Food Research International 97 (2017) 178–183

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Food Research International

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Kinetics of aflatoxin degradation during peanut roasting MARK

a,b,⁎ b a a
Ligia M. Martins , Anderson S. Sant'Ana , Beatriz T. Iamanaka , Maria Isabel Berto ,
John I. Pittc, Marta H. Taniwakia
a
Food Technology Institute — ITAL, Campinas, SP, Brazil
b
Department of Food Science, Faculty of Food Engineering, University of Campinas, Campinas, SP, Brazil
c
CSIRO Agriculture and Food, P.O. Box 52, North Ryde, NSW 1670, Australia

A R T I C L E I N F O A B S T R A C T

Keywords: This study investigated aflatoxin degradation during peanut roasting. First, peanuts contaminated with three
Peanut initial aflatoxin concentrations (35, 332 and 695 μg/kg) were roasted at 180 °C for up to 20 min. The percentage
Mycotoxin of aflatoxin degradation after 20 min were 55, 64 and 81% for peanuts contaminated with aflatoxin at 35, 332
Inactivation kinetics and 695 μg/kg, respectively. This difference was statistically significant (p < 0.05), showing that initial
Aspergillus section Flavi
concentration influences aflatoxin reduction. Thereafter, peanut samples contaminated with an initial aflatoxin
Food processing
concentration of 85 μg/kg were roasted at 160, 180 and 200 °C for 5, 10, 15, 20 and 25 min, then residual
concentrations of aflatoxin were determined. Roasting at 160, 180 and 200 °C resulted in an aflatoxin reduction
of 61.6, 83.6 and 89.7%, respectively. This study has provided quantitative data reinforcing the fact that roasting
alone is not enough to control aflatoxins in peanuts.

1. Introduction
Aflatoxins are secondary metabolites produced principally by
Aspergillus flavus, A. parasiticus and A. nomius (Pitt & Hocking, 2009).
These mycotoxins are the most potent liver carcinogens known and are
currently classified by the International Agency for Research on Cancer
as Group 1 carcinogens, i.e. known human carcinogens (IARC, 2002).
Moreover, aflatoxins have acute, chronic, genotoxic and immunosup-
pressive properties (ICMSF, 2002; Williams et al., 2004).
Aflatoxins have been found in a wide range of food commodities,
including cereals (EFSA, 2013), dairy products (Campagnollo et al., 2016)
spices (Hammami et al., 2014) and even in meat and eggs (Abd-
Elghany & Sallam, 2015; Herzallah, 2009; Oliveira et al., 2000). However,
oilseeds including peanuts (Martins et al., 2017; Mutegi, Ngugi,
Hendriks, & Jones, 2009; Mutegi et al., 2013; Oliveira, Gonçalves,
Rosim, & Fernandes, 2009), pistachios (Georgiadou, Dimou, & Yanniotis,
2012; Molyneux, Mahoney, Kim, & Campbell, 2007; Set & Erkmen, 2010),
hazelnuts (Baltaci, Ilysaglu, & Cavrar, 2012; Ozay, Seyhan, Pembeci,
Saklar, & Yilmaz, 2008), walnuts (Molyneux et al., 2007), cocoa
(Copetti, Iamanaka, Pereira, Fungaro, & Taniwaki, 2011; Turcotte,
Scott, & Tague, 2013), almonds (Gürses, 2006; Molyneux et al., 2007),
and sesame (Hosseininia, Vahabzadeh, Rashedinia, Riahi-
Zanjani, & Karimi, 2014; Kollia, Tsourouflis, & Markaki, 2016) are more
likely to be highly contaminated. Peanuts are one of the most susceptible
crops because peanut kernels can be invaded by A. flavus or A. parasiticus
in soil (Horn, Greene, & Dorner, 1995). After fungal invasion, aflatoxin can
formed in peanuts before harvest, during drying and during storage under
poor conditions (Horn et al., 1995; Pettit & Taber, 1968; Pitt,
Taniwaki, & Cole, 2012).
Peanuts are therefore an important source of aflatoxin in the human
diet (Mutegi et al., 2009, 2013; Oliveira et al., 2009). Various
approaches have been studied to control the contamination of peanuts
by aflatoxins (Pitt et al., 2012). However, for control to be effective,
quantitative data on the effects of processing are needed to assist in
effective management of aflatoxins. Aflatoxin degradation has been
reported by microwaves (Luter, Wyslouzil, & Kashyap, 1982; Mobeen,
Aftab, Asif, & Zuzzer, 2011), heating in oil (Lee, Cucullu, Franz, & Pons,
1969) and roasting (Arzandeh & Jinap, 2011; Ogunsanwo, Faboya,
Idowu, Lawal, & Bankole, 2004). However, the roasting temperatures
studied by these authors were different from those commercial roasting
conditions in Brazil and other countries that usually are from 160 to
200 °C. Additionally, Ogunsanwo et al. (2004) and Arzandeh and Jinap
(2011) did not focus on the analysis of the degradation kinetics of
aflatoxins. Therefore, the aims of this study were to investigate the
influence of initial aflatoxin concentration on aflatoxin degradation in
raw peanuts, evaluate the aflatoxin degradation kinetics during peanut
roasting and to evaluate the effect of roasting time and temperature on
peanut colour.

⁎
Corresponding author at: 2880 Brasil Avenue, Campinas, SP Cep 13070-178, Brazil.
E-mail addresses: ligmmartins@gmail.com (L.M. Martins), marta@ital.sp.gov.br (M.H. Taniwaki).

http://dx.doi.org/10.1016/j.foodres.2017.03.052
Received 18 September 2016; Received in revised form 15 March 2017; Accepted 19 March 2017
Available online 04 April 2017
0963-9969/ © 2017 Elsevier Ltd. All rights reserved.
L.M. Martins et al.
Fig. 1. Spouted bed roaster equipment.The figure shows the air dryer scheme set for
roasting peanuts. The sample was placed in the dryer bed (ϕ 104, 1 mm) on a perforated
screen. The fan air speed is controlled by a frequency inverter (Inv-F). The air velocity,
provided by centrifugal fan (BB), was previously set with the amount of peanut sample
(100 g) in order to establish a spouted bed, without dragging off the sample to the cyclone
(C), which directs possible entrained particles into the container (R). A proportional
control strategy tuned through a PLC controller maintained the drying temperature in the
set point values. Air-1 represents cold air before passing through the resistor, and Air-2
represents the warm air after passing through the sample.
2. Material and methods
2.1. Peanut samples
Approximately 20 kg of raw shelled peanuts with intact skins were
obtained directly from a peanut cooperative in São Paulo State, Brazil.
The nuts were surface disinfected and plated on Dichloran 18%
Glycerol agar (Pitt & Hocking, 2009) to assess infection by species from
Aspergillus section Flavi. After five days at 25 °C, 100% of the fifty nuts
examined were infected, so this batch provided samples naturally
contaminated with aflatoxins.
2.2. Roasting conditions
Peanut samples (100 g) were roasted in a vertical spouted bed
roaster (104 mm diameter, 590 mm height, 20 mm screen, 45°) (Fig. 1),
designed in the Process Engineering Laboratory at the Institute of Food
Technology.
2.3. Influence of initial concentration on aflatoxin degradation
A first study was conducted to determine whether initial concentra-
tion of aflatoxin influenced its degradation. The water activity of
peanuts (10 kg) were increased to 0.9 by addition of 10 mL/g of sterile
water and incubated at 25 °C. In order to obtain samples with different
initial aflatoxin concentrations, subsamples were removed over several
days and dried to 0.5 water activity in an incubator at 55 °C for 10 h.
After skins were removed, peanuts were homogenized and portions
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Food Research International 97 (2017) 178–183
(100 g) roasted at 180 °C for 5, 10, 15 and 20 min in duplicate.
2.4. Kinetic of aflatoxin degradation in peanuts during roasting as
influenced by temperature
Samples (100 g) were roasted for 5, 10, 15, 20 and 25 min at 160,
180 and 200 °C. The roaster was preheated until the roasting tempera-
ture (160, 180 or 200 °C) was achieved and after the roasting time the
heater was turned off and air passed through each sample to accelerate
cooling. The temperature and time values used were defined according
to the range used by Brazilian industries for roasted peanut production.
The roasted peanut samples were ground using a laboratory mill (Model
A11, IKA, Staufen, Germany) for colour and aflatoxin analyses. Each
time and temperature study was replicated, and means and standard
deviations calculated.
The aflatoxin degradation kinetic parameters were determined by
fitting the Weibull model to the data as shown in Eq. (1) (Mafart,
Couvert, Gaillard, & Leguerinel, 2002):
C ⎛ t ⎞p
log10 = −⎜ ⎟
C0 ⎝δ⎠ (1)
where C0 and C represent the initial and final concentrations of
aflatoxin (μg/kg), t the roasting time (min), δ treatment time for the
first decimal reduction, and p represents the shape parameter.
2.5. Aflatoxin analysis
Aflatoxin analyses were carried out according to Stroka, Anklam,
Jorissen, and Gilbert (2000) with modifications, as follows.
2.5.1. Clean-up
Twenty-five grams of ground peanuts were added to 2.5 g of NaCl
and extracted with 100 mL of methanol: water (8:2, v/v) for 30 min
using a horizontal shaker (New Brunswick Scientific Company, New
Brunswick, NJ, USA). After filtration through quantitative filter paper
(Nalgon, Brazil), and a glass microfiber filter (Vicam, Milford, MA,
USA), filtrate (4 mL) was diluted in phosphate buffered saline (60 mL;
pH 7.0) and applied to an immunoaffinity column (Aflatest WB, Vicam)
at a flow rate of 2–3 mL/min. The column was then washed with 30 mL
of distilled water and aflatoxins eluted with methanol (1250 μL)
followed by Milli Q water (1750 μL).
2.5.2. Chromatographic conditions
The HPLC system (Model 1260 Infinity, Agilent, Santa Clara, CA,
USA) was used with fluorescence detection (362 nm excitation and
455 nm emission), C18 column, and an isocratic and reverse phase
system, associated with a Kobracell electrochemical reactor (R-
Biopharm, Darmstadt, Germany) connected to a current of 100 μA for
post-column derivatization of aflatoxins B1 and G1. The mobile phase
was water:acetonitrile:methanol (6:2:3, v/v/v), with potassium bro-
mide (119 mg/L and nitric acid (4 M, 350 μL) at a flow rate of 1 mL/
min. The injection volume was 20 μL and the limit of detection (LOD)
and limit of quantification (LOQ) were 0.05 and 0.17 μg/kg respec-
tively. Aflatoxin standards were obtained from Sigma-Aldrich (St Louis,
MO, USA). Each 100 g peanut sample was analyzed as a single lot.
2.6. Colour analysis
Samples were ground and the colour reflectance values (CIELAB L*)
were measured in duplicate at ambient temperature using a Konica
Minolta CM-5 spectrophotometer (Konica Minolta, Chiyoda, Japan). A
reflectance specular method was used with a 3 cm area view, illuminant
D65 and 10° standard observer angle (Smith, Perry, Marshall,
Yousef, & Barringer, 2014). Two samples of roasted peanuts were
purchased in the market for comparison with colours of the peanuts
roasted in this work. Colour values were checked for significant
L.M. Martins et al. Food Research International 97 (2017) 178–183

Table 1 Table 2
Aflatoxin concentration, % of reduction obtained after roasting at 180 °C for up to 20 min. Temperature and time conditions of roasting, aflatoxin concentration, aflatoxin degrada-
tion and colour analysis in peanuts.
Roasting times (min) at 180 °C Aflatoxins (μg/kg)
Roasting conditions Aflatoxins (μg/kg) Aflatoxin Colour analysis⁎⁎
0 35.2 332.1 695.0 degradation
5 30.1 ± 1.9 311.6 ± 55.0 625.4 ± 2.3 Temperature (°C) Time (%) L*
10 28.2 ± 2.0 320.5 ± 19.2 420.1 ± 6.3 (min)
15 20.5 ± 0.8 265.8 ± 21.8 312.0 ± 2.4
20 15.9 ± 0.8 119.1 ± 1.2 130.4 ± 3.4 Standard 1 68.4 ± 0.07b
Aflatoxin degradation (%) 54.7 64.1 81.2 Standard 2 71.61 ± 0.83a
Raw peanut 84.77 ± 52.39
160 5 71.5 ± 50.2 15.7 69.21 ± 0.57b
statistical differences (p ≤ 0.05). 10 41.9 ± 4.9 50.6 64.0 ± 0.09c
15 36.2 ± 29.7 57.4 61.50 ± 0.33d
20 38.2 ± 0.3 54.9 60.23 ± 1.20e
2.7. Statistical analysis 25 32.6 ± 9.9 61.6 63.2 ± 0.02c
180 5 51.3 ± 32.9 39.5 60.79 ± 0.07e
The results were compared using one-factor analysis of variance 10 26.6 ± 13.8 68.6 54.08 ± 0.16g
15 32.2 ± 14.3 62.0 52.06 ± 0.70h
(ANOVA) followed by the Scott-Knott test. Statistical analyses were 20 21.1 ± 2.4 75.1 51.21 ± 1.04h
carried out in Assistat version 7.5 software (Silva & Azevedo, 2002). 25 13.9 ± 2.7 83.6 48.41 ± 0.08i
200 5 46.6 ± 2.1 45.1 57.53 ± 0.03f
3. Results and discussion 10 10.7 ± 1.9 87.4 46.94 ± 0.04j
15 9.5 ± 0.1 88.8 44.12 ± 0.02l
20 8.8 ± 0.4 89.7 41.98 ± 0.17m
3.1. Effect of initial aflatoxin concentration on degradation 25 8.7 ± 0.0 89.7 39.29 ± 0.06n

⁎⁎
The effect of initial aflatoxin concentration on percentage aflatoxin Different letter in the same column represents statistically significant difference
degradation can be observed in Table 1. The higher the initial aflatoxin (p < 0.05).

concentration the higher the reduction percentage. After 20 min

roasting, the aflatoxin degradation was 55, 64 and 81% when initial
aflatoxin contamination was 35, 332 and 695 μg/kg, respectively. This
difference was statistically significant (p < 0.05), showing that initial
concentration influences aflatoxin reduction. These results are in
agreement with the results of Lee et al. (1969) who studied the
aflatoxin degradation in peanuts during oil and dry roasting and also
found that the highest aflatoxin degradation was observed in the
peanuts with the highest initial concentration. The study by Lee et al.
(1969) was very similar to the present study, i.e. water activity of
samples was raised to stimulate aflatoxin production before redrying,
the only difference being that Lee et al. (1969) inoculated peanuts with
aflatoxin producing strains, while in the present study the peanuts
sample was naturally contaminated.
The results obtained in this study differ from those reported by
Pluyer, Ahmed, and Wei (1987), who used both artificially and
naturally contaminated peanuts to study aflatoxin destruction by oven
and microwave roasting. Their results on naturally contaminated
samples after roasting showed no correlation between aflatoxin initial
concentration and the percentage of destruction. On the other hand,
Arzandeh and Jinap (2011) found a higher percentage reduction with
increasing time and initial concentration in samples containing up to
200 μg/kg aflatoxin, but a decrease of the percentage reduction
occurred above this concentration. Arzandeh and Jinap (2011) used
naturally contaminated ground peanut samples. Although the use of
ground samples will result in greater homogeneity of aflatoxin con-
centration, roasting ground peanuts does not reflect the reality of the
industry. This may explain the differences between the study of
Arzandeh and Jinap (2011) and the present study. None of these
studies evaluated the degradation of different initial concentrations
under the same time and temperature conditions, so our study adds
novel information to the literature.
The lowest initial aflatoxin concentration studied (35 μg/kg) was
15 μg/kg above the maximum limit established by Brazilian legislation
(20 μg/kg). Under these conditions, roasting for 20 min at 180 °C led to
reduction in aflatoxin levels below the regulated limit. However,
peanut colour (Table 2) after this roasting time and temperature was
darker and statistically different (p < 0.05) from the colour of market
samples, and would likely not be acceptable to consumers. Depending
on the initial levels in the raw materials for roasting, other measures
may need to be implemented to reduce aflatoxin levels in peanuts and
to meet regulatory requirements.
3.2. Kinetics of aflatoxin degradation in peanuts during roasting as
influenced by temperature
Determined from 12 assays, the average aflatoxin concentration in
the raw peanuts used was 84 ± 77 μg/kg. The high standard deviation
in these analyses is not unusual. Data available in the literature indicate
that in addition to natural variability of A. flavus strains to produce
aflatoxins, the high standard deviation observed in the case of peanuts
can be explained by the fact that only a few kernels in any sample are
highly contaminated (Whitaker, 2003; Whitaker & Wiser, 1969). Even
with strict sample preparation approaches and 12 aflatoxin assays, a
high standard deviation was obtained. This was a constant source of
uncertainty in the measurement of contamination level in samples used
in the experiments.
The effect of temperature on aflatoxin concentration, percentage
aflatoxin degradation and colour analysis are depicted in Table 2. After
25 min at 160, 180 and 200 °C, aflatoxin reductions were 61.6, 83.6
and 89.7%, respectively.
Aflatoxin degradation during roasting was not always linear. The
Weibull model was used to fit the data obtained on aflatoxin degrada-
tion at 160, 180 and 200 °C (Fig. 2). The δ and p values for treatments
at 160, 180 and 200 °C were 92.6 ± 59.9 min and 0.61 ± 0.28,
38.6 ± 9.8 min and 0.70 ± 0.24, 19.7 ± 9.0 min and
0.49 ± 0.26, respectively. As δ represents the time for the first decimal
reduction, it can be seen that the higher the temperature, the shorter
the time to get the first decimal reduction. Only the temperature of
200 °C gave a δ value within the time interval used in this study.
Peanut colour after roasting (Fig. 3) was analyzed using the CIELAB
method (Table 2). L* values decreased with increasing roasting time for
all temperatures studied, i.e. at 160, 180 and 200 °C, after 5 to 25 min,
L* values ranged from 69.2 to 63.2, 60.8 to 48.4 and 57.5 to 39.3,
respectively. The colour of roasted peanuts is usually considered a
control parameter as it is related to flavour development (Smyth et al.,
1998). Colour analysis permitted comparison between roasted peanuts
from the market and samples from different roasting times and
temperatures. Although standard L* values obtained ranged from
68.4 to 71.61, only roasting at 160 °C for 5 min gave a statistically
similar L* value compared to the market samples. Under these

180
L.M. Martins et al.
Fig. 2. Aflatoxin degradation during roasting fitted using Weibull model at 160 °C (A), at 180 °C (B) and at 200 °C (C). Where N is the aflatoxin concentration (μg/kg).
conditions initial aflatoxin concentration was reduced by only 15%.
However, it is worth noting that the colour values obtained between the
market samples was also significantly different, showing that the
peanut roasting point in the market does not always follow the same
pattern.
We found that the same colour was achieved through different time
and temperature combinations, for example, roasting for 20 min at
160 °C resulted in the same L* value as roasting for 5 min at 180 °C.
However that does not guarantee that nut quality is the same, as
McDaniel, White, Dean, Sanders, and Davis (2012) suggested some
quality differences between peanuts roasted at different times and
temperatures that showed equivalent colours. Even so, the data
reported in the present study shows a possible alternative which results
in a shorter process time for the industry and yet a greater reduction of
aflatoxin, resulting in similar colour parameters after roasting.
The temperature and the time of process, as well as the aflatoxin
contamination in the peanuts used in this work were determined in
order to obtain results with practical and direct application by
industries. Our study used only naturally contaminated peanut samples,
as other studies have sometimes shown differences in aflatoxin reduc-
tion during roasting when contamination was artificial. For example,
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Food Research International 97 (2017) 178–183
Pluyer et al. (1987) studied the effects of roasting in an oven (150 °C for
30 min) and by microwave (8.5 min at 0.7 kW) on both naturally
occurring and artificially added aflatoxin in peanuts. Oven or micro-
wave roasting reduced aflatoxin B1 by 30–45% for artificially con-
taminated peanuts and 48–61% for naturally contaminated nuts.
Arzandeh and Jinap (2011) observed a maximum aflatoxin B1 and B2
reduction of 78.4 and 57.3% respectively for artificially contaminated
peanuts, compared with 80.2% for aflatoxin B1 and 69.7% for B2 for
naturally contaminated samples.
A similar study conducted by Lee et al. (1969) also used naturally
contaminated peanut samples and the average of all results in oil and
dry roasting with four different levels of aflatoxin (2200, 960, 90 and
50 μg/kg for aflatoxin B1 and 4100, 1600, 150 and 80 μg/kg ppb for
aflatoxin G1) were 64.5% (B1) 61.5% (G1), 68.8% (B1) and 66.8% (G1)
respectively. Although Lee et al. (1969) did not conduct a kinetic study,
their samples with initial contamination of 50 μg/kg aflatoxin B1 and
80 μg/kg aflatoxin B2 (a total of 130 μg/kg aflatoxins in the sample)
was the closest value to that used in the present study, and when
roasted at 204 °C for 5 min, obtained 56 and 24% of aflatoxin
degradation, a result close to that observed in the present study,
showing the same tendency in naturally contaminated samples.
L.M. Martins et al.
Fig. 3. Peanut colour as affected by time and temperature of roasting. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this
article.)
Other roasting processes than those used in our study have shown a
higher percentage of aflatoxin degradation. Luter et al. (1982) esti-
mated aflatoxin reduction through microwave roasting and found a
minimum reduction of 20.8% total aflatoxin at 140 °C for 4.5 min, a
maximum aflatoxin reduction of 100% at 137 °C for 20 min and 185 °C
for 7 min, but both temperature and time conditions resulted in
excessively dark peanut.
4. Conclusions
The results presented in this study reinforce the need for greater
control throughout the peanut production chain, so that, at the roasting
stage, the aflatoxin concentration lies close to the maximum permitted
limit. Colour sorting, blanching and roasting are processing stages used
for aflatoxin reduction in the peanut production chain (Pitt et al.,
2012). This study has provided quantitative data reinforcing the fact
that roasting alone is not enough to control aflatoxins in peanuts. These
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Food Research International 97 (2017) 178–183
data are useful for risk based management of consumer exposure to
aflatoxin.
Acknowledgements
The authors acknowledge the financial support of “Fundação de
Amparo a Pesquisa do Estado de São Paulo” (FAPESP) (Grant # 2011/
50136-0), “Conselho Nacional de Desenvolvimento Científico e
Tecnológico” (CNPq) (Grant # 302763/2014-7 and 305649/2014-0),
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
and CAP Agroindustrial.
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