uji aktivitas Antioksidan

13th ASEAN Food Conference, 9-11 September 2013, Singapore

Meeting Future Food Demands: Security & Sustainability

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NANOENCAPSULATED OF NATURAL ANTIOXIDANT BASED ON

KECOMBRANG (Nicolaia speciosa Horan ) FRUIT USING

CYCLODEXTRIN-GELATIN AS FILLER INGREDIENT

Rifda Naufalin1,*

Herastuti S. Rukmini1

1 Food Science and Technology, Jenderal Soedirman University, Purwokerto, Indonesia

* Corresponding author: [email protected]

Abstract

Introduction: Kecombrang (Nicolaia speciosa Horan ) fruit contains bioactive substances that can be used

as antioxidants. Unfortunately, extract of fruit kecombrang is very volatile and not stable upon exposures to

light and oxygen. This research used fruit kecombrang peel extract for making formula kecombrang with

fillers. Therefore, formulation is needed to develop nanoencapsulated kecombrang extract to make this

volatile extract more stable and easy to use for preservative in food products. This research was aimed of

production of nanoencapsulated antioxidant which is practical, stable and can be used for food products.

Methods: Method used for this research was extraction and formulation for extract encapsulation. Factors

tested were the proportion of filler ingredient cyclodextrin : gelatin, i.e.; 1:1, 2:1, 3:1 (w/w) and proportion

of extract : filler ingredient, i.e.; 1:1, 1:2, 1:3 (w/w).

Results: The result showed that filler material cyclodextrin-gelatin (2 : 1) gives highest antioxidant

activity (58.10%) total phenolic (287 mg/100 g) than other proportionstal; extract proportion with filler

material (1:2) gives highest antioxidant activity (61,45%) total phenolic (341 mg/100g) than other

proportion.

Conclusion: Nanoencapsulan formula of fruit kecombrang extract with cyclodextrin and gelatin (2:1) and

filler material and exctract (2 : 1), as the formulation is potential for natural antioxidant

Keywords antioxidant, extract, formulation, kecombrang, nanoencapsulation

1. Introduction

Food material is usually has the easiest nutrition damage such as fat, carbohydrate, and

protein. The damage caused from the activated microbe, enzyme and oxidation. Antioxidant is the

compounds that can clean off and provide the form or fuse the effect of reactive oxygen species.

These problems are related to quality of antioxidant as inhibitor oxidation reaction for reactive free

radical (Kuncahyo et al., 2007). The more antioxidant consumption will decrease the risk of

degenerative diseases such as cardiovascular, cancer, atherosclerosis and osteoporosis. Food

antioxidant consumption also increase immunology rate and obstruct degenerative diseases from

aging.

Kecombrang (Nicolaia speciosa Horan ) fruit contains bioactive substances that can be used

as antioxidants. Unfortunately, extract of fruit kecombrang is very volatile and not stable upon

exposures to light and oxygen (Naufalin et al., 2009). This research used fruit kecombrang peel

extract for making formula kecombrang with fillers. Therefore, formulation is needed to develop

nanoencapsulated kecombrang extract to make this volatile extract more stable and easy to use for

preservative in food products. This research was aimed of production of nanoencapsulated

antioxidant which is practical, stable and can be used for food products.



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Natural antioxidant in form of extractions have the lacking process, such as easier volatile

compound in processing in high temperature, easier oxidation process and not easier in dispersion

in dry material and hard process (Koswara, 2007). These lacking handed with adding the filling

and stabilizing in seed of fruit kecombrang being the stabilized formula. The coupled formulation is

for handling particles into small wrapping polymers. The general aim is as the medium to change

the active compounds into hard particle and it becomes easier to handle, restrains volatile

compound, flavors, and other reactive compounds. Otherwise, it protects the material from the light,

environment, and humidity.

This research studied the activity of antioxidant formula of peel fruit kecombrang with adding

the filler, in order to protect the active component of peel fruit kecombrang and stabilized the

formula. The filler raw used are cyclodextrin and gelatin, the aimed of this research is determine

cyclodextrin filler : gelatin into peel of fruit kecombrang extraction (b/v), that gives the highest

antioxidant activity and determine the proportion of gelatin : cyclodextrin (w/w) that produces the

highest antioxidant activity.

2. Methods

2.1. Preparation of Fruit kecombrang peel extraction

Kecombrang peel had weighed and added by sterilizing distilled water with a ratio of 1:1

(w/v) respectively. Then blended for 3 min and filtered using a filter cloth to separate the extract

and persipitat (residue) in order to obtain kecombrang seeds and peel extracts with concentration of

50%.

2.2. Formulation of the fruit kecombrang peel extract

The process of peel fruit kecombrang formula is by adding the filler such cyclodextrin and

gelatin with the different proportion and ratio. Using hotplate stirrer in 50C temperature, the filtrate

heated and add the filler and stirred on 15 minutes.

2.3. Design and Data Analyzed

This research used the experimental method. The trial design used is Completely

Randomized Design. The trial factors are filler proportion of cyclodextrin : gelatin, i.e.; 1:1, 2:1,

3:1 (w/w) and proportion of extract : filler ingredient, i.e.; 1:1, 1:2, 1:3 (w/w).. These trials are

in factorial trial design, reach 9 trial combinations and each trial repeated in 3 times and therefore it

gets 54 trial units. The observed variables were the fruit kecombrang extract antioxidant activity

using method of ferric tiosianate modificated (Yurttas et al., 2000), and total fenol (Othman et al.,

2005). Data were analyzed using analysis of variance (F test) that followed by Duncan's Multiple

Range Test (DMRT) at the level of 5 percent and regression test when it shows a significant effect

3. Result and Discussions

Antioxidant activity test from peel formula of fruit kecombrang used the ferric thiosianate

method in modification according to Yurttas et al., 2000. The principle of antioxidant activity using

FTC method is the value measurement of complex absorb in red color of ferric tiosianat. This color

complex is formed from peroxide radical that oxidized ion ferro (Fe2+

) into ion ferri (Fe3+

),

therefore, the formed ion ferri will be related to ion tiosianat make the complex red color of ferri



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tiosianat (Budijanto et al., 2000). Antioxidant activity was measured using a method based on the

ability feritiosianat formation of free radical compounds. Free radicals are formed due to the

oxidation of linoleic acid. Peroxide enhancement to this method is expressed as the number of

compounds that can oxidize Fe2+

into Fe3+

as expressed in the following equation: RO + Fe2+

RO - +

Fe3+

. Thiocyanate method of measuring the antioxidant activity based on the inhibition of the

formation of compounds that are reactive radicals. Fat oxidation processes produce primary

products peroxide. Expressed as peroxide compounds capable of oxidizing Fe2+

into Fe3+

, and Fe3+

ions with subsequent CNS produces red color was measured at a wavelength of 500 nm (Wahyudi,

2006).

2.1. Antioxidant Activity

The antioxidant activity of nanoencapsulan formula of fruit kecombrang in trials of filler

proportion and extraction proportion using filling raw were done in this research. In Figure 1. the

filler proportion of cyclodextrin : gelatin, i.e.; 1:1, 2:1, 3:1 (w/w) shows that the antioxidant

activity in encapsulant proportion 2:1 (w/w) gives higher antioxidant activity (58.10%) than other

proportions.

Figure 1. The effect of filler proportion cyclodextrin : gelatin (w/w) into nanoencapsulean

antioxidant activity.

Single factor of encapsulant proportion of cyclodextrin-gelatin, shows the average of

antioxidant activity in filler proportion 2:1 (w/w) highest (58,10 %) than other proportions. The

lowest antioxidant activity is shown in filler proportion of cyclodextrin-gelatin 3:1 (w/w) amount

54,17 %. The proportion of cyclodextrin-gelatin (2:1 w/w) shows the highest antioxidant activity

because it has the more cyclodextrin than gelatin. The proportion of the two components produces

an optimum interaction that stabilizes the emulsion formula. Proportion of 3:1. cyclodextrin

composition has three times more than the gelatin so the formula is less stable. The combination of

protein and carbohydrates are from the coating material role in the oil and water emulsion. In oil

and water emulsion, the protein which serves as the primary stabilizer in the absorption of protein

layer around the oil droplets, as it also serves as an emulsifying and film-forming agent (coating),

while carbohydrates as a secondary stabilizer to serve as a thickener or framework in solution phase



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(Montesqrit, 2007). Cyclodextrin is soluble in water because it has the hydrophilic characteristic

than gelatin. It can protect the exctract from the environment but soluble in water will make the

antioxidant compound extracted and obstruct and decrease the speed of peroxyde form as oxydated

reaction indicator. According to Lineback and Inlett (1982) xyclodekstrin has soluble characteristic

in cold and hot water, using low viscosity. Gelatin is soluble in water in temperature 71 C (Poppe,

1999), whereas, according to Wahyuni and Peranginangin (2007), gelatin has soluble characteristic

and make a gel in temperature 49C.

In Figure 2. Extract anf filler proportion, i.e.; 1:1,1:2, 1:3 (w/w) shows that the antioxidant

activity in encapsulant proportion 1:2 (w/w) gives higher antioxidant activity (61.45%) than other

proportions.

Figure 2. The effect of extract : filler proportion into nanoencapsulean antioxidant activity.

The peel formula of fruit kecombrang in extract proportion : filler raw 1:2 has the large activity than

others. It caused from filler proportion for twice were enough to protect the active component in peel of fruit

kecombrang, whereas the formula of filler proportion 1:1 the active component did not enough to be

protected and the antioxidant activity is little more. The proportion 1:3 is not significantly different on

concentrate 10% because on 15% the active component is being protected.

The couple formulation is the process to geminate particles into thin wrapping polymers. This is in

order to arrest volatile, flavor, aroma, and the other active components on food. It also to restrain their

characteristic release and protecting from the effect of environment such as ultra violet, air and humidity

(Naufalin dan Herastuti, 2012).

2.2. Total Phenolic

Antioxidant compound in peel of fruit kecombrang is phenolic or polyphenol in form of

flavonoid, cyanide acid descent. Phenolic content is analyzed by the Singleton and Rossi method

(Othman 1995). The principal is based on the reduction of phosphomolibdate-phosphotungstate

compound from Folin Ciocalteu reagent with blue color therefore it is determined using

spetrophotometer.



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Proportion of self-treatment encapsulant cyclodextrin : gelatin (w / w) showed the average total

phenol nanocapsules of proportion 1:1, 2:1 and 3:1 consecutively 287 mg/100g, 284,3 mg/100g and

199 mg/100g (Figure 3).

Figure 3. the filler proportion of cyclodextrin : gelatin : (w / w) of the total phenol

nanocapsules.

Analysis results of variance proportion of cyclodextrin-gelatin treatment greatly affect the value

of total phenol microcapsules. From the figure it can be seen that the proportion of

cyclodextrin-gelatin 2:1 (w/w) produce the highest value of total phenol (287 mg/100g). Formula

with the proportion of cyclodextrin-gelatin (w / w) of 2:1 had higher antioxidant activity may be

due to the proportion of the two components that produces an optimum interaction that stabilizes

the emulsion formula. Proportion of 3:1 cyclodextrin composition had three times more than the

gelatin so the formula is less stable. Aspect proportion of filler is best ratio between

cyclodextrin-gelatin (w / w) of 3:1 is formulation of peel fruit kecombrang extract with a value of the

highest total phenol. Comparison between the fill of protein and the right carbohydrat will produce a

stable formula. The combination of protein and carbohydrates are from the coating material role in

the oil and water emulsion. In the oil and water emulsion, the protein which serves as the primary

stabilizer in the absorption of protein layer around the oil droplets, as it also serves as an

emulsifying and film-forming agent (coating), while carbohydrates as a secondary stabilizer to

serve as a thickener or framework in solution phase (Montesqrit, 2007).

The extract and filler proportion showed the average total phenol nanocapsules of proportion

1:1, 1:2 and 1:3 consecutively 341 mg/100g, 249 mg/100g and 178 mg/100g (Figure 4).



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Figure 4. The effect of extract : filler proportion of the total phenol nanocapsules.

The filler material and exctract 2 : 1 is capable of protecting the active components in the

extract of fruit peel in kecombrang. The filler which is twice more than its extract has not been able

to completely protect the active components in the extract. This is because of the filler is twice more

than its extract, therefore the compounds bind bioactive compound in large numbers even in the

weak binding, therefore when doing the total phenol analysis the bioactive compound is released

and detected as total phenol. In 1:1 ratio the filler did not bind enough the bioactive compound,

while in proportion of 1:3 the bioactive compound is protected and bounded strongly therefore total

phenol is difficult to release and undetected.

Conclusion : Nanoencapsulant formula of fruit kecombrang extract with cyclodextrin and gelatin

(2:1) and filler material and exctract (2 : 1), as the formulation is potential for natural antioxidant

Acknowledgements

Thanks to the DP2M Higher Education which has provided research funding through Hibah

Kompetensi 2012

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uji aktivitas Antioksidan