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Method for separating and purifying puerarin by combination of double-aqueous phase extraction-ultrasonic wave

A technology of puerarin and two aqueous phases, which is applied in the field of two aqueous phase extraction-ultrasonic joint separation and purification of puerarin, can solve the problems of increasing the distribution coefficient and yield of target products, increasing operation difficulty and economic cost, and accelerating system equivalence. Achieve the effect of fast system phase separation, easy operation and high purity

Active Publication Date: 2016-10-26
苏州善淮堂生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing two-phase extraction technology needs to be supplemented by centrifugation and other operations after the phase separation of the system, thereby accelerating the phase separation of the system, and adding neutral salt KCl to the system to increase the distribution coefficient and yield of the target product increases the operational difficulty and economic cost

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] (1) Preparation of puerarin crude extract

[0024] Add 10g of kudzu root powder into 250g of 70% ethanol-water solution, reflux extraction and soaking in Soxhlet extractor for 12 hours, cool to room temperature, filter, and concentrate under reduced pressure to obtain 31.2g puerarin crude extract, the puerarin purity is 30.2%, the yield is 94.3%. The purity of puerarin was measured by high performance liquid chromatography, the fluidity was 50% methanol-water solution, the flow rate was 1 mL / min, and the column temperature was room temperature.

[0025] (2) Aqueous two-phase extraction

[0026] Take 14g of puerarin-containing crude extract, 3g of PEG600 and 3g of ammonium sulfate and add them to the Erlenmeyer flask. After 20 hours of dialysis in a dialysis membrane with a molecular weight cut-off of 5000, an aqueous solution of puerarin was obtained, and then freeze-dried at -15°C for 10 hours to obtain 13.2 g of puerarin powder, wherein the purity of puerarin was 97...

Embodiment 2

[0028] (1) Preparation of puerarin crude extract

[0029] Add 10g of kudzu root powder into 400g of 60% ethanol-water solution, reflux extraction and soaking in a Soxhlet extractor for 20 hours, cool to room temperature, filter, and concentrate under reduced pressure to obtain 31.4g of puerarin crude extract, the puerarin purity is 29.5%, the yield is 92.7%.

[0030] (3) Aqueous two-phase extraction

[0031] Take 12.8g of puerarin-containing crude extract, 3.6g of PEG 1000 and 3.6g of sodium sulfate and add them to the Erlenmeyer flask. After shaking and mixing, put them in an ultrasonic wave for 20 minutes, then separate the phases after standing for 20 minutes, and collect the upper phase. Place the upper phase in a dialysis membrane with a molecular weight cut-off of 5000 and dialyze for 25 hours to obtain an aqueous solution of puerarin, then freeze-dry at -15°C for 15 hours to obtain 12.3g of puerarin powder, wherein the purity of puerarin is 94.6%. The yield was 90.6%,...

Embodiment 3

[0033] (1) Preparation of puerarin crude extract

[0034] Add 10g of kudzu root powder into 500g of 50% ethanol-water solution, reflux extraction and soaking in a Soxhlet extractor for 24 hours, cool to room temperature, filter, and concentrate under reduced pressure to obtain 30.1g of puerarin crude extract, the puerarin purity is 30.7%, the yield is 92.3%.

[0035] (3) Aqueous two-phase extraction

[0036] Take 13.6g of puerarin-containing crude extract, 4g of PEG 600 and 2.4g of ammonium sulfate and add them to the Erlenmeyer flask. After shaking and mixing, put them in an ultrasonic wave for 40 minutes and then separate the phases after standing for 10 minutes. Collect the upper phase. Place the upper phase in a dialysis membrane with a molecular weight cut-off of 5000 and dialyze for 26 hours to obtain an aqueous solution of puerarin, then freeze-dry at -15°C for 18 hours to obtain 13.3g of puerarin powder, wherein the purity of puerarin is 95.6%. The yield was 93.2%, a...

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Abstract

The invention provides a method for separating and purifying puerarin by combination of double-aqueous phase extraction-ultrasonic wave. The method comprises the specific operation steps: extracting kudzu vine root powder with 50%-70% alcohol water solution under refluxing for 12-24 hours to obtain crude liquid; carrying out polyethylene glycol-inorganic salt double-aqueous phase extraction and putting the liquid into ultrasonic wave to be subjected ultrasonic treatment for 20-40 minutes; then standing for 10-20 minutes; taking an upper phase and dialyzing in a dialysis membrane with the molecular mass cut-off of 5000 for 20-26 hours; and freeze-drying to obtain puerarin powder. The method takes the 50%-70% alcohol water solution as an extracting solution, so that high yield and high purity can be obtained and solid and liquid phases can be rapidly separated; a double-aqueous phase extraction-ultrasonic wave combined method is used for purifying the crude liquid and the selectivity on the puerarin is high; a phase separation speed of the system is rapid and rapid phase separation can be realized without the help of equipment including a centrifuge and the like; and the purity of the puerarin in the prepared puerarin powder is up to 97.5% and the total yield is up to 86.6%.

Description

technical field [0001] The invention belongs to the technical field of biochemical separation, and relates to a puerarin separation and purification method, in particular to a two-phase extraction-ultrasonic combined separation and purification method for puerarin. Background technique [0002] Puerarin, also known as pueraria flavonoids, is a yellow crystal, soluble in water, methanol, ethanol, pyridine, easily soluble in hot water, insoluble in benzene, chloroform, ether, etc. Puerarin is the main active ingredient of kudzu root. It can reduce myocardial oxygen consumption, increase coronary and cerebrovascular blood flow, expand coronary and cerebrovascular blood vessels, and promote blood circulation. It is clinically used to treat hypertension and angina pectoris. , Cerebral insufficiency, cerebral infarction and sudden deafness. Our country is very rich in kudzu root resources. Therefore, the research and development of an optimized process route to obtain high-purity...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D407/04
CPCC07D407/04
Inventor 任新年
Owner 苏州善淮堂生物科技有限公司
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