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Method for preparation of icariin by enzymolysis

The technology of icariin and enzymatic hydrolysis method is applied in the field of preparing icariin by enzymatic hydrolysis method, which can solve the problems of many organic solvents, complicated operation and complicated process, and achieve the effects of improving yield, being environmentally friendly and simple to operate.

Inactive Publication Date: 2019-02-01
CHONGQING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The above-mentioned method is complex in process, consumes a lot of organic solvents, and the yield is low, which is not conducive to industrial production; while the Chinese patent CN102718819A mainly obtains icariin through water heating extraction, macroporous resin separation, and column separation and purification of Epimedium raw materials , the method is complicated to operate and needs to be processed twice, which is not conducive to mass production; Chinese patent CN103396463A first carries out microwave extraction of epimedium with ethanol aqueous solution, utilizes microporous membrane to remove particulate impurities therein, and then passes through simulated moving bed chromatography Carry out separation and purification, and finally get the finished product of icariin through concentrated spray drying. This method is applied to the simulated moving bed chromatography technology, and there are limitations of instruments, so it is not easy to popularize
The above existing icariin preparation methods generally use Epimedium plants as raw materials to obtain icariin, but the content of icariin in Epimedium plants is very low, only about 0.5%, so through complicated operations The yield of icariin prepared by the process is still low

Method used

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  • Method for preparation of icariin by enzymolysis
  • Method for preparation of icariin by enzymolysis
  • Method for preparation of icariin by enzymolysis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] (1) Preparation of Epimedium extract: take Epimedium powder, pass through a No. 3 sieve and add ethanol with a volume fraction of 70%, and the mass-volume ratio of the Wushan Epimedium powder to ethanol is 1:30, g : mL; Rotary evaporation extract obtains residual solid, and vacuum drying promptly obtains Epimedium extractum;

[0027] (2) Enzymolysis reaction: Add Epimedium extract and rhamnosidase in sequence according to the ratio of 1:1, mmol:U, and then add a phosphate buffer solution with a concentration of 0.2mol / L and a pH of 6.5 to stabilize the enzyme Bovine serum albumin, the mass-to-volume ratio of the epimedium extract to phosphate buffer solution and bovine serum albumin is 1:2:2, g:mL:mg, and the enzymolysis is carried out in a water bath at 40°C To react, extract a small amount of reaction solution to detect that the conversion rate of Epimedin C reaches 98%, and carry out boiling water inactivation to end the reaction.

[0028] (3) Purification of icarii...

Embodiment 2

[0030] (1) Preparation of Epimedium extract: get Epimedium Wushanii powder, add volume fraction after No. 3 sieve and be 70%-75% ethanol, the mass volume ratio of described Epimedium Wushanii powder and ethanol is 1 :25, g:mL; Rotary evaporation extract obtains residual solid, and vacuum drying obtains Epimedium Wushan extractum;

[0031] (2) Enzymolysis reaction: according to the ratio of 1:1, mmol: U, add Epimedium Wushanhu extract and rhamnosidase in sequence, then add phosphate buffer solution with a concentration of 0.2mol / L and a pH of 6.5 and enzyme Stabilizer bovine serum albumin, the mass-to-volume ratio of the epimedium extract to phosphate buffer solution and bovine serum albumin is 1:2:2, g:mL:mg, and the enzyme is carried out under a water bath condition of 50° C. Decomposition reaction, take a small amount of reaction solution and detect when the conversion rate of Epimedin C reaches 98%, carry out boiling water inactivation to end the reaction.

[0032] (3) Pur...

Embodiment 3

[0034] (1) Preparation of Epimedium extract: take Epimedium powder, pass through a No. 3 sieve and add ethanol with a volume fraction of 75%, and the mass-volume ratio of Wushan Epimedium powder to ethanol is 1:20, g : mL; Rotary evaporation extract obtains residual solid, and vacuum drying promptly obtains Epimedium extractum;

[0035] (2) Enzymolysis reaction: Add Epimedium extract and rhamnosidase in sequence according to the ratio of 1:1, mmol:U, and then add a phosphate buffer solution with a concentration of 0.2mol / L and a pH of 6.5 to stabilize the enzyme Bovine serum albumin, the mass-to-volume ratio of the epimedium extract to phosphate buffer solution and bovine serum albumin is 1:2:2, g:mL:mg, and the enzymolysis is carried out in a water bath at 60°C To react, extract a small amount of reaction solution to detect that the conversion rate of Epimedin C reaches 98%, and carry out boiling water inactivation to end the reaction.

[0036] (3) Purification of icariin: E...

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Abstract

Belonging to the field of drug preparation, the invention discloses a method for preparation of icariin by enzymolysis. The method provided by the invention mainly utilizes rhamnosidase to promote theenzymolysis reaction of epimedin C in epimedium so as to convert into icariin. The method has the advantages of simple operation, environmental friendliness and economic feasibility, can convert thepolyglycoside substance epimedin C with high content in epimedium into icariin, thus improving the yield of icariin and making up for the shortcomings in the prior art.

Description

technical field [0001] The invention belongs to the field of medicine preparation, and in particular relates to a method for preparing icariin by enzymatic hydrolysis. Background technique [0002] Icariin is a flavonoid with 8-prenyl flavone nucleus, molecular formula C 33 h 40 o 15 , with a molecular weight of 676.65 and a CAS number of 489-32-7, mainly found in plants of the genus Epimedium such as Epimedium, Epimedium Wushanense, Epimedium sagitta, Epimedium Korean and Epimedium pilosa stems and leaves Among the extracts, it has the effects of tonifying kidney and strengthening yang, strengthening muscles and bones, increasing blood flow in the cardiovascular system, promoting hematopoietic function, immune function and anti-aging. [0003] The current common preparation method of icariin is to obtain the main components of Epimedium Herba through hydroalcoholic extraction, and then separate them through various types of macroporous resin columns, polyamide columns, s...

Claims

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Application Information

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IPC IPC(8): C12P17/06
CPCC12P17/06
Inventor 夏之宁尹蔓妮张起辉
Owner CHONGQING UNIV
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