CN101940321B - Method for preparing nanoliposomes of medium chain fatty acids (MCFAs) by dynamic high-pressure microfluidization (DHPM)-freezing and thawing method - Google Patents
Method for preparing nanoliposomes of medium chain fatty acids (MCFAs) by dynamic high-pressure microfluidization (DHPM)-freezing and thawing method Download PDFInfo
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- CN101940321B CN101940321B CN2010102475030A CN201010247503A CN101940321B CN 101940321 B CN101940321 B CN 101940321B CN 2010102475030 A CN2010102475030 A CN 2010102475030A CN 201010247503 A CN201010247503 A CN 201010247503A CN 101940321 B CN101940321 B CN 101940321B
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Abstract
The invention relates to a preparation method of nanoliposomes of medium chain fatty acids (MCFAs), which is used for preparing nanoliposomes of MCFAs by taking liposoluble MCFAs as raw materials and lecithin and cholesterol as wall materials and adopting a dynamic high-pressure microfluidization (DHPM)-freezing and thawing method through the processes of dissolving, uniformly mixing, removing solvents in vacuum, washing a membrane by hydration, treating by DHPM, repeatedly freezing and thawing and the like. The nanoliposomes of the MCFAs, which are prepared by the invention, have the characteristics that the envelopment rate is 40%-50%, the mean particle size is 90-110nm, the dispersion coefficient is less than 0.10, and the zeta potential is (-30mV)-(-50mV); and the nanoliposomes have the advantages of uniform distribution, high bioavailability and good storage stability.
Description
Technical field
The present invention relates to the food nutrition field, be specifically related to a kind of preparation method of medium chain fatty acid nano liposomes.
Background technology
See that from the nutritional physiology viewpoint (medium-chain fatty acids MCFAs) is meant that carbon is 8 sad (octanoic acid to medium chain fatty acid; C8:0) and carbon be 10 capric acid (decanoic acid; C10:0).With the LCFA that in lymphatic system, absorbs (long-chain fatty acids; LCFAs) compare; Medium chain fatty acid (MCFAs) has many special physiological properties such as absorbing soon, be difficult for putting aside in vivo, very easily provide energy; Mainly show as: (1) medium chain fatty acid (MCFAs) fusing point is low, molecular weight is little, and normal temperature is down that liquid state, energy value are low; (2) do not rely on the L-carnitine and directly see through the mitochondria duplicature, get in the mitochondria and carry out oxidation, for body provides energy fast, can be used as sportsman, patients after surgery, the person's that has the fat malabsorption energy source; (3) medium chain fatty acid (MCFAs) is difficult in hepatic tissue and adipose tissue, accumulating, and can increase satiety, has the effect that keeps muscle and do not increase adipose tissue, can be used as obesity person's nutritional food therapy agent; (4) { scalable is to the immune response and the secreting, expressing that improves immunoglobulin A of lipopolysaccharides, and the protection alimentary canal plays anti-inflammatory effect, can be used for short bowel syndrome patient's treatment for medium chain fatty acid (MCFAs).But take in gastrointestinal discomfort symptoms such as a large amount of medium chain fatty acids (MCFAs) can cause feeling sick, vomiting, aerogastria, abdominal pain property spasm, diarrhoea in the short time, also can stimulate the secretion of cholecystokinin or other intestines internal hormone.
Liposome is the excellent carrier that a kind ofly has the cell membrane similar structures, can wrap up hydrophilic, lipophilic and amphipathic medicine and trophic factors.Liposome both can be protected medicine and trophic factors, reduced its toxic and side effect, reached the slowly-releasing purpose, can improve the target property and the bioavailability of medicine again.The preparation method that liposome is commonly used has film dispersion method, freeze-thaw method, reverse evaporation, alcohol injection, high pressure homogenization method etc.Preparation technology is simple for these conventional methods, but often introduces a large amount of toxic reagents, and is inappropriate for large-scale commercial production.
Dynamic high-pressure microjet (dynamic high-pressure microfluidization; DHPM) be a kind of high pressure homogenization technique; It utilizes hydraulic pump to make fluid produce high pressure, and in the microchannel in impact cavity, fluid is dispersed into two strands or multiply thread and carries out strong high-speed impact; The huge pressure of moment generation falls in this process; Realize comprehensive functions such as high-speed impact, highdensity plywood, cavitation erosion, dither, instantaneous pressure drop, under 100MPa, the time can reach purposes such as making material refinement, emulsification, homogeneous and modification less than 5s.
Summary of the invention
The objective of the invention is to the deficiency of prior art and the weakness of medium chain fatty acid (MCFAs); A kind of safety, medium chain fatty acid nano liposomes that functional character is good are provided, replenish for fat malabsorption person, patients after surgery, sportsman and obesity person provide special dietary.
The technical scheme that the present invention promptly adopts is: adopting dynamically is to press micro jetting technology to combine freeze-thaw method to prepare the medium chain fatty acid nano liposomes,
Processing step of the present invention is following:
1, each component and percentage by weight thereof are in the medium chain fatty acid nano-lipid raw material: medium chain fatty acid (MCFAs) 0.5%-2.0%; Lecithin 4.0%-8.0%; Cholesterol 0.8%-1.2%, Tween-80 1.5%-2.5%, vitamin E 0.1%-0.3%; Maltose 2.0%-5.0%, remaining is that concentration is that the PBS (PBS) of 0.05M is 81.0%-91.1%;
2, take by weighing medium chain fatty acid (MCFAs), lecithin, cholesterol, Tween-80 and vitamin E respectively by above-mentioned part by weight, under 40 ℃ of conditions, be dissolved in the 20ml absolute ethyl alcohol, dissolve each component fully by 1g lecithin;
3, step 2 gained solution is removed absolute ethyl alcohol on the rotary evaporation in vacuo appearance, form homogeneous film;
4, the maltose that in the PBS (PBS) of pH7.4,0.05M, adds 2.0%-5.0%; By 8% lipid concentration (lecithin and cholesterol total amount account for the percentage of volume of buffer solution) maltose solution is added the film that step 3 forms; Carry out hydration and wash film, form thick liposome suspension;
5, the thick liposome suspension that obtains with high pressure microjet homogenizer treatment step 4, pressure is 140MPa, number of processes is 4 times, gets medium chain fatty acid (MCFAs) liposome;
6, with step 5 gained medium chain fatty acid (MCFAs) liposome freezing 0.5h-1.5h under-80 ℃ of conditions; Take out the back and under 40 ℃ of water bath condition, melt 15min; Put under-80 ℃ of conditions freezingly once more, 3 times so repeatedly, promptly get medium chain fatty acid (MCFAs) nano liposomes.
The invention has the beneficial effects as follows:
Medium chain fatty acid (MCFAs) nano liposomes of preparation; Both overcome the weakness on medium chain fatty acid (MCFAs) function, and liposome property is stable, the liposome encapsulation that bioavailability is higher is 40%-50%; Particle mean size is 90-110nm; The coefficient of dispersion is less than 0.10, the zeta current potential be (30mV)-(50mV), storage-stable is good.
Description of drawings
Accompanying drawing is the process route view of preparation medium chain fatty acid (MCFAs) nano liposomes:
The specific embodiment
Embodiment 1
Take by weighing 0.45g medium chain fatty acid (MCFAs), 2.06g lecithin, 0.34g cholesterol, 0.62g Tween-80 and 0.04g vitamin E, be dissolved in fully in the 42ml absolute ethyl alcohol, absolute ethyl alcohol is removed in the vacuum rotation under 40 ℃ of water bath condition, forms homogeneous film.Getting 30ml concentration is that 0.05M, pH are 7.4 PBS (PBS), adds 0.9g maltose, gained maltose solution adding film is carried out hydration wash film, and the even suspension of formation is thick liposome.Thick liposome is joined among the DHPM, and Micro Fluid is handled 4 times under the 140MPa condition, obtains medium chain fatty acid (MCFAs) liposome.Medium chain fatty acid (MCFAs) liposome put under-80 ℃ of conditions take out behind the freezing 0.5h, under 40 ℃ of water bath condition, melt 15min, it is freezing to put into-80 ℃ of conditions once more, promptly obtains medium chain fatty acid (MCFAs) nano liposomes so repeatedly for 3 times.The medium chain fatty acid that makes (MCFAs) nano liposomes is a milky white solution, and envelop rate is 47.28%, and average grain diameter is 95.4nm, and the coefficient of dispersion is 0.075, and the zeta current potential is-43.79mV.
Embodiment 2
Take by weighing 0.90g medium chain fatty acid (MCFAs), 4.12g lecithin, 0.68g cholesterol, 1.23g Tween-80 and 0.08g vitamin E, be dissolved in fully in the 83ml absolute ethyl alcohol, absolute ethyl alcohol is removed in the vacuum rotation under 40 ℃ of water bath condition, forms homogeneous film.Getting 60ml concentration is that 0.05M, pH are 7.4 PBS (PBS), adds 1.8g maltose, gained maltose solution adding film is carried out hydration wash film, and the even suspension of formation is thick liposome.Thick liposome is joined among the DHPM, and Micro Fluid is handled 4 times under the 140MPa condition, obtains medium chain fatty acid (MCFAs) liposome.Medium chain fatty acid (MCFAs) liposome put under-80 ℃ of conditions take out behind the freezing 0.5h, under 40 ℃ of water bath condition, melt 15min, it is freezing to put into-80 ℃ of conditions once more, promptly obtains medium chain fatty acid (MCFAs) nano liposomes so repeatedly for 3 times.The medium chain fatty acid that makes (MCFAs) nano liposomes is a milky white solution, and envelop rate is 46.09%, and average grain diameter is 105.3nm, and the coefficient of dispersion is 0.084, and the zeta current potential is-44.07mV.
Embodiment 3
Take by weighing 0.45g medium chain fatty acid (MCFAs), 2.06g lecithin, 0.34g cholesterol, 0.84g Tween-80 and 0.08g vitamin E, be dissolved in fully in the 42ml absolute ethyl alcohol, absolute ethyl alcohol is removed in the vacuum rotation under 40 ℃ of water bath condition, forms homogeneous film.Getting 30ml concentration is that 0.05M, pH are 7.4 PBS (PBS), adds 0.9g maltose, gained maltose solution adding film is carried out hydration wash film, and the even suspension of formation is thick liposome.Thick liposome is joined among the DHPM, and Micro Fluid is handled 4 times under the 140MPa condition, obtains medium chain fatty acid (MCFAs) liposome.Medium chain fatty acid (MCFAs) liposome put under-80 ℃ of conditions take out behind the freezing 1.0h, under 40 ℃ of water bath condition, melt 15min, it is freezing to put into-80 ℃ of conditions once more, promptly obtains medium chain fatty acid (MCFAs) nano liposomes so repeatedly for 3 times.The medium chain fatty acid that makes (MCFAs) nano liposomes is a milky white solution, and envelop rate is 44.57%, and average grain diameter is 102.6nm, and the coefficient of dispersion is 0.072, and the zeta current potential is-40.63mV.
Embodiment 4
Take by weighing 0.90g medium chain fatty acid (MCFAs), 4.12g lecithin, 0.68g cholesterol, 1.64g Tween-80 and 0.16g vitamin E, be dissolved in fully in the 83ml absolute ethyl alcohol, absolute ethyl alcohol is removed in the vacuum rotation under 40 ℃ of water bath condition, forms homogeneous film.Getting 60ml concentration is that 0.05M, pH are 7.4 PBS (PBS), adds 1.8g maltose, gained maltose solution adding film is carried out hydration wash film, and the even suspension of formation is thick liposome.Thick liposome is joined among the DHPM, and Micro Fluid is handled 4 times under the 140MPa condition, obtains medium chain fatty acid (MCFAs) liposome.Medium chain fatty acid (MCFAs) liposome put under-80 ℃ of conditions take out behind the freezing 0.5h, under 40 ℃ of water bath condition, melt 15min, it is freezing to put into-80 ℃ of conditions once more, promptly obtains medium chain fatty acid (MCFAs) nano liposomes so repeatedly for 3 times.The medium chain fatty acid that makes (MCFAs) nano liposomes is a milky white solution, and envelop rate is 45.62%, and average grain diameter is 81.7nm, and the coefficient of dispersion is 0.084, and the zeta current potential is-39.58mV.
Claims (1)
1. dynamic high-pressure microjet-freeze-thaw method prepares the method for medium chain fatty acid nano liposomes, it is characterized in that, said each component of liposome raw material and percentage by weight thereof are: medium chain fatty acid (MCFAs) 0.5%-2.0%; Lecithin 4.0%-8.0%, cholesterol 0.8%-1.2%, Tween-80 1.5%-2.5%; Vitamin E 0.1%-0.3%; Maltose 2.0%-5.0%, remaining is that concentration is the PBS (PBS) of 0.05M, is 81.0%-91.1%; Preparation process is:
(1) takes by weighing medium chain fatty acid (MCFAs), lecithin, cholesterol, Tween-80 and vitamin E respectively by above-mentioned percentage by weight, under 40 ℃ of conditions, be dissolved in the 20ml absolute ethyl alcohol, dissolve each component fully by 1g lecithin;
(2) step 1 gained solution is removed absolute ethyl alcohol on the rotary evaporation in vacuo appearance, form homogeneous film;
(3) maltose of adding 2.0%-5.0% in the PBS (PBS) of pH7.4,0.05M; Account for 8% of volume of buffer solution by lecithin and cholesterol total amount; Maltose solution is added the film that step 2 forms, carry out hydration and wash film, form thick liposome suspension;
(4) the thick liposome that step 3 is obtained joins in the high pressure microjet homogenizer, and pressure is 140MPa, and number of processes is 4 times, gets medium chain fatty acid (MCFAs) liposome;
(5) with step 4 gained medium chain fatty acid (MCFAs) liposome freezing 0.5h-1.5h under-80 ℃ of conditions; Take out the back and under 40 ℃ of water bath condition, melt 15min; Put under-80 ℃ of conditions freezingly once more, 3 times so repeatedly, promptly get medium chain fatty acid (MCFAs) nano liposomes.
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| CN104012991A (en) * | 2014-05-23 | 2014-09-03 | 南昌大学 | Method for preparing eugenol nanoliposomes through alcohol injection-dynamic high-pressure microfluidization |
| CN109007779B (en) * | 2018-06-13 | 2021-07-23 | 福建农林大学 | Frozen minced fillet compound water-retaining agent and preparation method and application thereof |
| CN109007776B (en) * | 2018-06-13 | 2021-08-03 | 福建农林大学 | Compound water-retaining agent for Chinese meat dishes as well as preparation method and application of compound water-retaining agent |
| CN113208116A (en) * | 2021-02-10 | 2021-08-06 | 大连工业大学 | Nano liposome embedded with egg white peptide calcium compound and preparation method thereof |
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| US20060046962A1 (en) * | 2004-08-25 | 2006-03-02 | Aegis Therapeutics Llc | Absorption enhancers for drug administration |
| CN101019851B (en) * | 2007-02-12 | 2011-01-19 | 南昌大学 | Medium chain fatty acid liposome and its preparation |
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