Active part of Sambucus williamsii Hance for reducing risk of bone-related diseases of menopausal women and application thereof
An active part, elderberry technology, applied in the direction of bone diseases, organic active ingredients, medical preparations containing active ingredients, etc. Bone density, improving bone loss, increasing uterine weight
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Embodiment 1
[0046] Embodiment 1: the preparation method of elderberry active site
[0047] Take 50 kilograms of dry stems and branches of elderberry, after proper pulverization, heat and reflux extraction with 10 times the amount of 60% ethanol for 3 times, each time for 2 hours. The extracts were combined, and the solvent was evaporated under reduced pressure to obtain 2160 grams of the total extract of Elderberry. Get 1250 grams of elderberry total extract subsequently, dissolve with suitable amount of water, carry out macroporous resin open column chromatography, use the water of 5 times column bed volumes, 30%, 50%, 95% ethanol-water solution gradient elution successively, Collect each part eluent, reclaim solvent under reduced pressure respectively, obtain water elution part 712.5 grams, 30% ethanol elution part 160 grams, 50% ethanol elution part 125 grams and 95% ethanol elution part 111 grams, wherein 50 The fraction eluted with % ethanol is the active site of elderberry (for act...
Embodiment 2
[0048] Example 2: Isolation and Identification of Main Components in Elderberry Active Parts
[0049] Elderberry 50% ethanol elution part prepared in embodiment 1 has determined its fingerprint by HPLC analysis liquid phase, see figure 1 . Under the guidance of fingerprints, through ODS column chromatography, HW 40 column chromatography, RP-HPLC preparative liquid phase separation means, and through HPLC, MS, NMR and other analysis and identification methods, 7 compounds were identified, vanillic acid (1), coniferyl alcohol (2), L-threo-guaiacol-β-O-4'-coniferyl alcohol ether (3), D-erythro-1-(4-hydroxy-3-methoxyphenyl)- 2-[-4-(3-Hydroxypropyl)-2-hydroxyphenoxy]-1,3-propanediol (4), D-erythroguaiacol-β-O-4′- Coniferyl alcohol ether (5), dehydrodisconiferyl alcohol (6), dihydrodehydrodisconiferyl alcohol (7).
[0050] The compound structural formula obtained by separation is shown in figure 2 . Under the same chromatographic conditions as the Elderberry 50% ethanol elutio...
Embodiment 3
[0096] Example 3: The establishment of fingerprints and the identification of each compound
[0097] 3.1 Establishment of fingerprint
[0098] Take 40 mg of the 50% ethanol-eluted part of the above elderberry, dissolve it in 2 mL of 60% methanol-water, pass it through an ODS solid-phase extraction column, elute with 10 mL of methanol, evaporate the eluate to dryness, dissolve it in 2 mL of methanol, and filter through 0.45 μm film, to obtain the test solution of the active part of elderberry.
[0099] Precisely get 10 μL of the above-mentioned test solution, refer to the high-performance liquid phase analysis method specified in the appendix of the National Pharmacopoeia, inject high-performance liquid chromatography for chromatographic analysis, and record the chromatogram (see figure 1 ). The liquid chromatographic conditions are: using octadecylsiloxane bonded silica gel as the stationary phase; using methanol-water solution containing 0.1% acetic acid as the mobile phase...
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