Food Control 50 (2015) 858e863

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Food Control
journal homepage: www.elsevier.com/locate/foodcont

Occurrence of aatoxin B1 in conventional and organic our in Italy

and the role of sampling
Sara Armorini, Alberto Altani, Anna Zaghini, Paola Roncada*
Department of Veterinary Medical Sciences, School of Agriculture and Veterinary Medicine, Alma Mater Studiorum, University of Bologna, via Tolara di
Sopra 50, 40064 Ozzano Emilia, BO, Italy

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

Article history: Aatoxins (AFs) are mycotoxins produced by certain species of Aspergillus and aatoxin B1 (AFB1) is the
Received 1 August 2014 most toxic, consistently carcinogenic and genotoxic. In the present study, a total of 90 different samples
Received in revised form of organic and conventional ours (20 conventional wheat our, 20 organic wheat our, 42 conventional
14 October 2014
corn our, and 8 organic corn our) were analyzed for the presence of AFB1. The samples were purchased
Accepted 21 October 2014
Available online 25 October 2014
from local stores in the surrounding of Bologna during the period April to May 2013. For the quanti-
cation of AFB1 in ours, a fast and sensitive HPLCeFD method was developed and tested for several
validation parameters (linearity and range, specicity, accuracy and precision, LOD, LOQ and recovery).
Keywords:
Aatoxin B1
AFB1 was found in 13 samples of corn our, specically 4 organic and 9 conventional. These results
Conventional our conrm a higher incidence of contamination in corn compared to wheat, as reported in literature. No
Organic our signicant differences were observed in the concentration of AFB1 by comparing conventional corn our
Sampling samples and organic corn our samples (P > 0.05). The levels of contamination found in the positive
HPLC samples ranged between 0.17 and 3.75 mg kg
1 but only in one case the limit dened at Community level
(2 mg kg
1) was exceeded. This demonstrates the effectiveness of the network of monitoring before the
placing on the market of these food products. Furthermore, in this study efforts were made to evaluate
the minimum quantity of our related to a commercial package, which is reliable for the assessment of
contamination by AFB1. In fact, in sample preparation, the selection of the sample size for the laboratory
analysis is a very important step. The minimum amount to get a reliable result is 20 g and smaller
quantities can lead to incorrect results.
2014 Elsevier Ltd. All rights reserved.

1. Introduction known as aatoxin B1 (AFB1), aatoxin B2 (AFB2), aatoxin G1

Wheat our is one of the most important ingredient and food in
European and American culture. Bread, pasta, crackers, and many
cakes, among other foods and cooking recipes, are made using our
or including it as an ingredient (Gashgari, Shebany, & Gherbawy,
2010). In Italy, the cornmeal is a very versatile ingredient used to
prepare many recipes. The most important is polenta, prepared
by cooking cornmeal in boiling salted water, but it can be used for
breading various foods, and to prepare bread, cookies, and cakes.
Aatoxins (AFs) are mycotoxins produced by certain species of
Aspergillus, particularly Aspergillus avus, Aspergillus parasiticus and
Aspergillus nomius. There are more than 20 distinct but structurally
related aatoxin compounds e the four most commonly found are
* Corresponding author. Department of Veterinary Medical Sciences, School of
Agriculture and Veterinary Medicine, Alma Mater Studiorum, University of Bologna,
via Tolara di Sopra 50, 40064 Ozzano Emilia, BO, Italy. Tel.: 390512097511.
E-mail address: paola.roncada@unibo.it (P. Roncada).
(AFG1) and aatoxin G2 (AFG2) (Hernandez-Martnez & Navarro-
Blasco 2010; Tam et al., 2006). Aatoxin B1 (AFB1) is consistently
carcinogenic and genotoxic in vitro and in vivo (European Food
Safety Authority, 2007), and it was classied in the group 1 by
the International Agency for Research on Cancer (IARC, 2002).
Extensive contamination of food and drinks with mycotoxins is the
main problem over the world since they can also compromise the
safety of food and feed supplies and adversely affect health in
humans and animals (Rubert, Soler, & Man ~ es, 2011). In the Euro-
pean countries the usual content limit for AFB1 in our is 2 mg kg
1.
Tropical and sub-tropical regions (warm and humid weather)
provide optimal conditions for fungal growth (Aycicek, Aksoy, &
Saygi, 2005; Cavaliere et al., 2007; Herna ndez-Martnez &
Navarro-Blasco, 2010; Hussein & Brasel, 2001; Ramos Catharino
et al., 2005; Shenasi, Candlish, & Aidoo, 2002; Zinedine et al., 2006;
llner & Mayer-Helm, 2006). These fungi can grow on certain
Zo
foods and feeds under favorable conditions of temperature and
humidity and generate AFs before and/or during harvest, handling,

http://dx.doi.org/10.1016/j.foodcont.2014.10.031
0956-7135/ 2014 Elsevier Ltd. All rights reserved.
 S. Armorini et al. / Food Control 50 (2015) 858e863
shipment and storage. In Italy, in summer 2012 moisture and
temperature were particularly critical for aatoxin production. The
Ministry of Health trasmitted to the competent authorities and
operators in the food and animal feed sectors a guideline for the
prevention and the management of the risk aatoxin, entitled
Operating Procedures for the extraordinary prevention and risk
management of aatoxin contamination in the dairy industry and
in the production of corn for human and animal consumption, as a
result of extreme weather conditions.
The comparison between organic and conventional foods has
shown advantages in food composition by higher levels of nutri-
ents and lower levels of residues, such as pesticides (Huber,

Rembiakowska, Srednicka, Bgel, & Van De Vijver, 2011).
Organic and conventional production differ in many respects.
Pesticides and products of synthesis are not used. Crop protection
is implemented by selecting disease-resistant species and by
adopting specic cultivation techniques such as crop rotations,
intercropping, the planting of hedges and trees that host the
natural predators of pests, and create a natural physical barrier
against external pollutants. Moreover, natural fertilizing such as
dung and others organic substances (green manures) are used in
organic production. If necessary, vegetable, animals and mineral
substances are employed for crop protection: plant extracts, nat-
ural mineral or rock our to correct structure and chemical char-
acteristics of soil and to protect crops from cryptogams. Since no
fungicides are used during organic production, it has been sug-
gested that cereals may contain higher levels of mycotoxins
(Hoogenboom et al., 2008).
In a mycotoxin test procedure consisting of sampling, sample
preparation, and analysis, the main contribute to the total variance
of the test is given rstly by sampling and secondly by sample
preparation while analysis has a lesser weight. Because of the
variability associated with each step of the mycotoxin test proce-
dure, the true mycotoxin concentration of a bulk lot cannot be
determined with 100% certainty. Some good lots will be rejected by
the sampling plan (seller's risk or false positives) and some bad lots
will be accepted by the sampling plan (buyer's risk or false nega-
tives) (Whitaker, 2003).
Particularly in the case of large lots of cereals to be controlled for
mycotoxin contamination, it is important to follow dened sam-
pling protocols as the ones dened in the specic European
Regulation (European Commission, 2006b). The distribution of
mycotoxins is very heterogeneous with contamination patchy
even within the cereal storage silos.
Matrices, such as our and feed powder are generally consid-
ered more homogeneous than cereal grain, but in any case it must
be assessed the representativeness of the sample. Just after sam-
pling, sample preparation is the second source of error in a pro-
cedure for mycotoxin quantication.
The sample as received in the laboratory usually needs to be
further reduced and processed to what is frequently called the test
sample. This is a much smaller, but still representative, subsample
with an often ner particle size, from which test portions are
selected for specic analyte determinations.
In this context, the chose of the test portion size is very
important in that it must accurately reect the concentration of the
analyte in the entire laboratory sample.
The main aim of this study was to obtain data on the occurrence
of AFB1 from organic and conventional ours purchased from local
markets, supermarkets, and specialized shops for organic products,
intended for direct human consumption. A further aim was to
determine the minimum quantity of our related to a commercial
package which is reliable for the assessment of contamination by
aatoxin B1. This is an important aspect of the sample preparation
that may affect mycotoxin analysis.
859
2. Material and methods
2.1. Samples
From April to May 2013, 90 different samples were purchased in
a random manner from local stores (supermarkets, small retail
shops, small groceries, and specialized suppliers) located in the
surrounding of Bologna. All the different types of ours available
were purchased also in specialized shops for organic products. The
weight of the retail packs ranged between 250 and 1000 g. The
ours studied were either organic (n 28) or conventional (n 62).
In particular the sample were: 20 samples of conventional wheat
our and 20 of organic wheat our, 42 of conventional corn our
and 8 of organic corn our. The samples were stored at room
temperature until analysis. After purchase, the packs were regis-
tered and cataloged in the laboratory notebook.
Before analysis, the samples were mixed very thoroughly and
an aliquot of 200 g was separated from each original packaging
and putted into a new bag. From this sample, after thorough
mixing, 5 g of our were taken, extracted and analyzed as
described below. The choice to use 5 g of our was made based on
the indications suggested for many tests for screening analysis of
aatoxins.
In accordance with the purposes of the present work, after this
rst round of analysis of all the our samples, from the ones tested
positive, after thorough mixing, a subsample of 20 g was taken and
divided into 4 aliquots of 5 g to be submitted to the extraction and
analytical procedure.
Subsequently from ve of these samples, another portion of 20 g
was taken and divided into 4 aliquots of 5 g to be submitted to the
extraction and analytical procedure. Finally, the negative samples
(<LOD) were also considered. To assess the reliability of a negative
result of a screening test performed with an aliquot of 5 g of sample,
4 negative samples were selected and from each of them, a sub-
sample of 20 g was collected, divided in 4 aliquots of 5 g to be
submitted to the extraction and analytical procedure.
2.2. Solvents and reagents
The chemicals and solvents used for the extraction of AFB1 from
our samples (citric acid, dichloromethane) were ACS grade (Mal-
linckrodt Baker B.V., Deventer, The Netherlands). Triuoroacetic
acid used for the derivatization of AFB1 was HPLC grade (Sigma-
eAldrich Co., St Louis, MO, USA).
All solvents used for HPLC analysis (water, acetonitrile, isopropyl
alcohol, acetic acid), were HPLC grade (Mallinckrodt Baker B.V.,
Deventer, The Netherlands).
AFB1 standard was purchased from SigmaeAldrich Co. (St Louis,
MO, USA).
2.3. Chromatographic apparatus
The HPLC system consisted of a Beckman System Gold Pro-
grammable Solvent Module 126 pump equipped with an HTA HT
800 L autosampler tted with a 20 ml loop and a Jasco model 821 FP
uorescence detector (FD); uorescence excitation and emission
wavelengths were 365 and 418 nm respectively. The system was
computer-controlled by a Beckman Coulter 32 Karat Software. The
analytical column was a Phenomenex Onyx Monolithic C18
(100  4.6 mm).
Chromatographic separation was achieved in gradient
conditions.
The mobile phase consisted of water-acetonitrile-isopropyl
alcohol-acetic acid 1% mixtures in various ratios:
860 S. Armorini et al. / Food Control 50 (2015) 858e863
mobile phase A: water/acetonitrile/isopropyl alcohol/acetic acid
1% (91: 1: 1: 7 v/v).
mobile phase B: water/acetonitrile/isopropyl alcohol/acetic acid
1% (43: 25: 25: 7 v/v).
Gradient: 84% A and 16% B for 4.5 min, then in 4.5 mine52.5% A
and 47.5% B with linear increase. Flow: 1.3 ml/min for 6 min,
then to 1 ml/min in 1 min. The injection volume was 20 ml.
2.4. Extraction
The procedure involves extraction of aatoxin B1 with
dichloromethane and its conversion to the hydroxyl-derivative
aatoxin B2a using triuoroacetic acid (TFA) to increase his uo-
rescence intensity in HPLC-FD analysis.
Five grams of the different kinds of our were acidied with
2.5 ml of citric acid 20% and extracted with 25 ml of dichloro-
methane by means of ultrasound for 30 min. The sample was
centrifuged for 15 min (5400  g); an aliquot of 10 ml of the
dichloromethanic extract (equivalent to 2 g sample) was trans-
ferred into a centrifuge tube and evaporated to dryness. For tri-
uoroacetic acid derivatization, 20 ml of TFA were added to the
residue and allowed to react for 10 min; nally 1980 ml of injection
solvent (water-acetonitrile, 90:10) were added. The sample was left
in the dark for at least 20 min and then injected on column.
2.5. Quantication
The analytical method for HPLCeFD detection of aatoxins was
validated in terms of linearity and range, specicity, accuracy and
precision, limit of detection (LOD), limit of quantication (LOQ), and
recovery.
Linearity and range were evaluated through the response of
standards prepared from blank samples spiked directly, adding the
volume of AFB1 standard solution required to obtain six contami-
nation levels in the range 0.5e10 mg kg
1. The samples were
extracted and analyzed as described above.
Specicity was tested by comparing the chromatograms of
blank samples, standards and spiked ours samples at different
concentration levels.
Accuracy and precision were evaluated via analysis of spiked
samples at ve concentration levels in the range 1e10 mg kg
1; each
sample was prepared in quadruplicate over several days and
analyzed. The deviation of the mean from the true value was taken
as a measure of accuracy, while precision was expressed as relative
standard deviation (RSD) of the replicate measurements.
The LOD was calculated as three times the background noise
determined at the retention time of AFB1 in our unscathed (blank
sample). The mean value of ve determinations has been multi-
plied by 3 and compared with the signal of a reference standard.
The LOQ was established as the value corresponding to the
lowest standard on the calibration curve. The analyte response at
the LOQ should be at least ten times the blank response.
The recovery was obtained by analyzing spiked samples
extracted at ve different concentration levels in the range
1e10 mg kg
1 compared to the detector response obtained for pure
AFB1 standard solutions at the same concentration levels. Four
spiked samples for each considered concentration level were
prepared.
3. Results and discussion
3.1. Assay validation
In all regressions, the requirements for linearity were met. Four
calibration curves in the range 0.5e10 mg kg
1 were prepared and
analyzed in several days. The coefcient of determination (R2) for
each calibration curve was greater than 0.999.
A useful analytical method should permit resolution and
detection of the analytes of interests from others interfering sub-
stances and co-eluting sample compounds. Possible interferences
were evaluated by analyzing non-contaminated samples and
spiked samples at different concentration levels. No interfering
peaks were observed in the spiked samples and no signicant
peaks were found within the retention time window of AFB1 in the
non-contaminated samples. The retention time of AFB1 in naturally
contaminated samples and standard solutions was 8.5 0.5 min;
this value was stable (RSD calculated over four days: 6%). The run
time was 11 min. The Fig. 1 shows the chromatograms of a blank
sample (A), a standard calibration (7.5 ng ml
1) (B) and a our
sample naturally contaminated (C). With the mobile phase
composition described above, aatoxin B1 elutes as a sharp sym-
metrical peak with a very good resolution.
Accuracy and precision reect how well the test method per-
forms day to day in laboratory.
Five calibration standard at concentration of 1, 2.5, 5, 7.5 and
10 mg kg
1 were prepared and analyzed in four different days. The
relative standard deviations (RSDs) for each concentration assayed
never exceeded 3%.
The LOD and the LOQ of the method were 0.15 mg kg
1 and
0.50 mg kg
1, respectively.
Recovery experiments showed that the overall average recovery
in the tested range of concentrations was 84.6%. For each concen-
tration, four measurements were performed and the RSDs ranged
from 3 to 7.8%. The data are reported in Table 1.
As previously reported, the analytical method adopted has been
developed with some changes from the one used in our laboratory
for determination of aatoxin B1 in grains and forage by HPLC/FD.
The results were satisfactory, because they led to minimize the
volume of solvent used and also to reduce the processing times of
the unknown samples. The method has in fact guaranteed high rate
of recovery (about 85%), a LOD and LOQ respectively of 0.15 mg kg
1
and 0.50 mg kg
1, far below the legal limit established by European
Commission (2006c) equal to 2 mg kg
1.
It is important to underline the validity of the extraction pro-
cedure: it does not provide a passage of clean-up with SPE or
immunoafnity columns, but at the same time it has enabled us to
obtain analytical samples characterized by the absence of inter-
fering substances in the chromatogram.
The result for what concerns the linearity in the reference and in
the calibration curves was satisfactory, with a coefcient of deter-
mination always greater than 0.999. Finally the gradient developed
for conducting HPLC analysis, together with the use of a monolithic
column, allowed an improvement in efciency and a reduction in
the duration of the chromatographic run.
3.2. Occurrence of AFB1 in our samples
In this study, 90 samples of our were analyzed, 14 of which
were positive in the rst screening analysis made with aliquots of
5 g of sample. In order to evaluate the representativeness of 5 g, it
was decided to proceed with further investigations on these 14
positive samples. From each of them, after thorough mixing, it was
taken an aliquot of 20 g (accurately homogenized). These aliquots
were divided into four sub-aliquots of 5 g, which were subse-
quently analyzed (Table 2).
An additional aliquot of 20 g, then divided into 4 sub-aliquots,
was taken from 5 of these positive samples, and then analyzed
(Table 3).
For the reasons set out below, the average concentration
measured in the rst 4 aliquots of 5 g analyzed was taken as
 S. Armorini et al. / Food Control 50 (2015) 858e863 861

Fig. 1. Chromatogram of (A) blank sample, (B) standard calibration (7.5 mg kg
1) and (C) naturally contaminated sample.

Table 1 On the other hand, considering the concentrations of AFB1 in

Recovery and precision of the HPLC/FD method for determining AFB1 in our spiked the individual four aliquots of each sample (indicated by the letter
at ve concentration levels.
A, B, C, D), a higher variability can be noticed. Specically, referring
Conc. level (mg kg
1) Recovery (%)a RSD (%)a to Table 2, the concentration of some aliquots (sample 49 aliquots A
1 99.7 7.8 and C, sample 61 aliquot B, sample 103 aliquot C) were found to be
2.5 86.4 5.2 higher than the legal limit.
5 77.7 3.0 The same consideration can be made for the second four ali-
7.5 79.4 3.3
quots analyzed (especially in the sample 49 aliquot E and sample
10 79.7 3.5
103 aliquot E), as shown in Table 3.
a
Number of replicates: 4.
Considering all the aliquots analyzed for each sample and
averaging the results, all the concentration values of aatoxin B1
are largely within the legal limits, except for sample number 49.
reference for the concentration to be attributed to each sample of
These data would seem to support the thesis that an aliquot of 5 g,
our (Table 2). By comparing the concentrations of AFB1 in each
although a sub-aliquot of a total of 20 g, although thoroughly
sample measured in the rst round of analysis and the average of
mixed, does not guarantee a good representativeness of the sample.
the concentrations in the four aliquots of 5 g of each sample
On the other hand, by comparing for each sample the average
measured in the subsequent analysis, some variability has to be
concentration of AFB1 of the rst four aliquots analyzed with those
noticed (Table 2).

Table 2
AFB1 concentrations in the four aliquots of the samples found positive in the rst round of analysis.

Samplesa First analysis Aliquot A Aliquot B Aliquot C Aliquot D Mean SD aliquots AeDb RSD %
(screening) (mg kg
1) (mg kg
1) (mg kg
1) (mg kg
1) (mg kg
1) (mg kg
1)

19 (c) 0.59 0.53 0.32 0.31 0.45 0.40 0.10 26

21 (c) 0.22 0.75 0.23 <LOD <LOD 0.28 0.32 114
32 (o) 0.37 0.83 0.38 1.19 0.16 0.64 0.46 72
49 (c) 0.84 2.39 0.85 11.25 0.51 3.75 5.07 135
61 (o) 0.55 <LOD 2.34 <LOD <LOD 0.64 1.13 177
66 (c) 0.29 0.39 0.17 <LOD 0.22 0.21 0.13 62
69 (c) 0.57 <LOD <LOD <LOD <LOD <LOD /
70 (c) 0.75 <LOD <LOD <LOD <LOD <LOD /
72 (c) 0.20 <LOD 0.15 0.19 0.32 0.18 0.10 56
84 (c) 3.89 0.57 <LOD 0.15 0.38 0.29 0.23 77
86 (c) 0.81 1.04 0.26 0.92 0.29 0.63 0.41 65
103 (c) 1.31 1.66 1.39 2.19 1.51 1.69 0.35 21
108 (o) 0.19 <LOD <LOD 0.67 <LOD 0.22 0.30 132
111 (o) 0.18 0.17 0.18 0.18 0.17 0.17 0.00 2
a
(c) conventional our sample; (o) organic our sample.
b
In calculating the mean concentrations of AFB1, the values <LOD measured in the different aliquots were conventionally considered equal to LOD (0.075 mg kg
1)
, 2004).
(Istituto Superiore di Sanita
862 S. Armorini et al. / Food Control 50 (2015) 858e863

Table 3
AFB1 concentrations in the second four aliquots of ve samples found positive in the rst round of analysis.

Samplesa Aliquot E (mg kg
1) Aliquot F (mg kg
1) Aliquot G (mg kg
1) Aliquot H (mg kg
1) Mean SD aliquots EeHa (mg kg
1) RSD (%)

32 (o) 0.22 0.53 0.22 0.31 0.32 0.15 46

49 (c) 13.16 0.40 0.46 0.87 3.72 6.30 169
66 (c) 0.48 0.14 0.13 0.08 0.21 0.19 91
103 (c) 2.45 1.66 1.40 1.44 1.74 0.49 28
108 (o) 0.37 0.22 0.15 0.19 0.23 0.10 41
a
(c) conventional our sample; (o) organic our sample.

Table 4 These results conrm what has already been reported in liter-
Means of the analytical results of the rst and the second four aliquots analyzed. ature: a higher incidence of contamination in corn compared to
Samplea Mean aliquots AeD Mean aliquots Mean SD RSD (%) wheat. Considering only the samples of corn our, the percentage
(mg kg
1) EeH (mg kg
1) (mg kg
1) of positive samples, rises to 26%, a value that has to be evaluated. On
32 (o) 0.64 0.32 0.48 0.23 47 the other hand, focusing on the concentration of AFB1 in these
49 (c) 3.75 3.72 3.74 0.02 1 samples, the values do not exceed the legal limit (2 mg kg
1), except
66 (c) 0.21 0.21 0.21 0.00 0 one case (sample 49, average concentration of 3.75 mg kg
1), as
103 (c) 1.69 1.74 1.72 0.04 2
shown in Table 2. These results seem to suggest that the systematic
108 (o) 0.22 0.23 0.23 0.01 3
controls upstream of the marketing of these products, guarantee at
a
(c) conventional our sample; (o) organic our sample.
least levels of contamination below legal limits. Moreover, by
comparing conventional corn our samples and organic corn our
samples, no signicant differences can be observed in the con-
of the second four (Table 4), it is evident that variability is lower and centration of AFB1 (P > 0.05), while the percentage of positives in
this shows a better representativeness of 20 g in the analyses. conventional corn our samples (21,4%) is lower than in organic
Finally, even the samples negative for the presence of AFB1, corn our samples (50%). This last percentage is certainly high, but
were tested to evaluate if screening analysis made with aliquots of considering that just a small number in this category has been
5 g were reliable. The analytical results show that in two samples analyzed (8), it is not possible to say that in general the biological
(18, 31) all four aliquots were below LOD, while in the other two samples are more contaminated by AFB1.
(33, 63) the aliquot D for the rst and the aliquots A and D for the On the basis of these data, it can be assumed that the same good
second, were positive for the presence of AFB1, although with very farming practices promoted by organic farming such as soil fertility
low concentrations (respectively 0.21, 0.46 and 0.15 ppb, values all management, choice of species and varieties, multiannual crop
below the LOQ). It must be said that for these last two samples, the rotation, recycling organic materials and cultivation techniques,
mean concentration of the four aliquots is below LOD in one case present themselves as valid alternatives to the use of pesticides in
and just above LOD in the other, as shown in Table 5. the treatment of stress plants, and therefore also in the ght against
These additional results, however, are not sufcient to promote mycotoxins.
just 5 g as minimum amount of sample for reliable screening an- The number of samples positive for the presence of aatoxin B1
alyses because, even in the tests performed on samples positive is sufciently high to require the maintenance of a high control of
with four aliquots of 5 g, some concentration values were lower this toxic contaminant in cereal ours just for their fundamental
than the LOD. importance to human and animal nutrition, because used as food
In conclusion, on the basis of this data, it might be possible to base and raw material for the production of food and feed. Other
have a result <LOD in an aliquot sample of 5 g when in other parts studies demonstrate the importance to maintain a high level of
of the same sample, the mycotoxin may be present in quantities control of these foodstuffs, by implementing Hazard Analysis
exceeding the limits of law. On the other hand, it might be possible Critical Control Point (HACCP) from farm to fork to provide
to get a result out of law arising from the analysis of a single aliquot quality and safe food for enhancing food safety and global security
of 5 g when all the rest of the sample may not contain the (Ramesh, Sarathchandra, & Sureshkumar, 2013).
mycotoxin. It must be said that mycotoxins are not generally perceived as a
Examining the nal data on the occurrence of AFB1 from organic worrying danger by consumers, even though this toxins may be
and conventional ours, some considerations can be made. As re- present both in organic and in conventional unprocessed cereals at
ported above, 90 samples of our were analyzed, 13 of which (14.4% levels that could potentially reach a signicant portion or even
of total samples) were positive in the conrmation analyses made exceed the Tolerable Daily Intake (TDI) for average and high con-
on 20 g of sample (Table 6). It regards corn our, specically 4 sumers, respectively (Harcz et al., 2007).
organic (representing 30.8% of total positives) and 9 conventional The main nding is that people do not differentiate greatly be-
(representing 69.2% of total positives). tween the various types of risks, although they are more likely to

Table 5
AFB1 concentrations in the four aliquots of four samples found negative in the rst round of analysis.

Samplesa First analysis Aliquot A Aliquot B Aliquot C Aliquot D Mean SD Aliquots RSD (%)
(screening) (mg kg
1) (mg kg
1) (mg kg
1) (mg kg
1) (mg kg
1) AeD (mg kg
1)

18 (c) <LOD <LOD <LOD <LOD <LOD. <LOD /

31 (o) <LOD <LOD <LOD <LOD <LOD <LOD /
33 (o) <LOD <LOD <LOD <LOD 0.21 <LOD /
63 (o) <LOD 0.46 <LOD <LOD 0.15 0.19 0.18 96.5
a
(c) conventional our sample; (o) organic our sample.
 S. Armorini et al. / Food Control 50 (2015) 858e863
Table 6
Occurrence of AFB1 in organic and conventional our.
Samples N positive/N % positive
total samples
Flour (all types) 13/90 14.4
Wheat our (conventional organic) 0/40 0.0
Corn our (conventional organic) 13/50 26.0
Conventional corn our 9/42 21.4
Organic corn our 4/8 50.0
a
Values calculated considering only the positive samples.
worry about risks caused by external factors over which they have
no control. At the top end of the worry scale consumers express
concern regarding external factors that are clearly identied as
dangerous (pesticide residues, new viruses such as avian inuenza,
residues in meat, contamination of food by bacteria, unhygienic
conditions outside the home). In the mid-range, one nds other
external factors such as environmental pollutants (e.g. mercury),
GMOs, food additives, animal welfare and bovine spongiform en-
cephalopathy (European Commission, 2006a).
A result of our study was the analytical method developed for
the detection of aatoxin B1 in the examined our. This method
should always be accompanied by a good sampling procedure,
because sampling is always critical point in mycotoxin analysis. It is
important to understand the sources of error in the mycotoxin test
procedure so the errors can be effectively reduced (Whitaker,
2003). Sampling is one of the most crucial parts of the multifac-
eted and complex bulk of activities aimed at addressing and
managing food issues. The overall objective of good sampling is to
provide reliable samples to be analyzed that can represent the basis
for t for purpose investigations (Miraglia, De Santis, Minardi,
Debegnach, & Brera, 2005).
4. Conclusion
The analytical method made it possible to perform adequate
analysis, and at the same time to save materials, volumes of sol-
vents and processing times of the samples. Therefore, this could be
considered a good method to follow in conducting future similar
researches.
In sample preparation, the selection of the test sample from the
laboratory sample is a very important step. The present study
shows that the minimum amount to get a reliable result is 20 g and
smaller quantities can lead to incorrect results.
Comparing the results obtained from the analysis of organic and
conventional products, no signicant differences in aatoxin
contamination can be noticed between these two categories of
products. Except in one case, the levels of contamination found in
the tested samples did not exceed the limit dened at Community
level. This demonstrates the effectiveness of the network of
monitoring before the placing on the market of these food
products.
However, it should be stressed that the present study was only a
survey focusing on one set of samples produced in one year. It
would be interesting to continue this study examining a larger
number of samples.
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