR spectrum detection and structural analysis, and a nuclear magnetic resonance instrument (Bruker Avance III 500MHz NMR spectrometer) is used to analyze the compound 1 H-NMR (500MHz, CDCl 3 ), 13 C-NMR (CDCl 3 ,ppm,125MHz), HSQC, HMBC and 1 H- 1 H COZY detection, HRMS chart of the compound, 1 H-NMR (500MHz, CDCl 3 ), 13 C-NMR (CDCl 3 ,ppm,125MHz), HSQC, 1 H- 1 H COZY and HMBC are respectively as Figure 1-6 As shown, the spectral data obtained by it are shown in Table 1 below:
[0046]
[0047]
[0048] The obtained light yellow oily solid compound, HR-ESI-MS shows quasi-molecular ion peak [M-H] - It is m / z435.1794, (calculated value: [M-H] - 435.1808). The carbon spectrum shows a total of 26 carbon signals, which are consistent with the number of carbon atoms in the molecular...
Embodiment 3
[0057] Embodiment 3 compound Rhynchone A performance
[0058] 1. Effect of compound Rhynchone A on intracellular calcium content of human sperm in vitro
[0059] 1) Sperm sample processing
[0060] Human semen was passed upstream method (add 4mL sEBBS medium to a 50mL centrifuge tube, then add 1mL semen sample at the bottom, 37°C; 5% CO 2 Incubate for 1 hour, draw 3-4mL supernatant) After purification, adjust the sperm concentration to 1×10 7 individual / mL.
[0061] 2) Sperm staining
[0062] After mixing 494 μL upstream purified sperm suspension with 1 μL 2.5 mM Fluo-4 AM (Molecular Probes, Eugen, OR, USA, prepared with DMSO) and 5 μL 5% Pluronic F-127 (Molecular Probes, prepared with sEBBS medium), 37°C; 5% CO 2 Stain in the incubator for 30 minutes (staining at room temperature is also possible, but keep away from light, it is recommended to stain in a 37-degree incubator). Wash twice with sEBBS medium (without dye), and resuspend the pellet with 500 μL sEBBS medium. ...
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