CN103637989A - Preparation method of tea polyphenol nano-liposomes by ethanol injection-dynamic high-pressure microfluidization-enzymolysis - Google Patents
Preparation method of tea polyphenol nano-liposomes by ethanol injection-dynamic high-pressure microfluidization-enzymolysis Download PDFInfo
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Abstract
The invention discloses a preparation method of tea polyphenol nano-liposomes by combination of an ethanol injection method and dynamic high-pressure microfluidization-enzymolysis. Tea polyphenol, lecithin, tween-80 and a phosphate buffer solution are adopted as raw materials. The tea polyphenol nano-liposomes are prepared by utilization of an ethanol injection-dynamic high-pressure microfluidization-enzymolysis method. According to the prepared tea polyphenol nano-liposomes, the average particle size is 67.4 nm plus or minus 3.0 nm, the zeta potential is -6.07 mV plus or minus 0.59 mV, the distribution coefficient is 0.220 plus or minus 0.010, and the encapsulation efficiency is 79.7% plus or minus 5.4%. The prepared tea polyphenol nano-liposomes show good slow-release property. Only 30.3% plus or minus 2.9% of the tea polyphenol is released after 24 h. In a small-intestine simulation environment where pH is 7.4, the tea polyphenol nano-liposomes show good stability.
Description
Technical field
The present invention relates to a kind of preparation method of water soluble drug nanometer liposome, be specifically related to a kind of method that ethanol injection-dynamic high-pressure microjet is prepared tea polyphenol nano lipidosome, can further be applied to the fields such as food, medicine, cosmetics.
Background technology
Tea polyphenols has antioxidation, the function such as anticancer, be the potential use object of health food, medicine etc., but it is to light, oxygen sensitive, and its application is restricted.Liposome (liposome) is the vesicle that the inside that formed by phospholipid bilayer comprises water.Research shows, liposome has the advantages such as slow-releasing, cellular affinity, histocompatibility and targeting, has been successfully applied to the fields such as biological medicine, chemical industry agricultural.Liposome is used for wrapping up the materials such as nutrient, enzyme, food additive, food antibacterial and has demonstrated tempting prospect.Utilize liposome technology that tea polyphenols is encapsulated, can overcome the above-mentioned shortcoming of tea polyphenols.The shortcomings such as it is large that the standby liposome of ethanol injection-dynamic high-pressure microjet legal system can overcome thick liposome particle diameter, unstable, while alcohol injection has also been broken through membrane process and has been washed the obstacle of film difficulty and small lot batch manufacture.
Summary of the invention
The object of the invention is provides a kind of ethanol injection-dynamic high-pressure microjet to prepare the method for tea polyphenol nano lipidosome for the deficiencies in the prior art, to improve the stability of tea polyphenol liposomes, for the exploitation of tea polyphenol liposomes provides theoretical foundation and Technical Reference.
The method processing step that a kind of ethanol injection-dynamic high-pressure microjet of the present invention is prepared tea polyphenol nano lipidosome is as follows:
(1) mass volume ratio of the phosphate buffered solution that component tea polyphenols, lecithin, cholesterol and the tween 80 of tea polyphenol nano lipidosome and pH=6.0 concentration are 0.05M/L is: in the phosphate buffered solution that is 0.05M/L in 100mLpH=6.0 concentration: tea polyphenols 0.25-0.75g, lecithin 2.02-6.06g, cholesterol 0.30-0.90g, tween 80 0.51-1.53g;
(2) by above-mentioned mass volume ratio, take tea polyphenols, lecithin, cholesterol and tween 80, at 40 ℃-50 ℃, be dissolved in the dehydrated alcohol with phosphate buffered solution same volume;
(3) step (2) gained solution is joined in the phosphate buffered solution that the pH=6.0 concentration of the definite mass volume ratio of step (1) is 0.05M/L, stir and form emulsion;
(4) step (3) gained emulsion is evaporated on evaporimeter and remove ethanol and obtain the thick liposome of tea polyphenols;
(5) the thick liposome obtaining with dynamic high-pressure microjet treatment step (4), pressure is 110MPa, processes and once obtains tea polyphenol nano lipidosome.
The invention has the beneficial effects as follows:
(1) envelop rate: the standby tea polyphenol nano lipidosome of ethanol injection-dynamic high-pressure microjet legal system has higher envelop rate, and envelop rate is 79.7 ± 5.4%.
(2) particle diameter current potential: the mean diameter of the tea polyphenol nano lipidosome that ethanol injection-dynamic high-pressure microjet legal system is standby is 67.4 ± 3.0nm, breadth coefficient is 0.22 ± 0.010, current potential is-6.07 ± 0.59mV.
(3) microscopic appearance figure: the standby tea polyphenol nano lipidosome of ethanol injection-dynamic high-pressure microjet legal system, its atom is tried hard to (Fig. 1), can find out liposome globulate, and size is below 100nm, and it is more even to distribute.
(4) the standby tea polyphenol nano lipidosome of ethanol injection-dynamic high-pressure microjet legal system shows good sustained release performance, and after dialysis 24h, the release rate of tea polyphenols is only 30.3 ± 2.9%.
(5) stability experiment in alkaline environment: tea polyphenols solution and tea polyphenol nano lipidosome are placed under the alkaline environment of Gl tract, assess its stability, measure the content of its oxide under the wavelength of 430nm.Concrete steps are as follows: tea polyphenols solution and tea polyphenol nano lipidosome are used respectively to phosphate buffered solution (pH=7.4,0.05M/L) dilution, and the ultimate density that makes them is 200 μ g/mL, is then placed in 37 ℃ of insulations.Also it measures its absorbance under 430nm wavelength under the predefined time, to observe color.When 0h, the absorbance of tea polyphenols solution and tea polyphenol nano lipidosome is all very little, is respectively 0.083 and 0.062.After when the 24h, tea polyphenols solution reached respectively 1.304 and 0.827 with absorbance tea polyphenol liposomes.And under the identical time, the absorbance of tea polyphenol nano lipidosome is far below tea polyphenols solution.As when the 4h, the absorbance of tea polyphenol liposomes is 0.357, and the absorbance of tea polyphenols solution is 0.613.And As time goes on, difference increasing (Fig. 3) between the two, this shows that tea polyphenol nano lipidosome can significantly improve the stability of tea polyphenols.
Accompanying drawing explanation
Fig. 1: the particle size distribution of tea polyphenol nano lipidosome and atomic microgram;
Fig. 2: the release in vitro of tea polyphenols solution and tea polyphenol nano lipidosome;
Fig. 3: tea polyphenols solution and tea polyphenol nano lipidosome be stability observing in the environment of pH=7.4.
The specific embodiment
Embodiment 1
Take 121.2g lecithin, 18.1g cholesterol, 30.6g tween 80 and 15.0g tea polyphenols, at 45 ℃, be dissolved in 3L dehydrated alcohol completely, alcoholic solution is slowly injected in 3L phosphate buffered solution (pH=6.0,0.05M/L), form emulsion, then transferred in round-bottomed flask, under 45 ℃ of water bath condition, vacuum rotating is removed dehydrated alcohol, forms uniform suspension and is thick liposome.Thick liposome is joined in dynamic high-pressure microjet, and under 110MPa condition, Micro Fluid is processed 1 time, prepares nanometer liposome.The nanometer liposome making is compared with transparent milk white solution.The envelop rate of the tea polyphenol nano lipidosome making is 78.6%, and mean diameter is 68.5nm, and breadth coefficient is 0.223, and Zeta potential is-6.86mV that in release test, after 24h, the release rate of tea polyphenols is only 29.8% in vitro.
Embodiment 2
Take 82.5g lecithin, 10.6g cholesterol, 20.6g tween 80 and 10.2g tea polyphenols, at 50 ℃, be dissolved in 3L dehydrated alcohol completely, alcoholic solution is slowly injected in 3L phosphate buffered solution (pH=6.0,0.05M/L), form emulsion, then transferred in round-bottomed flask, under 45 ℃ of water bath condition, vacuum rotating is removed dehydrated alcohol, and the even suspension of formation is thick liposome.Thick liposome is joined in dynamic high-pressure microjet, and under 110MPa condition, Micro Fluid is processed 1 time, prepares nanometer liposome.The nanometer liposome making is compared with transparent milk white solution.The envelop rate of the tea polyphenol nano lipidosome making is 75.2%, and mean diameter is 71.2nm, and breadth coefficient is 0.219, and Zeta potential is-6.08mV that in release test, after 24h, the release rate of tea polyphenols is only 26.8% in vitro.
Embodiment 3
Take 182.1g lecithin, 28.4g cholesterol, 46.8g tween 80 and 22.5g tea polyphenols, at 50 ℃, be dissolved in 3L dehydrated alcohol completely, alcoholic solution is slowly injected in 3L phosphate buffered solution (pH=6.0,0.05M/L), form emulsion, then transferred in round-bottomed flask, under 45 ℃ of water bath condition, vacuum rotating is removed dehydrated alcohol, forms uniform suspension and is thick liposome.Thick liposome is joined in dynamic high-pressure microjet, and under 110MPa condition, Micro Fluid is processed 1 time, prepares nanometer liposome.The nanometer liposome making is compared with transparent milk white solution.The envelop rate of the tea polyphenol nano lipidosome making is 82.5%, and mean diameter is 65.4nm, and breadth coefficient is 0.232, and Zeta potential is-5.92mV that in release test, after 24h, the release rate of tea polyphenols is only 33.8% in vitro.
Take 138.4g lecithin, 20.9g cholesterol, 35.3g tween 80 and 17.0g tea polyphenols, at 45 ℃, be dissolved in 3L dehydrated alcohol completely, alcoholic solution is slowly injected in 3L phosphate buffered solution (pH=6.0,0.05M/L), form emulsion, then transferred in round-bottomed flask, under 45 ℃ of water bath condition, vacuum rotating is removed dehydrated alcohol, and the even suspension of formation is thick liposome.Thick liposome is joined in dynamic high-pressure microjet, and under 110MPa condition, Micro Fluid is processed 1 time, prepares nanometer liposome.The nanometer liposome making is compared with transparent milk white solution.The envelop rate of the tea polyphenol nano lipidosome making is 82.6%, and mean diameter is 64.5nm, and breadth coefficient is 0.208, and Zeta potential is-5.42mV that in release test, after 24h, the release rate of tea polyphenols is only 30.8% in vitro.
Claims (1)
1. alcohol injection is prepared a method for tea polyphenol nano lipidosome in conjunction with dynamic high-pressure microjet, it is characterized in that:
(1) mass volume ratio of the phosphate buffered solution that component tea polyphenols, lecithin, cholesterol and the tween 80 of tea polyphenol nano lipidosome and pH=6.0 concentration are 0.05M/L is: in the phosphate buffered solution that is 0.05M/L in 100mLpH=6.0 concentration: tea polyphenols 0.25-0.75g, lecithin 2.02-6.06g, cholesterol 0.30-0.90g, tween 80 0.51-1.53g;
(2) by above-mentioned mass volume ratio, take tea polyphenols, lecithin, cholesterol and tween 80, at 40 ℃-50 ℃, be dissolved in the dehydrated alcohol with phosphate buffered solution same volume;
(3) step (2) gained solution is joined in the phosphate buffered solution that the pH=6.0 concentration of the definite mass volume ratio of step (1) is 0.05M/L, stir and form emulsion;
(4) step (3) gained emulsion is evaporated on evaporimeter and remove ethanol and obtain the thick liposome of tea polyphenols;
(5) the thick liposome obtaining with dynamic high-pressure microjet treatment step (4), pressure is 110MPa, processes and once obtains tea polyphenol nano lipidosome.
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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 |
CN104382039A (en) * | 2014-12-19 | 2015-03-04 | 福州大学 | Nano lipidosome containing green tea extractive, and preparation method thereof |
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CN104382039A (en) * | 2014-12-19 | 2015-03-04 | 福州大学 | Nano lipidosome containing green tea extractive, and preparation method thereof |
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