Preparation method of Pollyalthia laui Merr roots

A technology of root extract and dark root is applied in the field of preparation of Hainan dark root extract, which can solve the problems such as no alkaloid biological activity, and achieve good broad-spectrum anti-tumor activity, high yield and low cost. Effect

Inactive Publication Date: 2014-08-13
HAINAN NORMAL UNIV
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Problems solved by technology

Preliminary studies have found that the ethanol extract of Annonaceae has good antitumor activity[Yuan Y, Huang G.J, Wang T.S et al.In vitro screening of five Hainan plants of Polyalthia(Annonaceae) against human cancer cell lines with MTT assay.Phytother.Res.,2010,27:1-15], but there are no research reports on alkaloid extraction and related biological activities at home and abroad

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  • Preparation method of Pollyalthia laui Merr roots
  • Preparation method of Pollyalthia laui Merr roots

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preparation example Construction

[0022] The steps of the preparation method of the extract of Rhizoma hainanensis are as follows:

[0023] 1), the crushed Rhizoma hainanensis medicinal material (through 20~80 mesh sieves) is washed with alkaline water (5~30% milk of lime solution, 5~30% Na 2 CO 3 solution or 10% ammonia water, etc.) after wetting, add 85-95% ethanol or methanol solution, soak at room temperature, heat or ultrasonically extract, the extraction time is 60 minutes to 7 days, and the solid-liquid ratio is 1:1-1:10 .

[0024] 2) Filtrating the root extract of Anthraca hainanensis, recovering under reduced pressure to obtain the total extract, adding water to suspend the extract, extracting with chloroform, recovering under reduced pressure to obtain the chloroform fraction, that is, lipophilic total alkaloids.

[0025] 3), repeated column chromatography on the chloroform part: petroleum ether-ethyl acetate (10:1, v / v) silica gel column chromatography to obtain compound (5); petroleum ether-chlor...

example 1

[0028] Example 1, the preparation of the lipophilic total alkaloids of the root of Anthera hainanensis

[0029] Get the root (10Kg) of Anthera hainanensis after being dried in the shade, pulverize, moisten with 10% ammonia water, soak 3 times at room temperature with 95% ethanol at room temperature, 7 days / time, combine the extracts, concentrate under reduced pressure to obtain the total extract (0.5Kg) , add water to suspend and disperse, and extract with chloroform to obtain the chloroform extraction fraction (25 g).

[0030] The chloroform extraction part was subjected to repeated column chromatography on silica gel and Sephdex LH-20, and 4 carbazoles, 3 aporphines and 1 amide alkaloids were obtained, respectively: petroleum ether-acetic acid Ethyl ester (10:1, v / v) silica gel column chromatography Liridine (5) 7.6mg; Petroleum ether-chloroform (5:1, v / v) silica gel column chromatography gave Liriodenine (6) 10.5mg; Petroleum ether- Ethyl acetate (4:1, v / v) was recrystalli...

Embodiment 2

[0032] Example 2. Antitumor activity research on the chloroform extraction part (lipophilic total alkaloid part) of the root of hainan root

[0033]1. Experimental method: four kinds of tumor cell lines K562, SPCA-1, SGC-7901, BEL-7402 were used in RPMI-1640 medium containing 10% calf serum, at 37°C, 5% CO 2 cultured in an incubator. The MTT method was used to carry out the cell proliferation inhibition test. The main operation was: take the tumor cell line in the logarithmic growth phase, digest it with 0.25% trypsin, and prepare 5×10 RPMI-1640 culture solution with 10% newborn calf serum 4 pcs / ml -1 The cell suspension was inoculated in a 96-well plate with 180 μL per well. At 37°C, 5% CO 2 Cultivate for 8-10 h under saturated humidity conditions, and when they adhere to the wall, add sample solutions prepared with PBS to each well so that the final concentrations of the samples are 1, 10, and 100 μg / mL, respectively. For each concentration, 3 wells were paralleled, and ...

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Abstract

The invention discloses a preparation method of Pollyalthia laui Merr roots. The preparation method comprises the following steps: wetting dry powder of Pollyalthia laui Merr roots with lye and adding a solvent for extracting; and filtering and concentrating the extracting solution at reduced pressure to obtain a total extract, adding water for suspending, extracting by using chloroform, and recycling under reduced pressure to obtain a chloroform extracting part which is the lipophilic total alkaloids part of the Pollyalthia laui Merr roots. The lipophilic total alkaloids part comprises four carbazole alkaloids, three aporphine alkaloids and an amide alkaloid. The extract prepared by the method disclosed by the invention is high in yield, simple in process flow, low in cost and suitable for preparation on a large scale, has good broad spectrum anti-tumor activity and can be applied to medicines for preventing and treating tumors.

Description

technical field [0001] The invention belongs to the technical field of extraction and separation of active ingredients of natural plants, and relates to a preparation method of an extract of Rhizoma hainanensis. Background technique [0002] Annonaceae (Annonaceae) plant of the genus Polyalthia, often erect shrubs. According to the Flora of China, there are about 200 species of this genus in the world, distributed in subtropical regions such as Malaysia, the Philippines, India, and Sri Lanka [South China Institute of Botany, Chinese Academy of Sciences. Flora of Hainan. Vol. 1. Beijing: Science Press. 1964, p247]. In Indian folk, Anthracite is used as a traditional antipyretic, abortifacient, antihypertensive, etc. At present, the research on chemical composition has obtained alkaloids, diterpenoids (mainly clodanes and en-kaurocanes), triterpenes and other structural types of chemical constituents from the plants of the genus Anthera, and pharmacological studies have foun...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D221/16C07D221/18C07D491/06C07C235/34C07C231/24A61P35/00
CPCC07D221/16C07C231/24C07C235/34C07D221/18C07D491/06
Inventor 宋小平陈文豪李小宝付艳辉陈光英
Owner HAINAN NORMAL UNIV
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