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Preparation method for extracting and purifying bruceine D form brucea javanica

A technology of Brucea Brucea and Brucea Brucea, which is applied in the field of traditional Chinese medicine preparation, can solve the problems of long contact time with toxic organic solvents, damage to the health of preparation personnel, and low efficiency of purifying Brucea Brucea D, so as to improve the carrying capacity and separation efficiency, The effect of short contact time and simple preparation method

Inactive Publication Date: 2013-02-13
FUJIAN AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to provide a more convenient and efficient preparation method for extracting and purifying brucein D from Brucea javanica to solve the problem of low efficiency, time-consuming, preparation personnel and toxic effects of extracting and purifying brucein D from plant Brucea javanica Long contact time with organic solvents will seriously damage the health of preparation personnel and other issues

Method used

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  • Preparation method for extracting and purifying bruceine D form brucea javanica

Examples

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Effect test

Embodiment 1

[0022] Example 1: Take 1.0 kg of sun-dried Bructus javanica seeds, pulverize them with a plant pulverizer and extract them 3 times with 10 L of 95% industrial ethanol (condition: reflux at 80°C for 1 hour), combine the extracts, and filter Concentrate under reduced pressure with a rotary evaporator to a viscous state to obtain a concentrated solution of about 165 mL, add distilled water to 1000 mL, heat and stir in a water bath at 80°C to fully dissolve, filter with suction, and apply the filtrate to D101 macroporous resin for 3 times. Each time about 300 mL of the suspension was passed through a chromatographic column (column diameter 6 cm, length 60 cm) filled with 1000 g of D101 macroporous resin, and the flow rate of the sample was about 10 mL / min. Then eluted with 3 column volumes of 80% ethanol, collected the alcohol eluate, combined the three eluates, concentrated under reduced pressure to form a paste, and obtained 46 g of paste. Dissolve the paste in methanol and add ...

Embodiment 2

[0023] Example 2: Weigh 1.0 kg of the skin of Brucea javanica plant dried at 55°C, pulverize it with a plant pulverizer, and extract it with 8 L of 95% industrial ethanol for 3 times under reflux (condition: reflux at 80°C for 1.5 hours), and combine the extraction Liquid, filtered and concentrated to about 180 mL with a rotary evaporator under reduced pressure, added distilled water to 1200 mL, stirred and heated in a water bath at 80°C to fully dissolve, filtered with suction, and the filtrate was applied to D101 macroporous resin for 3 times. Each time about 400 mL of the suspension was passed through a chromatographic column filled with 1300 g of D101 macroporous resin (column diameter 6 cm, length 80 cm), the sample loading flow rate was about 10 mL / min, and eluted with 3 column volumes of water first. Then elute with 4 column volumes of 80% ethanol, collect the alcohol eluate, combine the three eluates, concentrate under reduced pressure to form a paste, and obtain 41 g o...

Embodiment 3

[0024] Example 3: Weigh 1.5 kg of leaves of Brucea javanica plant dried at 45°C, pulverize them with a plant pulverizer, and then reflux them with 15 L of 95% industrial ethanol for 3 times (conditions: reflux at 80°C for 2 hours), and combine the extractions Liquid, filtered and concentrated to about 300 mL with a rotary evaporator under reduced pressure, added distilled water to 2000 mL, stirred and heated in a water bath at 80°C to fully dissolve, filtered with suction, and the filtrate was applied to D101 macroporous resin for 5 times. Each time about 400 mL of the suspension was passed through a chromatographic column filled with 1300 g of D101 macroporous resin (column diameter 6 cm, length 80 cm), the flow rate of the sample was about 10 mL / min, and 4 column volumes of water were used to elute first. Then eluted with 5 column volumes of 80% ethanol, collected the alcohol eluate, combined the 5 times of eluate, concentrated under reduced pressure to form a paste, and obta...

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Abstract

The invention provides a preparation method for extracting and purifying bruceine D form brucea javanica. The method comprises the steps of material smashing, reflux extraction through 95% of industrial alcohol, extracting solution concentration, extractive desugaring, silica-gel column chromatography and recrystallization, wherein D101 macroporous resin on water suspension and used for extracting a concentrated solution is adopted in the extractive desugaring step, the elution is first performed with water, then the elution is performed with 80% of alcohol, and alcohol eluant is collected. By means of the treatment, impurities including sugar and the like in an extractive can be effectively removed, and the bearing capacity and the separation efficiency of silica gel are improved. In the silica-gel column chromatography step, primary dry-column chromatography separation is first performed, and then wet-column chromatography separation is performed. By means of the dry-column chromatography separation, the separation time can be shortened, and human input is decreased. The purity of the bruceine D obtained by means of the preparation method can reach to more than 98%, the operation is simple, the using amount of a poisonous solvent can be saved, and manpower and time are saved.

Description

technical field [0001] The invention belongs to the technical field of preparation of traditional Chinese medicines, and relates to a method for extracting and preparing active ingredients of traditional Chinese medicines, in particular to a preparation method for extracting and purifying brucein D from Brucea javanica. Background technique [0002] Bruceine D (bruceine D) is derived from the plant Brucea javanica ( Brucea javanica (L) Merr), a tetracyclic triterpenoid bitternin, studies have shown that it not only has a strong inhibitory activity on a variety of tumor cells, but also has a strong inhibitory effect on the infection and proliferation of a variety of plant viruses effect. Bactanin D can not only inhibit the replication of tobacco mosaic virus (TMV) genomic RNA and subgenomic RNA, prevent the expression of virus-related proteins CP and MP, but also inhibit the movement of TMV in tobacco leaves to the stem and upper leaves of the plant , induce tobacco resista...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D493/10
Inventor 陈启建欧阳明安谭庆伟
Owner FUJIAN AGRI & FORESTRY UNIV
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