Food Chemistry 128 (2011) 1094–1099

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Catechin and epicatechin in testa and their association with bioactive compounds
in kernels of cashew nut (Anacardium occidentale L.)
Jennifer Trox a, Vellingiri Vadivel a, Walter Vetter b, Wolfgang Stuetz a, Dietmar R. Kammerer c,
Reinhold Carle c, Veronika Scherbaum a, Ute Gola a, Donatus Nohr a, Hans Konrad Biesalski a,⇑
a
Institute for Biological Chemistry and Nutrition, University of Hohenheim, Germany
b
Institute of Food Chemistry, University of Hohenheim, Germany
c
Institute for Food Science and Biotechnology, Chair of Plant Foodstuff Technology, University of Hohenheim, Germany

a r t i c l e i n f o a b s t r a c t

Article history: Catechins in testa and bioactive compounds in testa-free and testa-containing kernels of cashew nuts were
Received 13 August 2010 analysed. The cashew nut testa contained (+)-catechin and ()-epicatechin with concentrations of 5.70 and
Received in revised form 7 March 2011 4.46 g per kg DM, respectively. Testa-containing kernels revealed significantly higher levels of b-carotene
Accepted 7 April 2011
(218 vs. 89.6 lg/kg DM), lutein (525 vs. 292 lg/kg DM), and a-tocopherol (10.1 vs. 2.4 mg/kg DM), similar
Available online 16 April 2011
amounts of zeaxanthin (7.0 vs. 7.1 lg/kg DM), c-tocopherol (10.6 vs. 10.1 mg/kg DM), stearic acid (41 vs.
43 g/kg DM), oleic acid (214 vs. 219 g/kg DM) and linoleic acid (69 vs. 62 g/kg DM), but a lower concentra-
Keywords:
tion of thiamine (3.0 vs. 10.7 mg/kg DM) in comparison to testa-free samples. The testa-containing kernels
Anacardium occidentale L.
Cashew nut testa
provide high amounts of catechins and higher concentrations of b-carotene, lutein and a-tocopherol than
Catechin do testa-free cashew nut kernels. This could have potential health benefits for consumers.
Epicatechin Ó 2011 Elsevier Ltd. All rights reserved.
Carotenoids
Tocopherols
Thiamine
Fatty acids

1. Introduction
Cashew nut (Anacardium occidentale L.) kernels are regarded as
a nutritious food product, worldwide. The kernels of cashew nuts
are externally covered with a thin and reddish-brown-coloured
skin, known as testa. The testa constitutes about 1–3% of the total
weight of cashew nuts and is found to provide a rich source of
hydrolysable tannins with polymeric proanthocyanidins as major
polyphenols (Mathew & Parpia, 1970). Ethanolic extract of cashew
nut testa exhibited a significant level of antioxidant activity, which
was attributed to its phenolic composition (Kamath & Rajini, 2007).
Unfortunately, cashew nuts for human consumption are being
marketed without the testa. Usually after shelling, the testa is re-
moved from the kernels, in order to avoid bitter/astringent taste
and also to achieve a higher price in the market.
During conventional industrial shelling, the testa is exposed to a
very high processing temperature (75–200 °C) and ultimately
eliminated from the kernels. As an alternative, Nair (2005) pat-
ented (WO/2005/039322) a novel method to produce cashew nuts
with intact testa. The ‘‘Flores’’ hand-cracking method has recently
been introduced, to retain the testa with kernels, by PT. Profil Mitra
Abadi (PT. PMA), Tangerang, Indonesia. In this process, the cashew
nut kernels are separated from the shell by employing a specially
designed hand-cracking machine. Subsequently, the cashews are
dried for 3 h at a mild temperature (45 °C). Under these conditions,
the testa is not damaged and remains intact. Furthermore, we have
already reported the maximal retention of various bioactive com-
pounds in cashew nut kernels using the ‘‘Flores’’ hand-cracking
process (Trox et al., 2010).
Even though the nutritional value and health benefits of cashew
nut kernels were demonstrated by various research studies, the
nutritional importance of testa has been ignored. Currently, there
is no information available on the phenolic composition of testa
or its contribution to the nutritional quality of cashew kernels.
Hence, the present study was conducted in order to characterise
the polyphenolic compounds in cashew nut testa, and also to
determine the bioactive compounds in testa-containing and tes-
ta-free cashew nut kernels.
2. Materials and methods
2.1. Sample collection

⇑ Corresponding author. Tel.: +49 711 459 24112; fax: +49 711 459 23822. The cashew nut samples were collected from agricultural farms
E-mail address: Biesal@uni-hohenheim.de (H.K. Biesalski). located at four different villages (Rowa, Ile Padung, Ilenmedo and

0308-8146/$ - see front matter Ó 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2011.04.018
 J. Trox et al. / Food Chemistry 128 (2011) 1094–1099
Kringa) in central and eastern Flores Island, Indonesia, located at a
sea level of 10–45 m. To ensure the comparability between pro-
cessed samples, all the cashew nuts were harvested in October
2006, and also the same type of drying and storage conditions were
applied. To reduce water content from approximately 25% to 8–9%
the raw material was sun-dried for 2–3 days. It was stored in bags
under adequate ventilation. The collected samples were processed
as described below.
2.2. ‘‘Flores’’ hand-cracking
The hand-cracking of cashew nuts was conducted on the basis
of a detailed description of production (Rudolf Heering, President
Director, PT, PMA, Tangerang, Indonesia). After manual removal
of small samples, acceptable cashew nuts, with a mean weight of
6.42 g and a water content of 6.07% (±0.4%), were passed into the
breakers. It is inevitable that a minimum amount of cashew nut
shell liquid (CNSL) emerges from the mesocarp of the shell as a re-
sult of the cracking process. Thus, it is important to protect the tes-
ta of the cashew nut kernels from exiting the CNSL. The outer
surface of the cashew nut kernel was not damaged during the
cracking process, so that the cashew nuts were gained without vio-
lating the testa. Further, the cashew nut kernel, which is protected
by testa, was not in contact with the CNSL. The samples were
cracked and dried for 3 h at a temperature of 45 °C in a heating
block. Then the samples were frozen at 80 °C and freeze-dried
for 48 h.
2.3. Sample preparation
The processed samples were divided into two batches (each
containing 50 g). In one batch the testa was removed manually
from the kernels whereas, in the other batch, the cashew nut ker-
nels were stored without removing the testa. The manually re-
moved testa was used for the determination of phenolic
compounds, while the testa-containing and testa-free cashew nut
kernels were subjected to the analysis for bioactive compounds.
Testa-containing and testa-free cashew nut kernels were homoge-
nised by using a mortar and pestle, freeze-dried for 24 h and stored
at 9 °C prior to further use. Degradation of tocopherols was pre-
vented by working under a yellow light.
2.4. Polyphenolic compounds in cashew nut testa
In a preliminary experiment, approximately 70 mg of powdered
testa were mixed with 5 ml of methanol–water mixture (40:60 v/v)
and 50 ll of formic acid. This solution was centrifuged for 3 min at
2500 g and the supernatant was diluted (1:100, v/v) for subsequent
measurements by both HPLC with photodiode array (PDA) and
mass spectrometric (LC–MS) detection. For HPLC/PDA analysis,
20 ll were injected into a Varian HPLC (Pro Star 210) equipped with
a Shimadzu PDA (PD-M20A), applying the following chromato-
graphic conditions: Reprosil-Pur 120 C18 AQ column (5 lm,
250  4.6 mm, Trentec, Gerlingen, Germany) at 40 °C, and solvent
A of aqueous formic acid (5% v/v) and solvent B containing formic
acid, distiled water and acetonitrile (5:10:85, v/v/v). The gradient
programme was as follows: 5% B to 26% B (12.38 min), 26% B to
100% B (17.22 min), 100% B isocratic (10 min) with a flow-rate of
1 ml/min and a total run time of 40 min. Spectra were recorded
from 200 to 700 nm. Testa extracts were analysed by LC–MS
according to the method of Stintzing et al. (2004). The separation
of phenolic compounds was performed in an Agilent HPLC series
1100 (Agilent, Waldbronn, Germany), under the conditions de-
scribed above, applying the following gradient: 5% B to 15% B
(6 min), 15% B isocratic (4 min), 15% B to 26% B (6.38 min), 26% B
to 100% B (17.62 min), 100% B isocratic (10 min).
1095
This HPLC system was connected, in series, with a Bruker ion
trap mass spectrometer (Model: Esquire 3000+, Bremen, Germany)
fitted with an APcI source. The detector was operated in negative
ionisation mode (range: m/z 50–1200). Nitrogen was used as the
dry gas at a flow rate of 5.0 l/min and a pressure of 65.0 psi. The
nebuliser temperature was set at 350 °C, and the vaporiser temper-
ature at 400 °C. Using helium as the collision gas (4.1  106 mbar),
collision-induced dissociation spectra were obtained with a frag-
mentation amplitude of 1.0 V.
After peak assignment by LC–MS, the testa samples were
prepared as follows: 10 mg of powdered testa were vigorously
mixed with 1 ml of methanol in a 1.5 ml Eppendorf tube, and
centrifuged (11400 g for 5 min), and supernatants were finally di-
luted with water (1:25, v/v) prior to HPLC analysis. Aliquots of
20 ll were injected into a Varian Pro Star 210 and the phenolics
were separated with the same column and solvents as described
above, using the following gradient programme: 5% B to 15% B
(1 min), 15% B isocratic (4 min), 15% B to 26% B (6.38 min), 26%
B to 100% B (17.62 min), 100% B isocratic (10 min). ()-Epicate-
chin and (+)-catechin were identified at 278 nm, using a Waters
detector (Model: 2487). (+)-Catechin and ()-epicatechin were
quantified by using an external calibration, with respective stan-
dards diluted in methanol–water (50:50, v/v).
2.5. Carotenoids and tocopherols in cashew nut kernels
The b-carotene, lutein and zeaxanthin, as well as the a- and c-
tocopherol contents, in testa-containing and testa-free cashew
nut kernels were analysed by RP-HPLC after extraction and sapon-
ification as follows: 2 ml of ethanol, containing pyrogallol (2.5%)
and b-apo-80 -carotenal-methyloxime and 1 ml of 50% KOH solu-
tion, were added to 100 mg of the sample in a screw-capped glass
tube. The internal standard b-apo-80 -carotenal-methyloxime was
synthesised as described by Sommerburg, Zang, and Van Kuijk
(1997). For saponification, the suspension was covered by argon
and closed with screw caps, and then the mixture was stirred for
4 h in a water bath at 38 °C. After the addition of 2 ml of saline solu-
tion (15%), fat-soluble substances were extracted twice with l ml of
hexane. The combined hexane phases were washed (15% saline
solution), evaporated using nitrogen gas and finally re-dissolved
in 200 ll of ethanol–water (1:3, v/v) prior to analysis on a Varian
HPLC (Prostar-210) equipped with UV–VIS and fluorescence detec-
tors (Waters-2487, Waters-474), using the following chromato-
graphic conditions: Spherisorb ODS-2 analytical column (3 lm,
250  4.6 mm, Trentec, Germany) at 40 °C and a mobile phase
consisting of acetonitrile (82%), dioxane (15%) and methanol (3%,
containing 100 mM ammonium acetate and 0.1% triethylamine)
in a re-circulation mode with a flow rate of 1.6 ml/min. The carote-
noids were detected at 450 nm, while a- and c-tocopherols were
measured by fluorescence with an excitation/emission wavelength
of 298 and 328 nm, respectively.
2.6. Thiamine in cashew nut kernels
The determination of thiamine content of cashew nut samples
was performed by pre-column derivatization, RP-HPLC and fluores-
cence detection according to the method of Gerrits, Eidhof,
Brunnekreeft, and Hessels (1997) method with modifications. In
brief, 100 mg of the sample were mixed with 7.5 ml of 0.1 M HCl
solution and stirred for l h at 30 °C in the dark. Subsequently, an ali-
quot of 1.5 ml of this mixture was centrifuged at 5000 g and the
clear supernatant was derivatised as follows: 500 ll of the clear
solution were mixed with 100 ll of freshly prepared oxidation re-
agent (12.1 mM K3[Fe(CN)6] in 3.35 M NaOH solution). The ‘‘thio-
chrome reaction’’ was stopped by the addition of 20 ll of 6 M
orthophosphoric acid, and 20 ll of the aliquot were analysed on a
1096 J. Trox et al. / Food Chemistry 128 (2011) 1094–1099

Merck-Hitachi HPLC (LaChrom) equipped with a column oven (set
at 40 °C), fluorescence detector (L-7480) and Clarity chromato-
graphic station (DA-C50, DataApex Ltd., Praha). The separation
was achieved on a 5 lm analytical column (Grom-Sil 120 ODS-4
HE, 125  4 mm, Grom, Rottenburg-Hailfingen, Germany), using a
mobile phase consisting of methanol (27.5% v/v) and phosphate
buffer (pH 7.0) at a flow rate of 0.8 ml/min. Thiamine was detected
by excitation/emission set at 367/435 nm.
2.7. Fatty acids in cashew nut kernels
Determination of fatty acids (stearic acid, oleic acid and linoleic
acid) of cashew nut samples was carried out by following the
method of Thurnhofer, Lehnert, and Vetter (2008). Sample aliquots
(500 mg) were extracted by means of an ASE 200 system (Dionex,
Idstein, Germany), using 22 ml extraction cells. For this purpose,
22 ml extraction cells were used, which were filled with isolute-
HM-N. The cells were extracted twice with the azeotropic mixture
of cyclohexane and ethyl acetate (46:54, v/v) (Weichbrodt, Vetter,
& Luckas, 2000). The two combined extracts were concentrated,
using a rotary evaporator with vacuum controller at 60 °C and pre-
cisely adjusted to a definite volume. The lipid content was deter-
mined gravimetrically (Trox et al., 2010).
To determine fatty acids, the extracted oil was first transesteri-
fied. The first internal standard (10,11-dichloroundecanoic acid),
produced according to the method of Thurnhofer et al. (2008),
was pipetted into the mixture and 500 ll of 0.5 M methanolic
KOH solution were added for saponification at 80 °C. After a reac-
tion time of 5 min, the mixture was cooled in an ice bath. The sub-
sequent methylation was started by adding l ml of boron-trifluoride
in methanol and kept for 5 min at 80 °C. After cooling, 2 ml of hex-
ane and 2 ml of a saturated NaCl solution were added and shaken
well. The organic phase (180 ll) was mixed with 20 ll of the second
internal standard, oleic acid ethyl-ester, which was prepared
according to the method of Thurnhofer et al. (2008). Gas chroma-
tography, in combination with electron ionisation mass spectrom-
etry (GC–EI/MS) analyses, was performed with an HP 5890/5971
system (Agilent, Waldbronn, Germany). Helium (99.999% purity)
was used as the carrier gas with a flow rate of 1 ml/min. A CP-Sil
88 column (50 m  0.25 mm ID  0.20 lm film thickness) (Chrom-
pack, Middelburg, The Netherlands) was installed in the GC oven.
An injection volume of 1 ll was used at a temperature of 250 °C
and analysed for 38.81 min. In the selected ion monitoring (SIM)
mode, nine fragment ions were recorded and six of them were used
for quantification: (1) m/z 74 and (2) m/z 87 for the methyl esters of
saturated and monounsaturated fatty acids, (3) m/z 81 and (4) m/z
79 for the methyl esters of polyunsaturated fatty acids, (5) m/z 88
and (6) m/z 101 for the ethyl esters of saturated and monounsatu-
rated fatty acids.
3. Results and discussion
3.1. Polyphenolic compounds in cashew nut testa
The RP-HPLC analysis of phenolic extract of cashew nut testa re-
vealed two prominent peaks with absorption maxima at 278 nm
(data not shown). Based on the comparison of mass spectrometric
data of these peaks with an earlier report by Stintzing et al. (2004),
the phenolic compounds were identified as (+)-catechin and ()-
epicatechin and confirmed with authentic reference compounds.
To our knowledge, this is the first report to confirm the phenolic
profile of cashew nut testa by employing RP-HPLC and LC–MS
techniques.
Remarkable amounts of (+)-catechin and ()-epicatechin were
quantified with averages of 5.70 and 4.46 g per kg DM, respec-
tively, in the cashew nut testa (Fig. 1). In an earlier report, it has
been postulated that the catechins are mainly localised in the cash-
ew nut testa and not in the kernels (Mathew & Parpia, 1970), which
is in accordance with the results of the present research work. The
antioxidant and free radical-scavenging potential of cashew nut
extracts, demonstrated by Sajilatha and Singhal (2006), might be
attributed to the catechins found in testa, as deciphered by the
present investigation. Further, the antioxidant-rich extract from
cashew nut skin was found to have a protective effect against orga-
nophosphorus insecticide toxicity in rats (Kamath, Joshi, & Rajini,
2008).
It is important to mention that the polyphenolic content of
cashew nut testa was higher than those of green tea and dark choc-
olate (Khokhar & Magnusdottir, 2002; Natsume et al., 2000). In
comparison, the contents of (+)-catechin and ()-epicatechin in
cashew nut testa were 20 and 5-fold higher, respectively, than
those of the polyphenolic compounds reported in dark chocolate
(Natsume et al., 2000). The high levels of (+)-catechin and ()-epi-
catechin confer high nutritional quality to the cashew nut.
The results of the analysis of different varieties of green tea, from
a previous research work, showed average concentrations of
0.43 mg of (+)-catechin and 6.66 mg of ()-epicatechin per gramme
DM (Khokhar & Magnusdottir, 2002). The levels of phenolic

2.8. Statistical analysis

The statistical analysis was performed using SPSS for WINDOWS

(SPSS Inc., Chicago, IL, Version 11.0). 10 separate determinations
were carried out for catechins, carotenoids, tocopherols and thia-
mine and seven separate determinations were carried out for fatty
acids in testa-free and testa-containing cashew kernels; values
within each group were found to be normally distributed using Kol-
mogorov–Smirnov-test and were described by their means (SD).
Comparison of means between testa-free and testa-containing
cashew kernels was determined by independent Student’s t-test;
two-tailed p values <0.05 were considered statistically significant. Fig. 1. Catechin and epicatechin contents in cashew nut testa.
 J. Trox et al. / Food Chemistry 128 (2011) 1094–1099
compounds per cup of green tea were calculated on the basis of 2 g of
green tea in 200 ml of water, with extraction yields of 95% for (+)-
catechin and 85% for ()-epicatechin (extraction conditions:
5 min, 80 °C) (Khokhar & Magnusdottir, 2002). Based on data from
Khokhar and Magnusdottir (2002), an average cup of green tea con-
tains 0.82 and 11.3 mg of (+)-catechin and ()-epicatechin, respec-
tively. From the results of the present study, one single piece of
cashew nut, along with testa (’2 g), contains 11.4 and 8.92 mg of
(+)-catechin and ()-epicatechin, respectively. Hence, the (+)-cate-
chin content of one cashew nut kernel with testa is roughly equiva-
lent to the amount of (+)-catechin contained in 13 cups of green tea.
Further, the level of ()-epicatechin in one cashew nut containing
testa is equivalent to 75% of the ()-epicatechin amounts of an aver-
age cup of green tea.
Cabrera, Artacho, and Gimenez (2006) reviewed the properties of
green tea polyphenols (mainly catechins), revealing high antioxi-
dant, anti-mutagenic, anti-diabetic, anti-inflammatory, anti-bacte-
rial and anti-viral activities. Further, the results of some human
studies also indicate that the consumption of green tea reduces
the risk of cardiovascular diseases and some types of cancer (Cabrera
et al., 2006). However, it is important to note that the total amount of
polyphenolics in green tea is higher than that of cashew nut testa, as
green tea does not only contain (+)-catechin and ()-epicatechin,
but also high quantities of gallic acid esters, ()-epicatechin gallate,
()-epigallocatechin and ()-epigallocatechin gallate. These may
also be responsible for the above-mentioned health protective
effects of green tea.
In addition, the effect of the drying process at 45 °C, for 3 h in the
Flores hand-cracking method, on the level of polyphenolic com-
pounds of cashew nut testa has not yet been examined. A significant
reduction of polyphenolic content was noticed in millet as a result
of increasing the temperature from 65 to 75 °C, (Nwanguma & Eze,
1996). Nonetheless, no significant change of the level of polyphe-
nols or their antioxidant activity in grape pomace from red grapes
was observed at a drying temperature of 60 °C (Larrauri, Ruperez,
& Saura-Calixto, 1997). Consequently, it could be concluded that
drying at the mild temperature of 45 °C should not affect the poly-
phenolic compounds of cashew nut testa significantly.
3.2. Carotenoids in cashew nut kernels
The analysis of carotenoids exhibited significantly (p < 0.001)
higher contents of b-carotene and lutein in testa-containing ker-
nels (2.4- and 1.8-fold, respectively) than in testa-free samples (Ta-
ble 1). These significant differences in the carotenoids are most
likely a result of occurrence of high amounts of these compounds
in the testa. Additional analyses in testa samples showed approxi-
mately 2.5- to 3-fold higher levels of b-carotene and lutein, respec-
tively, in the testa than in kernel (data not shown). In contrast, the
content of zeaxanthin was virtually the same (7.1 vs. 7.0 lg/kg
Table 1
Carotenoids, tocopherols and thiamine in testa-free vs. testa-containing kernels of
cashew nut1,2.
Bioactive compounds Cashew nut samples
Testa-free kernels3 Testa-containing kernels
a
b-Carotene (lg/kg DM) 89.6 ± 3.9 218b ± 11.8
Lutein (lg/kg DM) 292a ± 26.7 525b ± 45.2
Zeaxanthin (lg/kg DM) 7.1a ± 2.3 7.0a ± 2.2
a-Tocopherol (mg/kg DM) 2.4a ± 0.4 10.1b ± 0.7
c-Tocopherol (mg/kg DM) 10.1a ± 0.6 10.6a ± 0.6
Thiamine (mg/kg DM) 10.7a ± 1.0 3.0b ± 0.5
1
Values are means ± standard deviation of 10 separate determinations (n = 10).
2
Values in the same row with different superscripts are significantly different
(p < 0.001).
3
Data taken from our previous report (Trox et al., 2010).
1097
DM) in testa-free and testa-containing cashew nut kernels. The
presence of such high amounts of these antioxidants in the testa
is of great importance for the development of cashew nuts during
germination, by protecting them from insects, microbial infection
and sun-light, through the hard CNSL-containing shell. Following
the breakdown of shell, the protective function of antioxidants
contained in the testa is activated (Chung, Wong, Wei, Huang, &
Lin, 1998).
In addition to the importance as provitamin A, b-carotene has a
protective function against both UV-light and oxidative stress,
through deactivation of singlet oxygen and by inhibition of lipid
peroxidation (Biesalski et al., 2009). Functions of carotenoids are
considerable as a precursor of vitamin A, which is required for
adaptive immunity and plays a significant role in the development
of both T-helper cells and B cells (Stephensen, 2001). Furthermore,
carotenoids are also known to play a role in the prevention of can-
cer and atherosclerosis (Krinsky & Johnson, 2005).
3.3. Tocopherols in cashew nut kernels
The levels of a- and c-tocopherols in testa-containing cashew
nut kernels were found to be 10.1 and 10.6 mg/kg DM, respectively
(Table 1). The testa-containing samples exhibited significantly
(p < 0.001) higher level of a-tocopherol (4.2-fold) but similar levels
of c-tocopherol in relation to testa-free kernels. The explanation is
the presence of a much higher concentration of a-tocopherol (23-
fold) in the testa than in kernels (data not shown).
Tocopherols are reported to constitute an essential part of bio-
logical membranes. It is well known that tocopherols exhibit a pro-
tective role against lipid peroxidation of membrane lipids,
lipoproteins and depot fats (Biesalski et al., 2009) and therefore
prevent atherosclerosis. Vitamin E is also able to induce apoptosis
in tumour cells and modulate oncogenes and this is probably the
reason for the low occurrence of cancer as a result of vitamin E-rich
diets, which was demonstrated in a variety of epidemiological
studies (Dutta & Dutta, 2003).
3.4. Thiamine in cashew nut kernels
Various enzymes of intermediary metabolism, including pyru-
vate dehydrogenase, a-ketoglutarate dehydrogenase and transke-
tolase, require thiamine pyrophosphate as an essential cofactor.
The testa-free cashew nut kernels were found to contain consider-
ably high amounts of thiamine (10.7 mg/kg DM) when compared
to testa-containing samples (3.0 mg/kg DM) (Table 1).
In testa-containing cashew nut samples, the thiamine content
was significantly reduced, by more than 70% with respect to testa-
free kernels, which is most likely due to the anti-thiamine activity
of phenolic compounds in testa, because phenolic compounds, such
as catechin, epicatechin and tannins, were reported to exhibit a
strong anti-thiamine activity (Hilker & Somogyi, 2006). Further, in
the present investigation, the testa-containing cashew nuts were
homogenised and stored for about 2 months prior to analysis. Dur-
ing this time thiamine seems to be partially degraded, by splitting
the thiazole ring of the thiamine, or accelerating the oxidation of
disulphide, due to the polyphenolic compounds of testa. The anti-
thiamine activity of phenolic compounds is closely related to the
number and position of OH groups on the benzene ring. Substances
with two vicinal OH groups were reported to exhibit high anti-thia-
mine activity (Somogyi & Bonicke, 1969) and, interestingly, both
()-epicatechin and (+)-catechin show such characteristic features.
Hence, it is recognised that the reduction of the thiamine level was
most likely due to the interaction with polyphenolic compounds of
cashew nut testa.
The thiamine concentrations of commercially available testa-
free and testa-containing cashew nuts are probably not much
1098 J. Trox et al. / Food Chemistry 128 (2011) 1094–1099
different, as shown in the present study, since they are stored for a
longer period and also offered as whole kernels and not as a homog-
enised powder. The anti-thiamine factors contained in the testa
may have less association in whole kernels whereas, in pulverised
form, there are more chances for close contact with polyphenolic
compounds, which might be responsible for the reduction of thia-
mine content. However, reduction of bioavailability of thiamine,
due to the presence of high concentrations of (+)-catechin and
()-epicatechin, still remains unclear upon consumption of whole
cashew nuts with testa. Nonetheless, it has been shown that the
consumption of tea containing high amounts of catechin polyphe-
nols causes a severe thiamine deficiency (Kositawattanakul, Tos-
ukhowong, Vimokesant, & Panijpan, 1977).
3.5. Fatty acids in cashew nut kernels
We observed no significant difference between the fatty acid
patterns of the testa-containing and testa-free samples (Table 2).
While the levels of oleic acid and stearic acid were comparable, lin-
oleic acid was, on average, 10% higher in testa-containing cashew
nut kernels than in testa-free samples. Since, the testa contributes
7% to the sample, the high concentration of linoleic acid in the
testa-containing kernels was attributed to testa. Subsequent anal-
yses of (separated) testa and testa-free kernels confirmed this
assumption. The relative contribution of linoleic acid to the fatty
acids in the separated testa was higher than those of stearic acid
and oleic acid. For instance, linoleic acid accounted more than
60% of oleic acid in the testa while the level in the testa-free nut
was <30%. Especially, linoleic acid is sensitive to oxidation, which
means that it must be protected from autoxidation. This would
also explain why the cashews with testa have higher levels of other
bioactive compounds, although they were processed similarly to
testa-free cashew nuts for 3 h at 45 °C. The samples were also pro-
tected against very high temperature, since on heating cytotoxic
products, such as trans-isomers, may be formed (Martin, Milinsk,
Visentainer, Matsushita, & De-Sauza, 2007).
The stearic acid level (41 g/kg DM) of testa-containing samples
was found to be higher than those in a previous report on cashew
nut (3.43 g/100 g), chestnut (0.02 g/100 g), peanut (1.30 g/100 g),
hazelnut (0.94 g/100 g), coconut (1.10 g/100 g), macadamia
(1.47 g/100 g), almond (0.55 g/100 g), pecan (1.51 g/100 g), pista-
chios (0.98 g/100 g) and walnut (1.37 g/100 g) (Souci, Fachmann,
& Kraut, 2008). The oleic acid content (214 g/ kg DM) was higher
than those of chestnut (0.98 g/100 g), coconut (2.10 g/100 g), Brazil
nut (18.5 g/100 g) and walnut (11.4 g/100 g), whereas the linoleic
acid level of testa-containing kernels (69 g/ kg DM) was higher
than those of coconut (0.68 g/100 g) and macadamia (1.74 g/
100 g) (Souci, Fachmann, & Kraut, 2008). Linoleic acid was assessed
as precursor of arachidonic acid and eicosapentaenoic acid in the
eicosanoid metabolism (Tapiero, Ba, Couvreur, & Tew, 2002). It is
also a component of phospholipids, which are responsible for the
structure and function of biological membranes. In recent years,
studies have demonstrated that dietary polyunsaturated fatty
acids have protective effects on metabolic syndrome diseases, such
Table 2
Fatty acids in testa-free vs. testa-containing kernels of cashew nut.1,2
Bioactive compounds (g/kg Cashew nut samples
DM)
Testa-free Testa-containing
kernels kernels
Stearic acid 42.8a ± 3.6 40.9a ± 6.3
Oleic acid 219a ± 18.5 214a ± 33.2
Linoleic acid 62.3a ± 5.1 68.6a ± 10.0
1
Values are means ± standard deviation of seven separate determinations (n = 7).
2
Values in the same row with similar superscripts are not significantly different
(p < 0.001).
as Type 2 diabetes, hyper-lipidemia or cardiovascular disease.
Equally promising results are now available regarding their posi-
tive effects on inflammatory diseases, cancer and osteoporosis
(Benatti, Peluso, Nicolai, & Menotti, 2004).
4. Conclusion
In the present study, the phenolic compounds in methanolic ex-
tract of cashew nut testa were clearly identified as (+)-catechin and
()-epicatechin by RP-HPLC and LC–MS. Further, it was demon-
strated that the levels of (+)-catechin and ()-epicatechin in the
testa of cashew nuts are higher than those reported for green tea
and chocolate. The testa-containing cashew nuts possess signifi-
cantly higher amounts of carotenoids and tocopherols, but a lower
level of thiamine when compared to testa-free kernels. Such higher
levels of carotenoids and tocopherols of cashew nut kernels were
contributed by the testa and the low thiamine content was likely
due to the diminishing effect of catechins in testa. The presence
of such potentially bioactive compounds in the testa-containing
cashew nut kernels could be of interest for the food and nutraceu-
tical industries, where it can be employed as an economic source of
natural antioxidants. While the cashew nut testa is heat-treated
and removed during various conventional processes, it is possible
to retain the testa by the newly developed ‘‘Flores’’ hand-cracking
method. Removal of cashew nut testa may significantly decrease
the uptake of essential bioactive compounds. Hence, the processing
of cashew nuts should be re-considered to avoid the loss of cashew
nut testa and its appreciable levels of catechins. In this context, the
recently developed ‘‘Flores’’ hand-cracking process could be rec-
ommended, on the industrial scale, in order to produce cashew
nut kernels along with testa at a mild temperature.
Acknowledgements
Authors are deeply grateful to Mr. Jochen Wolf, Managing
Director, Flores Farm GmbH, Germany, Rudolf Heering, President
Director, PT. PMA, Tangerang, Indonesia and Dr. Frank-Michael
Lange, Terra Fusca, Germany, for their immense help in providing
the cashew nut samples and continuous support. One of the
authors (VV) is thankful to the Alexander von Humboldt (AvH)
Foundation, Bonn, Germany for the award of a post doctoral re-
search fellowship.
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