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.

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Responsible editor：Xiaoxue WANG

Responsible proofreader：Xiaoyan WU

石榴皮多酚納米乳的制備

傅 力1*，王華斌2，李 遠2，章 斌1，蔡 麗2
（1.生命科學(xué)與食品科技學(xué)院，韓山師范學(xué)院，廣東潮州 521041；2.食品科學(xué)與藥學(xué)學(xué)院，新疆農(nóng)業(yè)大學(xué)，新疆烏魯木齊 830052）

［目的］對石榴皮多酚納米乳的制備工藝進行研究。［方法］采用滴定法結(jié)合偽三元相圖法，對影響石榴皮多酚納米乳形成的各種因素進行了研究，優(yōu)選了表面活性劑、助表面活性劑和油相。［結(jié)果］以蓖麻油聚氧乙烯醚（EL-40）為表面活性劑、無水乙醇為助表面活性劑、肉豆蔻酸異丙酯（IPM）為油相，石榴皮多酚納米乳的最優(yōu)配方為：石榴皮多酚4.4%（w/w）、EL-40 34.1%（w/w）、無水乙醇17.1%（w/w）、IPM 5.7%（w/w）、蒸餾水38.7%（w/w）。［結(jié)論］采用滴定法結(jié)合偽三元相圖法制備石榴皮多酚納米乳工藝可行，石榴皮多酚含量高，達到4.4%（w/w）。

石榴皮多酚；納米乳；偽三元相圖

新疆維吾爾自治區(qū)科技援疆項目（2013911072）。

傅力（1964-），女，天津人，教授，碩士生導(dǎo)師，從事農(nóng)產(chǎn)品加工方向研究，E-mail：fl1990@163.com。*通訊作者。

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