Preparation method and application of mulberry twig effective part with tyrosinase inhibiting effect
A technology of tyrosinase and active parts, applied in the preparation of organic compounds, medical preparations containing active ingredients, active ingredients of ketones, etc., can solve problems such as unclear ingredients
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Embodiment 1
[0019] Example 1 Preparation of effective fraction
[0020] 10 kg of mulberry branches were ultrasonically extracted with 2 times the amount of 70% ethanol for 3 times, 1 hour each time, concentrated to dryness under reduced pressure, and weighed. Dissolve an appropriate amount of dry matter in 95% ethanol, filter, add to macroporous resin D101, dry naturally or evaporate to dryness under reduced pressure, and then apply macroporous resin D101 chromatography column, using 20% and 40% of the volume of 8 columns respectively , 60%, 80%, and 95% ethanol were eluted, the eluent was collected, and the ethanol was recovered under reduced pressure to dryness, and the 40% eluted part was detected by high performance liquid chromatography, which contained five compounds oxidized resveratrol, grass The components of halberine, sangxinin M, 2,4,2',4'-tetrahydroxychalcone and mochalcone A are effective parts ( figure 1 ).
[0021] HPLC detection conditions:
[0022] Instrument: Water...
Embodiment 2
[0025] Example 2 Preparation of oxidized resveratrol and 2,4,2',4'-tetrahydroxychalcone
[0026] The extraction of mulberry branch, macroporous resin separation process is the same as embodiment 1, collects 8 column volumes of 40% ethanol eluting part, concentrates and dries to obtain sample, weighs, separates with 200-300 mesh silica gel column chromatography, uses volume ratio respectively For 50:1 and 30:1 chloroform-methanol elution, collect the eluate obtained by 30:1 chloroform-methanol elution by volume ratio, and then use Sephadex LH-20 gel column chromatography to separate, the volume ratio 1:1 methanol-water elution, that is.
[0027] The nuclear magnetic resonance spectrum data of oxidized resveratrol are as follows:
[0028] 1 H NMR (400MHz, CD 3 OD) δ: 7.32(1H,d,J=9.1Hz,H-6),7.26(1H,d,J=16.4Hz,H-7),6.81(1H,d,J=16.4Hz,H-8 ),6.45(2H,d,J=2.0Hz,H-2',6'),6.31(2H,overlap,H-3,5),6.14(1H,t,J=2.0Hz,H-4' ). 13 C NMR (100MHz, CD 3 OD)δ:159.8(C,C-3',5'),159.4(C,C-4),15...
Embodiment 3
[0035] Example 3 Preparation of Eugenin, Sangsin M and Mochalcone A
[0036]The extraction of mulberry branch, macroporous resin separation process is the same as embodiment 1, collects 8 column volumes of 40% ethanol eluting part, concentrates and dries to obtain sample, weighs, separates with 200-300 mesh silica gel column chromatography, uses volume ratio respectively For 50:1 and 30:1 chloroform-methanol elution, collect the eluate obtained by 50:1 volume ratio of chloroform-methanol elution, and then use Sephadex LH-20 gel column chromatography to separate, the volume Methanol-water ratio 1:1 elution, that is, eugenin, sangsin M and mochalcone A.
[0037] The nuclear magnetic resonance spectrum data of eugenin are as follows:
[0038] 1 H NMR (Acetone-d 6 ,400MHz)δ:12.210(1H,OH-5),9.019(2H,OH),7.290(1H,d,J=8.4Hz,H-6'),6.465(1H,d,J=2.4Hz,H -2'),6.415(1H,dd,J=8.4,2.4Hz,H-5'),5.955(1H,d,J=2.0Hz,H-8),5.936(1H,d,J=2.0Hz ,H-6), 5.692(1H,dd,J=13.2,2.8Hz,H-2),3.163(1H,dd,J=1...
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