Preparation on Nanoemulsion of Pomegranate Peel Polyphenol

Li FU，Huabin WANG，Yuan Ll，Bin ZHANG，Li CAl

1.School of Life Science and Food Technology，Hanshan Normal University，Chaozhou 521041，China；

2.College of Food and Pharmaceutical Science，Xinjiang Agricultural University，Urumqi 830052，China

Preparation on Nanoemulsion of Pomegranate Peel Polyphenol

Li FU1*，Huabin WANG2，Yuan Ll2，Bin ZHANG1，Li CAl2

1.School of Life Science and Food Technology，Hanshan Normal University，Chaozhou 521041，China；

2.College of Food and Pharmaceutical Science，Xinjiang Agricultural University，Urumqi 830052，China

［Objective］The aim was to obtain the preparation technology of the pomegranate peel polyphenol nanoemulsion.［Method］The pomegranate peel polyphenol nanoemulsion was prepared by the titration methods and the pseudo ternary diagram.The effect of various elements on the formation of the nanoemulsion was studied.［Results］The optimal prescription of the pomegranate peel polyphenol nanoemulsion was 4.4%（w/w）of pomegranate peel polyphenol，34.1%（w/w）of EL-40，17.1%（w/w）of anhydrous ethanol，5.7%（w/w）of IPM and 38.7%（w/w）of distilled water.［Conclusion］It is feasible to produce the pomegranate peel polyphenol nanoemulsion by the titration methods and the pseudo ternary diagram.It consists of pomegranate peel polyphenol，EL-40，anhydrous ethanol，IPM and distilled water.There is 4.4%（w/w）of the pomegranate peel polyphenol in the nanoemulsion.

Pomegranate peel polyphenol；Nanoemulsion；The pseudo ternary diagram

T he pomegranate resources are very rich in Xinjiang.The polyphenol extracted from the pomegranate peel is the by-product of the pomegranate production.About 30%of the pomegranate peel is not made full use of in the pomegranate processing.Pomegranate peel con tains polyphenols［1］，and the pomegranate peel polyphenol is a significant antioxidant.It can scavenge free ra dicals［2］，inhibit the oxidation of LDL cholesterol，prevent atherosclerosis，play the role of antilipedemic activity，inhibit angiotensin converting enzyme activity，fall blood pressure［3］，suppress hyperplasia of breast，prostate cancer and cervical cancer cells［4］，protect gastric mucosa，decrease incidence of gastric ulcer，etc［5-6］.

Nanoemulsion is also called the microemulsion，consisting of surfactant，cosurfactant，oil，water［7-8］.The nanoemulsion technology has been applied to the cosmetics，pharmaceutical，food，chemical synthesis，chemical analysis and other fields［9］. The nanoparticles granularity is small and uniform distribution，increasing the drug dispersion，promoting the absorption of drug in the body，and can improve the drug solubility and the bioavailability，enhance the stability of the unstable drug，low viscosity，form the protective effects of drugs and improve gastrointestinal absorption of the drug［10-11］.This paper reports the preparation technology ofpomegranate peel polyphenol nanoemulsion. It can not only make full use of pomegranate peel resources，but also provide theory basis for the development of pomegranate peel polyphenols for new oral preparations or cosmetic emulsions.

Materials and Methods

Reagents

The reagents included isopropyl myristate （IPM）（Japan TCI）），isopropyl palmitate （IPP）（Japan TCI），liquid paraffin，olive oil，soybean oil，peanut oil，sucrose fatty acid ester（SE） （Japan TCI），castor-oil plantsesame oil，polyoxyethylene（EL-40）（USA Acro），Tween-80，Span-80，anhydrous ethanol，glycerol， 1， 2-propanediol，1，3-butanediol，butanol and polyethylene glycol-400.

lnstrumentation

The instruments were a UV-Vis: TU-1810，Beijing，China，an ultrasonic Drug processor:JBT/C-YCL400T/3P（D），Jining，China，a rotary evaporator: RE-85C，Shanghai，China，a multipurpose water circulation pump:SHBIII，Zhengzhou，China，and a highspeed desktop centrifuge:TGL-10C，Shanghai.

Experiment methods

Screening ofsurfactants One gram of IPM （oil phase）was mixed with 6.000 g of Tween-80，EL-40 and Span-80 respectively，and then 3.000 g of anhydrous ethanol dissolved in pomegranate peelpolyphenolwas added.The mixture was shaken after distilled water was dropwise added and kept for 10d in warm bath at 70℃after it was centrifugated at 1 500-2 000 r/min for 10 min.The nanoemulsion formed if the mixture maintained limpid.If the surfactant can form the nanoemulsion dispersion system，the optimal surfactant was picked out by the pseudo-ternary phase diagram.

The pseudo-ternary phase diagram method:the mixture was prepared with IPM（oil phase），the different kinds of surfactants and the anhydrous ethanol dissolved with the pomegranate peel polyphenol as cosurfactant.The mass ratio of surfactant and the cosurfactant（anhydrous ethanol）was 3∶1 （Km=3∶1）.The mass ratio of both surfactant and cosurfactant to the oil phase（IPM）was 9∶1，8∶2，7∶3，6∶4，5∶5，4∶6，3∶7，2∶8 and 1∶9 respectively. The curves were drawn to determine nanoemulsion area after recording the mass ratio of critical point for the system from limpid to turbid.In the pseudo-ternary phase diagram，.The optimal surfactant was selected because it can form the largest nanoemulsion area.

Screening of cosurfactants The cosurfactants were screened according to Km 3∶1（w/w）.The cosurfactants supplied were:anhydrous ethanol，1，2-propylene glycol，glycerol，1，3-butanediol，butanol，polyethylene glycol-400.The experiment was conducted according to the method of screening of surfactants.If the cosurfactants individually formed nanoemulsion，the pseudo-ternary phase diagram could be used for optimization.

Screening of oil phase The oil phases supplied were IPM，IPP，liquid paraffin，olive oil，soybean oil and peanut oil.If the oil phases were suitable for nanoemulsion，the pseudoternary phase diagram could be used for optimization.

Table 1 The screening results of surfactants

Table 2 The screening results of cosurfactants

Results and Discussion

Results on screening surfactants

The results of non-ionic surfactants were screened shown in Table 1. Tween-80 and EL-40 can individually formed nanoemulsion at room temperature.

The pseudo-ternary phase diagram was applied for optimization surfactants （Tween-80，EL-40 and the mixture of Tween-80 and EL-40）at the base of table 1.The overdose was hard to dissolve in cosurfactant，the Km value was 3∶1，if the mass ratio of both surfactant-cosurfactantmixture and oil phase was 9∶1，the nanoemulsion formed could be infinitely diluted. Therefore，a group of mass ratio of 8.5:1.5 was added in test.

The results of the pseudo-ternary phase diagram were shown in Fig.1. Tween-80，EL-40 and the mixture of Tween-80 and EL-40 could all form thenanoemulsion dispersion system.The nanoemulsion area of EL-40 was obviously bigger than the others.Therefore，EL-40 was the optimal surfactant.

Table 3 The screening results of oil phase

Resultsonscreening cosurfactants

The results of six cosurfactant screened were shown in Table 2.The anhydrous ethanol could form stable nanoemulsion.

Results on screening oil phase

The results of the six kinds of oil screened were shown in Table 3.IPM and IPP can form nanoemulsion. Therefore，The results of the pseudo ternary phase diagram were shown in Fig.2.The nanoemulsion region of IPM was significantly bigger than IPP，so IPM was selected as the oil phase.

Conclusion

The optimal composition of pomegranate peelpolyphenolnanoemulsion was 4.4%（w/w）of pomegranate peel polyphenol，34.1%（w/w）of EL-40，17.1% （w/w）ofanhydrous ethanol，5.7%（w/w）of IPM and 38.7%（w/w）of distilled water.

［1］LI ZZ（李志洲）.Study on the optimal extraction conditions to polyphenols frompomegranate leaf with microwave assistance（微波辅助提取石榴叶中多酚类物质的工艺）［J］.Food and Machinery（食品与机械），2009，25（4）:72-75.

［2］YIN YL（尹燕雷），REN JH（任建辉），HE TM （何天明），et al.Cultivation condition of Xinjiang pomegranate（新疆的石榴栽培状况）［J］.Deciduous Fruit Trees（落叶果树），2006，（6）:15-16.

［3］LIU WJ（刘文江）.Resources of Punica granatum Linn.and its exploitationand utilization in Xinjiang（新疆石榴（Punica L.）资源及其开发利用）［J］.Arid Zone Research（干旱区研究），2007，24（2）:219-222.

［4］PALIDA ABLZ（帕丽达·阿布力孜），ZHANG XY（张笑颖），HAI LQ（海里茜），et al.The preliminary experiments of chemical composition of Xinjiang pomegranate peel and pomegranate seed（新疆石榴皮和石榴籽化学成分预试验）［J］.Strait Pharmaceutical Journal（海峡药学），2007，19（1）:52-53.

［5］The pharmacopoeia committee of the People’s Republic of China.Chinese pharmacopoeia 2005 edition（first）（中华人民共和国药典委员会.中国药典2005版（第一部）［S］.Beijing，Chemical industry press（北京化学工业出版社）.2005，medicinepieces:63.

［6］ZHANG J（张杰），ZHAN BY（詹炳炎），YAO XJ（姚学军），et al.Antiviral activity of tannin form the Pericarp of Punica granatum L.against Genital Herpes Virus in vitro（中药石榴果皮靴质成分抗生殖器疱疹病毒作用）［J］.China Journal of Chinese Materia Medica（中国中药杂志），1995，20（9）:556-558.

［7］ZHANG YD （张阳德）.Nanometer medicine（纳米药物学）［M］.Beijing: chemical industry press（北京:化学工业出版社），2006:163.

［8］FENG JF（奉建芳）.The research and application of nano drug delivery system in China（我国纳米给药系统的研究与应用）［J］.Journal of central south medicine（中南药学），2004，2（1）:29-33.

［9］AN HL（安红丽）.Prepartion on proanthocyanidins nanoemulsion（原花青素纳米乳的研制）［D］.Shanxi:Northwest agriculture and forestry university of science and technology（陕西:西北农林科技大学），2007.4.

［10］UCHEGBU I.F.non-ionic surfactant based vesicles（niosomes）in drug delivrery［J］.Int J Pharm，1998，172（1）: 33.

［11］CHENHB（陈华兵），WENG T（翁婷），YANG XL（杨祥良），et al.Progress of Microemulsions in Modern Pharmaceutics（微乳在现代药剂学中的研究进展）［J］.Chinese Journal of Pharmaceticals（中国医药工业杂志），2004，35（8）: 502.

［12］XIE C（谢成），ZHU HY（祝恒琛）.Applications of medical nanotechnology（纳米医药应用）［M］.Haikou:The press of Hainan（海口:海南出版社），2005:126-132.

Responsible editor：Xiaoxue WANG

Responsible proofreader：Xiaoyan WU

石榴皮多酚纳米乳的制备

傅 力1*，王华斌2，李 远2，章 斌1，蔡 丽2
（1.生命科学与食品科技学院，韩山师范学院，广东潮州 521041；2.食品科学与药学学院，新疆农业大学，新疆乌鲁木齐 830052）

［目的］对石榴皮多酚纳米乳的制备工艺进行研究。［方法］采用滴定法结合伪三元相图法，对影响石榴皮多酚纳米乳形成的各种因素进行了研究，优选了表面活性剂、助表面活性剂和油相。［结果］以蓖麻油聚氧乙烯醚（EL-40）为表面活性剂、无水乙醇为助表面活性剂、肉豆蔻酸异丙酯（IPM）为油相，石榴皮多酚纳米乳的最优配方为：石榴皮多酚4.4%（w/w）、EL-40 34.1%（w/w）、无水乙醇17.1%（w/w）、IPM 5.7%（w/w）、蒸馏水38.7%（w/w）。［结论］采用滴定法结合伪三元相图法制备石榴皮多酚纳米乳工艺可行，石榴皮多酚含量高，达到4.4%（w/w）。

石榴皮多酚；纳米乳；伪三元相图

新疆维吾尔自治区科技援疆项目（2013911072）。

傅力（1964-），女，天津人，教授，硕士生导师，从事农产品加工方向研究，E-mail：fl1990@163.com。*通讯作者。

2015-10-03

Supported by Scientific Research Projects Units of Science and Technology Department of Xinjiang Uygur Autonomous Region for Financial Support（Project No.2013911072）.

*Corresponding author.E-mail:fl1990@163.com

Received:October 3，2015 Accepted:November 15，2015

修回日期 2015-11-15