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A kind of preparation method and application of total alkaloids of bean root
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A technology of total alkaloids of bean root and bean root, which is applied in the field of natural medicinal chemistry, to achieve the effects of simple preparation process, broad application, and promotion of discovery
Active Publication Date: 2022-02-18
TAIZHOU GUOKEHUAWU BIOMEDICAL TECH CO LTD
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[0005] At present, there is no report on the activity of M3 receptors of total alkaloids of peticillium, therefore, it is very meaningful to discover and further clarify the active ingredients in total alkaloids
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Embodiment 1
[0029] A kind of preparation method of total alkaloids of bean root, see figure 1 ,Specific steps are as follows:
[0030] 1) Medicinal material extraction: Weigh 100g of pea root medicinal material, add 1000ml of 70% ethanol, soak for 24 hours, heat and reflux at 60°C for extraction twice, combine the extracts, concentrate, and dilute to 1L to obtain the pea root extract.
[0031] 2) Extraction with organic solvent: add 70% dilute sulfuric acid to the root extract to adjust the pH to 2-3, add the same volume of ethyl acetate to extract 3 times, and obtain the ethyl acetate layer and the acidic water layer. Add ammonia water to the acidic water layer to adjust the pH to between 9 and 10, add the same volume of n-butanol and extract three times to obtain the n-butanol layer and the alkaline water layer, and concentrate the n-butanol layer to obtain the crude bean root. The base is about 6.28g, accounting for 6.28% of the mass of the medicinal material.
[0032] 3) Refining of...
Embodiment 2
[0034] Activity test:
[0035] 1) Cell culture
[0036] HEK-293T-M3 cells were donated by Dalian Institute of Physical Chemistry, Chinese Academy of Sciences (Dalian, China). Culture HEK-293T-M3 in DMEM high glucose medium containing 1% P / S (penicillin 100U / ml, streptomycin 100ug / ml), 10% fetal bovine serum and selective antibiotic Geneticin (300ug / ml) Medium (GIBCO).
[0037] 2) FLIPR detection analysis
[0038] 2.1 Preparation of cell plate: Digest the cells with 0.25% trypsin EDTA and adjust the density of the cell suspension to 8.0×105 / mL, inoculate 100 microliters per well in a 96-well plate (the type of well plate is black around and transparent at the bottom) , placed in a 37°C, 5% CO2 incubator for 16-24 hours.
[0039]2.2 Prepare FLIPR buffer: add HEPES to 1X Hank’s Balanced Salt Solution, make the final concentration of HEPES 20mM, and adjust the pH to 7.4.
[0040] 2.3 Calcium flow detection: After the cells were cultured for 16-24 hours, 100 microliters of flu...
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Abstract
The present invention provides a preparation method and application of total alkaloids of Radix chinensis. The preparation method comprises the following steps: 1) extraction of medicinal materials: 6-10 L of ethanol with a volume fraction of 50-90% is added per kilogram of medicinal materials to soak, and soaked for 1 ~24h, heat to 50℃~90℃, extract 1~5 times under reflux, combine the extracts, and concentrate to get the extract of bean root; 2) Organic solvent extraction: add 30%~80% of the extract to the above extract After adjusting the pH to 1~4 with sulfuric acid, add 1~3 times the volume of organic solvent I to extract for 1~5 times, then add a weak base to the acid water layer to adjust the pH to 8~10, and then add 1~3 times the volume of organic solvent. Solvent II is extracted for 1 to 5 times, the organic layers are combined, and the layers are concentrated to obtain the crude alkali of bean root. 3) Refining of crude alkali: Decolorize and remove impurities by passing the obtained crude alkali from the bean root through an ion exchange chromatographic column to obtain the refined total alkali. Compared with the prior art, the advantages of the present invention are as follows: the preparation method of the present invention overcomes the disadvantages of traditional purification means such as complicated pretreatment procedures and low impurity removal efficiency.
Description
technical field [0001] The invention belongs to the field of natural medicinal chemistry, and in particular relates to a preparation method and application of total alkaloids of bean root. Background technique [0002] Beidougen is the dry rhizome of Menispermum dauricum, which is mainly produced in North China, Northeast China, Shaanxi and other places. Beidou root is bitter and cold in nature. It is a commonly used traditional Chinese medicine for clearing heat, detoxifying, expelling wind and relieving pain. It is often used in the treatment of sore throat, lung heat cough, tonsillitis, chronic bronchitis, enteritis dysentery, rheumatic arthralgia and other diseases. . Studies have shown that the main chemical components of bean root are alkaloids, which have anti-inflammatory, anti-bacterial, anti-arrhythmic, and anti-cancer effects. Alkaloids such as isoquinolines, morphinanes, and oxidized isoapomorphines. Bisbenzylisoquinoline alkaloids account for more than 85% of...
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