CN104961764B - Technology for extraction of phosphatidylcholine from clarias gariepinus leftover - Google Patents
Technology for extraction of phosphatidylcholine from clarias gariepinus leftover Download PDFInfo
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- CN104961764B CN104961764B CN201510293359.7A CN201510293359A CN104961764B CN 104961764 B CN104961764 B CN 104961764B CN 201510293359 A CN201510293359 A CN 201510293359A CN 104961764 B CN104961764 B CN 104961764B
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Abstract
The invention discloses a technology for extraction of phosphatidylcholine from clarias gariepinus leftover. The technology comprises 1, mincing and triturating head and internal organs of clarias gariepinus at a temperature less than 50 DEG C, 2, adding an acetic acid-sodium acetate buffer solution into the triturated materials obtained by the step 1 to adjust pH as protease enzymolysis pH, heating the material to a temperature of 37-45 DEG C, adding 0.015-0.075% of protease into the materials and carrying out enzymolysis for 60-100min, 3, adding extract into the enzymolysis product obtained by the step 2, carrying out stirring, carrying out extraction at a temperature of 35-45 DEG C for 60-100min and carrying out pressure reduction pumping filtration to obtain extract and 4, adding NaCl into the extract obtained by the step 3, carrying out stirring dissolution, carrying out standing for 3-4h to obtain precipitates and carrying out drying at a temperature less than 50 DEG C. The technology utilizes fish leftover, realizes waste recycle, prevents competition with human and animals in nutrition resources, reduces environmental pollution and has important ecological and economic meaning.
Description
Technical field
The present invention relates to technical field of lecithin, the technique that especially a kind of use clarias leather leftover bits and pieces extract lecithin.
Background technology
Lecithin, broad sense are lecithin, refer to the usual title of various commercially available organic phosphoric acid product salts.Its main component
There are phosphatidylcholine (PC), Phosphatidycholine (PE), phosphatidic acid and phosphoinositide (PI).And the lecithin of narrow sense refers to phosphatidyl
Choline, which is English for phosphati-dycholine, the one kind being made up of glycerolcholine phosphoric acid saturation and unsaturated fatty acid
Containing phospholipid substance.
At present, lecithin product is mainly from the brain and egg yolk of the aquation oil of the plants such as soybean oil, Oleum Brassicae campestriss or animal
Extract, especially with content highest in egg yolk, but it is higher that the cost of lecithin is extracted from egg yolk.According to the literature, in fish scale, fish
Containing abundant lecithin resource in the leftover bits and pieces such as dirty, fish head, and contain a large amount of unsaturated fatty acids, nutrition is more superior.
Research both at home and abroad to lecithin preparation method and technique is a lot, and new method continuously emerges.Mainly have at present with
Under it is several：
(1) solvent extraction method：Its principle is the different solubility using each phospholipid fraction in some solvents, to having
Effect ingredient solubility is big, to the solvent for not needing dissolved element dissolubility little, by the side of effective ingredient dissolution from raw tissue
Lecithin is separated by method with other components.Conventional at present is ethanol extraction, the method for acetone precipitation.
(2) Complex precipitation with inorganic salts：Lecithin can react to form coordination compound with metal ion, generate precipitation.Profit
This kind of property is used, lecithin can be separated from organic solvent, thus remove protein, the impurity such as fat, then with fitting
When solvent isolates inorganic salt and phospholipid impurity, the purity of lecithin can be so improved.CaCl2、ZnCl2、MgCl2、CdCl2
Complex reaction can occur with lecithin and form double salt.
(3) enzymatic isolation method：Lecithin belongs to complex lipid, and in Fish, lecithin is combined and is existed by materials such as protein.One
As solvent extraction method more difficult extract lecithin.Protease can be with the peptide bond of aminosal, so that protein cleavage is
Little peptide chain, destruction protein and liposome complexes.
(4) the homogeneous extraction of ultrasonic assistant：Using the cavitation of ultrasound wave, cause the alternate compression of medium particle and stretch
, increase the contact area of phospholipid and solvent (such as acetone), make the lubricant component in phospholipid particles be effectively dissolved in wherein, so as to
Improve the purity of effect of extracting and product.
(5) microwave loss mechanisms：Microwave radiation is mainly combined by Microwave Assisted Extraction Technique with extractant, to target
Active substance effectively extracts detached method, extracts separation process and takes that short, solvent load is few, selectivity is strong and extraction ratio is high,
The shortcomings of effectively overcoming traditional soxhlet extraction technique time-consuming, solvent load is big.
(6) emulsifying is extracted：At present useful emulsification and extraction equipment come purify high concentration phosphorus phosphatidylcholine (PC) (referring to：Peace
Red etc., in emulsification and extraction technology extracting soybean phospholipid phosphatidylcholine [J]. analytical chemistry, 31 (7)：824-827), mainly
Ethanol is added in extracted good crude lecithin, is extracted by emulsifying device stirring again.Lecithin is a kind of good
Emulsifying agent, therefore, in Extraction solvent, add surfactant play the effect of solubilising, and at present almost not over plus
Surfactant is by being extracted to lecithin solubilising.
In said extracted method, there is following technological deficiency：
(1) shortcoming of raw material：Lecithin is mainly extracted from the fowl egg such as the oil crop such as Semen sojae atricolor, egg yolk both at home and abroad at present,
Food resource is grabbed with the mankind on raw material；
(2) many Extraction solvents are poisonous：In extraction process, it is industrial often using organic solvent for example acetone, chloroform, just oneself
Alkane, ether, acetonitrile etc. are extracted, and product can be made to have certain organic solvent residual, and toxicity is very high, and this problem is never fine
Solution；
(3) in terms of inorganic salt is as precipitant：What research was more is to use ZnCl2And CdCl2, the two is respectively provided with certain poison
Property, this problem also never improves.
The content of the invention
The present invention is directed to the deficiencies in the prior art, proposes that a kind of use clarias leather leftover bits and pieces extract the technique of lecithin, behaviour
Make easy, effect is good.
In order to realize foregoing invention purpose, the present invention provides technical scheme below：A kind of clarias leather leftover bits and pieces are extracted
The technique of lecithin, comprises the following steps：
(1), the head and internal organs of clarias leather, temperature are carried out mincing below 50 DEG C, are smashed to pieces；
(2), the pH value of Acetic acid-sodium acetate buffer regulating step (1) stamping is added, and the pH value is protease hydrolyzed pH
Value, is heated to 37～45 DEG C, adds 0.015～0.075% protease, digests 60～100min；
(3) extracting solution is added after, (2) digesting to step in material, is stirred, 60～100min is extracted at 35～45 DEG C；Subtract
Pressure sucking filtration obtains extracting solution；
(4), NaCl is added in step (3) extracting solution, stirring and dissolving stands 3～4 hours, obtains precipitate, in less than 50 DEG C
Lower drying.
Preferably, the protease is papain, pepsin or trypsin.
Preferably, the extracting solution is sodium lauryl sulphate and the mixed solvent of ethanol, the quality of sodium lauryl sulphate
Volume ratio is 0.67%～6%, and concentration of alcohol is 25%～75%.
Preferably, (4) middle NaCl additions are that mass volume ratio is 1 to step:30.
Compared with prior art, the present invention has advantages below：
(1) fish pomace is utilized, carries out twice laid, nutrition money will not be grabbed with human and animal on raw material is extracted
Source, and environmental pollution can be reduced, with important ecological and economic implications；
(2) conventional solvent extraction method excessive use toxic organic solvents are solved the problems, such as, in whole technique, is only with the addition of
A kind of organic solvent of ethanol；And first passage adds Surfactant SDS (GB 2760-96 in ethanol
It is defined as food industry processing aid, such as foaming agent；Emulsifying agent etc.) and solubilization is played to lecithin；
(3) when salt is settled out product, NaCl harmless to the human body, i.e. Sal are employed by experiment, can be smooth
Obtain lecithin crude product.
Description of the drawings
Fig. 1 is the process chart that the present invention extracts lecithin from clarias leather leftover bits and pieces；
Ultraviolet spectrograms of the Fig. 2 for sample lecithin；
Standard curves of the Fig. 3 for potassium dihydrogen phosphate.
Specific embodiment
A kind of use clarias leather leftover bits and pieces as shown in Figure 1 extract the new technology of lecithin, belong to Separation of Natural Products neck
Domain, comprises the following steps：
(1) Feedstock treating
Clarias leather leftover bits and pieces refer mainly to fish head, fish internal organs, and this fish is without fish scale.Fish head is little, and bone is more, is less susceptible to place
Reason；Internal organs are red, more fatty than Ctenopharyngodon idellus etc. on the low side, easily smash process.Fish pomace can carry out freezen protective.Need to extract
When, taking-up is minced with knife in right amount, then is smashed with tissue mashing machine.It is noted that temperature control during smashing, temperature will be controlled
Less than 50 DEG C, temperature is too high, and lecithin can decomposes.Therefore, it can plus a small amount of ice cube carries out tissue mashing.
(2) enzymolysis processing
As phospholipid and protein are combined together, it is difficult to extract, therefore will first removes protein, can typically use
Papain, pepsin, trypsin.The raw material handled well in right amount is taken, adds NaAc_HAc buffer solution to adjust pH
For the optimum pH of protease, heating in water bath is then carried out, temperature is the enzymolysis preference temperature of protease, at 37～45 DEG C, when
When material temperature reaches, adding protease carries out enzymolysis processing.The protease enzyme amount of addition is unsuitable excessive, and general addition is
0.015～0.075% (referring to the quality of enzyme and the mass ratio of raw material), enzymolysis time are unsuitable long, in 60～100min；Such as wood
Melon protease (enzyme activity >=8 × 105U/g Optimal pH) is 7, and hydrolysis temperature is 50 DEG C, and enzymolysis time is 100min；Pepsin
Enzyme (enzyme activity 1：15000) Optimal pH is 3, and hydrolysis temperature is 37 DEG C, and enzymolysis time is 60min；Trypsin enzyme activity 1：
250) Optimal pH is 9, and hydrolysis temperature is 45 DEG C, and enzymolysis time is 80min；Through enzymolysis processing, fish pomace substantially becomes
It is rotten to the corn.
(3) extract, filter
The mixed solvent of sodium lauryl sulphate and ethanol is added in the fish pomace after enzymolysis, wherein, dodecyl
Sodium sulfate is 0.67%～6% (mass volume ratio：That is the volume ratio of the quality of sodium lauryl sulphate and ethanol) addition,
Concentration of alcohol is 25%～75%, is extracted after stirring, and extraction time is 60～100min, and Extracting temperature is 35～45 DEG C.Then
Decompression sucking filtration is carried out, residue after filtration, is discarded, the extracting solution containing lecithin is obtained.
The addition of anionic can play solubilization, addition species and addition
Impact will be produced on extraction ratio.
(4) salt precipitation
NaCl or Sal are added in filtrate, mass volume ratio (i.e. the volume ratio of the quality of salt and extracting solution) is 1:30,
Carry out salt precipitation.When salt is precipitated, it is to be ensured that NaCl solids are completely dissolved into, can be stirred, it is also possible to heat slightly.Salt
Dissolving after entering is placed on extracting solution in a low temperature of 5 DEG C and carries out, and such effect is preferable.The salt sedimentation time is 3～4h.Treat ovum
After phosphatide crude product is precipitated out, filtrate is discarded, dried or be spray-dried.Drying temperature is not above 50 DEG C.
Salt precipitation is carried out using other salt lecithin is also obtained, such as CaCl2、MgCl2Deng, but it is harmful, therefore
The present invention adopts NaCl or Sal, and finds that the extraction effect of NaCl is best in an experiment.
Describe the present invention with reference to specific embodiment, the description of this part is only exemplary and explains
Property, there should not be any restriction effect to protection scope of the present invention.
Embodiment 1
Raw material 3g is weighed, pH regulator to 7 adds papain (addition is 0.04%) to carry out enzymolysis processing about
100min, temperature are 50 DEG C, and (sodium lauryl sulphate is then to add the mixed solvent of sodium lauryl sulphate and ethanol
4% addition, concentration of alcohol are 33%) 30ml, and 60min is extracted at 35 DEG C, filter, discard residue, then in extracting solution
1g NaCl are added, salt precipitation is carried out, salt precipitation temperature is 5 DEG C, and the sedimentation time is 3 hours.Treat that lecithin crude product is precipitated out
Afterwards, dried or be spray-dried, its process chart such as Fig. 1.The extraction ratio minimum 20% or so of crude lecithin, highest can
Reach the lecithin crude product that 60% or so, i.e. 3g raw materials are at most proposed that 1.8g；Purification rate is close to 90%.
Embodiment 2
Raw material 3g is weighed, pH regulator to 3 adds pepsin (addition is 0.075%) to carry out enzymolysis processing about
60min, temperature are 37 DEG C, and (sodium lauryl sulphate is 6% then to add the mixed solvent of sodium lauryl sulphate and ethanol
Addition, concentration of alcohol is 25%) 30ml, and 60min is extracted at 40 DEG C, filters, discards residue, is then added in extracting solution
1g NaCl, carry out salt precipitation, and salt precipitation temperature is 5 DEG C, and the sedimentation time is 3.5 hours.After lecithin crude product is precipitated out,
Dried or be spray-dried.
Embodiment 3
Clarias leather leftover bits and pieces (predominantly internal organs) are minced, and is smashed to pieces with tissue mashing machine.Weigh raw material 3g, pH regulator
To 9, trypsin addition is added 0.015%) to carry out enzymolysis processing about 80min, temperature is 45 DEG C, then adds ten
(sodium lauryl sulphate is 0.67% addition to the mixed solvent of sodium dialkyl sulfate and ethanol, and 75%) concentration of alcohol is
30ml, extracts 60min at 45 DEG C, filters, discards residue, then adds 1g NaCl in extracting solution, carries out salt precipitation, salt
Precipitation temperature is 5 DEG C, and the sedimentation time is 4 hours.After lecithin crude product is precipitated out, is dried or be spray-dried.
Test example
1. the lecithin crude product for, obtaining above-described embodiment, carries out uv absorption after conventionally being dissolved with ethanol
Spectral scan, obtains Fig. 2.As can be seen from the figure its maximum absorption wavelength is consistent with document report (according to the Chinese people altogether
With state the Sanitation Ministry medicine standard second), ultraviolet maximum absorption wavelength all at 215nm, therefore, the product for being extracted is lecithin
Fat.
2., assay method
According to GB/T 5537-2008《The measure of grain and oil detection content of phospholipid》, phospholipid is determined according to molybdenum blue colorimetric method contain
Amount.At 650nm, the standard curve of potassium dihydrogen phosphate, such as Fig. 3 are made.
After the lecithin for being extracted is developed the color with sodium molybdate, absorbance is measured, the quality of phosphorus is calculated according to standard curve,
Then extraction ratio is calculated according to the following equation.
Extraction ratio (%)=[(P × 742.1)/30.97] × 100%
P- phosphorus contents；Natural phosphatidyl choline is C40H84NO6P.2H2O, molecular weight are 742.1；The atomic weight of phosphorus is 30.97.
3., the impact that enzymolysis processing is extracted to lecithin in clarias leather leftover bits and pieces
Impact-orthogonal design table that 1 enzymolysis processing of table is extracted to lecithin in clarias leather leftover bits and pieces
4., the impact that the species of salt is extracted to lecithin in clarias leather leftover bits and pieces
The impact that the species of 2 salt of table is extracted to lecithin in clarias leather leftover bits and pieces
| Salt species | NaCl | FeCl3 | ZnCl2 | MgCl2 | MnCl2 | CaCl2 | CdCl2 | K2SO4 |
| Extraction ratio (%) | 54.60 | 13.48 | 9.20 | 30.12 | 24.32 | 40.12 | 20.48 | 29.64 |
Show in table 2, when NaCl extracts lecithin in clarias leather leftover bits and pieces, effect is best, and no toxicity.
5., the impact that sodium lauryl sulphate and alcohol mixed solvent are extracted to lecithin in clarias leather leftover bits and pieces
The impact that 3 sodium lauryl sulphate of table and alcohol mixed solvent are extracted to lecithin in clarias leather leftover bits and pieces-
Orthogonal design table
Due to lecithin, in fish head, fish scale, internal organs especially fish roe, content is higher, and these in many fish market or
Catering companies exactly will be thrown away as the leftover bits and pieces of fish, and such as Beijing Xin La roads Catering Management company limited produces consumption every year
3000 tons of clarias leathers, about produce 30% leftover bits and pieces, mainly with fish head, based on fish internal organs, if all as garbage emission, will be right
Environment causes certain impact, if the leftover bits and pieces of Fish can be made full use of to extract lecithin, if the extraction ratio by 20% is calculated,
The annual leftover bits and pieces of the said firm can produce 180 tons of lecithin crude products.If pressing 50 yuan of 1kg lecithin calculates (Semen sojae atricolor ovum on the market
The price of phospholipid), then can sell 900,000 yuan within 1 year.If the leftover bits and pieces of Fish can be made full use of to extract lecithin, by change give up into
Treasured, greatly promotes expanding economy.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (3)
1. a kind of use clarias leather leftover bits and pieces extract the technique of lecithin, comprise the following steps：
(1), the head and internal organs of clarias leather, temperature are carried out mincing below 50 DEG C, are smashed to pieces；
(2), the pH value of Acetic acid-sodium acetate buffer regulating step (1) stamping is added, and the pH value is protease hydrolyzed pH value, plus
Heat adds 0.015～0.075% protease to 37～45 DEG C, digests 60～100min；
(3) extracting solution is added after, (2) digesting to step in material, is stirred, 60～100min is extracted at 35～45 DEG C；Decompression is taken out
Filter to obtain extracting solution；The extracting solution is sodium lauryl sulphate and the mixed solvent of ethanol, the quality volume of sodium lauryl sulphate
Than for 0.67%～6%, concentration of alcohol is 25%～75%；
(4), in step (3) extracting solution add NaCl, stirring and dissolving to stand 3～4 hours, obtain precipitate, do at less than 50 DEG C
It is dry.
2. the technique for extracting lecithin with clarias leather leftover bits and pieces as claimed in claim 1, it is characterised in that：The protease is wood
Melon protease, pepsin or trypsin.
3. the technique for extracting lecithin with clarias leather leftover bits and pieces as claimed in claim 1, it is characterised in that：Step (4) middle NaCl
It is 1 that addition is mass volume ratio:30.
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| CN109081850A (en) * | 2018-10-26 | 2018-12-25 | 浙江海洋大学 | A method of high bioactivity phosphatide is prepared from tuna eye |
| CN116585356A (en) * | 2023-06-08 | 2023-08-15 | 广州家化化学有限公司 | Preparation method and application of salmon roe extract |
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| JP2004002663A (en) * | 2002-03-29 | 2004-01-08 | Shizuoka Prefecture | Method for extracting lipid composition containing phospholipid with polyunsaturated fatty acid as constituent from fish internal organ |
| 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 |
| KR101258743B1 (en) * | 2010-10-06 | 2013-04-29 | 부경대학교 산학협력단 | Ａ ｍｅｔｈｏｄ ｆｏｒ ｅｘｔｒａｃｔｉｎｇ ｌｅｃｉｔｈｉｎ ｆｒｏｍ ｆｉｓｈ |
| CN104098600B (en) * | 2014-07-08 | 2017-01-25 | 汪华东 | Crocodile lecithin preparation method |
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