CN102641311B - Kiwi fruit seed oil liposome oral liquid and preparation method thereof - Google Patents
Kiwi fruit seed oil liposome oral liquid and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kiwi fruit seed oil liposome oral liquid and a preparation method thereof. The method comprises the following steps: (1) adding a phosphate buffer solution with pH of 6.8 in a constant temperature oscillator, heating in a water bath to 60 DEG C and keeping constant temperature; (2) based on anhydrous ethanol as a solvent, preparing a mixed solution containing15-25mg/ml of lecithin, 5-20mg/ml of cholesterol and 5-20mg/ml of kiwi fruit seed oil; and (3) injecting the mixed solution in the step (2) into the phosphate buffer solution in the step (1), placing in the constant-temperature water bath at the temperature of 60 DEG C, then performing ultrasonic treatment, adding distilled water to a certain volume, sealing and sterilizing to obtain the kiwi fruit seed oil liposome oral liquid. According to the kiwi fruit seed oil liposome oral liquid prepared by the method disclosed by the invention, kiwi fruit seed oil has high stability and safety, unsaturated fatty acid in the kiwi fruit seed oil can be prevented from being oxidized, and the purposes of convenience in transportation and long-term preservation can be further achieved.
Description
Technical field
The invention belongs to the health-care preparation field, be specifically related to a kind of Chinese gooseberry seed oil liposome oral fluid and preparation method thereof.
Background technology
Fructus actinidiae chinensis (
Actinidia chinensis) having another name called carambola, Fructus actinidiae chinensis, Prunus persica f. compressa, Radix Fici Hirtae etc., New Zealand Mi base dimension fruit (kiwifruit) is a kind of liana, is one of precious fruit of China's special product.Along with the development of Fructus actinidiae chinensis processing industry, the fruit level of residue increases.Mainly be fruit crude fibre and seed in the Fructus actinidiae chinensis residue.These factories produce a large amount of marcs in the processing process, wherein contain a large amount of Fructus actinidiae chinensis seed kernels.According to the notable big grade of Huang (Huang Zhuowei, Yi Minghua. Chinese gooseberry seed refines the research [J] of high-quality edible oil. oil and foodstuffs science and technology, 1992, (4): 11 ~ 12) report, more than 250 at least of the seed in each Fructus actinidiae chinensis wild fruit, nearly more than 800, average about 500.After measured, oil content is 28.85% in the seed, in the Chinese gooseberry seed oil unsaturated fatty acid content up to 90.37%, wherein linoleic acid, Caulis et Folium Lini
Acid content accounts for 74.83%; The results of animal demonstration, Chinese gooseberry seed oil has obvious effect for reducing blood fat, oral safety non-toxic.Yao Maojun (Yao Maojun, Li Jiaxing, Zhang Yongkang. [J] inquired in the development and use of Chinese gooseberry seed oil. food and fermentation industries, 2001,27(12): 28 ~ 30, ] Yao Maojun, Li Jiaxing, Zhang Yongkang. Chinese gooseberry seed oil physicochemical property and fatty acid form [J]. Wuxi Light Industry Univ.'s journal, 2002,2(3): 307 ~ 309), Zhang Yongkang (Zhang Yongkang, Jiang Jianbo, Chen Lihua. the mensuration of kiwifruit nut fatty acid oil and utilization thereof [J]. JOURNAL OF JISHOU UNIVERSITY (natural science edition), 2001,22(4): 37 ~ 38,82) etc. studied the seed of Xiangxi " Mi Liangyi number " Fructus actinidiae chinensis, find that oil content is 23.5%, the gas Chromatographic Determination fatty acid forms, and the a-linolenic acid content is up to 63.99%.Therefore Chinese gooseberry seed oil can be used as the important health-care oil resource of replenishing unsaturated fatty acid and is developed.
Containing linolenic acid and linoleic acid in the monkey peach seed oil, is the highest crude vegetal of linolenic acid content except Fructus Perillae oil of finding at present.Linoleic acid plus linolenic acid be two kinds to the essential fatty acid of health particular importance, can only be from external world picked-up and can not be synthetic voluntarily in human body.Alpha-linolenic acid is the precursor substance of EPA and DHA, thereby Chinese gooseberry seed oil has blood fat reducing, cholesterol and blood pressure, and angiocardiopathy preventing suppresses platelet aggregation, prevents thrombosis, prevents Carciuogenesis, the multiple effects such as antioxidation and slow down aging.So Chinese gooseberry seed oil is a kind of raw material of functional food, medicine and cosmetics of high-quality.
But content has brought problem up to 90% unsaturated fatty acid to the preservation of oils and fats in the Chinese gooseberry seed oil.Monounsaturated fatty acid molecule contains a plurality of pairs of keys, thus to oxygen, light and thermoae be responsive, oxidation deterioration very easily.The oxidation of oils and fats not only makes it lose due health protection function, but also can produce some harmful materials.
Liposome (liposome) is a kind of a kind of lipoid spherula that is made of phospholipid that Bangham found in nineteen sixty-five.It is the miniature spheroid (Bangham ad.difusion of univalent ions aeross the lamellae of swollen Phosphlipids [J] .Mol. Biol, 1965 13:238 ~ 252.) that drug encapsulation is made in the thin film that lipid (phospholipid and cholesterol) bilayer forms.Seal respectively fat-soluble and water soluble drug inside and outside the bilayer, its physicochemical property and similar cell membrane are called again artificial membrane.Liposome receives much concern as a kind of functional component preparation of novelty, and in recent years, Liposomal formulation research obtains very great development with application.The effect characteristics of Liposomal formulation mainly contain (Lu Bin. novel pharmaceutical formulation and new technique [M]. second edition. Beijing: People's Health Publisher, 2005.5):
(l) the defencive function composition improves stability
Some unsettled functional components can be subject to the protection of liposome duplicature by after liposomal encapsulated: wrapped functional component is not destroyed by enzyme and immune system in vivo in the transport process.
(2) reduce functional component toxicity
Liposome itself is nontoxic to human body, after functional component is wrapped in the heart, kidney cumulant specific ionization functional component low, the heart, the virose functional component of kidney sealed liposome after, can obviously reduce toxicity.
(3) slow releasing function
The rate of release of control functional component reaches long-acting slow-release.
(4) targeting
By reticuloendothelial system phagocytic, functional component is mainly accumulated in the histoorgans such as liver, spleen, lung and bone marrow after entering in the body, realized the passive target administration.
Liposome is used ripe in pharmaceuticals industry, but still belongs to the starting stage at field of food.Liposome by the coating of phospholipid, both can improve the stability of embedding substance as a good carrier, made again it have the target same sex, can improve the bioavailability of functional component.At present, the preparation method of liposome and example thereof are as follows:
1. mechanical dispersion method (mechanical dispersion) normally is dissolved in lipid in the organic solvent first, the decompression rotary evaporation makes it to form uniform lipid film at the inwall of glass container, then adds aqueous media and lipid is disperseed and the formation liposome by jolting.For example silybin oil lipidosome preparation: recipe quantity lecithin, cholesterol, Silybum Marianum Gaertn Seed Oil are dissolved in the 5ml chloroform, evaporated under reduced pressure organic solvent on Rotary Evaporators, after reaching colloidal state, the phosphate buffered solution (PBS) that adds an amount of volume, in 35 ℃ of constant temperature water bath vibrations to complete aquation, with 0.45 μ m filtering with microporous membrane 3 times, and get final product.Adopt the method to prepare liposome comparatively general, but the organic reagents such as employing chloroform, toxicity is large, is not suitable for field of food.
2. solvent dispersion method
Solvent dispersion method (solvent dispersion) is dissolved in lipid in the organic solvent first, join and contain the aqueous phase that is wrapped medicine, phospholipid is arranged with monolayer (be liposome bi-layer membrane half) on organic facies and water interface.
Preparation such as tea tree oil liposome: take by weighing lecithin, cholesterol, tea tree oil, VE, sodium deoxycholate, tween 80 of recipe quantity etc., ultrasonic even to suspension to wherein adding an amount of dehydrated alcohol, get the lipoid suspension; Hydration medium places on the constant temperature blender with magnetic force to keep temperature and the mixing speed of regulation.The lipoid suspension is slowly splashed in the hydration medium, get just suspension (wherein volume grade of alcohol is 10%) of liposome, this first suspension is placed ultrasonic 20 min of water-bath type Ultrasound Instrument, namely get the tea tree oil liposome suspension.The mass concentration of lecithin is 10 gL
-1, lecithin and cholesterol mass ratio be that the mass concentration of 5:1, tea tree oil is 1.0 gL
-1, VE mass concentration be 1.0 gL
-1, sodium deoxycholate mass concentration be 0.5 gL
-1, tween 80 mass concentration be 4.0 gL
-1, aqueous media is that pH value is 6.8 PBS buffer solution.Taking the method to prepare the case of liposome less, mainly be because technological parameter must be controlled accurately, otherwise the liposome that makes is unstable.
For the characteristics of the easy oxidation of Chinese gooseberry seed oil, the employing spray drying methods such as Feng Weihua carry out Research of Microencapsulation (research of Chinese gooseberry seed oil microencapsulation technology, Transactions of the Chinese Society of Agricultural Engineering, 2004,20(1) 234 ~ 236) to Chinese gooseberry seed oil.Yao Maojun etc. have prepared Soft Capsules of Kiwifruit Seed Oil, and (research of kiwifruit oil soft capsule is fermented and food industry, 29(12): 2003,58 ~ 61).Spray drying method still needs higher temperature (180 ℃ of air inlets, 80 ℃ of air outlets), therefore Chinese gooseberry seed oil is had loss.The open report that preparation Chinese gooseberry seed oil lipidosome is not yet arranged at present.
Summary of the invention
Technical problem to be solved by this invention provides a kind of Chinese gooseberry seed oil liposome oral fluid and preparation method thereof, the Chinese gooseberry seed oil liposome oral fluid for preparing with the inventive method, Chinese gooseberry seed oil stable high, safety, avoid the unsaturated fatty acid in the Chinese gooseberry seed oil oxidized, thereby reached the purpose of convenient transportation and long-term preservation.
Technical scheme provided by the invention is: a kind of preparation method of Chinese gooseberry seed oil liposome oral fluid, the method comprises the steps:
(1) measure phosphate buffer and place constant temperature oscillator, heating in water bath is to 55-65 ℃, and maintenance constant temperature;
(2) take dehydrated alcohol as solvent, compound concentration is the lecithin of 15-25 mg/ml, the cholesterol of 5 ~ 20mg/ml, the Chinese gooseberry seed oil mixed solution of 5 ~ 20mg/ml respectively;
(3) with in the phosphate buffer that is injected into behind three kinds of solution mix homogeneously described in the step (2) in the step (1), in water bath with thermostatic control 20-40min, remove residual ethanol, then carry out supersound process 20-40min, the adding distil water standardize solution, sealing namely gets Chinese gooseberry seed oil lipidosome oral administration solution after the sterilization.
Described preparation method, preferably, in the step (1), the phosphate buffer that measures pH6.8 places constant temperature oscillator, heating in water bath to 60 ℃, and under the 150r/min rotating speed, keep constant temperature; In the step (3), in 60 ℃ of water bath with thermostatic control 30min, remove residual ethanol, then carry out supersound process 30min(ultrasonic power 80W), the adding distil water standardize solution, sealing namely gets Chinese gooseberry seed oil lipidosome oral administration solution after the sterilization.
Described preparation method, preferably, in the step (1), preparation lecithin soln concentration is 20mg/ml; After three kinds of solution mixed in the step (3), lecithin wherein and the mass ratio of cholesterol were 2:1, and the mass ratio of lecithin and Chinese gooseberry seed oil is 4:1.。
Described preparation method, preferably, in the step (3), 60 ℃ of water bath with thermostatic control 30min, most residual ethanol is waved in evaporation, then carries out supersound process, and 30min, ultrasonic power are 80W.
Method of the present invention preferably adopts Hunan Laodie Agricultural Technology Development Co., Ltd. that Chinese gooseberry seed oil is provided, and for example batch number is 20111230.
The present invention also provides the Chinese gooseberry seed oil that is prepared by above-mentioned preparation method liposome oral fluid.
The present invention has following beneficial effect:
Chinese gooseberry seed oil is the important component that the Chinese gooseberry seed commonly used utilizes, because its contained linolenic health-care effect, improve the stability of Chinese gooseberry seed oil, realize linolenic targeted in the Chinese gooseberry seed oil, improve its absorption and utilize the focus that has become domestic and international research.Macaque seed oil is insoluble in water, oral administration biaavailability is low, limited greatly its clinical efficacy, because linolenic the containing up to more than 60% in the Chinese gooseberry seed oil, extremely easily oxidized, the present invention is in conjunction with targeting and the slow-releasing of liposome, Chinese gooseberry seed oil is prepared into the liposome oral administration solution, can improve Chinese gooseberry seed oil stability, safety, avoid the unsaturated fatty acid in the Chinese gooseberry seed oil oxidized, and can realize the targeting transportation of Chinese gooseberry seed oil, reach the effect that improves the Chinese gooseberry seed oil absorption rate.Thereby reach the purpose of convenient transportation and long-term preservation, promoted the commercial value of Chinese gooseberry seed oil liposome technology.The present invention takes lower temperature, has overcome the higher shortcoming of spray drying temperature in the microencapsulation process, and rate of release is slow than soft capsule simultaneously, can be further used as the raw material of slow releasing preparation.
The specific embodiment
Detailed description below by the specific embodiment is further illustrated the present invention, but is not limitation of the present invention, only does the example explanation.
Embodiment 1
1, the mensuration of Chinese gooseberry seed oil liposome encapsulation
Adopt the content of dialysis-determined by ultraviolet spectrophotometry Chinese gooseberry seed oil.Pipette 20ml Chinese gooseberry seed oil lipidosome and put into bag filter, behind the dialysis 17h, pour solution in the bag into the 100ml volumetric flask, selecting ethanol is demulsifier, in conjunction with ultrasonic, add Petroleum ether extraction, mixed solution is carried out centrifugal (3000r/min, 3min), get the 10ml supernatant in the 25ml volumetric flask, use the petroleum ether standardize solution, measure its absorbance A at the 233nm place
1; Measure the blank liposome absorbance A with method
0, calculate △ A, obtained the content of the Chinese gooseberry seed oil of sealing by standard curve.
Envelop rate %=△ A*100/A
1
2, prescription and optimal process
The key problem in technology of liposome oral administration solution is the preparation of liposome.Take envelop rate as index, four factors such as the mass ratio (L:O) of the mass ratio (L:C) of lecithin and cholesterol, lecithin and oil, water-bath time (t), heating-up temperature (T) are on sealing the impact of effect in the investigation prescription.According to the single factor experiment result, select L9 (4
3) orthogonal table carries out liposome prescription Orthogonal Experiment and Design.
Table 1 orthogonal design factor table
Table 2 orthogonal array and L(9 as a result) 3
4
| Grouping | L:C(w/w) | L:O(w/w) | The water-bath time (min) | T(℃) | Envelop rate (%) |
| 1 | 1 | 1 | 1 | 1 | 36.34 |
| 2 | 1 | 2 | 2 | 2 | 38.54 |
| 3 | 1 | 3 | 3 | 3 | 45.67 |
| 4 | 2 | 1 | 2 | 3 | 74.45 |
| 5 | 2 | 2 | 3 | 1 | 65.35 |
| 6 | 2 | 3 | 1 | 2 | 66.36 |
| 7 | 3 | 1 | 3 | 2 | 50.56 |
| 8 | 3 | 2 | 1 | 3 | 45.57 |
| 9 | 3 | 3 | 2 | 1 | 55.20 |
| K1 | 40.183 | 50.783 | 49.423 | 52.297 | |
| K2 | 68.720 | 49.820 | 56.063 | 51.820 | |
| K3 | 50.443 | 55.743 | 53.860 | 55.230 | |
| Optimal level | 2 | 3 | 2 | 3 | |
| R | 28.537 | 5.923 | 6.640 | 3.410 | |
| Significance | *(p<0.05) |
By intuitive analysis, best prescription and process conditions are for therefore as can be known, and the best preparation technology of Chinese gooseberry seed oil lipidosome is: 60 ℃ of the mass ratio of lecithin and cholesterol (2:1), lecithin and oily mass ratio (4:1), water-bath time 30min, heating-up temperatures.The ratio of lecithin and cholesterol is to having the greatest impact that liposome forms in the prescription.
Embodiment 2
The constant temperature oscillator temperature setting is set to 60 ℃, preheating.The phosphate buffer that accurately measures 100mlpH6.8 places constant temperature oscillator, heating in water bath to 60 ℃, and under the 150r/min rotating speed, keep constant temperature.Take dehydrated alcohol as solvent, preparing respectively lecithin (LC) concentration is 20mg/ml, and cholesterol (CH) concentration is 10mg/ml, and Chinese gooseberry seed oil (KFO) concentration is 15mg/ml, purity 99.0%.To evenly be injected in the phosphate buffer in the constant temperature oscillator with syringe behind each solution mix homogeneously.60 ℃ of water bath with thermostatic control 30min remove residual ethanol, supersound process 30min (ultrasonic power 80W), and adding distil water is settled to 250ml, and sealing namely gets Chinese gooseberry seed oil lipidosome oral administration solution behind the irradiation sterilization.
Method of the present invention preferably adopts Hunan Laodie Agricultural Technology Development Co., Ltd. that Chinese gooseberry seed oil is provided, and the used batch number of the present embodiment is 20111230.
Embodiment 3
Preparing respectively lecithin (LC) concentration with ethanol as solvent is 16mg/ml, and cholesterol (CH) concentration is 12mg/ml, and Chinese gooseberry seed oil (KFO) concentration is 10mg/ml, and preparation method is with embodiment 2.
Embodiment 4
Preparing respectively lecithin (LC) concentration with ethanol as solvent is 20mg/ml, and cholesterol (CH) concentration is 20mg/ml, and Chinese gooseberry seed oil (KFO) concentration is 5mg/ml, and preparation method is with embodiment 2.
Embodiment 5
Preparing respectively lecithin (LC) concentration with ethanol as solvent is 23mg/ml, and cholesterol (CH) concentration is 6mg/ml, and Chinese gooseberry seed oil (KFO) concentration is 18mg/ml, and preparation method is with embodiment 2.
Claims (4)
1. the preparation method of a Chinese gooseberry seed oil liposome oral fluid is characterized in that the method comprises the steps:
(1) phosphate buffer that measures pH6.8 places constant temperature oscillator, and heating in water bath is to 55-65 ℃, and keeps constant temperature;
(2) take dehydrated alcohol as solvent, compound concentration is the lecithin soln of 15 ~ 25 mg/ml, concentration is the cholesterol of 5 ~ 20mg/ml, concentration is the Chinese gooseberry seed oil mixed solution of 5 ~ 20mg/ml, after three kinds of solution mix, lecithin wherein and the mass ratio of cholesterol are 2:1, and the mass ratio of lecithin and Chinese gooseberry seed oil is 4:1;
(3) with in the phosphate buffer that is injected into behind three kinds of solution mix homogeneously described in the step (2) in the step (1), in water bath with thermostatic control 20-40min, remove residual ethanol, then carry out supersound process 20-40min, the adding distil water standardize solution, sealing namely gets Chinese gooseberry seed oil lipidosome oral administration solution after the sterilization.
2. preparation method according to claim 1 is characterized in that: in the step (1), and heating in water bath to 60 ℃, and under the 150r/min rotating speed, keep constant temperature; In the step (3), in 60 ℃ of water bath with thermostatic control 30min, remove residual ethanol, then carry out supersound process 30min, ultrasonic power is 80W, the adding distil water standardize solution, and sealing namely gets Chinese gooseberry seed oil lipidosome oral administration solution after the sterilization.
3. preparation method according to claim 1 is characterized in that: in the step (2), preparation lecithin soln concentration is 20mg/ml.
4. Chinese gooseberry seed oil liposome oral fluid, it is characterized in that: it is prepared by each described preparation method of claims 1 to 3.
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| CN103053906B (en) * | 2013-01-20 | 2014-03-19 | 湖南奇异生物科技有限公司 | Preparation method of kiwi fruit seed oil oral emulsion |
| CN103202810A (en) * | 2013-04-01 | 2013-07-17 | 浙江中医药大学 | Chinese actinidia root polysaccharide lipidosome and preparation method thereof |
| CN107951031A (en) * | 2018-01-01 | 2018-04-24 | 王艳萍 | A kind of preparation method of pine nut oral liquid |
| CN111638188A (en) * | 2020-05-27 | 2020-09-08 | 东北农业大学 | Method for measuring entrapment rate of lipid liposome |
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