CN105147734A - Euphausia superba oil nanostructure lipid carrier and preparing method thereof - Google Patents
Euphausia superba oil nanostructure lipid carrier and preparing method thereof Download PDFInfo
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Abstract
The invention discloses a euphausia superba oil nanostructure lipid carrier. The euphausia superba oil nanostructure lipid carrier is prepared from, by mass, 3.5-6.5% of euphausia superba oil, 3.5-6.5% of solid lipid, 0.9-1.1% of emulgator and the balance water. The invention further provides a preparing method of the euphausia superba oil nanostructure lipid carrier. The preparing method includes the following steps that 1, the solid lipid and the euphausia superba oil are fused and mixed; 2, the emulgator is added into water and stirred until the emulgator is dissolved; 3, under the magnetic stirring condition, the fused mixed lipid is dripped into a water solution obtained in the step 2 while the lipid is hot; 4, primary emulsion obtained in the step 3 is homogenized; 5, the micro emulsion obtained after homogenizing in the step 4 is naturally cooled to the room temperature to obtain an aqueous dispersion of the euphausia superba oil nanostructure lipid carrier. Storage stability and bioavailability of the euphausia superba oil can be improved, and application of the euphausia superba oil to water-soluble food is broadened.
Description
Technical field
The present invention relates to a kind of preparation method of antarctic krill oil nano structured lipid carrier.
Background technology
Antarctic krill oil takes from the wild Antarctic krill in pure marine site, containing a large amount of phospholipid, omega-3 polyunsaturated fatty acids, astaxanthin, also containing vitamin A. D. E, animal Protoapigenone, chitin and trace element isoreactivity material, and safe without toxic side effect.And large quantity research show antarctic krill oil be occurring in nature uniquely with phospholipid in conjunction with ω-3, a kind of natural activity multi-effect compound molecule having high bioavailability and superpower resistance to oxidation concurrently formed with superpower natural anti-reflecting oxide bond respectively again, and the phospholipid of its phospholipid and human body cell closely, the phospholipid being therefore rich in omega-3 polyunsaturated fatty acids is highly susceptible to absorption and the utilization of human body.The domestic and international research to antarctic krill oil at present mainly concentrates on medical value aspect, and comprise blood fat reducing, prevention cardiovascular and cerebrovascular disease, antiinflammatory, alleviates premenstrual syndrome, to control and the improving studing ability of type ii diabetes.Nearly 300,000,000 people in the current whole world are at edible antarctic krill oil.
Nano structured lipid carrier (NLC) is up-to-date nano-lipid carrier, using solid-liquid mixing lipid as the substrate of bioactive ingredients, by surfactant, bioactive substance is embedded in liquid phase, form nanoscale emulsion, the imperfections crystalline structure that then recrystallization is formed under greenhouse or low temperature.NLC can increase envelop rate, drug loading, physical stability, and is that the one improving the chemical stability of lipotropy functional molecular in food, bioavailability and Co ntrolled release is selected well.Medicine is fixed in solid granulates substrate by NLC, prevents combined bioactive molecule from degrading.Lipidic matrix can protect encapsulated sensitivity as physical barriers, active molecule is destroyed by the monkey wrench of aqueous phase.NLC is widely used in pharmacy and cosmetic industry at present as the novel delivery system of one, and the applied research in food rarely has report, is scarcely out of swaddling-clothes.
Antarctic krill oil, to light, oxygen, thermo-responsive, is easily oxidized, and its hydrophobic property limits its application in food.At present, antarctic krill oil great majority exist with the form of capsule, applies it in water-soluble food and also lack effective means.Therefore, current a kind of nanometer transport system of research of needing badly is to improve the stability of the functional mass such as DHA, EPA in antarctic krill oil in processing and storing process, and Shelf-life, ensures that oxidation deterioration does not occur before absorption of human body for it; Also these application of fat-soluble functional mass in water-soluble food are increased on the other hand.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of storage-stable, bioavailability, the antarctic krill oil nano structured lipid carrier widening its application in water-soluble food and preparation method thereof that can improve antarctic krill oil.
In order to solve the problems of the technologies described above, the invention provides a kind of nano structured lipid carrier of antarctic krill oil, be made up of the component of following mass content:
Antarctic krill oil 3.5-6.5%;
Solid lipid 3.5-6.5%;
Emulsifying agent 0.9-1.1%;
Surplus is water.
The present invention also provides the preparation method of the nano structured lipid carrier of above-mentioned antarctic krill oil simultaneously, it is characterized in that comprising the following steps:
1), by solid lipid and antarctic krill oil in 65 ~ 75 DEG C of (being preferably 70 DEG C) melting mixing;
2), by emulsifying agent be added in water and stir until emulsifiers dissolve, 65 ~ 75 DEG C of (being preferably 70 DEG C) preheatings;
3), under magnetic agitation condition, by step 1) molten state of gained mixing lipid (65 ~ 75 DEG C) drops to step 2) in the aqueous solution (65 ~ 75 DEG C) that obtains, obtain colostrum;
Remarks illustrate: above-mentioned time for adding is 20 ~ 40s (being preferably 30s); The rotating speed of magnetic agitation is 3500 ~ 4500rpm (being preferably 4000rpm); After dropwising, continue stirring 0.5 ~ 1.5 minute (being preferably 1 minute);
4), by step 3) colostrum of gained utilizes high pressure homogenization method or ultrasonic method to carry out homogenizing;
5), step 4) microemulsion (for high temperature microemulsion) of gained naturally cools to room temperature (that is, being placed in room temperature borehole cooling) after homogenizing, can obtain the aqueous dispersions of antarctic krill oil nano structured lipid carrier.
In the present invention, room temperature refers generally to 15 ~ 25 DEG C.
Improvement as the preparation method of the nano structured lipid carrier of antarctic krill oil of the present invention: described solid lipid is palm stearin (Glycerylpalmitostearate, fusing point 58.6, iodine number <17) or glyceryl tristearate.
Remarks illustrate: palm stearin full name: glyceryl palmitostearate.Inform in the following documents: Vivek, K., Reddy, H., & Murthy, R.S.R. (2007) .Investigationsoftheeffectofthelipidmatrixondrugentrapme nt, invitrorelease, andphysicalstabilityofolanzapine-loadedsolidlipidnanopar ticles.AAPSPharmaceuticalScienceandTechnology, 8 (4), 16 – 24.
Further improvement as the preparation method of the nano structured lipid carrier of antarctic krill oil of the present invention: described emulsifying agent is soybean lecithin or Tween 80.
Further improvement as the preparation method of the nano structured lipid carrier of antarctic krill oil of the present invention: described step 4) in,
High pressure homogenization method is: by step 3) to pour setting pressure into be carry out circulation homogenizing (circulation homogenizing can be 4 ~ 6 times) in the high pressure homogenizer of 70 ~ 90MPa (being preferably 80MPa) for the colostrum of gained;
Described ultrasonic method is: by step 3) colostrum of gained pulverizes 4 ~ 6 minutes (being preferably 5 minutes) through ultrasonic probe with 180 ~ 220W (being preferably 200W) power ultrasonic.
The present invention, using the carrier system of nano structured lipid carrier as the bioactive substance of antarctic krill oil, improves its dissolubility in water, improves bioavailability and provide good physical and chemical stability.Instant invention overcomes the difficult problem be applied to by antarctic krill oil in water-soluble food (beverage, milk product etc.), also a kind of effective new method is provided to the shelf life of the food extending antarctic krill oil and strengthening thereof.
The mean diameter of the antarctic krill oil nano structured lipid carrier in the aqueous dispersions that the present invention prepares and obtains is about 143.5nm (Fig. 1), antarctic krill oil nano structured lipid carrier globulate or elliposoidal (Fig. 2) under transmission electron microscope.In the antarctic krill oil nano structured lipid carrier of preparation, the envelop rate of DHA, EPA and astaxanthin is all greater than 96%.The antarctic krill oil nano structured lipid carrier physically stable of preparation is good, keeps mean diameter after 70 days to be no more than 250nm, and its aqueous dispersions under rotating speed 10000 revs/min after centrifugal 5 minutes mean diameter change not obvious, without obvious lamination.The antarctic krill oil nano structured lipid carrier of preparation can significantly improve the photooxidation stability (Fig. 3) of DHA, EPA and astaxanthin in antarctic krill oil.Preparation process of the present invention is simple, and repeatability is high.
The present invention has following technical characterstic:
1, on the basis of great many of experiments, suitable solid lipid of the present invention, emulsifying agent is obtained; Thus successfully prepare antarctic krill oil nano structured lipid carrier;
2, on the basis of great many of experiments, formula of the present invention is obtained, thus realize under the condition only adopting a kind of emulsifying agent, content is more than 50% (namely in TL for liquid fatty substance antarctic krill oil, ensure that the load capacity of antarctic krill oil is high), and still can keep high stability and high encapsulation rate.
3, the antarctic krill oil nano structured lipid carrier physical stability that obtains of the present invention is good, the holding time is long, can significantly improve the photooxidation stability of antarctic krill oil.
Accompanying drawing explanation
Below in conjunction with accompanying drawing, the specific embodiment of the present invention is described in further detail.
Fig. 1 is the particle diameter of the antarctic krill oil nano structured lipid carrier that the present invention obtains.
Fig. 2 is the transmission electron microscope photo of the antarctic krill oil nano structured lipid carrier that the present invention obtains.
Fig. 3 is that the antarctic krill oil nano structured lipid carrier that obtains of the present invention is to the design sketch improving antarctic krill oil photooxidation stability.Vertical coordinate is the content of DHA, EPA and astaxanthin, and abscissa is the time.
In Fig. 3, EPAinNLC represents the content of EPA in the embodiment of the present invention 1 gained antarctic krill oil nano structured lipid carrier; EPAinkrilloil represents EPA content in antarctic krill oil; DHAinNLC represents the content of DHA in the embodiment of the present invention 1 gained antarctic krill oil nano structured lipid carrier; DHAinkrilloil represents DHA content in antarctic krill oil.
AstaxanthininNLC represents the content of astaxanthin in the embodiment of the present invention 1 gained antarctic krill oil nano structured lipid carrier; Astaxanthininkrilloil represents the content of astaxanthin in antarctic krill oil.
Detailed description of the invention
The formula of embodiment 1, antarctic krill oil nano structured lipid carrier is made up of the component of following mass percent:
Antarctic krill oil 6.5%; Palm stearin 3.5%; Soybean lecithin 1.07%; Water 88.93%.
The preparation method of this antarctic krill oil nano structured lipid carrier is:
(1) melting mixing at 70 DEG C by solid lipid and antarctic krill oil;
(2) get emulsifying agent and be added to stirring and dissolving in water, 70 DEG C of preheatings;
(3) molten state mixing lipid step (1) obtained dropwise joins in the aqueous solution that step (2) obtains (above-mentioned time for adding is about 30s) while hot, and the process that drips adopts magnetic stirrer (rotating speed is 4000rpm); After dropwising, continue magnetic agitation 1 minute, make colostrum;
(4) colostrum that step (3) obtains is poured into the high pressure homogenizer circulation homogenizing 5 times that setting pressure is 80MPa.
(5) the high temperature microemulsion that step (4) obtains is placed in room temperature borehole cooling, the aqueous dispersions of the nano structured lipid carrier taking antarctic krill oil as substrate can be obtained.
The mean diameter of the antarctic krill oil nano structured lipid carrier that embodiment 1 is prepared in the aqueous dispersions of gained is about 143.5nm.
Embodiment 2, antarctic krill oil nano structured lipid carrier formula are made up of the component of following mass percent:
Antarctic krill oil 5%; Glyceryl tristearate 5%; Tween 1%; Water 89%.
The preparation method of this antarctic krill oil nano structured lipid carrier is:
(1) melting mixing at 70 DEG C by solid lipid and antarctic krill oil;
(2) get emulsifying agent and be added to stirring and dissolving in water, 70 DEG C of preheatings;
(3) molten state mixing lipid step (1) obtained dropwise joins in the aqueous solution that step (2) obtains (above-mentioned time for adding is about 30s) while hot, and the process that drips adopts magnetic stirrer (rotating speed is 4000rpm); After dropwising, continue magnetic agitation 1 minute, make colostrum;
(4) colostrum that step (3) obtains is pulverized 5 minutes through ultrasonic probe with 200W power ultrasonic;
(5) the high temperature microemulsion that step (4) obtains is placed in room temperature borehole cooling, the aqueous dispersions of the nano structured lipid carrier taking antarctic krill oil as substrate can be obtained.
The mean diameter of the antarctic krill oil nano structured lipid carrier that embodiment 2 is prepared in the aqueous dispersions of gained is about 154.2nm.
Embodiment 3, antarctic krill oil nano structured lipid carrier formula are made up of the component of following mass percent:
Antarctic krill oil 3.5%; Palm stearin 6.5%; Tween 0.9%; Water 89.1%.
Preparation method is equal to embodiment 1.
The mean diameter of the antarctic krill oil nano structured lipid carrier that embodiment 3 is prepared in the aqueous dispersions of gained is about 160.5nm.
Embodiment 4: antarctic krill oil nano structured lipid carrier formula is made up of the component of following mass percent:
Antarctic krill oil 6.5%; Glyceryl tristearate 3.5%; Soybean lecithin 1.1%; Water 88.9%.
Preparation method is equal to embodiment 2.
The mean diameter of the antarctic krill oil nano structured lipid carrier that embodiment 4 is prepared in the aqueous dispersions of gained is about 148.6nm.
Experiment 1, adopt GB28404 method (surveying DHA, EPA) and UV-VIS spectrophotometry (survey astaxanthin) to prepare above-described embodiment 1 ~ embodiment 4 antarctic krill oil nano structured lipid carrier in the envelop rate of DHA, EPA and astaxanthin detect, acquired results is respectively as following table 1.
Table 1
| Envelop rate | DHA(％) | EPA(％) | Astaxanthin (%) |
| Embodiment 1 | 96.79±0.79 | 96.66±0.54 | 97.29±0.42 |
| Embodiment 2 | 96.51±0.56 | 96.48±0.42 | 96.83±0.53 |
| Embodiment 3 | 98.24±0.47 | 98.67±0.53 | 99.11±0.33 |
| Embodiment 4 | 96.03±0.54 | 96.20±0.39 | 97.14±0.46 |
Remarks illustrate: the assay method that UV-VIS spectrophotometry analyzes astaxanthin is mixed by a certain amount of sample and 60% (v/v) normal hexane/aqueous isopropanol, carry out 3 extractions.Then nitrogen dries up solvent, and the dry extract obtained is dissolved in be determined in the normal hexane of volume.Its absorbance is measured under 472nm wavelength, last basis is by S á nchez-Camargo, Martinez-Correa, Paviani, andCabral (2011) and Tolasa, the standard curve that the method for Cakli, andOstermeyer (2005) is done, calculates the content of astaxanthin.
Experiment 2, the antarctic krill oil nano structured lipid carrier of above-described embodiment 1 ~ embodiment 4 gained is preserved 70 days after mean diameter be no more than 250nm, and its aqueous dispersions under rotating speed 10000 revs/min after centrifugal 5 minutes mean diameter change not obvious, without obvious lamination.
This experiment shows that antarctic krill oil nano structured lipid carrier physically stable of the present invention is good.
The photooxidation stability experiment of DHA, EPA and astaxanthin is carried out in experiment 3, the antarctic krill oil nano structured lipid carrier (NLC) embodiment 1 prepared, 5gNLC (containing 0.325g krill oil) and 0.325g antarctic krill oil (in contrast) are placed in teat glass respectively, in ventilation, are exposed in 110W ultraviolet lighting (dual wavelength 253.7nm+185nm) 60 hours under room temperature condition.Every 10 hours, measure the content of bioactive substance (DHA, EPA, astaxanthin).Acquired results is as Fig. 3.
Calculate reaction rate constant by figure, the reaction rate constant of each material accurately sorts as follows: the DHA (0.02435 ± 0.00108h in antarctic krill oil
-1) EPA (0.02332 ± 0.00095h in > antarctic krill oil
-1) astaxanthin (0.01290 ± 0.00067h in > antarctic krill oil
-1) DHA (0.00581 ± 0.00028h in >NLC
-1) EPA (0.00463 ± 0.00019h in >NLC
-1) astaxanthin (0.00358 ± 0.00020h in >NLC
-1).Visible, when being exposed under ultraviolet light, be encapsulated in the degradation speed of bioactive substance degradation rate well below the bioactive molecule in krill oil former state of antarctic krill oil in NLC.Show that antarctic krill oil NLC of the present invention significantly can strengthen antarctic krill oil photooxidation stability, reduce its degradation rate.
Comparative example 1, make the palm stearin in embodiment 1 into stearic acid, Palmic acid, glyceryl monostearate, trilauryl glyceride, three palmitic acid fat respectively; Content is constant, and all the other are equal to embodiment 1.
Final phenomenon is: aqueous dispersions generation layering, generation precipitate or flocculate after standing a period of time; That is, the result finally caused cannot successfully prepare antarctic krill oil nano structured lipid carrier.
Comparative example 2, in order to elected with three palmitic acid fat described in above-mentioned comparative example 1 as solid lipid time, corresponding carrier can be prepared into, therefore, need several emulsifying agent with the use of, and antarctic krill oil ratio (otherwise cannot obtain) need be reduced, that is, its formula is as follows:
Antarctic krill oil 2.5%; Three palmitic acid fat 7.5%; Lecithin 1.07%, poloxamer1880.8%, sucrose ester 1.2%, surplus is water.
In this comparative example 2, liquid fatty substance antarctic krill oil content in TL is only 25%; And embodiments of the invention 1, liquid fatty substance antarctic krill oil in TL content up to 65%.(TL=Solid lipid+liquid fatty substance)
The mean diameter of the antarctic krill oil nano structured lipid carrier that comparative example 2 is prepared in the aqueous dispersions of gained is about 186.9nm.
Contrast test, above-mentioned comparative example 2 detected according to method described in above-mentioned experiment 1 ~ 3, acquired results is as follows:
| Envelop rate | DHA(％) | EPA(％) | Astaxanthin (%) |
| Comparative example 2 | 88.47±0.87 | 90.63±0.71 | 93.11±0.78 |
Preserving mean diameter after 60 days is 320nm, within 70 days, occurs flocculation phenomenon afterwards; And its aqueous dispersions under rotating speed 10000 revs/min after centrifugal 5 minutes mean diameter be changed to 201nm, there will be obvious lamination.
The reaction rate constant of this comparative example 2 correspondence is as follows:
DHA in NLC is 0.00747 ± 0.00034h
-1;
EPA in NLC is 0.00663 ± 0.00015h
-1;
Astaxanthin in NLC is 0.00517 ± 0.00024h
-1.
Finally, it is also to be noted that what enumerate above is only several specific embodiments of the present invention.Obviously, the invention is not restricted to above embodiment, many distortion can also be had.All distortion that those of ordinary skill in the art can directly derive from content disclosed by the invention or associate, all should think protection scope of the present invention.
Claims (5)
1. the nano structured lipid carrier of antarctic krill oil, is characterized in that: be made up of the component of following mass content:
Antarctic krill oil 3.5-6.5%;
Solid lipid 3.5-6.5%;
Emulsifying agent 0.9-1.1%;
Surplus is water.
2. the preparation method of the nano structured lipid carrier of antarctic krill oil as claimed in claim 1, is characterized in that comprising the following steps:
1), by solid lipid and antarctic krill oil in 65 ~ 75 DEG C of melting mixing;
2), by emulsifying agent be added in water and stir until emulsifiers dissolve, 65 ~ 75 DEG C of preheatings;
3), under magnetic agitation condition, by step 1) molten state of gained mixing lipid drops to step 2) in the aqueous solution that obtains, obtain colostrum;
4), by step 3) colostrum of gained utilizes high pressure homogenization method or ultrasonic method to carry out homogenizing;
5), step 4) microemulsion of gained naturally cools to room temperature after homogenizing, can obtain the aqueous dispersions of antarctic krill oil nano structured lipid carrier.
3. the preparation method of the nano structured lipid carrier of antarctic krill oil according to claim 2, is characterized in that: described solid lipid is palm stearin or glyceryl tristearate.
4. the preparation method of the nano structured lipid carrier of antarctic krill oil according to claim 3, is characterized in that: described emulsifying agent is soybean lecithin or Tween 80.
5. the preparation method of the nano structured lipid carrier of the antarctic krill oil according to claim 2,3 or 4, is characterized in that:
Described step 4) in,
High pressure homogenization method is: by step 3) to pour setting pressure into be carry out circulation homogenizing in the high pressure homogenizer of 70 ~ 90MPa for the colostrum of gained;
Ultrasonic method is: by step 3) colostrum of gained pulverizes 4 ~ 6 minutes with 180 ~ 220W (being preferably 200W) power ultrasonic through ultrasonic probe.
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| CN107006842A (en) * | 2016-01-28 | 2017-08-04 | 中国海洋大学 | A kind of preparation method of fucoxanthin microcapsules |
| CN107874257A (en) * | 2017-11-09 | 2018-04-06 | 大连工业大学 | The preparation method of high embedding rate small particle antarctic krill oil nanoparticle |
| CN110897026A (en) * | 2019-11-30 | 2020-03-24 | 华南理工大学 | Krill oil beverage with chocolate flavor and preparation method thereof |
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| CN110897026A (en) * | 2019-11-30 | 2020-03-24 | 华南理工大学 | Krill oil beverage with chocolate flavor and preparation method thereof |
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