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Agriculture and Agricultural Science Procedia 9 (2016) 297 – 303

International Conference on Food, Agriculture and Natural Resources, IC-FANRes 2015

Antioxidant Activity and Physicochemical Properties of Nicolaia

speciosa Flower Extract
Rifda Naufalin*, Herastuti Sri Rukmini
Agricultural Technology, Agricultural. Faculty Jenderal Soedirman University, dr Soeparno street, Purwokerto 53123, Indonesia

Abstract

Flower of Nicolaia speciosa contain some bioactive compounds which are potential as natural antioxidant agent. N. speciosa
flowers contains bioactive compounds such as polyphenols, alkaloids, flavonoids, steroids, saponins and essential oils.
Unfortunately, extract of N. speciosa flower is very volatile and not stable upon exposures to light and oxygen. Therefore the
extract should be modified into nanoencapsulan powder. This research aimed 1) to study the best encapsulant proportion of
gelatin-cyclodextrin (w/w) and N. speciosa flower extract proportion that affected into antioxidant activity. The results showed
that the proportion of encapsulan gelatin-cyclodextrin (w/w) 1:1 (w/w), the proportion of extract:encapsulan 1:1 (w/w), with
nanoencapsulating method resulted the highest a total phenolic 635.8 mg/100g and the highest antioxidant activity that is
59.34%.

©
© 2016
2015 Published by Elsevier
The Authors. PublishedB.V.
by This is anB.V.
Elsevier open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
Peer-review under responsibility of the organizing committee of IC-FANRes 2015.
Peer-review under responsibility of the organizing committee of IC-FANRes 2015
Keywords: Nicolaia speciosa flower, nanoencapsulan, antioxidant activity, physicochemical properties

1. Introduction

Nicolaia speciosa flowers contains bioactive compounds such as polyphenols, alkaloids, flavonoids, steroids,
saponins and essential oils. N. speciosa flower content of bioactive compounds, phenolic and triterpenoid (Naufalin
et al., 2010). Bioactive components of N. speciosa flower is found by solvent extractions, becoming a concentrate if
the solvent is reduced. Qi & Hedges (1995) stated that most of bioactive compounds are sensitively on air, rays, or

* Corresponding author. Tel.: +6281548875448; fax: +62281626945.

E-mail address: rnaufalin@yahoo.co.id

2210-7843 © 2016 Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
Peer-review under responsibility of the organizing committee of IC-FANRes 2015
doi:10.1016/j.aaspro.2016.02.132
298 Rifda Naufalin and Herastuti Sri Rukmini / Agriculture and Agricultural Science Procedia 9 (2016) 297 – 303

heat. Otherwise, bioactive compounds should be made in nanoencapsulant. Nanoencapsulant has smaller size than
microencapsulant. Nanoparticle is solid colloid particle on 1 – 1000 nm (1µm) (Donbrow, 2000).
Encapsulant or other fillers were customary used in many research for antibacterial or antioxidant activity.
According to Naufalin & Herastuti (2012a), antioxidant activity formula of peel and seed N. speciosa fruit up to
22,49% with gelatin-maltodextrin filler 6% into concentrate. N. speciosa fruit formula with gelatin-maltodekstrin
filler in proportion filler 1:1 (b/b) and concentration 6% (b/b) become the highest antioxidant activity amount
29,22%.
This research studied about encapsulan proportion of gelatin-cylodextrin, encapsulan concentrate, effect of
nanoencapsulation and self-life to antioxidant activity during storage. According to Naufalin & Herastuti (2012c),
maltodextrin as filler has the lower stability emulsion though it has higher stability oxidative. Moreover, the used
encapsulants are gelatin and cyclodextrin. Kim et al. (1996) stated that protein and some of carbohydrate are
interacted in water, ion and some of oil or water that stabilized droplet emulsion partly during homogenization. This
research aimed 1) to study the best encapsulant proportion of gelatin-cyclodextrin (w/w) and N. speciosa flower
concentrate proportion that affected into antioxidant activity.

2. Research Method

2.1. Place and Time of Research

The research was conducted at the Laboratory of Agricultural Technology, Faculty of Agriculture UNSOED. The
research held from March until July 2015.

2.2. Materials

Materials needed research consisting of: Nicolaia speciosa flower. Chemicals used among other technical
solvents, namely ethanol 96 %; encapsulant (gelatin and cyclodextrin), ethanol 99.8 % (PA), linoleic acid, distilled
water, ammonium thiocyanate, 0.02 M FeCl2 tetrahydrate, concentrated HCl, N2 gas. Other materials needed
include a coarse filter paper, Whatman No. 41 paper, plastic, paper label, paper towels and aluminum foil.

2.3. Equipment

The tools used in this research that cabinet dryer dryers, blenders, analytical scales, rotary evaporator (Bibby
RE 200), shaker, oven (Memmert, Japan), desiccators, pipettes (Pyrex, Germany), pH meter. Tool for the analysis of
total phenol and antioxidant activity consists of spectrophotometers, centrifuges, glass tools (Pyrex, Germany),
vortex, Fial tubes, micropipette (Gilson), and bath tobacco leaf waste, ie shaker, rotary, tube N2. Tool to extract
formulation, a set of tools for testing antimicrobial activity glasses, vortex, micropipettes, incubator 37oC,
desiccator, refrigerator and vacuum pump.

2.4. Undertaking Experiment

N. speciosa flower powder sample preparation (Naufalin, 2013), N. speciosa flower are selected. Material
selection results cleaned with water, then dried in a dryer at a temperature of 500C until the moisture content of 8-
10%. Furthermore simplicia dry milled to obtain a homogeneous powder.
The preparation of N. speciosa flower extraction and nanoencapsulation (Naufalin and Herastuti, 2013). N.
speciosa flowert powder was extracted twice with ethanol (1:4 w / v). The extraction process is done by maceration
at 37°C, with a rotation speed of 150 rpm for 24 hours every level. Filtrat separated from the solvent by evaporation
in a rotary evaporator until no solvent dripping again. The solvent was evaporated at a temperature of 50oC.
Residual solvent is removed with nitrogen gas to produce an essential oil (Naufalin et al., 2013).
Separated N. speciosa flower extractt 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 N. speciosa flower extracts with concentration of 50%. The process of N. speciosa
 Rifda Naufalin and Herastuti Sri Rukmini / Agriculture and Agricultural Science Procedia 9 (2016) 297 – 303 299

flower nanoencapsulation is by adding the filler such gelatin and cyclodextrin with the different proportion and
ratio. Using hotplate stirrer in 50°C temperature, the filtrate heated and add the filler and stirred on 15 minutes.
Furthermore done diminution emulsion particles into nanoparticles by using a dispersing machine (Ultra-Turrax)
22,000 rpm for 30 minutes (Naufalin & Herastuti, 2013).

2.5. Experimental design

The research was conducted in experimental method. Experimental design used was Randomized Completely
Block Design (RCBD). The tested factor contains 4 factors, are encapsulant of gelatin:cyclodextrin proportion (w/w)
1:1, 1:2, 1:3; encapsulant concentrate proportion (w/w) 1:1, 1:2, 1:3; using nanoencapsulation, without
nanoencapsulation; self-life 0, 2, 4, 6 weeks. These experiments are made by factorial trial designs, so it found 72
combinations of treatment unit and each unit repeated in twice so that found 144 units.

2.6. Determination Procedure

Measurement of Total Phenol Othman et al. (2007) were modified. Activity of antioxidant formulas were
analyzed using modified iron thiocyanate method (Yurttas et al., 2000). This determination is based on the
formation of peroxide as a product of oxidation of linoleic acid. Percent oxidation was calculated by comparing the
absorbance value of the test sample (As) with control absorbance value (A ctrl) multiplied by 100%, which is
expressed by the formula:

Oxidation Percent (%) =

Antioxidants activity = 100% - oxidation percent

2.7. Statistical Analysis

The observed variables in this research are antioxidant activity of nanoencapsulant powder involved total
phenol measurement, physicochemical properties including extracted fraction content noncapsulated, and
photomicrograph. Quantitative data was analyzed used variance test (F test) in 5%, continued in Duncan’s Multiple
Range Test (DMRT) if it finds the significant difference.

3. Result and Discussion

3.1. Total Phenol

Total phenol content of powder nanoencapsulant N. speciosa flower is analyzed through describing method
from Othman et al. (2007) that is modificated. This method is performed to determine the phenolic content in fruit
powder nanoenkapsulan N. speciosa flower quantitatively. The principles based on the reduction ability of
fosfomolibdat-fosfotungstat compounds from blue colour of Folin-Ciocalteu reagen, so it can be determined by
spectrophotometer (Suryanto et al., 2005).
Self-life give the significant effect on total phenol content of powder nanoencapwsulant kecombrang fruit.
Average value of total phenol content of powder nanoencapsulant in self-life on 0, 2, 4 and 6 weeks are 447.8
mg/100g; 353.2 mg/100g; 270.9 mg/100g dan 292.7 mg/100g (Figure 1). Based on Figure 1, nanoencapsulant
powder on week 0 self-life describes the highest total phenol 447.8 mg/100g than others.
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nanoencapsulation). Encapsulant proportion 1:2 are the proper combination, than encapsulant proportion 1:3,
because it had three times more in cyclodextrin ingredient than gelatin, therefore it is unstable formula. The
combination of protein and carbohydrate from encapsulated properties role as oil and water emulsifier. During water
and oil emulsion, protein is function as primer stabilizer where protein layer is absorbed around oil particles,
otherwise it is function as emulsifier and film agent, whereas carbohydrates as a secondary stabilizer to serve as a
thickener or framework in solution phase (Montesqrit, 2007). The particle becomes smaller, and encapsulant could
protect the fractions, futhermore the non-capsulated fraction content will be low.

3.3. Relationship between Antioxidant Activity and Total Phenol

Relationship between antioxidant activity and total phenol nanoencapsulan N. speciosa flower indicated by the
equation y = 1.001 x + 48.639 and the value of correlation r = 0.026. This shows that there is no significant
correlation between total phenol and antioxidant activity of nanoencapsulant N. speciosa flower (Figure 4). The
highest total phenol did not show high antioxidant activity, but the high antioxidant activity may be related to some
particular classes of phenol. This is consistent with the results of research conducted by Naufalin and Herastuti
(2013b), that there is no correlation between total phenol and antioxidant activity of fruit kecombrang. The results
Othman et al. (2007), also found that the lack of correlation between the phenol content and antioxidant activity of
berry fruit and wine.

Figure 4. The correlation between total phenol and antioxidant activity of nanoencapsulant N.speciosa flower

4. Conclusion

Nanoencapsulant N. speciosa flower with encapsulant proportion (gelatin-cylodextrin) 1:1 (w/w) give the highest
antioxydant activity (49.54%), concentrate proportion encapsulant 1:2 (b/b) give the highest antioxydant activity
(49.53%), and total phenol (454.5mg/100g). Nanoencapsulation gave the highest total phenol (368.8 mg/100g) and
antioxydant activity (49.77%) than without nanoencapsulation, moreover will push the decreased value of total
phenoln 15.02% and antioxydant activity 3,17%. Storage at week 0, resulted the highest total phenol and
antioxydant (447.8 mg/100g and 53.1%). Antioxydant activity on 6 weeks will be descreased amount 19.38% than
0 week.

Acknowledgements

Thanks to the DP2M Higher Education, which has provided research funding through Hibah Kompetensi 2015

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