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Method for separating compounds from bergenia purpurascens in bulk and oxidation resistance detection method

A large-scale separation and anti-oxidative technology, which is used in the separation/purification of carboxylic acid compounds, chemical instruments and methods, material separation, etc., and can solve the problems of high operator requirements and low efficiency.

Active Publication Date: 2020-10-23
SHANDONG ANALYSIS & TEST CENT +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The prior art provides a high-speed counter-current chromatography method for the separation and purification of compounds from medicinal materials such as tea leaves, but the inventors found that the high-speed counter-current chromatography in the prior art often separates one compound at a time, and the efficiency is low, and each separation For a compound, the parameters must be changed again, which requires high requirements for operators
And the high-speed countercurrent chromatography in the prior art has not been applied to the compound extraction and separation of rock cabbage

Method used

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  • Method for separating compounds from bergenia purpurascens in bulk and oxidation resistance detection method
  • Method for separating compounds from bergenia purpurascens in bulk and oxidation resistance detection method
  • Method for separating compounds from bergenia purpurascens in bulk and oxidation resistance detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] This example provides a method for large-scale separation of compounds from rock cabbage.

[0051] 1. Extraction of samples

[0052] Soak 5 kg of rock cabbage medicinal material (collected from Pakistan), and extract with 2×12 L of 80% ethanol for 24 hours, and repeat 3 times. All the extracts were combined and evaporated to dryness at 40° C. to obtain 256 g of crude rock cabbage, which was stored in a refrigerator at 4° C. for further purification.

[0053] 2. Countercurrent Chromatographic Separation and Preparation of Monomer Components

[0054] Prepare the two-phase solvent system tert-butyl methyl ether / n-butanol / methanol / water (1:3:1:5, v / v) according to the above-mentioned solvent ratio, put it in a separatory funnel, shake it up and let it stand for separation. After a period of equilibrium, separate the upper and lower phases, take 1.0 g of the crude product, dissolve it in 5 mL and 5 mL of the lower phase and set aside. A high-speed countercurrent chromatog...

Embodiment 2

[0064] This example provides the lower K under different solvent components D value.

[0065] The K of the target compound of table 1 different solvent systems D value

[0066]

[0067] Choosing the right two-phase solvent system for countercurrent chromatographic separation is a very critical step, and a suitable two-phase solvent system can successfully complete the separation experiment. In this study, various biphasic mixed solvent systems including two, three or four solvent mixtures were tested at different ratios. A good solvent system can provide ideal partition coefficients (K D )value. K D The value describes the ratio of solute distribution between two-phase solvent systems in mutual equilibrium. The biphasic solvent systems studied in our experiments included n-hexane / ethyl acetate / methanol / water (1:9:1:9, v / v), ethyl acetate / n-butanol / water (1:4: 5, v / v). In addition, different ratios of biphasic solvent system tert-butyl methyl ether / n-butanol / methanol...

Embodiment 3

[0069] This example provides the anti-oxidation cell activity test of the six monomer compounds described in Example 1.

[0070] The crude extract and monomeric compounds, namely β-arbutin (1), petogenin (2), 6-O-galloyl arbutin (3), gallic acid (4), 11-O-galloyl Cytotoxicity of petracenin (5) and (-)-epicatechin 3-O-gallate (6) was determined in HepG2 (human hepatoma cell line) cells according to the MTT assay.

[0071] First, HepG2 cells were cultured in DMEM supplemented with penicillin (100 U / mL) / streptomycin (100 μg / mL) and 10% FBS. Then place the cells in 5% CO 2 Incubate in the presence at 37°C. Trypsin solution was used to digest HepG2 cells in log phase. The cell density was then adjusted to 5 × 10 5 / mL. Incubate the cells in a volume of 100 µL / well at 37 °C and 5% CO 2 seeded into 96-well cell culture plates. Afterwards, the seeded cells were treated with appropriate concentrations of test samples for 24 hours. The wavelength of absorbance was kept at 570nm ...

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Abstract

The invention relates to a production method of effective components of a traditional Chinese medicine, and particularly provides a method for separating compounds from bergenia purpurascens in bulk and an oxidation resistance detection method. The method for separating the compounds from the bergenia purpurascens in bulk comprises the following step: conducting extraction on the bergenia purpurascens to obtain an extract, and conducting counter-current chromatography separation on the extract to obtain monomer components of beta-arbutin, bergenin, 6-O-galloylarbutin, gallic acid, 11-O-galloylbergenin and (-)-epigallocatechin-3-O-gallate. The problems that in the prior art, a compound is often separated at a time through a high-speed counter-current chromatography method, the efficiency islow, and high-speed counter-current chromatography is not yet applied to extraction and separation of compounds in bergenia purpurascens are solved.

Description

technical field [0001] The invention relates to a preparation method of active ingredients of traditional Chinese medicines, and specifically provides a method for separating compounds from rock cabbage in large quantities and an anti-oxidation detection method. Background technique [0002] The statements herein merely provide background information related to the present disclosure and may not necessarily constitute prior art. [0003] Rock cabbage (Saxifrageaceae) is a perennial herb located in the Himalayan region at an altitude of 2700-4700 meters. In the Gilgit region of Pakistan, the plant has been used as a herbal remedy for diabetes. An herbal remedy containing rock cabbage has been reported to treat acne, diabetes, obesity, and viral infections. In addition, it has also been reported that the plant is useful for treating kidney stones, wounds, etc., and has antifungal, antioxidant, antibacterial, and immunomodulatory effects. The main chemical constituents repor...

Claims

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Application Information

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IPC IPC(8): C07H1/08C07H15/203C07D493/04C07C51/47C07C65/03C07D311/62A61P3/10A61P9/00A61P25/00A61P35/00A61P39/06G01N30/88G01N30/06A23L33/105
CPCA23V2002/00A23L33/105A61P3/10A61P9/00A61P25/00A61P35/00A61P39/06C07C51/47C07D311/62C07D493/04C07H1/08C07H15/203G01N30/06G01N30/88G01N2030/065G01N2030/8809G01N2030/884C07C65/03A23V2200/30A23V2250/21A23V2300/14
Inventor 王岱杰伊夫蒂哈尔·阿里沙希德·阿齐兹施树云崔莉郭莹李晓骄阳吕其刚
Owner SHANDONG ANALYSIS & TEST CENT
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