Preparation method of active ingredients in pleurospermum plant and application thereof in preparation of anti-inflammatory drugs
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A technology of active ingredients and ridges, which is applied in the field of separation and purification of active ingredients in natural plants, can solve problems such as the separation and research of phytochemical separation components of P.
Active Publication Date: 2021-08-20
QILU UNIV OF TECH
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Embodiment 1
[0036] 1.1 Preparation of crude extract
[0037] Crush 3Kg of P. candollei plant, soak in 2×8L of 95% ethanol for 48h, repeat 3 times, combine the filtrates, concentrate in vacuo at 40°C to obtain 124g of crude extract, and store it at 4°C for future use.
[0038] 1.2 Screening and KD value determination of two-phase solvent system
[0039] Choosing an appropriate two-phase solvent system is the key to the successful separation of HSCCC. Several different ratios of n-hexane / ethyl acetate / methanol / water solvent systems were tested in this experiment.
[0040] Configure 10mL of n-hexane / ethyl acetate / methanol / water solvent system with different ratios, add about 2mg of crude extract, shake vigorously, and fully dissolve. Take 1 mL each of the upper and lower phases, blow dry with nitrogen, dissolve the residue in 1 mL methanol, and analyze by HPLC. According to the formula K D =A U / A L Calculate the partition coefficient K of the compound D value, where A U and A L are...
Embodiment 2
[0066] Anti-inflammatory activity assay:
[0067] The MTT assay was used to assess cytotoxicity. The macrophage Raw 264.7 cell line was obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). will total 2×10 4 cells / mL were plated in 96-well plates and incubated overnight at 37°C in DMEM containing 10% FBS. After removing the cell culture medium, Raw 264.7 cells were washed 3 times with PBS buffer. Then, the cells were treated with different concentrations of the six compounds and incubated at 37°C for an additional 24 h. At the end of each incubation period, each well was washed 3 times with PBS buffer, and 150.0 μL of fresh MTT solution (0.5 mg mL –1 ). Subsequently, remove the supernatant, add 150.0 μL DMSO, shake for 15 min, and incubate at 37 °C for 4 h. Finally, the optical density of each well was measured in triplicate at 490 nm.
[0068] The anti-inflammatory effects of the six compounds were evaluated by inhibiting LPS-induced nitric ...
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Abstract
The invention relates to the technical field of separation and purification of active ingredients in natural plants, in particular to a preparation method of active ingredients in umbelliferae pleurospermum plant P.candollei. P.candollei is firstly extracted by adopting alcohol extraction to obtain a crude extract, the crude extract is then separated by adopting high-speed counter-current chromatography, a solvent system adopted by the high-speed counter-current chromatography is an n-hexane/ethyl acetate/methanol/water two-phase solvent system, an upper phase and a lower phase are taken before the high-speed counter-current chromatography is used, the upper phase is used as a stationary phase, and the lower phase is used as a mobile phase, and thefollowing compound monomers as described in the specification are obtained. The separation and purification method established by the invention realizes successful separation of nine monomer ingredients in the P.candollei plant, the purity of each obtained monomer is greater than 98%, and the separation and purification effects are good; the compounds 7 and 8 obtained through separation and purification can effectively inhibit NO generation.
Description
technical field [0001] The invention relates to the technical field of separation and purification of active ingredients in natural plants, in particular to a method for preparing the active ingredients of plants of the genus Piperia and its application in the preparation of anti-inflammatory drugs. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily to be regarded as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Celery belongs to the Umbelliferae family and has analgesic, anti-inflammatory and anti-malarial properties, and is also used as a condiment and herbal perfume. In Chinese folk medicine, celery is used as a diaphoretic, analgesic, and antipyretic. A variety of chemical constituents have been found from plants of the gen...
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