CN109603195A - A kind of degreasing method of animal tissue and its application - Google Patents

A kind of degreasing method of animal tissue and its application Download PDF

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Publication number
CN109603195A
CN109603195A CN201811478365.XA CN201811478365A CN109603195A CN 109603195 A CN109603195 A CN 109603195A CN 201811478365 A CN201811478365 A CN 201811478365A CN 109603195 A CN109603195 A CN 109603195A
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animal tissue
degreasing
carbon dioxide
fat
degreasing method
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CN201811478365.XA
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CN109603195B (en
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张伟
胡康
唐容
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Guan Hao Biotech Inc
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Guan Hao Biotech Inc
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    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01DSEPARATION
    • B01D11/00Solvent extraction
    • B01D11/02Solvent extraction of solids
    • B01D11/0203Solvent extraction of solids with a supercritical fluid
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01DSEPARATION
    • B01D11/00Solvent extraction
    • B01D11/02Solvent extraction of solids
    • B01D11/0288Applications, solvents

Abstract

The invention discloses a kind of degreasing method of animal tissue and its application, include the following steps: that (1) pre-processes: animal tissue being cleaned, disinfection, be cut into sheet, bulk or smash into powder to pieces;(2) be dehydrated: animal tissue is through ethanol solution dehydration;(3) it extracts: animal tissue is packed into progress supercritical carbon dioxide degreasing in closed extraction kettle;(4) separate: fat and oil-soluble impurities are as CO 2 fluid enters separating still, fat and oil-soluble impurities and carbon dioxide separation, carbon dioxide participates in extraction cycle next time, and fat and oil-soluble impurities are removed in separating still, recycle by least reextraction to obtain the final product.After animal tissue's degreasing method of the invention uses supercritical carbon dioxide extracting processing, the degreasing rate of animal tissue can completely remove the fat and oil-soluble impurities in animal tissue 99% or more;On the other hand, after supercritical carbon dioxide treatment, the potential animal virus removal rate of animal tissue reaches 99% or more.

Description

A kind of degreasing method of animal tissue and its application
Technical field
The present invention relates to defatting technology field, the degreasing method of especially a kind of animal tissue and its application.
Background technique
Currently, the degreasing method of animal tissue includes the methods of chemical immersion extracting and supercritical fluid extraction.Chemistry leaching Bubble extracting mainly use lye (such as sodium hydrate aqueous solution, by with fat occur saponification), organic reagent (such as acetone, The organic reagents such as petroleum ether, ethyl acetate, are extracted by similar compatibility principle) etc..Chemical method degreasing is general time-consuming long, chemistry examination Agent dosage is big, pollutes environment, and reagent easily remains in animal tissue.
Supercritical fluid extraction is to utilize some substances (such as carbon dioxide, water, nitrous oxide) in the supercritical state , the characteristics such as density big small with viscosity, have many advantages, such as that dissolubility is strong, diffusivity is good, easily controllable, pass through control temperature and pressure Extraction and separation of the supercritical fluid to fat can be realized in power, efficiently can thoroughly slough the objects such as the fat in animal tissue Matter.Meanwhile supercritical carbon dioxide extracting is low with supercritical temperature, cleaning noresidue, carbon dioxide source is wide, cheap easy , it is safe and non-toxic, non-ignitable the advantages that, have a extensive future.But supercritical fluid extraction the high requirements on the equipment, overcritical item The maintenance energy consumption of part needs to consider with safety.
Common animal tissue virus inactivating method is mostly that strong acid and strong base solution is handled, ion irradiation radiation treatment etc., but These methods will cause chemical bond rupture so as to cause the decrease of mechanical strength, and to animal tissue to animal tissue itself Excess processes will lead to albuminous degeneration, influence the application effect etc. of material in vivo.
Summary of the invention
Based on the above issues, a kind of animal is provided it is an object of the invention to overcome above-mentioned the deficiencies in the prior art place The degreasing method of tissue, this method degreasing rate can reach 99% or more, while can remove potential virus in animal tissue, removal rate 99% or more can be reached;Meanwhile without chemical agent residue in organizing after this method degreasing, the mechanics that will not weaken animal tissue is strong Degree, not will lead to protein denaturation in animal tissue.
To achieve the above object, the technical scheme adopted by the invention is as follows: a kind of degreasing method of animal tissue, including as follows Step:
(1) it pre-processes: animal tissue being cleaned, disinfection, be cut into sheet, bulk or smash into powder to pieces;
(2) be dehydrated: animal tissue is through ethanol solution dehydration;
(3) it extracts: animal tissue is packed into progress supercritical carbon dioxide degreasing in closed extraction kettle;
(4) separate: fat and oil-soluble impurities as CO 2 fluid enters separating still, fat and oil-soluble impurities with Carbon dioxide separation, carbon dioxide participate in extraction cycle next time, and fat and oil-soluble impurities are removed in separating still, pass through At least reextraction recycles to obtain the final product.Wherein, animal tissue from cortex bone, cancellous bone, cartilage, peritonaeum, pericardium, rouge nethike embrane, The tissues such as submucous layer of small intestine, bladder base, corium or organ.
Preferably, the length of sheet and width are 10~300 millimeters in the step (1), the block-like volume is 0.5~ 10.0 cubic centimetres.
Preferably, animal tissue is dehydrated 0.5~5.0 hour using 75~100% ethanol solutions in the step (2).
Preferably, dynamic extraction degreasing is used in the step (3).Preferably, in the step (3), carbon dioxide stream Body has been subjected to purified treatment before entering extraction kettle contact animal tissue, and the purified treatment is passed through with certain ultra microstructure Ultrafiltration filter membrane.
Preferably, 40~60 DEG C of skimming temp in the step (3), pressure is 20~40MPa.
Preferably, carbon dioxide flow is 18~20Hz in the step (3).
Preferably, the dynamic extraction degreasing time is 0.5~20.0 hour.Preferably, extraction kettle volume be 0.5~ 150.0 liters, it is interior be equipped with stainless steel filter screen, 5~25 microns of filtering accuracy.
Preferably, in the step (3) when supercritical carbon dioxide degreasing, entrainer is added in extraction kettle.It needs Bright, entrainer effect is to mix with extractant (such as carbon dioxide), changes the polarity of solvent, and it is molten to improve solvent dissolution The ability of matter is remarkably improved and is difficult to extract originally or the extraction yield of substance that extraction yield is low, or can improve separating effect.
Preferably, the entrainer is at least one of methanol, ethyl alcohol, acetone, ethyl acetate, benzyl carbinol, entrainer Mass concentration be 0.1~10%.
Preferably, separating still temperature is 30~70 DEG C in the step (4).
On the other hand, the present invention provides above-mentioned degreasing methods to be used for animal sources soft tissue, the fat of sclerous tissues and liposoluble The removal or abjection of property substance.
The present invention also provides above-mentioned degreasing methods for removing animal source material fat and liposoluble substance, while will dive Animal virus be removed or deviate from.
In conclusion the invention has the benefit that
After animal tissue's degreasing method of the invention is using supercritical carbon dioxide extracting processing, the degreasing rate of animal tissue 99% or more, the fat and oil-soluble impurities in animal tissue can be completely removed;On the other hand, at through supercritical carbon dioxide After reason, the potential animal virus removal rate of animal tissue reaches 99% or more, after treatment by using the treatment method, examines in animal tissue's material The presence of animal virus is not detected;The processing method can also be used in combination with acid-base solution, and the effect for removing animal virus is more preferable.
Specific embodiment
The present invention relates to a kind of using supercritical carbon dioxide extracting to the method for animal tissue's degreasing, and the present invention is using only Special technology controlling and process, degreasing rate can reach 99% or more, efficiently quickly can carry out ungrease treatment, animal tissue to animal tissue Including the biomembranes such as bone, cartilage, pericardium class, corium etc..Meanwhile method of the invention can be also used for it is potential in animal tissue The removal of animal virus, animal virus removal rate reach 99% or more, after treatment by using the treatment method, detect not in animal tissue's material To the presence of animal virus;This method can be used in combination with acid-base solution simultaneously, and treatment effect is more preferable.
In some embodiments, the processing steps such as degreasing method of the invention, including pretreatment, dehydration, extraction, separation, Specifically, the following steps are included:
A. it pre-processes: animal tissue being cleaned, disinfection, be cut into sheet, bulk or smash into powder to pieces, wherein sheet length and width 10~300 millimeters, blocky volume is 0.5~10.0 cubic centimetre;
B. be dehydrated: animal tissue is dehydrated 0.5~5.0 hour through 75~100% ethanol solutions;
C. it extracts: after ethanol dehydration is handled, animal tissue being packed into closed extraction kettle and carries out supercritical carbon dioxide Degreasing, 40~60 DEG C of skimming temp, pressure is 20~40MPa;Blocky or sheet animal tissue uses dynamic extraction degreasing, dioxy Change carbon flow is 18~20Hz, and dynamic extraction degreasing time is 0.5~20.0 hour;It is equal during supercritical carbon dioxide extracting Added with entrainer;
D. separate: as CO 2 fluid enters separating still, adjusting temperature is 30~70 DEG C for fat and oil-soluble impurities, The substances such as lipid and carbon dioxide separation, extraction cycle, the substances such as fat are gone in separating still next time for carbon dioxide participation It removes, i.e. completion degreasing.
To better illustrate the object, technical solutions and advantages of the present invention, below in conjunction with specific embodiment to the present invention It is described further.
Embodiment 1
A kind of embodiment of the degreasing method of animal tissue of the invention, includes the following steps:
(1) it pre-processes: animal tissue's cartilage being cleaned, disinfection, be cut into sheet, 10 millimeters × 300 millimeters of sheet length and width;
(2) be dehydrated: animal tissue is dehydrated 0.5 hour through 75% ethanol solution;
(3) it extracts: after ethanol dehydration is handled, animal tissue being packed into closed extraction kettle and carries out supercritical carbon dioxide Degreasing, 40 DEG C of skimming temp, pressure 20MPa;Sheet animal tissue uses dynamic extraction degreasing, and carbon dioxide flow is 20Hz, dynamic extraction degreasing time are 0.5 hour;Added with entrainer methanol (quality during supercritical carbon dioxide extracting Concentration 0.1%);
(4) separate: fat and oil-soluble impurities are as CO 2 fluid enters separating still, and adjusting temperature is 30 DEG C, rouge The substances such as matter and carbon dioxide separation, carbon dioxide gas participate in second extraction cycle, and the substances such as fat are in separating still quilt Removal to get.
Embodiment 2
A kind of embodiment of the degreasing method of animal tissue of the invention, includes the following steps:
(1) it pre-processes: animal tissue's pericardium being cleaned, disinfection, be cut into bulk, blocky volume is 3 cubic centimetres;
(2) be dehydrated: animal tissue is dehydrated 2 hours through 90% ethanol solution;
(3) it extracts: after ethanol dehydration is handled, animal tissue being packed into closed extraction kettle and carries out supercritical carbon dioxide Degreasing, 50 DEG C of skimming temp, pressure 40MPa;Blocky animal tissue uses dynamic extraction degreasing, and carbon dioxide flow is 18Hz, dynamic extraction degreasing time are 7 hours;Added with entrainer during supercritical carbon dioxide extracting, entrainer is second The mixed solution (mass concentration 5%) of pure and mild ethyl acetate, wherein the mass ratio of ethyl alcohol and ethyl acetate is 1:5;
(4) separate: fat and oil-soluble impurities are as CO 2 fluid enters separating still, and adjusting temperature is 50 DEG C, rouge The substances such as matter and carbon dioxide separation, extraction cycle, the substances such as fat are gone in separating still next time for carbon dioxide gas participation Except to get.
Embodiment 3
A kind of embodiment of the degreasing method of animal tissue of the invention, includes the following steps:
(1) it pre-processes: animal tissue's corium being cleaned, disinfection, smash into powder to pieces.
(2) be dehydrated: animal tissue is dehydrated 5.0 hours through 100% ethanol solution
(3) it extracts: after ethanol dehydration is handled, animal tissue being packed into closed extraction kettle and carries out supercritical carbon dioxide Degreasing, 60 DEG C of skimming temp, pressure 30MPa;Powdery animal tissue uses dynamic extraction degreasing, and carbon dioxide flow is 19Hz, dynamic extraction degreasing time are 20.0 hours;Added with entrainer acetone (quality during supercritical carbon dioxide extracting 10%) concentration is;
(4) separate: fat and oil-soluble impurities are as CO 2 fluid enters separating still, and adjusting temperature is 70 DEG C, rouge The substances such as matter and carbon dioxide separation, carbon dioxide gas participate in second of extraction cycle, and the substances such as fat are gone in separating still Except to get.
Embodiment 4
A kind of embodiment of the degreasing method of animal tissue of the invention, includes the following steps:
1) it takes fresh Pigs Hearts encapsulation tissue to clean, disinfection, is cut into sheet, 150 millimeters × 150 millimeters of length and width;
2) it is dehydrated: being dehydrated 2 hours through ethanol solution;
3) it extracts: pericardial tissue being put into extraction kettle after dehydration, temperature is 45 DEG C, pressure 30MPa, dioxy Change carbon flow be 18Hz, entrainer be mass concentration 5% ethyl alcohol, dynamic extraction 8 hours;
4) temperature 60 C in separating still isolates the substances such as fat, and carbon dioxide enters next extraction cycle, through super To get the animal tissue after degreasing after supercritical fluid extraction.
The measure of merit of the degreasing method of the invention of embodiment 5
(1) fat content detection is referring to the second method of GB 5009.6-2016 acid hydrolysis process.
Pericardium weighing after the degreasing of Example 4 is placed in test tube, and hydrochloric acid solution is added in 70~80 DEG C of water-baths Heat hydrolysis in pot, until digestion completely, is then added ethanol solution, moved into after cooling in tool plug graduated cylinder, be added anhydrous ether into Row extraction, after extraction take out upper layer ether layer in conical flask, then heating remove ether, to remaining solid content be dried to Constant weight is to get content fatty in product.
The results show that pericardial tissue degreasing rate reaches 99.5%.
(2) virus titer test uses cytopathy political reform (CPE).
Method particularly includes: the weighing of coring coating is separately added into corresponding indicator virus [including bovine viral diarrhea virus (BVDV), porcine pseudorabies virus (PRV), encephalomyocarditis virus (EMCV), pig parvoviral (PPV)] culture after, using embodiment 4 method processing, is then added serum free medium grinding, supernatant is sterile filtered, by filtered liquid, using 96 orifice plates Cytopathy (CPE) method, observes the cytopathy situation of each gradient dilution liquid, according to Reed-Muench method calculate test group and The lgTCID50/0.1 milliliter of 4 kinds of indicator virus in control group.
CPE method: logarithmic growth phase cell is inoculated with 96 orifice plates (every hole 104It is a), 5%CO2, 37 DEG C of incubator cultures 24 it is small When, cell is adherent and forms uniform monolayers;Gradient dilution virus is to 10-5~10-139 gradients, every hole add 100 microlitres, each column One dilution 9 arranges totally;Culture solution is added in remaining 3 column, makees blank control;37 DEG C, 5%CO2Culture 10 days.Daily microscope Lower observation cytopathy situation records each epidemy and becomes situation, according to Reed-Muench formula:
The viral dilution of TCID 50=CPE>50%+(>50% percentage -50)/(>50% percentage -<50% Percentage) * log10;
△ lgTCID50 indicates the order of magnitude of viral reduction after processing, is greater than 4, that is, shows that removal rate is greater than 99.99%;
Virus titer is calculated, is as a result indicated with infectious unit/milliliter (IFU/ml).
The results are shown in Table 1, and method of the invention removes viral 99% or more rate, 4 kinds of indicator virus titre logarithm variations It is all larger than 4logs, meets the requirement of national relevant laws and regulations and standard.
The method of the invention of table 1 handles the result after 4 kinds of indicator virus
Indicator virus BVDV PRV EMCV PPV
△lgTCID50 ≥7.5 ≥6.9 ≥6.7 ≥4.2
Finally it should be noted that the above embodiments are merely illustrative of the technical solutions of the present invention rather than protects to the present invention The limitation of range, although the invention is described in detail with reference to the preferred embodiments, those skilled in the art should be managed Solution, can with modification or equivalent replacement of the technical solution of the present invention are made, without departing from technical solution of the present invention essence and Range.

Claims (12)

1. a kind of degreasing method of animal tissue, which comprises the steps of:
(1) it pre-processes: animal tissue being cleaned, disinfection, be cut into sheet, bulk or smash into powder to pieces;
(2) be dehydrated: animal tissue is through ethanol solution dehydration;
(3) it extracts: animal tissue is packed into progress supercritical carbon dioxide degreasing in closed extraction kettle;
(4) separate: fat and oil-soluble impurities are as CO 2 fluid enters separating still, fat and oil-soluble impurities and dioxy Change carbon separation, carbon dioxide participates in extraction cycle next time, and fat and oil-soluble impurities are removed in separating still, by least Reextraction recycles to obtain the final product.
2. degreasing method according to claim 1, which is characterized in that in the step (1) length of sheet and width be 10~ 300 millimeters, the block-like volume is 0.5~10.0 cubic centimetre.
3. degreasing method according to claim 1, which is characterized in that molten using 75~100% ethyl alcohol in the step (2) Liquid is dehydrated animal tissue 0.5~5.0 hour.
4. degreasing method according to claim 1, which is characterized in that use dynamic extraction degreasing in the step (3).
5. degreasing method according to claim 1, which is characterized in that 40~60 DEG C of skimming temp in the step (3), pressure It is by force 20~40MPa.
6. degreasing method according to claim 1, which is characterized in that in the step (3) carbon dioxide flow be 18~ 20Hz。
7. degreasing method according to claim 4, which is characterized in that the dynamic extraction degreasing time is 0.5~20.0 Hour.
8. degreasing method according to claim 1, which is characterized in that supercritical carbon dioxide degreasing in the step (3) When, extraction kettle is interior to be added with entrainer.
9. degreasing method according to claim 8, which is characterized in that the entrainer is methanol, ethyl alcohol, acetone, acetic acid At least one of ethyl ester, benzyl carbinol, the mass concentration of entrainer are 0.1~10%.
10. degreasing method according to claim 1, which is characterized in that separating still temperature is 30~70 in the step (4) ℃。
11. degreasing method according to claim 1 is for animal sources soft tissue, the fat of sclerous tissues and liposoluble substance Removal or abjection.
12. degreasing method according to claim 1 will dive for removing animal source material fat and liposoluble substance Animal virus be removed or deviate from.
CN201811478365.XA 2018-12-04 2018-12-04 Animal tissue degreasing method and application thereof Active CN109603195B (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110179830A (en) * 2019-07-10 2019-08-30 山东中泰药业有限公司 A kind of compound degreasing method of fresh animal medicinal material and defatted animal medicinal material

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US20160158763A1 (en) * 2013-08-02 2016-06-09 Hitachi, Ltd. Solid/liquid separation apparatus, and method for same
CN106244740A (en) * 2016-08-30 2016-12-21 张玉斌 A kind of employing supercritical CO2abstraction technique carries out the method for Corii Sus domestica defat

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Publication number Priority date Publication date Assignee Title
US3966981A (en) * 1974-11-26 1976-06-29 The United States Of America As Represented By The Secretary Of Agriculture Process for removing residual solvents
CN101139543A (en) * 2007-10-29 2008-03-12 中国农业大学 Method for extracting sheep placental extract grease by over-critical carbon dioxide
CN101812087A (en) * 2010-05-07 2010-08-25 青岛海恩赐生物工程有限公司 Method for extracting high-purity phospholipid from sleeve-fish-processing waste
CN102771620A (en) * 2012-07-14 2012-11-14 李世泰 Method for producing hydrolyzed brain protein powder and cephalin by grease removal of supercritical carbon dioxide
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110179830A (en) * 2019-07-10 2019-08-30 山东中泰药业有限公司 A kind of compound degreasing method of fresh animal medicinal material and defatted animal medicinal material

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