CN107629871A - A kind of preparation method of swimming crab crab oil active phospholipid - Google Patents
A kind of preparation method of swimming crab crab oil active phospholipid Download PDFInfo
- Publication number
- CN107629871A CN107629871A CN201710866649.5A CN201710866649A CN107629871A CN 107629871 A CN107629871 A CN 107629871A CN 201710866649 A CN201710866649 A CN 201710866649A CN 107629871 A CN107629871 A CN 107629871A
- Authority
- CN
- China
- Prior art keywords
- crab
- crab oil
- oil
- swimming
- lipid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Fats And Perfumes (AREA)
Abstract
A kind of preparation method of swimming crab crab oil active phospholipid, the preparation method comprise the following steps：（1）Fresh swimming crab is chosen, is cleaned, removes shell on the back, draws crab oil part；（2）Crab oil is divided, dries, obtains dry crab oil；（3）Dried crab oil is crushed with low speed pulverizer, is digested under Papain enzyme effect, and batch (-type) is aided with ultrasound-assisted enzymolysis per hour；（4）After enzyme digestion reaction terminates, impurity is removed, takes upper strata oil layer, anhydrous sodium sulfate is added and stands overnight to remove unnecessary moisture；（5）100 mesh silica gel add methanol, and drying is activated after at 100 DEG C, and dichloromethane is added after cooling, add stirring to install additional into chromatographic column, treat that post layer is stable after filling post；（6）Crab oil lipid is dissolved in dichloromethane, instills in chromatographic column, does not have with dichloromethane eluent neutral fats；Phosphatide is eluted with methanol；Methanol component is collected, is concentrated with Rotary Evaporators, 50 DEG C of dryings to constant weight, obtains crab oil active phospholipid.
Description
Technical field
The present invention relates to a kind of preparation method of swimming crab crab oil phosphatide, belong to the crab oil processing technique field of swimming crab.
Background technology
Phosphatide is animals and plants and microbial cell film, the main constituents of nerve fiber, because it has stronger biology
Activity and increasingly favored by people, be widely used to food, health products, medicine and cosmetic industry.Marine organisms
Active phospholipid, because of the living environment difference residing for it, has different compounds compared with plant phosphatide and terrestrial animal phosphatide
26S Proteasome Structure and Function activity.At present, the phospholipid prod on domestic and international market, still based on soybean lecithin and yolk phospholipid, by terrestrial
It is stronger that the limitation of animal, plant fatty acid compositing characteristic, these plant phosphatide and terrestrial animal phosphatide do not include bioactivity
EPA and DHA, and the EPA and DHA content in marine organisms are higher, are the good sources for developing natural EPA/DHA phosphatide.
Swimming crab is the important economic crab in China, and year, amount of fishing was in the trend being gradually increasing.The swimming crab product in China
Listing is concentrated, measures that big valency is low, and shelf life is short, and product category is single, based on the primary product such as the crab that lives, added value is not high.Shuttle
Sub- crab crab oil is the accessory substance in swimming crab primary process, there are about hundreds of tons of swimming crab crab oil accessory substance every year and produces.
Lipid content enriches in swimming crab crab oil, including substantial amounts of active phospholipid composition, in phosphatide, is combined with abundant unsaturated lipid
Fat acid, good EPA, DHA etc..These phosphatide are to repairing the brain cell of human body, improving memory, prevention alzheimer's disease with very
Good bioactivity.Therefore, these swimming crab crab oil accessory substances possess higher value.
At present, swimming crab crab oil accessory substance is mainly used to Fodder making, its deep processing and utilization rate is relatively very low, added value not
It is high.On the other hand, the research to swimming crab crab oil active phospholipid not yet finds relevant report.Develop swimming crab crab oil active phospholipid
Product can both make full use of the accessory substance that swimming crab is processed, and promote the benign development of swimming crab aquatic products processing industry, again can be with
The market space of abundant ocean bioactive lipid matter, the development of health and marine health food industry to the mankind have product
Pole meaning.
The content of the invention
Present invention aims to overcome that the shortcomings of the prior art, and provide that a kind of method is simple and direct, controllability is stronger, energy
The comprehensive utilization of swimming crab processing waste resource is realized, improves the swimming crab crab oil active phospholipid of swimming crab crab oil application value
Preparation method.
The purpose of the present invention is completed by following technical solution, a kind of preparation side of swimming crab crab oil active phospholipid
Method, comprise the following steps：
(1) crab oil is taken：Fresh swimming crab is chosen, is cleaned, removes shell on the back, draws crab oil part；
(2) dry：Crab oil is divided, thickness is 0.8~1.2cm, dries, obtains dry crab oil；
(3) batch (-type) ultrasound-assisted enzymolysis：Dried crab oil is crushed with low speed pulverizer, in Papain enzyme effect
Under digested, and batch (-type) is aided with ultrasound-assisted enzymolysis per hour；
(4) lipid is separated：After enzyme digestion reaction terminates, 7,500r/min 15~20min of centrifugation take upper strata oily to remove impurity
Lipid layer, add anhydrous sodium sulfate and stand overnight to remove unnecessary moisture, crab oil lipid part can be obtained；
(5) preparative separation silicagel column：100-200 mesh silica gel adds methanol, and its mass volume ratio is 1:3 cleaning 25-
35min, dry after activating 80-100min at 100-120 DEG C, dichloromethane is added after cooling, its mass volume ratio is 1:
3, add stirring to install additional into chromatographic column, treat that post layer is stable after filling post；
(6) phosphatide separates：Crab oil lipid is dissolved in dichloromethane, its mass volume ratio is 1:3, instill in chromatographic column,
Do not have with dichloromethane eluent neutral fats, wherein crab oil lipid and dichloromethane mass volume ratio are 1:250；Phosphorus is eluted with methanol
Fat, wherein crab oil lipid are 1 with methanol quality volume ratio:300；Elution flow rate is 2.5~3ml/min；Methanol component is collected, is used
Rotary Evaporators are concentrated, 50 DEG C of dryings to constant weight, obtain crab oil active phospholipid.
As preferred：In step (3), dry Xie Huang ︰ water be 1 ︰ 5 in mass ratio, papain addition for 5000~
The dry crab oils of 7000U/g, 40~50 DEG C of hydrolysis temperature, pH value 7.5~8.5,3~4h of enzyme digestion reaction time；What is be ultrasonically treated is specific
Parameter is：Processing time is 10~30min, and ultrasonic sound field intensity is 40~60W/cm2, temperature is 50~70 DEG C, supersonic frequency
For 20~25kHz.
The present invention is to be reacted under ultrasonic assistant effect using papain enzymolysis, separates swimming crab to greatest extent
Crab oil lipid, using silica gel column chromatography Separation of Neutral fat and phosphatide, to obtain swimming crab crab oil active phospholipid.
Compared with prior art, it is of the invention to have the prominent advantages that：
1st, swimming crab crab oil lipid is separated using ultrasonic assistant papain enzymolysis method, enzymolysis efficiency is high, method letter
Just, production control is easily carried out.
2nd, compared with the extraction separation method of plant phosphatide and terrestrial animal phosphatide, neutral fats elution is substituted with dichloromethane
Toxicity stronger chloroform, the security of operating process and obtains the security of product and is improved.
Embodiment
Below by specific embodiment, the invention will be further described.A kind of swimming crab crab oil of the present invention is lived
The preparation method of property phosphatide, the preparation method comprise the following steps：
(1) crab oil is taken：Fresh swimming crab is chosen, is cleaned, removes shell on the back, draws crab oil part；
(2) dry：Crab oil is divided, thickness is 0.8~1.2cm, dries, obtains dry crab oil；
(3) batch (-type) ultrasound-assisted enzymolysis：Dried crab oil is crushed with low speed pulverizer, in Papain enzyme effect
Under digested, and batch (-type) is aided with ultrasound-assisted enzymolysis per hour；
(4) lipid is separated：After enzyme digestion reaction terminates, 7,500r/min 15~20min of centrifugation take upper strata oily to remove impurity
Lipid layer, add anhydrous sodium sulfate and stand overnight to remove unnecessary moisture, crab oil lipid part can be obtained；
(5) preparative separation silicagel column：100-200 mesh silica gel adds methanol, and its mass volume ratio is 1:3 cleaning 25-
35min, dry after activating 80-100min at 100-120 DEG C, dichloromethane is added after cooling, its mass volume ratio is 1:
3, add stirring to install additional into chromatographic column, treat that post layer is stable after filling post；
(6) phosphatide separates：Crab oil lipid is dissolved in dichloromethane, its mass volume ratio is 1:3, instill in chromatographic column,
Do not have with dichloromethane eluent neutral fats, wherein crab oil lipid and dichloromethane mass volume ratio are 1:250；Phosphorus is eluted with methanol
Fat,
Wherein crab oil lipid and methanol quality volume ratio are 1:300；Elution flow rate is 2.5~3ml/min；Collect methanol group
Point, concentrated with Rotary Evaporators, 50 DEG C of dryings to constant weight, obtain crab oil active phospholipid.
In step (3), in enzymolysis, dry Xie Huang ︰ water be 1 ︰ 5 in mass ratio, papain addition for 5000~
The dry crab oils of 7000U/g, 40~50 DEG C of hydrolysis temperature, pH value 7.5~8.5,3~4h of enzyme digestion reaction time；What is be ultrasonically treated is specific
Parameter is：
Processing time is 10~30min, and ultrasonic sound field intensity is 40~60W/cm2, temperature is 50~70 DEG C, supersonic frequency
For 20~25kHz.
Embodiment 1
(1) crab oil is taken：Fresh swimming crab is chosen, is cleaned, removes shell on the back, draws crab oil part；
(2) dry：Crab oil is divided, thickness is 0.8~1.2cm, dries, obtains dry crab oil；
(3) batch (-type) ultrasound-assisted enzymolysis：Dried crab oil is crushed with low speed pulverizer, takes the dry crab cream powders of 500g,
Add in 5L distilled water, add the dry crab oils of papain 7000U/g, 40 DEG C of hydrolysis temperature, pH value 7.5, enzyme digestion reaction time
3h；Period is aided with ultrasound-assisted enzymolysis, processing time 20min per hour, and ultrasonic sound field intensity is 50W/cm2, temperature is
60 DEG C, supersonic frequency is 20~25kHz；
(4) lipid is separated：After enzyme digestion reaction terminates, 7,500r/min centrifugation 20min take upper strata grease to remove impurity
Layer, add anhydrous sodium sulfate and stand overnight to remove unnecessary moisture, crab oil lipid part can be obtained.To obtain crab oil lipid than dry
Actual lipid calculates in crab oil, yield 87.8%；
(5) preparative separation silicagel column：50g 100-200 mesh silica gel adds 150ml methanol cleaning 30min, dries after 110
90min is activated at DEG C, 150m dichloromethane is added after cooling, adds stirring to install additional into chromatographic column, treats that post layer is stable after filling post；
(6) phosphatide separates：2g crab oil lipids are dissolved in 6ml dichloromethane, instilled in chromatographic column, with 500ml dichloromethanes
Alkane elutes neutral fats；Phosphatide is eluted with 600ml methanol；Elution flow rate is 3ml/min；Methanol component is collected, uses Rotary Evaporators
Concentrated, 50 DEG C of dryings to constant weight, obtain crab oil active phospholipid, by obtain active phospholipid than actual phosphatide in crab oil lipid in terms of
Calculate, yield 91.7%.
Embodiment 2
To verify influence of the enzymatic hydrolysis condition to preparation swimming crab crab oil lipid yield in the step of embodiment 1 (3),
Enzyme concentration, hydrolysis temperature, pH value, four factors of enzymolysis time in step (3), design the water of four factor three
Flat orthogonal experiment, remaining is with embodiment 1, so as to swimming crab crab oil lipid yield corresponding to obtaining, as shown in table 1.
1 different enzymatic hydrolysis conditions of table are to preparing swimming crab crab oil lipid yield
By table 1 it can be found that enzyme concentration is in the dry crab oils of 7000U/g, 45 DEG C of hydrolysis temperature, pH value 7.5, enzymolysis time 4h
When, crab oil lipid yield highest in orthogonal experiment, about 88.2%.
Embodiment 3
It is ultrasonication factor in the checking step of embodiment 1 (3) to the swimming crab crab oil lipid yield for preparing
Influence, three factors of temperature, processing time and ultrasonic sound field intensity in step (3), design Three factors-levels are just
Experiment is handed over, remaining is with embodiment 1, so as to swimming crab crab oil lipid yield corresponding to obtaining, as shown in table 2.
Swimming crab crab oil lipid yield obtained by the ultrasonication of the different temperatures of table 2, processing time and ultrasonic sound field intensity
By table 2 it can be found that temperature 60 C, processing time 20min, sound field intensity 50W/cm2Under ultrasonic pretreatment,
Swimming crab crab oil lipid yield highest, it is 87.8%.
Embodiment 4
To obtain the aliphatic acid composition in phosphatide in the checking step of embodiment 1 (6), gas phase is carried out to the phospholipid composition of acquisition
Chromatography/Mass Spectrometry connection usage analysis aliphatic acid composition.As shown in table 3.
The swimming crab crab oil fatty acids in phospholipids of table 3 is analyzed
Note：NA is represented and is not detected by
By table 3 it can be found that unsaturated fatty acid content accounts for the 53.7% of total lipid content, wherein EPA content is higher, accounts for
The 50.9% of unsaturated fatty acid content, but DHA is not detected.The swimming crab crab oil phosphatide for showing to obtain is a kind of high EPA content
Active phospholipid.
Embodiment described above is a kind of preferable scheme of the present invention, not the present invention is made any formal
Limitation, there are other variants and remodeling on the premise of without departing from the technical scheme described in claim.
Claims (2)
1. a kind of preparation method of swimming crab crab oil active phospholipid, it is characterised in that the preparation method comprises the following steps：
（1）Take crab oil：Fresh swimming crab is chosen, is cleaned, removes shell on the back, draws crab oil part；
（2）Dry：Crab oil is divided, thickness is 0.8 ~ 1.2cm, dries, obtains dry crab oil；
（3）Batch (-type) ultrasound-assisted enzymolysis：Dried crab oil is crushed with low speed pulverizer, is entered under Papain enzyme effect
Row enzymolysis, and batch (-type) is aided with ultrasound-assisted enzymolysis per hour；
（4）Separate lipid：After enzyme digestion reaction terminates, 7,500 r/min centrifuge 15 ~ 20min to remove impurity, take upper strata grease
Layer, add anhydrous sodium sulfate and stand overnight to remove unnecessary moisture, crab oil lipid part can be obtained；
（5）Preparative separation silicagel column：100-200 mesh silica gel adds methanol, and its mass volume ratio is 1:3 25-35min of cleaning, do
Dry to add dichloromethane after cooling after activating 80-100min at 100-120 DEG C, its mass volume ratio is 1:3, add stirring
Install additional into chromatographic column, treat that post layer is stable after filling post；
（6）Phosphatide separates：Crab oil lipid is dissolved in dichloromethane, its mass volume ratio is 1:3, instill in chromatographic column, with two
Chloromethanes elution neutral fats does not have, and wherein crab oil lipid and dichloromethane mass volume ratio are 1:250；Phosphatide is eluted with methanol, its
Middle crab oil lipid is 1 with methanol quality volume ratio:300；Elution flow rate is 2.5 ~ 3ml/min；Methanol component is collected, is steamed with rotation
Hair instrument is concentrated, and 50 DEG C of dryings to constant weight, obtains crab oil active phospholipid.
2. the preparation method of swimming crab crab oil active phospholipid according to claim 1, it is characterised in that：Step（3）In, enzyme
Xie Zhong, dry crab Huang ︰ water are 1 ︰ 5 in mass ratio, and papain addition is the dry crab oils of 5000 ~ 7000 U/g, hydrolysis temperature 40 ~
50 DEG C, pH value 7.5 ~ 8.5,3 ~ 4h of enzyme digestion reaction time；The design parameter of supersound process is：Processing time is 10 ~ 30min, is surpassed
Several field intensities are 40 ~ 60W/cm2, temperature is 50 ~ 70 DEG C, and supersonic frequency is 20 ~ 25kHz.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710866649.5A CN107629871A (en) | 2017-09-22 | 2017-09-22 | A kind of preparation method of swimming crab crab oil active phospholipid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710866649.5A CN107629871A (en) | 2017-09-22 | 2017-09-22 | A kind of preparation method of swimming crab crab oil active phospholipid |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107629871A true CN107629871A (en) | 2018-01-26 |
Family
ID=61103432
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710866649.5A Pending CN107629871A (en) | 2017-09-22 | 2017-09-22 | A kind of preparation method of swimming crab crab oil active phospholipid |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107629871A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115043872A (en) * | 2022-06-17 | 2022-09-13 | 哈尔滨工业大学 | Method for extracting deer blood phospholipid and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103773828A (en) * | 2012-10-23 | 2014-05-07 | 李丹榕 | Method for extraction of collagen from fish scale |
| CN106588977A (en) * | 2016-12-13 | 2017-04-26 | 山东省科学院生物研究所 | Method for extracting high-purity marine phospholipids from Euphausia superba |
| CN107099368A (en) * | 2017-04-10 | 2017-08-29 | 浙江海洋大学 | High EPA, DHA content a kind of swimming crab crab oil oil producing method |
-
2017
- 2017-09-22 CN CN201710866649.5A patent/CN107629871A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103773828A (en) * | 2012-10-23 | 2014-05-07 | 李丹榕 | Method for extraction of collagen from fish scale |
| CN106588977A (en) * | 2016-12-13 | 2017-04-26 | 山东省科学院生物研究所 | Method for extracting high-purity marine phospholipids from Euphausia superba |
| CN107099368A (en) * | 2017-04-10 | 2017-08-29 | 浙江海洋大学 | High EPA, DHA content a kind of swimming crab crab oil oil producing method |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115043872A (en) * | 2022-06-17 | 2022-09-13 | 哈尔滨工业大学 | Method for extracting deer blood phospholipid and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103320217A (en) | Method for extracting krill oil rich in phospholipid from euphausia superba | |
| CN103981021A (en) | Method for refining krill oil from Antarctic krill powder | |
| CN102311882B (en) | Extraction method for unsaturated fatty acid | |
| CN105601666B (en) | Method of phosphatide and products thereof is extracted in a kind of fish head from silver carp | |
| CN1904008A (en) | Method of deeply developing and utilizing seashore cheeseflower | |
| CN102277230B (en) | Method of optimum rapid solvent extracting treatment for raising extraction amount of microalgae grease | |
| CN103130909A (en) | Preparation method of selenium-rich Morchella polysaccharide | |
| CN102358865A (en) | Method of extracting Euphausia superba oil by using supercritical carbon dioxide | |
| CN104186705A (en) | Enzymatic acidolysis-based method for synthesizing structured lipids from palmitic acid triglycerides | |
| CN103666750A (en) | Refining method of peony seed oil and soapmaking technology | |
| CN101564080A (en) | Phospholipid with coordinative fatty acid ratio and preparation method thereof | |
| CN106349133A (en) | Extraction method of sulforaphane | |
| CN102942987A (en) | Method for extracting fish oil from head and internal organ of Scomber japonicus by using compound protease | |
| CN105018210A (en) | Method for enzymaticly digesting porcine fat tissue and applications thereof | |
| CN101057873A (en) | Method for preparing hepatitis B virus resisting maggot protein powder and its maggot oil and fatty acid | |
| CN102320953B (en) | Method for preparing natural alpha-linolenic acid from crude oil of idesia polycarpa var.vestita diels | |
| CN103045349A (en) | Method for extracting grease from barley pests | |
| CN107586593A (en) | A kind of method that peony seeds essential oil is extracted from peony seeds | |
| CN106832021A (en) | A kind of preparation method of Chinese cassia tree slag Thick many candies | |
| CN107629871A (en) | A kind of preparation method of swimming crab crab oil active phospholipid | |
| CN106701311A (en) | Method for preparing microalgae oil and protein polysaccharide by adopting wet algae and one-step method | |
| CN109666709A (en) | A method of diglyceride is prepared by raw material of high-acid value grease | |
| CN102311881B (en) | Method for extracting mixed aliphatic acid from abdominal fat of foie gras | |
| CN105400835B (en) | A kind of method and DHA preparing DHA using bean dregs | |
| CN106833886A (en) | A kind of method of free fatty in utilization ionic liquid removing vegetable crude oil |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180126 |
|
| RJ01 | Rejection of invention patent application after publication |