CN103816187B

CN103816187B - Preparation method for fat-soluble extract of common mussels

Info

Publication number: CN103816187B
Application number: CN201410049970.0A
Authority: CN
Inventors: 邓尚贵; 霍健聪
Original assignee: Zhejiang Ocean University ZJOU
Current assignee: Zhejiang Ocean University ZJOU
Priority date: 2014-02-13
Filing date: 2014-02-13
Publication date: 2017-04-26
Anticipated expiration: 2034-02-13
Also published as: CN103816187A

Abstract

The invention discloses a preparation method for a fat-soluble extract of common mussels. The preparation method comprises the following steps: (1) pre-treating the common mussels, specifically, after the common mussels are fished out, cleaning, unshelling, separating meat, and removing byssus; then mincing the mussel meat to obtain minced mussel meat; (2) performing enzymolysis by a complex enzyme, specifically, putting the minced mussel meat into water, adding the complex enzyme, stirring, adjusting the pH value, keeping the temperature and performing enzymolysis to obtain water-insoluble crude fat, wherein Vc is added before enzymolysis, and the liquid is abandoned after enzymolysis; (3) dissolving the crude fat in an organic solvent while adding VE, freezing, filtering at the temperature of 5 below zero to 10 below zero to obtain the fat-soluble extract crude product of the common mussels, and then evaporating the organic solvent through water bath reflux to obtain the fat-soluble extract of the common mussels. The fat-soluble extract of the common mussels, obtained through the method, has the characteristics of higher purity, low cost and low oxygenation rate, is relatively high in fluidity, contains 26 unsaturated fatty acids, found out through gas chromatography, and has purity reaching 95.21 percent.

Description

A kind of preparation method of Mytilus eduliss liposoluble extract

Technical field

The present invention relates to a kind of preparation method of the Mytilus eduliss liposoluble extract with anti-inflammatory activity.

Background technology

Shengsi County is the maximum production base of China mussel, there is the good reputation in " township of Chinese mussel ".Shengsi mussel mainly with Based on cultivation Mytilus eduliss and Mytilus crassitesta Lischke, but because Mytilus crassitesta Lischke growth cycle is long, go back main based on Mytilus eduliss at present, 2013 About 11.2 ten thousand tons of annual production.The current major consumers mode of Shengsi mussel is that marketing fresh and processing gelation mussel or dry mussel are sold, but Because mussel economic worth is relatively low, fresh goods is only 2 yuan/jin or so in terminal sale price.Mussel attraction is excavated, mussel Jing is improved Ji value is extremely urgent.Inflammation is the general name of the big class disease for perplexing the mankind, and arthritis, asthma etc. belong to such disease. And containing the anti-inflammatory substance with Lyprinol activity in mussel.Therefore, with Shengsi mussel as raw material, exploitation anti-inflammatory drug tool There are great economic implications and social meaning.

With regard to " the thick-shell mussel fat-soluble that the existing forefathers' research of extraction of anti-inflammatory activity thing in mussel, Yuan Gaofeng etc. are delivered Extract antiinflammatory bioactivity research "（Zhejiang University reports（Agricultural and life sciences version） 33（2）：169～173,2007）Refer to Go out：With Mytilus crassitesta Lischke as raw material, through lyophilization and chloroform: methanol（2: 1, V: V）It is extracted thick-shell mussel fat-soluble extraction Thing, and demonstrate its anti-inflammatory activity；Chinese Patent Application No. is 200910101136.0, and publication date is on December 23rd, 2009 Also disclose that by lyophilization in " thick-shell mussel fat-soluble extract and its production and use " patent application The thick-shell mussel fat-soluble extract prepared with the method such as supercritical carbon dioxide extraction, but due to fat-soluble anti-inflammatory substance it is main For unsaturated fatty acid, and these methods or to there is extracting method excessively simple, cause fatty acid to aoxidize, or exist and extract Method is excessively complicated, the drawbacks of extraction cost is high.

The content of the invention

The present invention is directed to disadvantages mentioned above, on the basis of fat-soluble anti-inflammatory substance activity is ensured, there is provided a kind of low cost The preparation method of the Mytilus eduliss liposoluble extract with anti-inflammatory activity.

A kind of preparation method of Mytilus eduliss liposoluble extract, it is characterised in that comprise the following steps：

1）Mytilus eduliss pretreatment：Cleaned after Mytilus eduliss are fished for up, shelled, adopted meat, remove byssus；Then by mussel Meat carries out rubbing process, obtains Mytilus eduliss minced meat；

2）Complex enzyme zymohydrolysis：Mussel minced meat are added to the water, compound enzyme are subsequently adding and are stirred, adjust pH value, insulation, enzyme Solution, adds Vc before enzymolysis, liquid is given up after enzymolysis, obtains water-fast crude fat；

3）Crude fat organic solvent is dissolved, adds VE during dissolving, it is then chilled, under the conditions of -5～-10 DEG C Mytilus eduliss extract crude product is filtrated to get, then obtains that Mytilus eduliss are fat-soluble to be carried after water-bath refluxed evaporator falls organic solvent Take thing.

Preferably, step 1）Described in rubbing process be adopt blender with 10000r/min speed stirring 30s, stop Machine, then the 30s that starts shooting, shut down, and so on 3 times.

Preferably, step 2）Described in compound enzyme be by food flavor enzyme, papain, neutral protease, alkaline egg White enzyme, animal compound protease two or more carry out compounding and form.

Preferably, step 2）In, it is 5.0～6.5 that the compound enzyme adding proportion is 0.5%～1%, pH, and temperature keeps 40 DEG C～45 DEG C, enzymolysis time is 0.5～1.5 hour.

Preferably, step 3）Described in organic solvent be ethyl acetate or acetone or ether.

Preferably, step 3）Described in cryogenic temperature be -20 DEG C～-30 DEG C, cooling time be 5～8 hours, water-bath Return time 2～3 hours.

Preferably, the step 2）1% Vc, the step 3 are added before middle enzymolysis）The VE of addition 1% during middle dissolving.

The Mytilus eduliss liposoluble extract for obtaining is extracted by the method for the invention there is higher purity, low cost, oxygen The characteristics of rate is low, with stronger mobility, Jing gas chromatographic analysiss contain 26 kinds of unsaturated fatty acids, and purity reaches 95.21%。

Specific embodiment

The present invention is described in further detail with reference to embodiments.

Embodiment 1：

The first step：Mytilus eduliss pretreatment：Cleaned after Mytilus eduliss are fished for up, shelled, adopted meat, remove byssus；Then Mussel meat is carried out into rubbing process, Mytilus eduliss minced meat are obtained.

Second step：Complex enzyme zymohydrolysis：First Mytilus eduliss minced meat are adopted into blender and 30s are stirred with 10000r/min speed, Shut down, then the 30s that starts shooting, shut down, and so on 3 times；Then 1 part of mussel minced meat is added and add compound enzyme, compound enzyme after 9 parts of water Food flavor enzyme 10000IU/g, alkaline protease 5000IU/g and animal compound protease 100000IU/g are selected as compound enzyme, it is multiple It is 1: 1: 0.1 with ratio, compound enzyme adding proportion 0.5%, while adding the Vc that mass fraction is 1%；Tune pH value is after stirring 5.0, temperature is maintained at 40 DEG C, enzymolysis time 0.5 hour；Enzymolysis is filtered after terminating, and gives up liquid, takes solid crude fat standby.

3rd step：The crude fat ethyl acetate dissolving of 5 times of quality, while adding the VE of crude fat quality 1%, is placed in -20 After freezing 5 hours at a temperature of DEG C, obtain Mytilus eduliss extract crude product after filtering at a temperature of -5 DEG C, then Jing after water-bath backflow it is 2 little When obtain Mytilus eduliss extract.

Embodiment 2：

Second step：Complex enzyme zymohydrolysis：First Mytilus eduliss minced meat are adopted into blender and 30s are stirred with 10000r/min speed, Shut down, then the 30s that starts shooting, shut down, and so on 3 times；Then 1 part of mussel minced meat is added and add compound enzyme, compound enzyme after 9 parts of water Food flavor enzyme 10000IU/g, alkaline protease 5000IU/g and animal compound protease 100000IU/g are selected as compound enzyme, it is multiple It is 1: 1: 0.1 with ratio, compound enzyme adding proportion 0.75%, while adding the Vc that mass fraction is 1%；Tune pH value is after stirring 5.5, temperature 45 C, enzymolysis time 1 hour；Enzymolysis is filtered after terminating, and gives up liquid, takes solid crude fat standby.

3rd step：By the crude fat ether dissolution of 5 times of quality, while adding the VE of crude fat quality 1%, -25 DEG C are placed in At a temperature of after freezing 6 hours, obtain Mytilus eduliss extract crude product after filtering at a temperature of -8 DEG C, then Jing after water-bath backflow it is 2.5 little When obtain Mytilus eduliss extract.

Embodiment 3：

Second step：Complex enzyme zymohydrolysis：First Mytilus eduliss minced meat are adopted into blender and 30s are stirred with 10000r/min speed, Shut down, then the 30s that starts shooting, shut down, and so on 3 times；Then 1 part of mussel minced meat is added and add compound enzyme, compound enzyme after 9 parts of water Food flavor enzyme 10000IU/g, alkaline protease 5000IU/g and animal compound protease 100000IU/g are selected as compound enzyme, it is multiple It is 1: 1: 0.1 with ratio, compound enzyme adding proportion 1%, while adding the Vc of mass fraction 1%；PH value is adjusted to be 6.0 after stirring, temperature 45 DEG C of degree, enzymolysis time 1.5 hours；Enzymolysis is filtered after terminating, and gives up liquid, takes solid crude fat standby.

3rd step：By the crude fat acetone solution of 5 times of quality, while adding the VE of crude fat quality 1%, -30 DEG C are placed in At a temperature of after freezing 8 hours, Mytilus eduliss extract crude product is obtained after filtering at a temperature of -10 DEG C, then Jing after water-bath backflow 3 hours Time obtains Mytilus eduliss extract.

The Mytilus eduliss fat-extraction thing for obtaining is extracted by the method for the invention there is higher purity, low cost, oxidation The characteristics of rate is low, extract is presented transparent fawn oily, and with stronger mobility, Jing gas chromatographic analysiss contain 26 Unsaturated fatty acid is planted, purity reaches 95.21%.

Claims

1. a kind of preparation method of Mytilus eduliss liposoluble extract, is cleaned after first Mytilus eduliss are fished for up, is shelled, is adopted Meat, remove byssus；Then mussel meat is carried out into rubbing process, obtains Mytilus eduliss minced meat, its feature is further comprising the steps of：

1) complex enzyme zymohydrolysis：Mussel minced meat are added to the water, the ratio that is initially charged before enzymolysis is 1% Vc, is subsequently adding ratio Compound enzyme for 0.5%～1% is stirred, adjusts pH value to be 5.0～6.5, is persistently incubated under the conditions of 40 DEG C～45 DEG C, digests 0.5～1.5 hour time, liquid is given up after enzymolysis, obtains water-fast crude fat, described compound enzyme be by food flavor enzyme, Papain, neutral protease, alkaline protease, animal compound protease two or more carry out compounding and form；

2) by the crude fat with organic solvent dissolve, during dissolving addition 1% VE, then Jing temperature be -20 DEG C～-30 DEG C, Time is the freezing processing of 5～8 hours, and Mytilus eduliss extract crude product is filtrated to get under the conditions of -5～-10 DEG C, little through 2～3 When water-bath refluxed evaporator to fall obtain after organic solvent Mytilus eduliss liposoluble extract, described organic solvent is ethyl acetate Or acetone or ether.

2. the preparation method of a kind of Mytilus eduliss liposoluble extract according to claim 1, it is characterised in that：Step 1) in It is to adopt blender to stir 30s with 10000r/min speed that described rubbing is processed, and is shut down, then the 30s that starts shooting, and is shut down, so past It is multiple 3 times.