CN105105118B - A kind of preparation method rich in EPA sea cucumber intestine lipids - Google Patents
A kind of preparation method rich in EPA sea cucumber intestine lipids Download PDFInfo
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- CN105105118B CN105105118B CN201510551605.4A CN201510551605A CN105105118B CN 105105118 B CN105105118 B CN 105105118B CN 201510551605 A CN201510551605 A CN 201510551605A CN 105105118 B CN105105118 B CN 105105118B
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- sea cucumber
- cucumber intestine
- homogenate
- solution
- chloroform
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- 210000000936 intestine Anatomy 0.000 title claims abstract description 92
- 241000251511 Holothuroidea Species 0.000 title claims abstract description 88
- 150000002632 lipids Chemical class 0.000 title claims abstract description 28
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims abstract description 66
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 63
- 239000000243 solution Substances 0.000 claims abstract description 51
- 239000007853 buffer solution Substances 0.000 claims abstract description 23
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims abstract description 20
- 230000009514 concussion Effects 0.000 claims abstract description 14
- 229910052757 nitrogen Inorganic materials 0.000 claims abstract description 10
- CBMPTFJVXNIWHP-UHFFFAOYSA-L disodium;hydrogen phosphate;2-hydroxypropane-1,2,3-tricarboxylic acid Chemical compound [Na+].[Na+].OP([O-])([O-])=O.OC(=O)CC(O)(C(O)=O)CC(O)=O CBMPTFJVXNIWHP-UHFFFAOYSA-L 0.000 claims abstract description 4
- 238000000605 extraction Methods 0.000 claims description 14
- 238000005119 centrifugation Methods 0.000 claims description 8
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 claims description 3
- 239000000460 chlorine Substances 0.000 claims description 3
- 229910052801 chlorine Inorganic materials 0.000 claims description 3
- 239000000872 buffer Substances 0.000 claims description 2
- 239000004519 grease Substances 0.000 abstract description 14
- 238000000034 method Methods 0.000 abstract description 14
- 238000007664 blowing Methods 0.000 abstract description 9
- 239000012190 activator Substances 0.000 abstract description 2
- 238000000926 separation method Methods 0.000 abstract 1
- 239000007788 liquid Substances 0.000 description 10
- 239000008367 deionised water Substances 0.000 description 7
- 229910021641 deionized water Inorganic materials 0.000 description 7
- -1 disodium hydrogen Chemical class 0.000 description 7
- 239000011521 glass Substances 0.000 description 7
- 239000001257 hydrogen Substances 0.000 description 7
- 229910052739 hydrogen Inorganic materials 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Substances OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 6
- 238000011084 recovery Methods 0.000 description 5
- 238000010521 absorption reaction Methods 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 235000019784 crude fat Nutrition 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 238000003754 machining Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 241000965254 Apostichopus japonicus Species 0.000 description 1
- 241000208340 Araliaceae Species 0.000 description 1
- 241000941624 Holothuria atra Species 0.000 description 1
- 206010020466 Hunger Diseases 0.000 description 1
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 1
- 235000003140 Panax quinquefolius Nutrition 0.000 description 1
- 239000011149 active material Substances 0.000 description 1
- 125000001931 aliphatic group Chemical group 0.000 description 1
- 230000003139 buffering effect Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
- 235000001497 healthy food Nutrition 0.000 description 1
- 235000003642 hunger Nutrition 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000003760 tallow Substances 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
Abstract
The invention discloses a kind of preparation method rich in EPA sea cucumber intestine lipids, including step：Sea cucumber intestine is pre-processed, homogenate, obtains sea cucumber intestine homogenate, the McIIvaines buffer solutions of disodium hydrogen phosphate citric acid solution are added in homogenate, then add the concussion of chloroform/methanol solution to extract, mixed liquor is put into centrifuge separation lipid layer, nitrogen obtains sea cucumber intestine grease after blowing.The present invention has technique simple, simple to operate, the characteristics of efficiency high, and extracting method mild condition can preserve original physiological activator in sea cucumber intestine lipid to greatest extent, product is had trophism and feature concurrently.
Description
Technical field
The present invention relates to a kind of preparation method of sea cucumber intestine lipid, EPA sea cucumber tripe tallows are rich in more specifically to one kind
The preparation method of matter.
Background technology
Sea cucumber intestine is the accessory substance during Holothurian machining, and its fat content is up to 7.80 ± 0.49% or so (with dry weight
Meter), unrighted acid relative amount is up to 84.62% or so, and EPA content is higher, therefore, and sea cucumber intestine lipid is many insatiable hungers
With the good source of aliphatic acid, there is good development prospect in healthy food field.In China, sea cucumber intestine is more by as discarded
It is extremely low that thing loses utilization rate, causes the wasting of resources and environmental pollution.
On the document report of sea cucumber intestine grease, it is concentrated mainly on and lipid content is studied.South Korea scholar is to from Yu Ling
Island and the Red sea of Dokdo ginseng and Holothuria atra crude fat are determined, and as a result show that wall of sea cucumber Stichopus japonicus and internal organ crude fat content are accounted for
0.14~2.12%.However, the preparation for grease in sea cucumber intestine is there is not yet relevant report.
This patent uses McIIvaines buffer solution combination organic solvents in the method for extracting sea cucumber intestine grease, with technique
Simply, easy to operate, the characteristics of efficiency high, extracting method mild condition can be preserved original in sea cucumber intestine grease to greatest extent
Physiological activator, makes product have trophism and feature concurrently, and the exploitation for sea cucumber intestine resource have great importance.
The content of the invention
Unrighted acid relative amount is up to 84.62% in sea cucumber intestine, and EPA content is higher, and oily in sea cucumber intestine
The method that is prepared by of fat there is no report.The preparation problem of grease, EPA is rich in the invention provides one kind in unresolved sea cucumber intestine
The preparation method of sea cucumber intestine lipid, the characteristics of the method for the invention has mild condition, production efficiency is high is made by this method
Sea cucumber intestine lipid be rich in EPA.
A kind of preparation method rich in EPA sea cucumber intestine lipids of the present invention, it is characterised in that comprise the following steps：
S1, sea cucumber intestine pretreatment, homogenate obtain sea cucumber intestine homogenate；
S2, the addition McIIvaines buffer solutions into sea cucumber intestine homogenate described in step S1, are well mixed, obtain sea cucumber intestine
Mixed liquor；The McIIvaines pH of buffer is 3~8；
S3, chloroform/methanol solution concussion 1~2min of extraction into sea cucumber intestine mixed liquor described in step S2, obtain sea cucumber intestine
Extract solution；The imitative volume ratio with methanol of the chloroform/methanol Chlorine in Solution is 2:1；
S4, sea cucumber intestine extract solution described in step S3 centrifuged, collect lower floor's chloroform layer solution；The chloroform layer solution is through nitrogen
It is sea cucumber intestine lipid after blowing；
The volume ratio of the sea cucumber intestine homogenate, McIIvaines buffer solutions and chloroform/methanol solution is 1:1~3:1~
3。
Under preferred embodiment, McIIvaines buffer solutions described in step S2 are disodium hydrogen phosphate-citric acid solution.It is optimal
Under mode, McIIvaines pH of cushioning fluid described in step S2 is 3；The volume that the McIIvaines buffer solutions are added is step
3 times of sea cucumber intestine homogenate described in S1.
Under preferred embodiment, described in step S3 after the concussion extraction of chloroform/methanol solution is added, chloroform concussion extraction is added
1~2min；The volume ratio of the sea cucumber intestine homogenate and chloroform/methanol solution, chloroform is 1:1~3:1~3.Under optimum way,
Step S3 adds chloroform concussion 1min first to add the concussion 1min of chloroform/methanol solution；The sea cucumber intestine homogenate and chlorine
Imitative/methanol solution, the volume ratio of chloroform are 1:3:1.
Under preferred embodiment, the condition centrifuged described in step S4 is that 2000~5000r/min centrifuges 5~10min.
The technological innovation of the present invention is：
1st, the utilization rate of Holothurian machining byproduct in process thing is improved, its active material contained is fully developed profit
With while reducing the pollution to environment.Sea cucumber intestine lipid after working process is rich in EPA, can be used to open as function base-material
Send out several functions product.
2nd, present invention extraction grease process temperature is low, and some original physiology in sea cucumber intestine lipid can be preserved to greatest extent and are lived
Property material, makes product have trophism and feature concurrently；
3rd, operating process of the present invention is simple, it is not necessary to complicated equipment, and sea cucumber intestine lipids extraction rate is high, reaches
5.4%, EPA relative amount improve production efficiency up to 30.97%, are adapted to industrialized production.
Brief description of the drawings
Fig. 1 is the recovery rate that different pH extract sea cucumber intestine lipid.
Embodiment
The inventive method is concretely comprised the following steps：
S1, take fresh sea cucumber intestine, remove silt in intestines, rinsed well with deionized water, homogenate obtains sea cucumber intestine homogenate
Liquid；
PH3 is added in S2, sea cucumber intestine homogenate, pH4, pH5, pH6, pH7, pH8 disodium hydrogen phosphate-lemon acid buffering is molten
The McIIvaines buffer solutions (1~3 of liquid:1)；
The chloroform/methanol (2 of 1~3 times of volume is added in mixed liquor obtained by S3, step S2：1) solution shakes 1~2min, so
The chloroformic solution for adding 1~3 times of volume afterwards shakes 1~2min；
Mixed liquor obtained by S4, step S3 is put into centrifuge and centrifuges 5~10min with 2000~5000r/min, treats that sample is mixed
After liquid centrifugation layering, draw lower floor's chloroform layer solution and be transferred in glass tube, nitrogen obtains sea cucumber intestine grease after blowing.
Under above-mentioned preferred embodiment, optimal situation is：
The McIIvaines pH of cushioning fluid of disodium hydrogen phosphate-citric acid solution is 3 in step S2, and adding proportion is
1:3。
3 times of volume of chloroform/methanol (2 are first added in step S3：1) the concussion 1min of solution, then adds 1 times of volume
Chloroformic solution shakes 1min.
Embodiment 1
1st, fresh sea cucumber intestine is taken, silt in intestines is removed, is rinsed well with deionized water, homogenate obtains sea cucumber intestine homogenate
Liquid；
2nd, sea cucumber intestine homogenate 1ml, adds the McIIvaines buffer solutions of pH3 disodium hydrogen phosphates-citric acid solution
3ml；
3rd, 3ml chloroform/methanol (2 is added in mixed liquor：1) solution concussion 2min, then adds 1ml chloroformic solution
Shake 1min；
4th, mixed liquor is put into centrifuge and centrifuges 10min with 4000r/min, after the centrifugation of sample mixed liquor is layered, under absorption
Layer chloroform layer solution is transferred in glass tube, and nitrogen obtains sea cucumber intestine grease, extraction rate reached 5.4% after blowing, EPA relative amounts are
31.13%.
Embodiment 2
1st, fresh sea cucumber intestine is taken, silt in intestines is removed, is rinsed well with deionized water, homogenate obtains sea cucumber intestine homogenate
Liquid；
2nd, sea cucumber intestine homogenate 1ml, adds the McIIvaines buffer solutions of pH4 disodium hydrogen phosphates-citric acid solution
3ml；
3rd, 2ml chloroform/methanol (2 is added in mixed liquor：1) solution concussion 2min, then adds 1ml chloroformic solution
Shake 1min；
4th, mixed liquor is put into centrifuge and centrifuges 10min with 2000r/min, after the centrifugation of sample mixed liquor is layered, under absorption
Layer chloroform layer solution is transferred in glass tube, and nitrogen obtains sea cucumber intestine grease, extraction rate reached 4.3% after blowing, EPA relative amounts are
29.87%.
Embodiment 3
1st, fresh sea cucumber intestine is taken, silt in intestines is removed, is rinsed well with deionized water, homogenate obtains sea cucumber intestine homogenate
Liquid；
2nd, sea cucumber intestine homogenate 1ml, adds the McIIvaines buffer solutions of pH5 disodium hydrogen phosphates-citric acid solution
2ml；
3rd, 2ml chloroform/methanol (2 is added in mixed liquor：1) solution concussion 2min, then adds 2ml chloroformic solution
Shake 1min；
4th, mixed liquor is put into centrifuge and centrifuges 5min with 5000r/min, after after the centrifugation layering of sample mixed liquor, draws lower floor
Chloroform layer solution is transferred in glass tube, and nitrogen obtains sea cucumber intestine grease, extraction rate reached 4.1% after blowing, and EPA relative amounts are
26.02%.
Embodiment 4
1st, fresh sea cucumber intestine is taken, silt in intestines is removed, is rinsed well with deionized water, homogenate obtains sea cucumber intestine homogenate
Liquid；
2nd, sea cucumber intestine homogenate 1ml, adds the McIIvaines buffer solutions of pH6 disodium hydrogen phosphates-citric acid solution
3ml；
3rd, 2ml chloroform/methanol (2 is added in mixed liquor：1) solution concussion 1min, then adds 1ml chloroformic solution
Shake 2min；
4th, mixed liquor is put into centrifuge and centrifuges 10min with 3000r/min, after the centrifugation of sample mixed liquor is layered, under absorption
Layer chloroform layer solution is transferred in glass tube, and nitrogen obtains sea cucumber intestine grease, extraction rate reached 3.3% after blowing, EPA relative amounts are
28.86%.
Embodiment 5
1st, fresh sea cucumber intestine is taken, silt in intestines is removed, is rinsed well with deionized water, homogenate obtains sea cucumber intestine homogenate
Liquid；
2nd, sea cucumber intestine homogenate 1ml, adds the McIIvaines buffer solutions of pH7 disodium hydrogen phosphates-citric acid solution
1ml；
3rd, 2ml chloroform/methanol (2 is added in mixed liquor：1) solution concussion 2min, then adds 3ml chloroformic solution
Shake 1min；
4th, mixed liquor is put into centrifuge and centrifuges 10min with 4000r/min, after the centrifugation of sample mixed liquor is layered, under absorption
Layer chloroform layer solution is transferred in glass tube, and nitrogen obtains sea cucumber intestine grease, extraction rate reached 2.6% after blowing, EPA relative amounts are
30.24%.
Embodiment 6
1st, fresh sea cucumber intestine is taken, silt in intestines is removed, is rinsed well with deionized water, homogenate obtains sea cucumber intestine homogenate
Liquid；
2nd, sea cucumber intestine homogenate 1ml, adds the McIIvaines buffer solutions of pH8 disodium hydrogen phosphates-citric acid solution
1ml；
3rd, 1ml chloroform/methanol (2 is added in mixed liquor：1) solution concussion 2min, then adds 3ml chloroformic solution
Shake 2min；
4th, mixed liquor is put into centrifuge and centrifuges 5min with 3000r/min, after after the centrifugation layering of sample mixed liquor, draws lower floor
Chloroform layer solution is transferred in glass tube, and nitrogen obtains sea cucumber intestine grease, extraction rate reached 2.9% after blowing, and EPA relative amounts are
29.71%.
Table 1 is influences of the pH to EPA in sea cucumber intestine lipid, when the pH of McIIvaines buffer solutions is 3, sea cucumber intestine lipid
EPA relative amount highests, reach 31.13%, with pH rise, EPA relative amounts present and first reduce and raises afterwards in lipid
Trend.When the pH of McIIvaines buffer solutions is 5, EPA relative amounts minimum 26.02% in sea cucumber intestine lipid；When
When the pH of McIIvaines buffer solutions rises to 6,7 and 8, EPA relative amounts gradually go up in sea cucumber intestine lipid.
Table 1
The state that sea cucumber intestine lipid is extracted under condition of different pH is different, and when cushioning liquid pH is 3, the color of lipid layer is most
Deep, emulsion layer is most thin.In the range of pH4~8, with gradually stepping up for pH, lipid layer color gradually becomes shallower as, and emulsion layer is gradually
Increase.
Influences of the pH to sea cucumber intestine lipids extraction as seen from Figure 1, when the pH of McIIvaines buffer solutions is 3, sea
Join the recovery rate highest of tripe tallow matter, reach 5.40 ± 0.15%, with the pH neutrality that is increased to, lipids extraction rate is gradually reduced.
When the pH of McIIvaines buffer solutions is 4 and 5, sea cucumber intestine recovery rate has no significant difference (p>0.05)；Work as McIIvaines
When the pH of buffer solution rises to 6 and 7, sea cucumber intestine recovery rate is remarkably decreased, and is reached minimum.When the pH of McIIvaines buffer solutions is
When 8, slightly has rise compared with pH7 recovery rates.
The foregoing is only a preferred embodiment of the present invention, but protection scope of the present invention be not limited thereto,
Any one skilled in the art in the technical scope of present disclosure, technique according to the invention scheme and its
Inventive concept is subject to equivalent substitution or change, should all be included within the scope of the present invention.
Claims (1)
1. a kind of preparation method rich in EPA sea cucumber intestine lipids, it is characterised in that comprise the following steps：
S1, sea cucumber intestine pretreatment, homogenate obtain sea cucumber intestine homogenate；
S2, addition McIIvaines (disodium hydrogen phosphate-citric acid) buffer solution into sea cucumber intestine homogenate described in step S1, mixing
Uniformly, sea cucumber intestine mixed liquor is obtained；The McIIvaines pH of buffer is 3；
The volume that the McIIvaines buffer solutions are added is 3 times of sea cucumber intestine homogenate described in step S1；
S3, addition chloroform/methanol solution concussion 1~2min of extraction into sea cucumber intestine mixed liquor described in step S2, obtain sea cucumber intestine
Extract solution；The imitative volume ratio with methanol of the chloroform/methanol Chlorine in Solution is 2:1；
The volume ratio of the sea cucumber intestine homogenate, McIIvaines buffer solutions and chloroform/methanol solution is 1:3:1~3；
S4, sea cucumber intestine extract solution described in step S3 centrifuged, collect lower floor's chloroform layer solution；After the chloroform layer solution blows through nitrogen
As sea cucumber intestine lipid；
The condition of the centrifugation is that 2000~5000r/min centrifuges 5~10min.
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| CN201510551605.4A CN105105118B (en) | 2015-09-01 | 2015-09-01 | A kind of preparation method rich in EPA sea cucumber intestine lipids |
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| CN201510551605.4A CN105105118B (en) | 2015-09-01 | 2015-09-01 | A kind of preparation method rich in EPA sea cucumber intestine lipids |
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| CN105105118B true CN105105118B (en) | 2017-09-01 |
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| CN109793150A (en) * | 2019-03-02 | 2019-05-24 | 大连棒棰岛海产股份有限公司 | Sea cucumber intestine extract solid seasoning and its preparation method |
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| US6055936A (en) * | 1997-01-21 | 2000-05-02 | Collin; Peter Donald | Sea cucumber carotenoid lipid fractions and process |
| CN1293175C (en) * | 2004-11-10 | 2007-01-03 | 杨玉赞 | Method for extracting gut tallow from alaska pollack |
| CN103141872A (en) * | 2011-12-22 | 2013-06-12 | 大连工业大学 | Method for preparing active substance by sea cucumber intestine autolysis |
| CN102660377A (en) * | 2012-05-28 | 2012-09-12 | 杨玉赞 | Method for extracting animal offal oil |
| CN103749940B (en) * | 2014-01-10 | 2016-08-17 | 上海海洋大学 | A kind of saury internal organs method of comprehensive utilization |
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