CN102641311B - Kiwifruit seed oil liposome oral liquid and preparation method thereof - Google Patents

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

Translated fromChinese

猕猴桃籽油脂质体口服液及其制备方法Kiwifruit seed oil liposome oral liquid and preparation method thereof

技术领域technical field

本发明属于保健品制剂领域，具体涉及一种猕猴桃籽油脂质体口服液及其制备方法。The invention belongs to the field of health product preparations, and in particular relates to a kiwifruit seed oil liposome oral liquid and a preparation method thereof.

背景技术Background technique

猕猴桃(Actinidia chinensis)又名杨桃、藤梨、仙桃、毛桃等，新西兰猕基维果(kiwifruit)，是一种藤本植物，是我国特产珍贵水果之一。 随着猕猴桃加工业的发展，水果残渣量增多。猕猴桃残渣中主要是水果粗纤维和种子。这些工厂在加工生产过程中，产生大量果渣，其中含有大量的猕猴桃种子籽粒。据黄倬伟等（黄倬伟，易明华.猕猴桃籽提炼优质食用油的研究[J].粮油食品科技，1992，（4）：11~12）报道，每个猕猴桃野果中的籽粒少则250多粒，多达800余粒，平均500粒左右。经测定，籽粒中含油率为28.85%，猕猴桃籽油中不饱和脂肪酸含量高达90.37%，其中亚油酸、亚麻Kiwifruit (Actinidia chinensis ), also known as carambola, vine pear, fairy peach, hair peach, etc., New Zealand kiwifruit, is a vine and one of the special and precious fruits in China. With the development of kiwi fruit processing industry, the amount of fruit residues has increased. Kiwi fruit residues are mainly fruit crude fiber and seeds. During the processing and production of these factories, a large amount of pomace is produced, which contains a large amount of kiwifruit seed grains. According to Huang Zhuowei et al. (Huang Zhuowei, Yi Minghua. Research on the Extraction of High-quality Edible Oil from Kiwifruit Seeds [J]. Cereals, Oils and Food Science and Technology, 1992, (4): 11~12), the number of seeds in each wild fruit of kiwifruit ranges from as little as 250 grains, as many as 800 grains, with an average of about 500 grains. It has been determined that the oil content in the seeds is 28.85%, and the content of unsaturated fatty acids in kiwifruit seed oil is as high as 90.37%, of which linoleic acid, flax

酸含量占74.83%；动物实验结果显示，猕猴桃籽油具有明显的降血脂作用，口服安全无毒。姚茂君（姚茂君，李加兴，张永康.猕猴桃籽油的开发利用探讨[J].食品与发酵工业，2001，27（12）：28~30，]姚茂君，李嘉兴，张永康.猕猴桃籽油理化特性及脂肪酸组成[J].无锡轻工大学学报，2002，2（3）：307~309）、张永康（张永康，蒋剑波，陈莉华.猕猴桃果仁油脂肪酸的测定及其利用[J].吉首大学学报(自然科学版),2001，22（4）：37~38，82）等研究了湖南湘西“米良一号”猕猴桃的种子，发现含油率为23.5%，气相色谱测定脂肪酸组成，a-亚麻酸含量高达63.99%。因此猕猴桃籽油可作为补充不饱和脂肪酸的重要保健油资源加以开发利用。 The acid content accounts for 74.83%. Animal experiment results show that kiwi seed oil has obvious blood lipid-lowering effect, and it is safe and non-toxic when taken orally. Yao Maojun (Yao Maojun, Li Jiaxing, Zhang Yongkang. Discussion on the development and utilization of kiwifruit seed oil [J]. Food and Fermentation Industry, 2001, 27 (12): 28~30,] Yao Maojun, Li Jiaxing, Zhang Yongkang. Physicochemical properties and fatty acids of kiwifruit seed oil Composition [J]. Journal of Wuxi University of Light Industry, 2002, 2 (3): 307~309), Zhang Yongkang (Zhang Yongkang, Jiang Jianbo, Chen Lihua. Determination and utilization of fatty acids in kiwi fruit kernel oil [J]. Journal of Jishou University (Nature Science Edition), 2001, 22 (4): 37~38, 82) and others studied the seeds of "Miliang No. 1" kiwifruit in Xiangxi, Hunan, and found that the oil content was 23.5%. The fatty acid composition was determined by gas chromatography, and the a-linolenic acid content was as high as 63.99%. Therefore, kiwifruit seed oil can be developed and utilized as an important health oil resource for supplementing unsaturated fatty acids. the

猴桃籽油中含亚麻酸与亚油酸，是目前发现的除苏子油外亚麻酸含量最高的天然植物油。亚油酸和亚麻酸是两种对人体健康特别重要的必需脂肪酸，只能从外界摄取而不能在人体内自行合成。α-亚麻酸是EPA和DHA的前体物质，因而猕猴桃籽油具有降低血脂、胆固醇和血压，预防心血管疾病，抑制血小板凝集，防止血栓形成，防止癌症发生，抗氧化和延缓衰老等多种作用。所以猕猴桃籽油是一种优质的功能食品、药品和美容化妆品的原料。Kiwi seed oil contains linolenic acid and linoleic acid, and is the natural vegetable oil with the highest content of linolenic acid found so far except perilla oil. Linoleic acid and linolenic acid are two essential fatty acids that are particularly important to human health. They can only be ingested from the outside and cannot be synthesized in the human body. α-linolenic acid is the precursor of EPA and DHA, so kiwi seed oil has many functions such as lowering blood lipid, cholesterol and blood pressure, preventing cardiovascular disease, inhibiting platelet aggregation, preventing thrombosis, preventing cancer, anti-oxidation and delaying aging. effect. So kiwi seed oil is a high-quality raw material for functional food, medicine and beauty cosmetics.

但是，猕猴桃籽油中含量高达90%的不饱和脂肪酸给油脂的保存带来了问题。不饱和脂肪酸分子含有多个双键，因而对氧气、光线和热极为敏感，极易氧化变质。油脂的氧化不仅使其失去了应有的保键功能，而且还会产生一些对人体有害的物质。However, the unsaturated fatty acids in kiwifruit seed oil, which contain as much as 90%, pose problems for oil preservation. Unsaturated fatty acid molecules contain multiple double bonds, so they are extremely sensitive to oxygen, light and heat, and are easily oxidized and deteriorated. The oxidation of oil not only makes it lose its proper key function, but also produces some harmful substances.

脂质体(liposome)是Bangham在1965年发现的一种由磷脂构成的一种类脂小球体。是将药物包封在脂质(磷脂与胆固醇)双分子层形成的薄膜中制成的微型球状体（Bangham ad.difusion of univalent ions aeross the lamellae of swollen Phosphlipids [J].Mol. Biol，1965 13:238~252.）。双分子层内外分别包封脂溶性和水溶性药物，其理化性质和结构类似细胞膜，又称为人工生物膜。 脂质体作为一种新颖的功能成分制剂而备受关注，近年来，脂质体制剂研究与应用取得很大发展。脂质体制剂的作用特点主要有（陆彬.药物新剂型与新技术[M].第二版.北京:人民卫生出版社，2005.5）：Liposome is a small lipid sphere composed of phospholipids discovered by Bangham in 1965. It is a microsphere made by encapsulating drugs in a film formed by a lipid (phospholipid and cholesterol) bilayer (Bangham ad.difusion of univalent ions aeross the lamellae of swollen Phosphlipids [J]. :238~252.). The fat-soluble and water-soluble drugs are encapsulated inside and outside the bimolecular layer, and its physical and chemical properties and structure are similar to cell membranes, also known as artificial biomembranes. As a novel functional ingredient preparation, liposome has attracted much attention. In recent years, the research and application of liposome preparation have made great progress. The main features of liposome preparations are (Lu Bin. New dosage forms and new technologies of drugs [M]. Second Edition. Beijing: People's Medical Publishing House, 2005.5):

(l)保护功能成分提高稳定性(l) Protect functional components to improve stability

一些不稳定的功能成分被脂质体包封后，可受到脂质体双层膜的保护:被包裹的功能成分在体内转运过程中不受酶和免疫系统所破坏。After some unstable functional ingredients are encapsulated by liposomes, they can be protected by liposome bilayer membranes: the encapsulated functional ingredients are not destroyed by enzymes and immune system during in vivo transport.

(2)降低功能成分毒性(2) Reduce the toxicity of functional components

脂质体本身对人体无毒，功能成分被包裹后在心、肾中累积量比游离功能成分低，对心、肾有毒性的功能成分包封脂质体后，可明显降低毒性。The liposome itself is non-toxic to the human body, and the accumulation of functional components in the heart and kidney after encapsulation is lower than that of free functional components. The toxicity of functional components that are toxic to the heart and kidney can be significantly reduced after encapsulating liposomes.

(3)缓释作用(3) Sustained release effect

控制功能成分的释放速率，达到长效缓释。Control the release rate of functional ingredients to achieve long-acting sustained release.

(4)靶向性(4) Targeting

进入体内后被网状内皮系统吞噬，使功能成分主要在肝、脾、肺和骨髓等组织器官中蓄积，实现被动靶向给药。After entering the body, it is phagocytosed by the reticuloendothelial system, so that the functional components mainly accumulate in tissues and organs such as liver, spleen, lung and bone marrow, realizing passive targeted drug delivery.

脂质体在医药行业应用成熟，但在食品领域尚属起步阶段。脂质体作为一个优良的载体，通过磷脂的包覆，既可提高包埋物质的稳定性，又使其具有靶同性，可提高功能成分的生物利用率。目前，脂质体的制备方法及其例子如下：Liposomes are well-established in the pharmaceutical industry, but still in their infancy in the food industry. As an excellent carrier, liposomes can not only improve the stability of embedded substances, but also make them have target homogeneity through the coating of phospholipids, which can improve the bioavailability of functional components. At present, liposome preparation methods and examples thereof are as follows:

1. 机械分散法 (mechanical dispersion) 通常是先将脂质溶于有机溶剂中，减压旋转蒸发使之在玻璃容器的内壁形成均匀的脂质膜，然后加入水相介质通过振摇将脂质分散并形成脂质体。例如水飞蓟油脂质体制备：将处方量卵磷脂、胆固醇、水飞蓟油溶解于5ml氯仿，在旋转蒸发仪上减压蒸干有机溶剂，达胶态后，加入适量体积的磷酸盐缓冲溶液(PBS)，于35℃水浴恒温振荡至完全水化，用0.45μm微孔滤膜过滤3次，即得。采用此方法制备脂质体较为普遍，但采用氯仿等有机试剂，毒性大，不适合用于食品领域。1. The mechanical dispersion method (mechanical dispersion) usually first dissolves the lipid in an organic solvent, and rotates it under reduced pressure to form a uniform lipid film on the inner wall of the glass container, and then adds an aqueous medium to dissolve the lipid by shaking. Disperses and forms liposomes. For example, the preparation of milk thistle liposomes: dissolve the prescribed amount of lecithin, cholesterol, and milk thistle oil in 5ml of chloroform, and evaporate the organic solvent to dryness under reduced pressure on a rotary evaporator. After reaching a colloidal state, add an appropriate volume of phosphate buffer The solution (PBS) was shaken in a water bath at 35°C until it was completely hydrated, and filtered through a 0.45 μm microporous membrane for 3 times. It is more common to prepare liposomes by this method, but organic reagents such as chloroform are used, which are highly toxic and not suitable for use in the food field.

2. 溶剂分散法2. Solvent dispersion method

溶剂分散法(solvent dispersion) 是先将脂质溶于有机溶剂中，再加入到含有被包裹药物的水相中，在有机相与水相交界面上磷脂质以单分子层(即脂质体双分子层膜的一半) 排列。The solvent dispersion method (solvent dispersion) is to dissolve the lipid in the organic solvent first, and then add it to the aqueous phase containing the encapsulated drug, and the phospholipid forms a monomolecular layer at the interface between the organic phase and the aqueous phase (i.e. liposome bilayer). Half of the molecular layer membrane) arrangement.

例如茶树油脂质体的制备：称取处方量的卵磷脂、胆固醇、茶树油、VE、去氧胆酸钠、吐温-80等,向其中加入适量无水乙醇,超声至悬液均匀,得类脂混悬液;水合介质置于恒温磁力搅拌器上以保持规定的温度和搅拌速度。将类脂混悬液慢慢滴入水合介质中,得脂质体初悬液(其中乙醇的体积分数为10%),将此初悬液置于水浴式超声仪中超声20 min,即得茶树油脂质体悬液。卵磷脂的质量浓度为10 g·L^-1、卵磷脂与胆固醇的质量比为5:1、茶树油的质量浓度为1.0 g·L^-1、VE的质量浓度为1.0 g·L^-1、去氧胆酸钠的质量浓度为0.5 g·L^-1、吐温-80的质量浓度为4.0 g·L^-1、水相介质是pH值为6.8的PBS缓冲溶液。采取此方法制备脂质体的案例较少，主要是因为工艺参数必须控制精确，否则制得的脂质体不稳定。For example, the preparation of tea tree oil liposomes: Weigh the prescribed amount of lecithin, cholesterol, tea tree oil, VE, sodium deoxycholate, Tween-80, etc., add an appropriate amount of absolute ethanol to it, and ultrasonically until the suspension is uniform, to obtain Lipid suspension; hydration medium is placed on a constant temperature magnetic stirrer to maintain the specified temperature and stirring speed. The liposome suspension was slowly dropped into the hydration medium to obtain the liposome primary suspension (the volume fraction of ethanol was 10%), and the primary suspension was placed in a water-bath ultrasonic instrument for 20 min to obtain Tea tree oil liposomal suspension. The mass concentration of lecithin is 10 g·L^-1 , the mass ratio of lecithin to cholesterol is 5:1, the mass concentration of tea tree oil is 1.0 g·L^-1 , the mass concentration of VE is 1.0 g·L^-1 , The mass concentration of sodium deoxycholate was 0.5 g·L^-1 , the mass concentration of Tween-80 was 4.0 g·L^-1 , and the aqueous medium was PBS buffer solution with a pH value of 6.8. There are few cases in which liposomes are prepared by this method, mainly because process parameters must be controlled accurately, otherwise the liposomes obtained are unstable.

针对猕猴桃籽油容易氧化的特点， 冯卫华等采用喷雾干燥法对猕猴桃籽油进行微胶囊化研究（猕猴桃籽油微胶囊化技术研究,农业工程学报，2004,20（1）234~236）。 姚茂军等制备了猕猴桃籽油软胶囊 （猕猴桃油软胶囊的研究，发酵与食品工业，29（12）：2003,58~61）。喷雾干燥法仍需较高温度（进风口180℃，出风口80℃），因此对猕猴桃籽油有损失。目前尚未有制备猕猴桃籽油脂质体的公开报道。In view of the easy oxidation of kiwifruit seed oil, Feng Weihua et al. used the spray drying method to study the microencapsulation of kiwifruit seed oil (Research on microencapsulation technology of kiwifruit seed oil, Journal of Agricultural Engineering, 2004, 20 (1) 234~236). Yao Maojun and others prepared kiwi seed oil soft capsules (Research on kiwi oil soft capsules, Fermentation and Food Industry, 29 (12): 2003, 58~61). The spray-drying method still requires a higher temperature (180°C at the air inlet and 80°C at the air outlet), so there is a loss of kiwifruit seed oil. At present, there is no public report on the preparation of kiwifruit seed oil liposomes.

发明内容Contents of the invention

本发明所要解决的技术问题是提供一种猕猴桃籽油脂质体口服液及其制备方法，用本发明方法制备得到的猕猴桃籽油脂质体口服液，猕猴桃籽油的的稳定性高，安全，避免了猕猴桃籽油中的不饱和脂肪酸被氧化，从而达到方便运输和长期保藏的目的。The technical problem to be solved by this invention is to provide a kind of kiwifruit seed oil liposome oral liquid and preparation method thereof, the kiwifruit seed oil liposome oral liquid prepared by the inventive method, the stability of kiwifruit seed oil is high, safe, avoid The oxidation of unsaturated fatty acids in kiwifruit seed oil is achieved, so as to achieve the purpose of convenient transportation and long-term preservation.

本发明提供的技术方案是：一种猕猴桃籽油脂质体口服液的制备方法，该方法包括如下步骤：The technical scheme provided by the invention is: a kind of preparation method of kiwifruit seed oil liposome oral liquid, the method comprises the steps:

（1）量取磷酸盐缓冲液置于恒温振荡器中，水浴加热至55-65℃，并保持恒温；(1) Measure the phosphate buffer solution and place it in a constant temperature oscillator, heat it in a water bath to 55-65°C, and keep it at a constant temperature;

（2）以无水乙醇为溶剂，分别配制浓度为15-25 mg/ml的卵磷脂、5~20mg/ml的胆固醇、5~20mg/ml的猕猴桃籽油混合溶液；(2) Using absolute ethanol as a solvent, prepare a mixed solution of lecithin with a concentration of 15-25 mg/ml, cholesterol with a concentration of 5-20 mg/ml, and kiwifruit seed oil with a concentration of 5-20 mg/ml;

（3）将步骤（2）中所述的三种溶液混合均匀后注入到步骤（1）中的磷酸盐缓冲液中，于恒温水浴20-40min，除去残留乙醇，然后进行超声处理20-40min， 加蒸馏水定容，密封，灭菌后即得猕猴桃籽油脂质体口服溶液。(3) Mix the three solutions described in step (2) evenly and inject them into the phosphate buffer solution in step (1), place in a constant temperature water bath for 20-40 minutes to remove residual ethanol, and then perform ultrasonic treatment for 20-40 minutes , add distilled water to constant volume, seal, and sterilize to obtain the kiwi seed oil liposome oral solution.

所述的制备方法，优选地，步骤（1）中，量取pH6.8的磷酸盐缓冲液置于恒温振荡器中，水浴加热至60℃，并在150r/min转速下保持恒温；步骤（3）中，于60℃恒温水浴30min，除去残留乙醇 ，然后进行超声处理30min（超声功率80W），加蒸馏水定容，密封，灭菌后即得猕猴桃籽油脂质体口服溶液。In the preparation method, preferably, in step (1), measure the phosphate buffer solution with pH 6.8 and place it in a constant temperature oscillator, heat the water bath to 60°C, and keep the constant temperature at 150r/min; step ( 3) in a constant temperature water bath at 60°C for 30 minutes to remove residual ethanol, then perform ultrasonic treatment for 30 minutes (ultrasonic power 80W), add distilled water to constant volume, seal, and sterilize to obtain kiwifruit seed oil liposome oral solution.

所述的制备方法，优选地，步骤（1）中，配制卵磷脂溶液浓度为20mg/ml；步骤（3）中三种溶液混合后，其中的卵磷脂与胆固醇的质量比为2:1，卵磷脂与猕猴桃籽油的质量比为4:1。。In the preparation method, preferably, in step (1), the concentration of the prepared lecithin solution is 20 mg/ml; after the three solutions are mixed in step (3), the mass ratio of lecithin to cholesterol is 2:1, The mass ratio of lecithin to kiwi seed oil is 4:1. .

所述的制备方法，优选地，步骤（3）中，60℃恒温水浴30min，蒸发挥尽残留乙醇，然后进行超声处理，30min,超声功率为80W。In the preparation method, preferably, in step (3), 60°C constant temperature water bath is used for 30 minutes to evaporate residual ethanol, and then ultrasonic treatment is performed for 30 minutes, and the ultrasonic power is 80W.

本发明的方法优选采用湖南老爹农业科技开发股份有限公司提供猕猴桃籽油，例如产品批号为20111230。The method of the present invention preferably adopts the kiwifruit seed oil provided by Hunan Laodi Agricultural Science and Technology Development Co., Ltd., for example, the product batch number is 20111230.

本发明还提供由上述制备方法制备得到的猕猴桃籽油脂质体口服液。The present invention also provides the kiwifruit seed oil liposome oral liquid prepared by the above preparation method.

本发明具有以下有益效果：The present invention has the following beneficial effects:

猕猴桃籽油是常用的猕猴桃籽利用的重要组分，由于其所含的亚麻酸的保健作用，提高猕猴桃籽油的稳定性，实现猕猴桃籽油中亚麻酸的靶向输送，提高其吸收和利用已经成为国内外研究的热点。猕猴籽油难溶于水，口服生物利用度低，极大的限制了其临床疗效，由于猕猴桃籽油中的亚麻酸的含高达60%以上，极其容易被氧化，本发明结合脂质体的靶向性和缓释性，将猕猴桃籽油制备成脂质体口服溶液，可以提高猕猴桃籽油的的稳定性，安全性，避免猕猴桃籽油中的不饱和脂肪酸被氧化，并且可以实现猕猴桃籽油的靶向运输，达到提高猕猴桃籽油吸收率的作用。 从而达到方便运输和长期保藏的目的，推广了猕猴桃籽油脂质体技术的商业价值。本发明采取较低的温度，克服了微囊化过程中喷雾干燥温度较高的缺点，同时释放速度较软胶囊慢，可进一步作为缓释制剂的原料。Kiwi fruit seed oil is an important component commonly used in kiwi fruit seed utilization. Due to the health care effect of linolenic acid contained in it, it improves the stability of kiwi fruit seed oil, realizes the targeted delivery of linolenic acid in kiwi fruit seed oil, and improves its absorption and utilization It has become a research hotspot at home and abroad. Kiwi seed oil is insoluble in water, and its oral bioavailability is low, which greatly limits its clinical curative effect. Since the content of linolenic acid in kiwi seed oil is as high as 60%, it is extremely easy to be oxidized. The present invention combines liposome Targeting and sustained release, the preparation of kiwifruit seed oil into liposome oral solution can improve the stability and safety of kiwifruit seed oil, avoid the oxidation of unsaturated fatty acids in kiwifruit seed oil, and realize the Targeted transport of oil to achieve the effect of improving the absorption rate of kiwifruit seed oil. Thereby the purpose of convenient transportation and long-term preservation is achieved, and the commercial value of kiwifruit seed oil liposome technology is promoted. The invention adopts lower temperature, overcomes the disadvantage of higher spray drying temperature in the microencapsulation process, and at the same time releases slower than soft capsules, and can be further used as a raw material for sustained-release preparations.

具体实施方式Detailed ways

下面通过具体实施方式的详细描述来进一步阐明本发明，但并不是对本发明的限制，仅仅作示例说明。The present invention will be further clarified through the detailed description of specific embodiments below, but it is not intended to limit the present invention, but only for illustration.

实施例1Example 1

1、 猕猴桃籽油脂质体包封率的测定1. Determination of Encapsulation Efficiency of Kiwifruit Seed Oil Liposomes

采用透析-紫外分光光度法测定猕猴桃籽油的含量。移取20ml猕猴桃籽油脂质体放入透析袋中，透析17h后，将袋中溶液倒入100ml容量瓶，选用乙醇为破乳剂，结合超声，加入石油醚提取，将混合溶液进行离心(3000r/min，3min)，取10ml上清液于25ml容量瓶中，用石油醚定容，在233nm处测定其吸光度A₁;同法测定空白脂质体吸光度A₀ ，计算△A，由标准曲线求出所包封猕猴桃籽油的含量。 The content of kiwifruit seed oil was determined by dialysis-ultraviolet spectrophotometry. Pipette 20ml kiwifruit seed oil liposomes into the dialysis bag, after dialysis for 17h, pour the solution in the bag into a 100ml volumetric flask, select ethanol as the demulsifier, combine ultrasound, add sherwood oil to extract, the mixed solution is centrifuged (3000r/ min, 3min), get 10ml supernatant in 25ml volumetric flask, constant volume with sherwood oil, measure its absorbance A₁ at 233nm place; Measure blank liposome absorbance A₀ with the same method, calculate △ A, obtain by standard curve The content of encapsulated kiwi fruit seed oil is shown.

                  包封率%=△A*100/A₁Encapsulation rate%=△A*100/A₁

2、配方与工艺优选2. Formula and process optimization

脂质体口服溶液的技术关键在于脂质体的制备。以包封率为指标，考察配方中卵磷脂与胆固醇的质量比(L:C)、 卵磷脂与油的质量比(L:O)、水浴时间（t）、加热温度(T)等四个因素对包封效果的影响。根据单因素试验结果，选用L9(4³)正交表进行脂质体配方正交试验设计。The technical key of liposome oral solution lies in the preparation of liposome. Taking the encapsulation rate as an index, four factors including the mass ratio of lecithin to cholesterol (L:C), the mass ratio of lecithin to oil (L:O), water bath time (t), and heating temperature (T) were investigated. The influence of factors on the encapsulation effect. According to the single factor test results, the L9(4³ ) orthogonal table was chosen to carry out the orthogonal test design of liposome formulation.

表1   正交设计因素表Table 1 Orthogonal design factor table

表2  正交设计表及结果 L（9）3⁴Table 2 Orthogonal design table and results L(9) 3⁴

分组groupL:C(w/w)L:C(w/w)L:O(w/w)L:O(w/w)水浴时间（min）Water bath time (min)T(℃)T(°C)包封率（%）Encapsulation rate (%)    1 11111111136.3436.34    2 21122222238.5438.54    331133333345.6745.67    442211223374.4574.45    552222331165.3565.35    662233112266.3666.36    773311332250.5650.56    8 83322113345.5745.57    9 93333221155.2055.20K1K140.18340.18350.78350.78349.42349.42352.29752.297 theK2K268.72068.72049.82049.82056.06356.06351.82051.820 theK3K350.44350.44355.74355.74353.86053.86055.23055.230 the最优水平optimal level22332233 theRR28.53728.5375.9235.9236.6406.6403.4103.410 the显著性significant*(p<0.05)*(p<0.05) the the the the

通过直观分析，可知最佳处方及工艺条件为因此，猕猴桃籽油脂质体最佳制备工艺为: 卵磷脂与胆固醇的质量比(2:1)、 卵磷脂与油的质量比(4:1)、水浴时间30min、加热温度60℃。配方中卵磷脂与胆固醇的比例对脂质体形成的影响最大。Through intuitive analysis, it can be seen that the best prescription and process conditions are therefore, the best preparation technology of kiwifruit seed oil liposomes is: the mass ratio (2:1) of lecithin to cholesterol, the mass ratio (4:1) of lecithin to oil , water bath time 30min, heating temperature 60 ℃. The ratio of lecithin to cholesterol in the formulation has the greatest effect on liposome formation.

实施例2Example 2

将恒温振荡器温度设置为60℃，预热。准确量取100mlpH6.8的磷酸盐缓冲液置于恒温振荡器中，水浴加热至60℃，并在150r/min转速下保持恒温。以无水乙醇为溶剂 ，分别配制卵磷脂(LC)浓度为20mg/ml，胆固醇(CH)浓度为10mg/ml，猕猴桃籽油(KFO) 浓度为15mg/ml，纯度99.0%。将各溶液混合均匀后用注射器均匀注入到恒温振荡器中的磷酸盐缓冲液中。 60℃恒温水浴30min，除去残留乙醇，超声处理30min(超声功率80W) ，加蒸馏水定容至250ml，密封，辐照灭菌后即得猕猴桃籽油脂质体口服溶液。 Set the constant temperature oscillator temperature to 60 °C and preheat. Accurately measure 100ml of pH 6.8 phosphate buffer solution and place it in a constant temperature shaker, heat it in a water bath to 60°C, and keep it at a constant temperature at a speed of 150r/min. Using absolute ethanol as solvent, respectively prepare lecithin (LC) concentration of 20mg/ml, cholesterol (CH) concentration of 10mg/ml, kiwifruit seed oil (KFO) concentration of 15mg/ml, purity 99.0%. After the solutions were mixed evenly, they were uniformly injected into the phosphate buffer solution in the constant temperature shaker with a syringe. 60°C constant temperature water bath for 30 minutes, remove residual ethanol, ultrasonic treatment for 30 minutes (ultrasonic power 80W), add distilled water to make up to 250ml, seal, irradiate and sterilize to obtain kiwifruit seed oil liposome oral solution. the

本发明的方法优选采用湖南老爹农业科技开发股份有限公司提供猕猴桃籽油，本实施例所用产品批号为20111230。The method of the present invention preferably adopts kiwifruit seed oil provided by Hunan Laodi Agricultural Science and Technology Development Co., Ltd., and the batch number of the product used in this embodiment is 20111230.

实施例3Example 3

用乙醇作为溶剂分别配制卵磷脂(LC)浓度为16mg/ml，胆固醇(CH)浓度为12mg/ml，猕猴桃籽油(KFO) 浓度为10mg/ml，制备方法同实施例2。Use ethanol as solvent to prepare lecithin (LC) concentration respectively as 16mg/ml, cholesterol (CH) concentration as 12mg/ml, kiwifruit seed oil (KFO) concentration as 10mg/ml, preparation method is with embodiment 2.

实施例4Example 4

用乙醇作为溶剂分别配制卵磷脂(LC)浓度为20mg/ml，胆固醇(CH)浓度为20mg/ml，猕猴桃籽油(KFO) 浓度为5mg/ml，制备方法同实施例2。Use ethanol as solvent to prepare respectively lecithin (LC) concentration to be 20mg/ml, cholesterol (CH) concentration to be 20mg/ml, kiwifruit seed oil (KFO) concentration to be 5mg/ml, preparation method is with embodiment 2.

实施例5Example 5

用乙醇作为溶剂分别配制卵磷脂(LC)浓度为23mg/ml，胆固醇(CH)浓度为6mg/ml，猕猴桃籽油(KFO) 浓度为18mg/ml，制备方法同实施例2。Use ethanol as solvent to prepare respectively lecithin (LC) concentration to be 23mg/ml, cholesterol (CH) concentration to be 6mg/ml, kiwifruit seed oil (KFO) concentration to be 18mg/ml, preparation method is with embodiment 2.

Claims (4)

Translated fromChinese

1.一种猕猴桃籽油脂质体口服液的制备方法，其特征在于该方法包括如下步骤：1. a preparation method of kiwifruit seed oil liposome oral liquid, is characterized in that the method comprises the steps:（1）量取pH6.8的磷酸盐缓冲液置于恒温振荡器中，水浴加热至55-65℃，并保持恒温；(1) Measure the phosphate buffer solution with pH 6.8 and place it in a constant temperature oscillator, heat the water bath to 55-65°C and keep it at a constant temperature;（2）以无水乙醇为溶剂，配制浓度为15~25 mg/ml 的卵磷脂溶液，浓度为5~20mg/ml的胆固醇，浓度为5~20mg/ml的猕猴桃籽油混合溶液，三种溶液混合后，其中的卵磷脂与胆固醇的质量比为2:1，卵磷脂与猕猴桃籽油的质量比为4:1；(2) Using absolute ethanol as a solvent, prepare a lecithin solution with a concentration of 15-25 mg/ml, a mixed solution of cholesterol with a concentration of 5-20 mg/ml, and a mixed solution of kiwifruit seed oil with a concentration of 5-20 mg/ml, three kinds After the solution is mixed, the mass ratio of lecithin and cholesterol is 2:1, and the mass ratio of lecithin and kiwifruit seed oil is 4:1;（3）将步骤（2）中所述的三种溶液混合均匀后注入到步骤（1）中的磷酸盐缓冲液中，于恒温水浴20-40min，除去残留乙醇，然后进行超声处理20-40min，加蒸馏水定容，密封，灭菌后即得猕猴桃籽油脂质体口服溶液。(3) Mix the three solutions described in step (2) evenly and inject them into the phosphate buffer solution in step (1), place in a constant temperature water bath for 20-40 minutes to remove residual ethanol, and then perform ultrasonic treatment for 20-40 minutes , add distilled water to constant volume, seal, and sterilize to obtain the kiwifruit seed oil liposome oral solution.2.根据权利要求1所述的制备方法，其特征在于：步骤（1）中，水浴加热至60℃，并在150r/min转速下保持恒温；步骤（3）中，于60℃恒温水浴30min，除去残留乙醇，然后进行超声处理30min，超声功率为80W，加蒸馏水定容，密封，灭菌后即得猕猴桃籽油脂质体口服溶液。2. The preparation method according to claim 1, characterized in that: in step (1), the water bath is heated to 60°C and kept at a constant temperature at a speed of 150r/min; in step (3), the water bath is heated at 60°C for 30 minutes , remove residual ethanol, then carry out ultrasonic treatment for 30min, the ultrasonic power is 80W, add distilled water to constant volume, seal, and promptly obtain kiwifruit seed oil liposome oral solution after sterilization.3.根据权利要求1所述的制备方法，其特征在于：步骤（2）中，配制卵磷脂溶液浓度为20mg/ml。3. The preparation method according to claim 1, characterized in that in step (2), the concentration of the prepared lecithin solution is 20 mg/ml.4.一种猕猴桃籽油脂质体口服液，其特征在于：其由权利要求1至3任一项所述的制备方法制备得到。4. A kiwifruit seed oil liposome oral liquid, characterized in that: it is prepared by the preparation method described in any one of claims 1 to 3.