IOP Conference Series: Earth and Environmental Science

PAPER • OPEN ACCESS Related content

- Antioxidant and Cytotoxic Activity of
Pineapple peel wastes as a potential source of Phyllanthus acidus Fruit Extracts
D Andrianto, W Widianti and M Bintang
antioxidant compounds - The Effect of Fermentation and Drying on
Clicanthus Nutans Lindai Extract Chemical
Constituents and Bioactivity
To cite this article: V Saraswaty et al 2017 IOP Conf. Ser.: Earth Environ. Sci. 60 012013 Y Y Lee, P S Teh, S Kumaresan et al.

- Biohydrogen Production from Pineapple

Waste: Effect of Substrate Concentration
and Acid Pretreatment
K Cahyari, A M Putri, E D Oktaviani et al.
View the article online for updates and enhancements.

Recent citations
- Antibacterial activity of phytogenic silver
nanoparticles using domestic herbs plant
extract
Dimas Pradhasumitra Mahardika et al

- Bioassay Studies on the Reaction of

Aedes aegypti & Aedes albopictus
(Diptera: Culicidae) on Different
Attractants
M.H. Nur Athen et al

- Rohayu Ramli et al

This content was downloaded from IP address 124.105.45.201 on 31/08/2021 at 02:21

 1st International Symposium on Green Technology for Value Chains 2016 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 60 (2017) 012013 doi:10.1088/1755-1315/60/1/012013

Pineapple peel wastes as a potential source of antioxidant

compounds

V Saraswaty1*, C Risdian1, I Primadona1, R Andriyani1, D G S Andayani1 and T

Mozef2
1
Research Unit for Clean Technology, Indonesian Institute of Sciences (LIPI)
Jalan Cisitu 21/154 D, Bandung 40135, Indonesia
2
Research Centre for Chemistry, Indonesian Institute of Sciences, Kawasan Puspiptek Serpong,
Tangerang Selatan, Banten, Indonesia, 15314
*Email : [email protected]

Abstract. Indonesia is a large pineapple (Ananas comosus) producing country. Food

industries in Indonesia processed this fruit for new products and further resulted wastes of
which cause an environmental problems. Approximately, one pineapple fruit total weight is
400 gr of which 60 g is of peel wastes. In order to reduce such pineapple peel wastes (PPW),
processing to a valuable product using an environmentally friendly technique is indispensable.
PPW contained phenolic compound, ferulic acid, and vitamin A and C as antioxidant. This
study aimed to PPW using ethanol and water as well as to analyze its chemical properties. Both
dried and fresh PPW were extracted using mixtures of ethanol and water with various
concentrations ranging from 15 to 95% (v/v) at room temperature for 24 h. The chemical
properties, such as antioxidant activity, total phenolic content (Gallic acid equivalent/GAE),
and total sugar content were determined. The results showed that the range of Inhibition
Concentration (IC)50 value as antioxidant activity of extracts from dried and fresh PPW were in
the range of 0.8±0.05 to 1.3±0.09 mg.mL-1 and 0.25±0.01 to 0.59±0.01 mg.mL-1, respectively,
with the highest antioxidant activity was in water extract. The highest of total phenolic content
of 0.9 mg.g-1 GAE, was also found in water extract.

1. Introduction
Pineapple (Ananas comosus) is one of the main agricultural commodities from Subang, West Java,
Indonesia. The city produces over 135.000 tons of pineapple in the year of 2014 [1]. As the impact of
pineapple food industries, pineapple peel wastes (PPW) are the important issue of waste management
of which urgently to be overcame. PPW is therefore converted into highly valuable product, since
contains considerable content of antioxidant property, sugar, phenolic compound, high fiber, and
protein. PPW also provides high potential bromelain enzyme as functional material [2-4].
Generally, PPW has been used for animal feed or land fertilizer [5], as reducing agent or
antioxidant, and substrate for production of bio-ethanol [6-7]. Antioxidant or reducing agent plays an
important role both in health sector and chemical industry. Unfortunately, chemically synthesis
antioxidant such as butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) are not safe
for health. It is found that at high dose both BHA and BHT induced tumor growth [8]. Therefore we
aimed to explore PPW as a potential of natural antioxidant. One of interesting pineapple applications
as reducing agent is in biosynthesis of metal nanoparticles. It is found that biosynthesis of metal
nanoparticles can be carried out by aqueous pineapple extract [9-10]. The work had successfully

Content from this work may be used under the terms of the Creative Commons Attribution 3.0 licence. Any further distribution
of this work must maintain attribution to the author(s) and the title of the work, journal citation and DOI.
Published under licence by IOP Publishing Ltd 1
 1st International Symposium on Green Technology for Value Chains 2016 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 60 (2017) 012013 doi:10.1088/1755-1315/60/1/012013

produced silver and cooper nanoparticles. Chlorogenic acid and ferulic acid in aqueous pineapple
extract is considered having an important role as reducing agent or antioxidant [9].
Study of antioxidant activity showed that methanol was the best organic solvent for
pineapple extract [2,11]. However, methanol is a toxic organic solvent which caused several health
problems [12]. Mixtures of ethanol and water are an environmentally preferable solvent [13]. Study on
the pineapple as antioxidant by ethanol was considerable aginst DPPH radical and achieved at
concentrations of 50 and 70% v/v. , and is considerable against DPPH radical [11]. The study aimed to
investigate the effect of water-ethanol at various concentrations for PPW as antioxidant activity.

2. Experimental
2.1. Material
PPW was collected from UKM Alam Sari, Subang, West Java Indonesia. PPW was washed under tap
water and dried in a blower oven at 55°C for 48 hr. The dried PPW was grinded mechanically and
further used for extraction. Meanwhile for fresh material, PPW was cut into small pieces, then directly
macerated in organic solvent. Chemical reagents sulphuric acid, anthrone, methanol, Na2CO3, Follin
Ciocalteu are pro analysis grade from Merck, meanwhile DPPH radical, ascorbic acid, gallic acid are
from SIGMA-ALDRICH, and ethanol is technical grade from Bratachem.

2.2.Method
2.2.1. Extraction
Ten g of fresh and dried PPW were macerated in 100 mL of ethanol:water at various concentration
(0:100, 15:85, 35:65, 55:45, 75:25 and 100:0 for 24 hr at ambient temperature). The filtrates were
evaporated under rotary vacuum evaporator (Heidolph, Germany). The yield of extraction was
determined as percentage of concentrated extract compare to weight of sample before extraction.

2.2.2. Total sugar content

Total sugar content was determined according to anthrone method [19] with a slight modification. One
g of anthrone was dissolved in 500 mL 72% v/v sulphuric acid. One mL of sample was put in a test
tube and added by 5 mL of anthrone reagent. The reaction was incubated in a water bath at 80°C for
11 min. The absorbance was then measured at 630 nm by a UV-VIS Spectrophotometer (Hitachi,
U2800) with glucose as standard. Total sugar content was calculated based on glucose standard curve.
The analysis was performed triplicate.

2.2.3 Antioxidant activity

Free radical scavenging effect was performed according to the following protocol: 50 l of samples at
various concentrations (0.5, 1, 2, 4 and 8 mg.mL-1) was put into 96-micro wells plate and added with
200 l of 0.077 mM DPPH. The reaction was incubated at room temperature for 15 min. The
absorbance was then measured at 520 nm by ELISA reader (Multiskan, BioRad). Percentage of
inhibition was calculated by using the following formula:

% Inhibition =


 (1)

C is absorbance of DPPH, and S is absorbance of sample. Ascorbic acid was used as positive control.
The procedure was performed triplicate [14].

2.2.4. Total phenolic content

Total phenolic content was determined spectrophotometrically according to Follin-Ciocalteu method.
Sample of 300 l of at concentration of 1000
g/mL was mixed with 1200 l Folin Ciocalteau (10%
v/v) and 1500 l Na2CO3 (7.5% w/v). The reaction was incubated at 55oC for 15 min. Absorbance was
then measured at λ 760 nm by UV-VIS Spectrophotometer (Hitachi, U2800). Gallic acid was used as

2
 1st International Symposium on Green Technology for Value Chains 2016 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 60 (2017) 012013 doi:10.1088/1755-1315/60/1/012013

standard and total phenolic content was calculated based on gallic acid standard curve. The total of
phenolic content was expressed as gallic acid equivalent (GAE). The procedure was performed
triplicate [15]

3. Result and discussion

3.1 Yield of extraction
PPW is highly containing water of which will cause a rapid rottening process and significantly
pollutes environment. Air contamination occures due to putrid odor whereas land contamination due to
microbial growth. Water content of PPW is shown in Table 1 It shows that water content of fresh PPW
is higher than dried PPW. This results revealed that high water content of PPW was reduced through
drying process.
Yield of PPW in various ethanol concentration is shown in Table 1. It shows that yield of dried
PPW extraction is higher than fresh PPW. This is due to the effect of drying process in which
significantly reduced water content. Moreover, water content in fresh PPW directly diluted ethanol
concentration and increased sample weight. Meanwhile, effect of ethanol concentration on either dried
or fresh PPW extraction is similar. This result indicated that compounds of PPW were majorly soluble
in mixture of ethanol and water, in which 55% v/v of ethanol/water is the most approriate solvent for
extraction. This process of extraction followed the principle of like-dissolves-like [16]. Total sugar
content determination is important since sugar plays important role in promotion of microbial growth
as the carbon source. However, since microbial growth was not found, total sugar content in dried
PPW was not evaluated. In addition, water content of dried PPW was less than 10% for which is not
promote for microbial growth. Table 1 shows that PPW water extract of PPW reveals lower sugar
content compared with ethanol extract. This is due to the sugar in PPW was more easily extracted by
the mixture of ethanol-water rather than by water solely. Fresh PPW extract contains sugar so that
easily promote the growth of microbes. Therefore, mixtures of ethanol and water has an advantage. In
extraction process, presence of ethanol will reduce the growth of microbes due to that ethanol is an
organic solvent of which exhibits a rapid and broad spectrum antimicrobial activity [17].

Table 1. Yield of extraction, antioxidant activity and total sugar content.

Water content [Ethanol] Yield Antioxidant activity Total Sugar*

(% w/w) (% v/v ) (% w/w) (IC50, mg.mL-1) (% w/w)
Fresh PPW 85.5 0 5.16 0.2±0.009 3.69
15 6.48 0.4±0.004 7.51
35 6.57 0.4±0.003 7.16
55 6.94 0.5±0.020 7.40
75 5.70 0.6±0.040 6.23
95 4.18 0.6±0.001 5.65
Dried PPW 5.43 0 24.67 0.8±0.050 n.d
15 30.95 1.3±0.090
35 28.93 1.2±0.001
55 45.38 0.9±0.070
75 30.09 0.9±0.070
95 30.37 1.2±0.110
n.d = not determined, PPW : Pineapple peel wastes, *based on dry extract.

3
 1st International Symposium on Green Technology for Value Chains 2016 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 60 (2017) 012013 doi:10.1088/1755-1315/60/1/012013

3.2 Antioxidant activity of PPW

Antioxidant activity of PPW extract in various ethanol concentrations was evaluated. The lower the
IC50 value the more the active extract. Table 1 shows, the lowest IC50 value of antioxidant activity was
found in water extract, both for fresh and dried PPW. These results indicated that water extract
contains polar compound as phenolic compound and serves as antioxidant. Moreover, the increase of
total sugar content in mixture of ethanol:water affected the decrease of PPW antioxidant activity. On
the other hand, IC50 value of fresh PPW was smaller than dried PPW. This result indicated that drying
of PPW at 50°C affected antioxidant activity in which significantly increased IC50 value. The lower
antioxidant activity in dried extract may due to bioactive compound degradation which is sensitive to
heat. Other antioxidant activity of different variety of pineapples have been carried out of which IC50
value of antioxidant activity (DPPH free radical scavenger) were in the range of 12.4 – 29.0 mg.mL-1
[11]. It was found that current IC50 of PPW extract was low both for fresh and dried PPW (Table 1).
This result indicated that currently, PPW shows as high potential source of antioxidant for DPPH free
radical scanvenger.

3.3 Total phenolic content of PPW

Phenolic compounds has strong correlation with antioxidant activity. Therefore, total of phenolic
content from both dried and fresh PPW was evaluated. The results are shown in Figure 1. It shows that
in ethanol 75% fresh PPW contained higher total phenolic content than dried PPW. We assumed, the
drying process of PPW would degrade compounds and significantly affect the antioxidant activity.

1.4
Total Phenolic Content/GAE of extract (%, w/w)

1.2

0.8

Dried PPW
0.6
Fresh PPW
0.4

0.2

0
0 15 35 55 75 95
Ethanol concentration (%)

Figure 1. Total Phenolic content/GAE (%, w/w) of PPW extract at various ethanol concentrations
in water.

From Figure 1 it shows that the highest total phenolic content of fresh PPW was found in water
extract (0% ethanol), whereas for dried PPW was of 75% v/v ethanol extract. The results of total
phenolic content in dried PPW almost the same with selected Malaysian pineapple [18].

4
 1st International Symposium on Green Technology for Value Chains 2016 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 60 (2017) 012013 doi:10.1088/1755-1315/60/1/012013

The different of total phenolic content from fresh and dried PPW was due to heat processing. Fresh
PPW was not processed and phenolic compound has yet decomposed. Whereas total phenolic content
in water extract is the highest among other. This due to water extract of PPW contain polar compound
of which more soluble in water than in aqueous ethanol,

From Figure 1 shows that PPW extract contained 540 – 1260 mg GAE/100 g PPW, nearly about 10
times higher than previous study was of 34.7-54.7 [18] and 22.7-53.8 mg GAE/100 g fruit [11].

4. Conclusion
PPW from Subang, West Java, has succesfully been converted into a potential source of antioxidant.
Ethanol/Water of 55% (v/v) provided high yield of antioxidant and avoided microbial contamination.

Acknowledgement
Author thanks to Research Unit for Clean Technology, Indonesian Institute of Sciences, for support
this research under Riset Mandiri 2016 and Ms. Rossy Choerun Nissa for her technical help on this
research.

References
[1] BPS 2015 Subang in Figure BPS Kabupaten Kota Subang hal. 76
[2] Hossain M A and Rahman M M S 2011 Food Res. Int. 44 672-6
[3] Ramalingan C, Srinath R and Islam M M 2012 Elixir Food Scie. 45 7822-26
[4] Ketnawa S, Chaiwut P and Rawdkuen S 2011 Food Bioprod. Process. 90 385-91
[5] Makinde O A, Odeyinka S M and Ayandiran S K 2011 Livest. Res. Rural Dev. 23
[6] Pornpunyapat J, Chotigeat W and Chetpattananondh P 2014 Adv. Mat. Res. 875-7 242–5
[7] Erukainure O L, Ajiboye J A, Okafo O Y, Kosok R S B, Owolab O F O and Adenekan I S O
2012 J. Food Biochem. 36 643–7
[8] Kahl R and Kappus H 1993 Z. Lebensm. Unters. Forsch., 196 329–38
[9] Ahmad N and Sharma S 2012 Green Sustain.Chem. 2 141-7
[10] Ranjitham A M, Ranjani G S, and Caroling G 2015
Int.J. Pharm Tech Res. 8 750–69
[11] Yuris A and Siow L F 2014 J. Food Stud. 3 40
[12] Kraut J A and Kurtz I 2008 Clinical J. the American Soc. of Nephrology. 3 208–25
[13] Capello C, Fischer U, and Hungerbühler K 2007 Green Chem. 9 927–34
[14] Saraswaty V, Risdian C, Budiwati T A and Tjandrawati M, 2013 Pros. Teknol. Untuk
Mendukung Pembang. Nas. 1 196–200
[15] Saraswaty V, Risdian C, Lelono R A A and Mozef T 2015 Oxid. Antioxid. Med. Sci. 4 97
[16] Chew K K, Khoo M Z, Ng S Y, Thoo Y Y, Aida W M W, and Ho C W 2011 Int. Food Res. J.
18 1427–35
[17] Mcdonnell G and Russell A D 1999 Clinical Microbiol. Rev. 12 147–79
[18] Alothman M, Bhat R, and Karim A A 2009 Food Chem. 115 785–8
[19] Yemm E W and Willis A J 1954 Biochem J. 57 508-14