986 International Journal of Food Science and Technology 2016, 51, 986–993

Original article
Sacha inchi (Plukenetia volubilis L.) shell: an alternative source of
phenolic compounds and antioxidants

Rosana Chirinos,1 Ornella Necochea,1 Romina Pedreschi2 & David Campos1*

1 Instituto de Biotecnolog
ıa (IBT), Universidad Nacional Agraria La Molina – UNALM, Av. La Molina s/n, Lima, Peru
2 School of Agronomy, Pontificia Universidad Cat
olica de Valpara
ıso, Calle San Francisco s/n, La Palma, Casilla 4-D, Quillota, Chile
(Received 18 August 2015; Accepted in revised form 10 December 2015)

Summary Sacha inchi seed (SI) is known as a rich source of oil with high content of polyunsaturared fatty acids of
the x-3 and x-6 type (~85% of total fatty acids). However, few studies have focused on the use of by-
products from the seed. The aim of this study was to characterise the main phenolic families present in SI
shell and to evaluate the best extraction solvent for the extraction of phenolic compounds (PC) and
antioxidant capacity (AOXC). The PC content corresponded to 74.5
5.1 mg g1 of which 93.1% were
condensed tannins and the remaining compounds corresponded to free and bound phenolic acids,
hydrolyzable tannins, flavonoids and flavanoids. Protocatechic and p-coumaric acids but also hydroxyci-
nammic acid derivatives of ferulic and o-coumaric type and lignan derivatives were identified. Acetone
containing solvents favoured the extraction of higher amounts of total PC and AOXC. This study high-
lights the potential use of SI shell as a novel and alternative source of PC antioxidants for the nutraceuti-
cal and/or functional food industries.
Keywords Antioxidant capacity, phenolic compounds, Plukenetia volubilis, shell.

2009), triticale bran and straw (Hosseinian & Mazza,

Introduction
Sacha inchi (Plukenetia volubilis L.) is an oleaginous
plant that belongs to the Euphorbiaceae family. It grows
in the lowlands of the Peruvian Amazon having been
cultivated for centuries by the indigenous population
(Hamaker et al., 1992; Guti
errez et al., 2011). Sacha
inchi (SI) seeds are composed of~33–35% of shell and
~65–67% of kernel. Kernel contains approximately
54% oil and 33% proteins (Hamaker et al., 1992). SI
seeds are mainly industrialised into oil due to its unique
fatty acid composition, comprising approximately 34%
linoleic acid (x-6) and 51% linolenic acid (x-3), respec-
tively (Hamaker et al., 1992; Guti
errez et al., 2011;
Chirinos et al., 2015). However, SI seed industrialisa-
tion generates by-products such as the shell and cake
and both are discarded without any further use.
Several investigations state that the by-products gen-
erated from different plant sources after processing could
become important sources of antioxidant bioactive com-
pounds of the phenolic type. For instance, antioxidant
bioactive compounds have been reported in almond shell
(Garrido et al., 2007), hazelnut shell (Shahidi et al.,
2007; Contini et al., 2008), sunflower shell (Weisz et al.,
*Correspondent: E-mail: dcampos@lamolina.edu.pe
2009), chesnut fruit pericarp and integument (de Vascon-
celos et al., 2010), pecan nut shell (do Prado et al., 2014),
rice brand and rice husk (Wanyo et al., 2014) to cite a
few plant sources. Previous studies carried out in our lab-
oratory showed that the phenolic compounds (PC) con-
tent of the shell is eight fold higher that the PC content of
the cake (D. Campos, R. Chirinos and R. Pedreschi, un-
published results). Up to date, to our knowledge, there
are no investigations reported related to the PC content
and antioxidant properties of the SI shell as an alterna-
tive by-product source of antioxidant compounds. Thus,
the main aims of this study were: to characterise the main
phenolic families present in SI shell (by-product) and to
evaluate the best extraction solvent for the extraction of
total phenolic compounds (TPC) and antioxidant capac-
ity (AOXC). Results from this study will allow to add-
value to this by product as a source of natural antioxi-
dant bioactive compounds.
Materials and methods
Sample material
Seeds of Plukenetia volubilis, ecotype Pinto Redondo
were obtained from Tarapoto city, province of San

doi:10.1111/ijfs.13049
© 2016 Institute of Food Science and Technology

Martin (Region of San Mart
ın) from Peru. Seeds
(5 kg) were manually cleaned and selected. The shell
was manually removed using a small hammer. The
shell was ground in a disc mill (≤1 mm) and kept at
20 °C until further phenolics characterisation.
Chemicals
Trolox (6-hydroxy-2,5,7,8-tetramethyl chroman-2-car-
boxylic acid), fluorescein sodium salt, 2,20 -azinobis (2-
amidinopropane) dihydrochloride (AAPH), 2,20 -azino-
bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS),
2,4,6-tripyridryl-s-triazine (TPTZ), 2N Folin-Ciocal-
teau reagent, q-dimethyl amino cinnamaldehyde
(DMACA), ethylene-diamine-tetra-acetic acid (EDTA)
were purchased from Sigma Chemicals Co. (St. Louis,
MO, USA). Phenolic acid standards (p-coumaric,
o-coumaric, protocatechuic, ferulic, gallic, caffeic,
chlorogenic, p-hydroxybenzoic) were purchased from
Sigma Chemicals Co. Cyanidin was purchased from
ChromaDex (Santa Clara, CA, USA). HPLC-grade
acetonitrile, methanol and other solvents and reagents
were purchased from Merck (Darmstadt, Germany)
and Fischer Scientific (Fair Lawn, NJ, USA).
Extraction of phenolic compounds from SI shell
Different solvents for the extraction of PC from SI shell
were tested in this investigation. The selection of sol-
vents, combinations and extraction conditions were
based on frequently reported data in the literature and
based on preliminary trials of our group. Six different
solvents were evaluated: methanol/water/acetic acid
(80/19/1, v/v/v, pH 4.32), ethanol/water/acetic acid (80/
19/1, v/v/v, pH 4.28), acetone/water/acetic acid (80/19/
1, v/v/v, pH 4.39), methanol/ethanol/water/acetic acid
(40/40/19/1, v/v/v/v, pH 4.26), methanol/acetone/water/
acetic acid (40/40/19/1, v/v/v/v, pH 4.46) and ethanol/
acetone/water/acetic acid (40/40/19/1, v/v/v/v, pH 4.46).
Extraction conditions corresponded to: 60 °C, sample/
solvent ratio of 1/10, orbital agitation of 300 r.p.m.,
two extraction steps of 60 and 30 min, respectively. The
resulting mixtures were centrifuged at 4000 g for
10 min at 4 °C and the supernatants were recovered
and combined to evaluate the PC content and AOXC
using three methods: ABTS (2,2 azinobis (3-ethylben-
zothiazoline 6-sulfonate)), FRAP (ferric reducing ability
of plasma) and ORAC (oxygen radical absorbance).
Determination of free and bound phenolic acids
Free and bound phenolic acids were extracted as
described by Hosseinian & Mazza (2009) with slight
modifications. Free phenolic acids were extracted by
mixing 1 g of sample with 80% ethanol (v/v) in a 1/50
ratio. Extraction was carried out at room temperature,
© 2016 Institute of Food Science and Technology
Antioxidant phenolics from sacha inchi shell R. Chirinos et al. 987
under 300 r.p.m. agitation and two extraction steps of
60 and 30 min were used. Then, the extracts were cen-
trifuged at 4000 g for 10 min, and the supernatants
were pooled together and vacuum evaporated (≤40 °C)
until dryness. Finally, the residue was reconstituted in
MilliQ water and ready for HPLC analysis. Bound
phenolic acids were extracted from the residue of the
free phenolic acids extraction. The residue was mixed
with 50 mL of 2M NaOH and 100 lL of 0.1% EDTA
(v/v) at room temperature, 300 r.p.m. agitation during
4 h. After the extraction, the resulting mixture was
acidified to pH 2 with 2 M HCl and transferred to a
separatory funnel to which 50 mL of diethyl ether was
added and mixed. The supernatant was recovered and
the whole procedure repeated one more time. Both
supernatants were pooled together and vacuum con-
centrated (≤40 °C) until dryness and finally reconsti-
tuted in MilliQ water and ready for HPLC analysis.
Phenolic acid profiles were determined according to
the procedure proposed by Chirinos et al. (2008). This
analysis was performed with a Waters 2695 Separation
Module (Waters, Milford, MA, USA) with an autoin-
jector and a 2996 photodiode array detector. Chro-
matographic separation was carried using an X-terra
RP18 (5 lm, 250 9 4.6 mm) column (Waters). The
mobile phase consisted of water: acetic acid (94:6, v/v,
pH 2.27) (solvent A) and acetonitrile (solvent B). Gra-
dient condition was as follows: 0–15% B in 40 min,
15–45% B in 45 min, and 45–100% B in 10 min. The
flow rate was kept at 0.5 mL min1 and the injection
volume was 20 lL. Phenolic acids were identified and
quantified by comparing their retention time and UV–
visible spectral data to known previously injected stan-
dards. Results were expressed as mg of phenolic
acid g1 or mg/100 g of sample.
Determination of total flavanoids and total flavonoids
Total flavanoid (compounds containing flavan 3-ols,
flavan 4-ols, flavan 3,4-diols, flavanones and deriva-
tives) were extracted and estimated using the chro-
mogen DMACA reagent following the method
proposed by Delcour & Devarebeke (1985). Results
were expressed in mg of catechin equivalents (CE) per
g of sample FW. Flavonoids (compounds containing
flavonol and flavone families) extraction and quantifi-
cation was carried out by means of the aluminium
chloride colorimetric method proposed by Chang et al.
(2002). Results were expressed as mg of quercetin
equivalents (QE) per g of sample.
Determination of hydrolysable (gallotannins) and
condensed (proanthocyanidins) tannins
Hydrolysable tannins were extracted and quantified
using the procedure reported by Inoue & Hagerman
International Journal of Food Science and Technology 2016
988 Antioxidant phenolics from sacha inchi shell R. Chirinos et al.
(1988). Hydrolysable tannin (gallotannin) contents were
determined using the rhodanine assay. Results were
expressed as mg of gallic acid equivalents (GAE) g1 of
sample. The extraction of the condensed tannins is based
on the protocol proposed by Adamson et al. (1999).
Condensed tannin quantification was carried out follow-
ing the method reported by Julkunen-Tiitto (1985) and
results were expressed as cyanidin equivalents (CYE) per
g of sample.
Determination of lignans
Lignans were extracted by direct alkaline hydrolysis
using a modified procedure described by Johnsson
et al. (2000). A 0.5 g defatted sample was hydrolysed
with 10 mL of 2 M NaOH under 300 r.p.m. agitation
for 2 h at 60 °C. After hydrolysis, the mixture was
centrifuged and the supernatant was transferred to a
20 mL erlenmeyer flask, acidified to pH 3 using 2 M
sulphuric acid, adjusted to 25 mL and filtered before
HPLC analysis. The separation of lignans was accom-
plished using a SymmetryÒ C18 (5 lm, 250 9 4.6 mm)
column (Waters) and a guard column
(4.6 mm 9 2.0 mm) at 30 °C. The mobile phase was
composed of solvent (A) aqueous 25 mM phosphoric
acid and solvent (B) acetonitrile. Gradient conditions
were: 10–40% B in 25 min, 40–100% B in 1 min,
100% B in 5 min and 100–10% B in 5 min. A flow
rate of 0.6 mL min1 was used and the injection vol-
ume was 10 lL. Lignans were identified (at 280 nm)
and quantified by comparing their retention time and
UV–visible spectral data to known previously injected
standards: secoisolaricirecinol diglucoside (SDG) and
matairesinol (MAT). Results were expressed as mg of
lignans g1 sample.
Total phenolic compounds and antioxidant capacity
Total phenolic compounds and AOXC were deter-
mined in the extracts obtained from SI shell using dif-
ferent extraction solvents. Total phenolics were
determined following the method of Singleton & Rossi
(1965). The results were expressed as mg of GAE/g
sample FW. The AOXC was determined with the
ABTS, FRAP and ORAC assays. Three different
methods to measure AOXC were tested based on their
particular mechanism of action (substrate, assay condi-
tions, etc.) as to evaluate the performance of SI shell
antioxidants under different conditions.
For ABTS assay, the procedure followed was the
same procedure described by Campos et al. (2006).
Samples (150 lL) were allowed to react with 2850 lL
of ABTS+ solution in ethanol until a steady absor-
bance was reached at room temperature and dark con-
ditions. The decrease in absorbance due to
antioxidants was recorded at 734 nm. The FRAP
International Journal of Food Science and Technology 2016
assay was conducted according to Benzie & Strain
(1996) with minor modifications. The FRAP reagent
consists of acetate buffer (300 mM, pH 3.6), TPTZ
(10 mM in HCl 40 mM) and FeCl36H2O (20 mM)
(10:1:1, v/v/v). A total of 2850 lL of FRAP reagent
was mixed with 150 lL of sample at 37 °C. Then, the
absorbance at 593 nm decreased to a stable value,
implicating that Fe3+-2,4,6-tri-pyridyl-S-triazine was
reduced by the samples. The ORAC assay was per-
formed in a 96-well microplate fluorometer (Bio Tek
Instruments), essentially as described by Ou et al.
(2001). Peroxyl radical was generated using AAPH and
fluorescein was used as the substrate. To all experimen-
tal wells, 250 lL of sodium fluorescein solution 48 nM
were added. In addition, blank, standard and samples
received 25 lL of 75 mM phosphate buffer (pH 7.4),
Trolox and extract dilutions respectively, and were
incubated for 10 min at 37 °C. Reactions were initiated
by the addition of 25 lL of AAPH. Fluorescence filters
were used at an excitation wavelength of 485 nm and
an emission wavelength of 520 nm, taking measure-
ments from each sample at 60 s intervals for 55 min.
AOXC for the three assays was calculated as lmol of
Trolox equivalents (TE)/g of sample from a standard
curve developed with Trolox.
Statistical analyses
All quantitative analysis were performed in triplicate.
Values of different results were expressed as the
mean
standard deviation (SD). Results were tested
for statistical significance by one-way ANOVA. A Dun-
can’s post-hoc test was used to assess statistical signifi-
cant differences among treatments (P < 0.05). The
Statgraphics Centurion XV software 15.2.06 (Stat
Point Inc., Virginia, USA) was used for all statistical
calculations.
Results and discussion
The mechanical extraction of SI seed resulted in yields
of oil, shell and cake of 27.0, 34.8 and 38.2 g/100 g
seed, respectively. The water content of the shell corre-
sponded to 8.4%.
Characterisation of phenolic compounds from sacha inchi
shell
Phenolic contents by family type are presented in
Table 1. The main phenolic family present in SI shell
were condensed tannins (69.4 mg CYE g1 or 75.7
CYE g1, dry weight DW) representing 93.1% of the
PC content. Naczk & Shahidi (2004) indicated that
proanthocyanidins (condensed tannins) are localised in
the out layer of the seed coat and the endosperm, hav-
ing much higher contents of these compounds. The
© 2016 Institute of Food Science and Technology
 Antioxidant phenolics from sacha inchi shell R. Chirinos et al. 989

Table 1 Phenolic compounds in sacha inchi shell
Phenolic compounds
Free phenolic acids (mg g ) 1
Bound phenolic acids (mg g1)
Flavonoids (mg QE g1)
Flavanoids (mg CE g1)
Hydrolysable tannins (mg GAE g1)
Condensed tannins (mg CYE g1)
Lignans (mg SDGE g1)
Total (mg g1)
Results correspond to the average
standard deviation of three repli-
cates.
condensed tannin content of the SI shell was higher
than the values reported for different by-products
derived from the industrial processing such as: skin
and brown hull of almonds that ranged from 16.5 to
48.8 and from 0.14 to 0.40 mg CYE g1, respectively
(Garrido et al., 2007), pecan nut shell with a value of
36.9 mg CE g1 DW reported by do Prado et al.
(2014) and triticale, wheat, rye and oat bran with
reported values of 2.65, 5.09, 2.12 and 1.48 mg g1,
respectively (Hosseinian & Mazza, 2009). The content
of hydrolyzable tannins in SI shell corresponded to
3.28 mg GAE g1 (or 3.58 mg GAE g1, DW), repre-
Shell
senting, 4.3% of the PC content. Lower contents of
0.11
0.02 hydrolysable tannins have been reported in the peri-
0.40
0.01 carp and integument of four chestnut cultivars (ranged
0.36
0.03 from 0.98 to 1.86 and from 0.20 to 0.49 mg g1,
0.15
0.01 respectively) (de Vasconcelos et al., 2010). The pres-
3.28
0.01 ence of important quantities of tannins (condensed
69.42
4.93 and hydrolysable) in SI shell might be indicative of
0.84
0.12
potential high antioxidant properties compared to the
74.56
5.13
tannin monomers (Khanbabaee & van Ree, 2001).
Condensed tannins have been reported to display a
wide range of biological and pharmacological activities
including antioxidative, cardioprotective, antitumour,
antibacterial, antiviral, antiinflammatory effects (Liu
et al., 2009). These bioactive properties associated to
the presence of tannins in SI shell might be indicative
of the potential high added-value of this novel by
product. The remaining phenolic families corresponded
to phenolic acids (free and bound), flavonoids, flava-
noids and lignans that accounted all together for only
2.6% of the PC content in SI shell.
The phenolic acid profile (free and bound) and the
spectral characteristics of the compounds are presented

(a) 3
0.25

0.20
6
0.15 1
AU

0.10
2 5
4
7
0.05 8

0.00

(b)
3
1.00

0.80

0.60
AU

0.40 9

13
0.20 11
2 12 14
4
0.00
0.00 10.00 20.00 30.00 40.00 50.00 60.00 70.00 80.00 90.00 100.00
Retention time (min)

Figure 1 HPLC-DAD chromatograms of free (a) and bound (b) phenolic acids from sacha inchi shell recorded at 280 nm. Different peak
numbers correspond to the ones reported in Table 2.

© 2016 Institute of Food Science and Technology International Journal of Food Science and Technology 2016
990 Antioxidant phenolics from sacha inchi shell R. Chirinos et al.

Table 2 Chromatographic and spectral characteristics of free and biaceae family as SI, phenolic acids of the p-coumaric,
bound phenolic acids from sacha inchi shell ferulic, o-coumaric, siringique and cinammic types;
being the most predominant the two-first ones. This
Retention Amount Phenolic compound
Peak # time (min) k max (nm) (mg/100 g) assignment
reported profile is very similar to the one found for SI
seed in this study. The content of flavonoids and flava-
Free phenolic acids noids in SI shell corresponded to 0.36 mg QE g1 and
1 13.1 285.5 1.2
0.21 Cinnamic acid 0.15 mg CE g1, respectively. Lower amounts of flavo-
derivative* noids have been reported for rice husk (25.8 lg g1,
2 20.8 260.6, 295.0 2.8
0.67 Protocatechuic acid DW) (Wanyo et al., 2014).
3.9
0.61
3 23.2 280.8, 311.6 Hydroxycinnamic
A lignans content of 0.84 mg g1 (Table 1) or
acid derivative†
4 39.8 341.4 1.5
0.11 Hydroxycinnamic
0.91 mg g1 (DW) was obtained for SI shell. The
acid derivative‡
chromatograms and absorption spectra for the SDG
5 45.7 276.0 0.3
0.05 Cinnamic acid and MAT standards are presented in Fig. 2a. Hos-
derivative* seinian & Mazza (2009) indicated that SDG and MAT
6 52.5 281.9 0.6
0.06 Cinnamic acid are present in vegetables, grains, seeds and fruits. Only
derivative* two peaks presented similar spectra to lignans but
7 65.6 290.2, 321.2 0.5
0.18 Hydroxycinnamic none of them corresponded to SDG and MAT stan-
acid derivative† dards (Fig. 2b) and the results were expressed as SDG.
8 67.6 337.9 0.4
0.03 Hydroxycinnamic The content of SDG reported by Hosseinian & Mazza
acid derivative‡
(2009) for triticale straw corresponded to 0.27 mg/
Bound phenolic acids
9 15.0 284.3 2.8
0.32 Cinnamic acid
100 g and the contents of MAT for triticale bran and
derivative*
straw corresponded to 0.01 and 0.20 mg/100 g, respec-
2 21.0 260.6, 295.0 1.5
0.06 Protocatechuic acid tively. Higher contents of lignans have been reported
3 23.4 280.8, 310.4 33.2
0.30 Hydroxycinnamic for flax (2.8–3.69 mg g1) and it is considered as the
acid derivative† main source of lignans (Raffaelli et al., 2002).
11 31.9 285.5 0.5
0.09 Cinnamic acid The sum of PC in SI shell (sum of all quantified
derivative* phenolic families) corresponded to 74.56 or
12 38.2 279.6, 310.4 0.6
0.10 Hydroxycinnamic 81.20 mg g1 (DM). Weisz et al. (2009) reported total
acid derivative† PC (by adding up the individual amounts of all fami-
4 40.1 341.4 0.7
0.01 Hydroxycinnamic
lies) for sunflower seed within the range of 0.40–
acid derivative‡
13 50.9 310.4 1.6
0.01 p-coumaric acid
0.86 mg g1 (DM) being these values lower than the
14 53.7 321.2 0.7
0.07 Hydroxycinnamic
value found for SI shell in this study.
acid derivative‡

*
Quantified at 280 nm as cinnamic acid.
†
Quantified at 320 nm as o-coumaric acid.
‡
Quantified at 320 nm as ferulic acid. Results correspond to the aver-
age
standard deviation of three replicates.
in Fig. 1 and Table 2, respectively. Eight different
peaks were identified corresponding to free phenolics
(Fig. 1a). Peak # 2 was identified as protocatechuic
acid (hydroxybenzoic acid) and peaks # 3, 4, 7 and 8
were identified as hydroxycinnamic acid derivatives
with spectral characteristics similar to ferulic and
o-coumaric acid and peaks # 1, 5, and 6 presented
spectral characteristics similar to cinnamic acid. In
addition, eight bound phenolic acids were detected
(Fig. 1b).The identified compounds corresponded to
protocatechuic acid (peak # 2), p-coumaric acid (peak
# 13) and derivatives of hydroxycinnamic acids of the
o-coumaric (peaks # 3 and 12), ferulic (peaks 4 and
14) and cinnamic (peaks 9 and 11) types. Similarly,
Srinivas & Nagaraj (2007) identified in castor seed
(Ricinus communis), that also belongs to the Euphor-
Extraction of phenolic compounds from SI shell and their
antioxidant capacities
The selection of the extraction solvent is a key factor
in any extraction process since it influences the quan-
tity and quality of the recovered metabolites. Results
of TPC (determined with the Folin Ciocalteau method)
and AOXC for the three assays (ABTS, FRAP and
ORAC) using different combinations of extraction sol-
vents from SI shell is presented in Table 3. Acetone/
water/acetic acid (80/19/1) and methanol/acetone/
water/acetic acid (40/40/19/1) displayed a significant
higher efficiency in the extraction of TPC (P < 0.05).
Higher values of TPC in SI seed were obtained in
presence of acetone possibly due to the presence of
condensed tannins as main PC in SI shell. Similarly,
the mixture acetone/water/acetic acid (70/29.5/0.5, v/v/
v) was successfully used to recover phenolic com-
pounds from cacao with a high content of procyani-
dins (Adamson et al., 1999). Garrido et al. (2007)
recovered higher amounts of TPC and proanthocyani-
dins from almond skin using acetone as part of the
solvent extraction mixture. Higher contents of TPC

International Journal of Food Science and Technology 2016 © 2016 Institute of Food Science and Technology
 Antioxidant phenolics from sacha inchi shell R. Chirinos et al. 991

197.1 197.1
(a)
2.00 4.00
0.60

MAT

AU
AU
2.00
1.00
281.9
281.9
0.00
0.40 0.00

SDG
200.00 400.00
200.00 400.00
AU

nm
nm

0.20

0.00

(b) 0.30
201.8
0.25 0.40

AU
0.20 0.20

281.9
AU

0.15 0.00

200.00 400.00
nm
0.10
1
0.05 2

0.00
0.00 5.00 10.00 15.00 20.00 25.00 30.00 35.00
Retention time (min)

Figure 2 HPLC-DAD chromatograms of (a) lignan standards: secoisolaricirecinol diglucoside (SDG), matairesinol (MAT) and (b) lignans
from sacha inchi shell recorded at 280 nm.

Table 3 Total phenolic compounds and antioxidant capacity ABTS, FRAP and ORAC of sacha inchi shell

Total phenolic
compounds ABTS FRAP ORAC
Extraction solvent (mg GAE g1) (lmol TE g1) (lmol TE g1) (lmol TE g1)

Methanol/water/acetic acid (80/19/1, v/v/v) 9.5
0.34c 46.0
2.4d 45.0
3.2e 104.1
6.9d
Acetone/water/acetic acid (80/19/1, v/v/v) 15.3
0.2a 93.9
3.4a 114.0
10.3a 145.0
8.9b
Ethanol/water/acetic acid (80/19/1, v/v/v) 9.7
0.1c 34.6
0.9e 51.3
2.0d 116.7
12.9c
Methanol/acetone/water/acid acetic (40/40/10/1, v/v/v/v) 15.1
0.2a 70.5
5.8b 68.5
1.6b 189.4
18.4a
Ethanol/acetone/water/acid acetic (40/40/10/1, v/v/v/v) 13.7
0.3b 65.2
4.2c 68.8
1.4b 192.6
12.8a
Ethanol/methanol/water/acid acetic (40/40/10/1, v/v/v/v) 9.7
0.5c 34.1
2.4e 57.8
2.4c 92.5
5.4d

Results are means
SD (n = 3). Means within a column with the different letter are significantly different (P < 0.05) as revealed by a Duncan test.

from SI shell (15.1–15.3 mg GAE g1) were obtained
in relation to the values reported for almond skin
extracted with different solvents with reported values
that ranged from 0.82 to 2.06 mg g1 (Garrido et al.,
2007); hazelnut skin with reported TPC values of 1.5,
1.6 and 2.1 mg GAE g1 using as extraction solvents
80% methanol, 80% ethanol and 80% acetone,
respectively (Contini et al., 2008). Similar amounts of
TPC to the ones obtained from SI shell in this study
have been reported by John & Shahidi (2010) for Bra-
zil nut with values of 17.1 and 18.6 mg GAE g1
using 80% methanol and 70% acetone as extraction
solvents, respectively.
Significant differences (P < 0.05) were found in the
ABTS, FRAP and ORAC AOXC of SI shell related
to the extraction solvent (Table 3). ABTS, FRAP and
ORAC AOXC were within the range of 34.1 and 93.9;
45.0 and 114.0 and, 92.5 and 192.6 lmol TE g1,

© 2016 Institute of Food Science and Technology International Journal of Food Science and Technology 2016
992 Antioxidant phenolics from sacha inchi shell R. Chirinos et al.
respectively. Similarly as for the TPC, higher AOXC
was obtained with the mixture of solvents containing
acetone in its composition. Different AOXC values for
different agro-industrial by products have been previ-
ously reported. For instance, Wanyo et al. (2014)
reported for rice husk a FRAP AOXC within the
range of 13.5 and 18.2 lmol TE g1 being this range
of values lower than the FRAP AOXC obtained for
SI shell extracted with the solvent mixtures containing
acetone (68.8–114.0 lmol TE g1; Table 3). Values of
ABTS AOXC obtained from SI shell (70.5–93.9 lmol
TE g1) are similar to the ABTS range of values
reported for Brazil nut skin (John & Shahidi,
2010).The ORAC AOXC obtained for SI shell extracted
with acetone containing mixtures (145–192 lmol
TE g1) were much higher than the value reported by
Garrido et al. (2007) for almond shell using 50% acetone
as extraction solvent (50.4 lmol TE g1) and similar to
the value obtained for the brown skin of Brazil nut
extracted with 70% acetone (189.3 lmol TE g1; John &
Shahidi, 2010).
Conclusions
Results obtained in this study indicate that SI shell is
an important source of antioxidant phenolic com-
pounds. Condensed tannins (93.1%) are the main
family of phenolic compounds present in SI shell.
The other phenolic families present in SI shell
(hydrolyzed tannins, free and bound phenolic acids,
flavanoids, flavonoids and lignans) were present in
amounts either similar or higher than reported for
other agro-industrial by products of the seed type.
The best extraction results to obtain a high PC and
AOXC contents were obtained with acetone/water/
acetic acid (80/19/1, v/v/v) and methanol/acetone/wa-
ter/acid acetic (40/40/10/1, v/v/v/v) possibly due to
the important amounts of condensed tannins present
in SI shell. Thus, our results indicate that SI shell has
a high potential as an alternative and novel source of
antioxidant PC from an agro-industrial by product
that could be derived to the nutraceutical and func-
tional food industry.
Acknowledgment
This research was supported by the PIC project of the
Belgian Coop
eration Universitaire au D
eveloppement
(CUD, Belgium).
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