Behavior of Aatoxin M

1
in dairy wastes subjected to different technological
treatments: Ricotta cheese production, ultraltration and spray-drying
Tiziana Maria Piera Cattaneo
a,
*
, Laura Marinoni
a
, Stefania Iametti
b
, Lucia Monti
a
a
Consiglio per la ricerca e la sperimentazione in agricoltura - Centro di ricerca per le produzioni foraggere e lattiero-casearie, Via A. Lombardo 11, 26900 Lodi, Italy
b
DISMA-University of Milan, Via Celoria 2, 20133 Milan, Italy
a r t i c l e i n f o
Article history:
Received 7 March 2012
Received in revised form
8 August 2012
Accepted 3 November 2012
Keywords:
Aatoxin M
1
Dairy wastes
Ricotta cheese
Ultraltration
Spray-drying
a b s t r a c t
Although considered by-products of cheese production, milk whey, deproteinized whey and valuable
components extracted from them, have several commercial uses as nutritional supplements and pro-
cessing aids. In order to verify the safety of these products, destiny of AFM
1
contaminating whey and
deproteinized whey subjected to different technological treatments was investigated. During Ricotta
cheese production the main part of AFM
1
, on average 94%, was lost in the liquid portion, while only 6%
remained in the curd. Whey and deproteinized whey remaining from Ricotta cheese production were
then subjected to ultraltration and spray-drying. In the rst case, ultraltration coupled with dial-
tration allowed to remove more than 90% of toxin without losing proteins. Spray-drying was efcient in
reducing AFM
1
contamination, too: in whey, toxin recovery was around 60%, aligned with powder
recovery, while in deproteinized whey AFM
1
recovery was around 39%, by far lower than powder
recovery, conrming a loss.
2012 Elsevier Ltd. All rights reserved.
1. Introduction
Aatoxin M
1
(AFM
1
) is a toxic and carcinogenic compound
which is excreted in milk when lactating animals are fed with
aatoxin B
1
(AFB
1
) contaminated feed. Results of several investi-
gations showed a preferential binding of AFM
1
to casein (Brackett &
Marth, 1982a; Galvano, Galofaro, & Galvano, 1996) and thus an
enrichment in cheese, ranging from 1.7 to 8.0 times if compared to
the original milk contamination, according to cheese technology
and shelf-life (Brackett, Applebaum, Wiseman, & Marth, 1982;
Brackett & Marth, 1982b,c; Cattaneo et al., 2008; Kiermeier &
Buchner, 1977; Motawee & McMahon, 2009; Oruc, Cibik, Yilmaz,
& Kalkanli, 2006; Pietri, Bertuzzi, Bertuzzi, & Piva, 1997). Despite
AFM
1
ecasein interaction, during cheese production part of toxin
content is drained from the curd together with whey.
Whey is considered to be a by-product of cheese production but
it still contains valuable components such as proteins and peptides
(i.e. a-lactalbumin, b-lactoglobulin, serum albumin, lactoferrin and
lactoperoxidase, glycomacropeptide, phosphopeptides and other
enzymes and hydrolysis products), lactose, vitamins and minerals
along with fat traces (Chianese et al., 1997; De Wit, 1998; Outinen,
Rantamki, & Heino, 2010), the exact composition depending on
the cheese-making process. Until some years ago, whey was used
only for Ricotta cheese production and as animal feed. Nowadays it
has several commercial uses. So far, a great number of whey-based
beverages has been presented to the markets (zer & Kirmaci,
2010). Moreover, whey proteins are ingredients widely used in
the food industry due to their excellent nutritive and functional
properties. Whey proteins isolates are often used as a nutritional
supplement. In addition, their ability to form gels capable of
holding water, lipids, and other components while providing
textural properties made it indispensable in the formulation of
many foods such as processed meat, dairy and bakery products
(Burrington, 1999; De Wit, 1998; Gsta Bylund, 1995; Johnson,
2000; Kantor, 1990; Outinen et al., 2010; Thompson, 1990).
Although AFM
1
afnity for milk caseins is well known, very little
information is available on AFM
1
ewhey proteins interaction.
Consequently, due to the wide use of whey in products for human
consumption, the aim of this work was to investigate the destiny of
AFM
1
accidentally contaminating whey subjected to different
technological treatments. In particular, AFM
1
distribution during
Ricotta cheese production was investigated; the fate of toxin in
whey and deproteinized whey (named scotta) subjected to
ultraltration and spray-drying, two of the most used techniques
to fractionate, concentrate and stabilize liquid whey and its
by-products, was also considered.
* Corresponding author. Present address: Consiglio per la ricerca e la sper-
imentazioneinagricoltura- Unitdi ricercaper i processi dell'industriaagroalimentare,
Via Venezian 26, 20133 Milan, Italy. Tel.: 39 037145011; fax: 39 037135579.
E-mail address: tiziana.cattaneo@entecra.it (T.M.P. Cattaneo).
Contents lists available at SciVerse ScienceDirect
Food Control
j ournal homepage: www. el sevi er. com/ l ocat e/ f oodcont
0956-7135/$ e see front matter 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.foodcont.2012.11.007
Food Control 32 (2013) 77e82
2. Materials and methods
2.1. Whey and deproteinized whey samples
Whey used in these experiments derived from Mozzarella
cheese production, both by chemical acidication and biological
fermentation of AFM
1
naturally contaminated milk. After cheese
making, whey at three different levels of contamination (nearly 20,
100 and 200 ng kg
1
) was recovered, divided into aliquots and
frozen at 20

C until use. Deproteinized whey (named scotta)
was the liquid part remaining after Ricotta cheese production.
2.2. Ricotta cheese production
Ricotta cheese was produced on a laboratory scale using 8e9 kg
of whey for each test. When necessary, whey was neutralized to
pH 6.7 using 2 N NaOH (Riedel-de Han, SigmaeAldrich Laborche-
mikalien Gmbh, Seelze), then it was heated under magnetic stirring
up to 85

C: at this temperature 1% NaCl (w/v) (Riedel-de Han,
SigmaeAldrich Laborchemikalien Gmbh, Seelze) was added. Heating
was continued to 90

C, when a citric acid solution (5 g each 10 L of
whey, diluted 1:10 in water) was added. After 2 min, stirring was
stopped and after 15 min the remaining proteins in the whey curdled
out. A skimmer was used to dip out oating curd and liquid in excess
was drained in a ne meshed colander. Both Ricotta cheese and
deproteinized whey were sampled to be analyzed for AFM
1
deter-
mination. Deproteinized whey was also used to perform ultraltra-
tion and spray-drying. All samples were frozen at 20

C until
analysis.
2.3. Ultraltration of whey and deproteinized whey
Ultraltration was performed in duplicate on four aliquots at
different concentration levels both for whey and deproteinized
whey. The process was carried out on a laboratory scale using an
Amicon apparatus with an Ultracel YM10 membrane (cut-off
10 kDa; Millipore, Billerica, MA, USA) at a pressure of 2.5 bar. Fifty
mL of sample were forced through the ultraltration unit until
40 mL of permeate (PERMEATE 1) were collected. Two of the 10 mL
of retentate (RETENTATE 1) were used to perform AFM
1
analysis,
while the rest was re-diluted by adding 32 mL of double-distilled
water and it was subjected to a further ultraltration step under
the same experimental conditions (dialtration). The process was
run until 8 mL of retentate (RETENTATE 2) and 32 mL of permeate
(PERMEATE 2) were collected. Volumes and AFM
1
concentrations of
permeates and retentates were recorded to calculate toxin distri-
bution in both fractions during ultraltration process.
2.4. Spray-drying of whey and deproteinized whey
Spray-drying was performed on both whey and deproteinized
whey using a Mini Spray Dryer Be191 apparatus (Bchi Labor-
technik AG Flawil, Switzerland), working at 180

C, with high
aspirator ow rate (nearly 100%) and pump speed not higher than
25% to avoid the obstruction of the nozzle. Four pools at different
concentration levels were processed and each drying was repeated
three times. Powder obtained from spray-drying was re-hydrated
using distilled water to get a total solid concentration equal to
that of the starting product, and it was analyzed to determine the
AFM
1
content after the drying process.
2.5. Chemical analyses
Chemical parameters determined on whey, Ricotta cheese and
deproteinized whey were:
- Total solids: dry matter analysis was performed in duplicate,
according to ISO-IDF Standard Method (ISO-IDF, 2004).
- Protein content: protein content analysis was performed
according to Kjeldahl method (ISO-IDF, 2008b).
- Fat: fat content analysis was performed in duplicate according
to GerbereVan Gulik method (ISO-IDF, 2008a).
2.6. AFM
1
detection by ELISA
ELISA method was performed by using a commercial kit (Iscreen
Aa M
1
,Tecna S.r.l. Trieste, Italy; range of detection: 5e250 ng L
1
),
following the procedures given in the instruction booklet. Liquid
samples were tested directly and, when necessary, they were
neutralized by means of 1 N NaOH and/or diluted with the buffer
enclosed in the kit. Solid samples were analyzed after applying an
extraction procedure using solvents (dichloromethane, hexane,
methanol), as recommended by the producer. AFM
1
detection was
performed at 450 nm in a microplate reader (Model 680, BIO-RAD,
Richmond, CA, USA) and toxin concentrations were calculated
referring to the response of the standard solutions enclosed in the
kit. All samples were analyzed in duplicate.
2.7. Evaluation of AFM
1
percentage of distribution
The amount of AFM
1
in each fraction was calculated considering
toxin concentration ([AFM
1
]) and mass (M) of raw material used or
product obtained in each experiment
AFM
1
ng AFM
1

ng kg
1

*Mkg: (1)
AFM
1
percentage of distribution was calculated from the
following formula:
%distribution AFM
1product
ng=AFM
1rawmaterial
ng*100 (2)
Finally, the Enrichment Factor (EF) was calculated
EF AFM
1

product

ng kg
1
.
AFM
1

raw material

ng kg
1

(3)
3. Results and discussion
3.1. Fate of AFM
1
during Ricotta cheese production
Ricotta cheese was produced from whey coming from Mozza-
rella cheese production, both with chemical acidication and
biological fermentation of naturally AFM
1
contaminated milk.
Three different levels of contamination were considered. All
samples were analyzed for toxin content by means of an ELISA test
which fullled ISO guidelines (UNI EN ISO, 2003) on the use of
competitive enzyme immunoassays for the quantitative determi-
nation of AFM
1
in milk and milk products. The performances of
the chosen ELISA method had been previously compared against
other ELISA kits (Cattaneo, Panarelli, Iametti, Pietri, & Monti,
2007) and against an HPLC method (Cattaneo et al., 2008).
Comparison of the two techniques, both for liquid and solid
samples, indicated very similar results, with a linear relationship
of data, good slope of the regression line and good coefcient of
determination. Moreover, the reliability of the applied ELISA
method was veried by prociency test rounds organized by Milk
Standard Laboratory of A.I.A. (Italian Association of Breeders). The
z-scores obtained in the A.I.A. ring test, always below 2, indicated
good performance of the applied method and technical compe-
tence of the laboratory.
T.M.P. Cattaneo et al. / Food Control 32 (2013) 77e82 78
In Table 1 i) mass of whey processed and ii) Ricotta cheese and
iii) deproteinized whey produced, iv) total solids, v) proteins and vi)
fat content of each sample, together with vii) AFM
1
concentration
and viii) toxin distribution during the process are reported.
Mass balances expressed on dry matter produced recoveries
ranging from101.3 to 122.5%: during the process there was a loss of
material caused especially by evaporation of liquid during pro-
longed heating at high temperatures. These losses did not inuence
toxin recoveries which ranged from 97.2 to 107.8% (Table 1): in all
cases it was evident that the main part of AFM
1
, on average 94%,
was lost in the liquid portion while only 6% remained in Ricotta
cheese. Anyway, considering the Enrichment Factors (EF) and the
percentage of toxin recovered in Ricotta cheese, there was not an
evident proportionality with the contamination level of whey used
in the process.
Very few data could be found in literature about AFM
1
contam-
ination in Ricotta cheese. In a review on aatoxins in the dairy
industry, Applebaum, Brackett, Wiseman, and Marth (1982) quote an
old experiment in which Ricotta cheese was made by curdling arti-
cially contaminated whole milk with bacterially produced lactic
acid and heat. Thirty percent of the AFM
1
was recovered in curd and
70% appeared in whey. The use of whole milk, with the strong
inuence of casein in AFM
1
binding, and the use of articially
contaminated materials could be responsible of the discrepancy of
results between the two experimental Ricotta cheese makings.
Anyway, the AFM
1
content of Ricotta cheese in that experiment was
not lowered by the severe heating process used inits manufacture, in
agreement with our results. Palomba, Battacone, Brundu, and Pulina
(2003) studied AFM
1
concentration in ovine milk, cheese and Ricotta
and found a toxin concentration in curd and Ricotta 5e8 fold higher
than in milk. Also in this case, a direct comparison of results is
difcult for the use of milk of different species, and consequently for
differences in protein composition.
Ricotta production is a low yield process since a high amount of
whey is required to produce it: small quantities of toxin concentrate
in small quantities of material, giving concentration values which are
equal and in some cases higher than the starting contamination
values registered in whey. Toxin content in Ricotta cheese can partly
be explained by whey inclusion in the coagulum, but a selective
binding with whey proteins, as already evidenced with casein during
cheese making, cannot be excluded. Ricotta cheese is mainly
constituted of a-lactalbumin and b-lactoglobulin, the main proteins
in cheese whey. Whey proteins are highly susceptible to heat-
induced denaturation: thermal treatments cause signicant alter-
ation of secondary structures and interactions between milk
proteins. Consequently, when exposed to high temperatures, whey
proteins can display reactive groups capable of interacting withother
components, like probably AFM
1
. This could also explain why during
cheese making, when whey proteins are in a native state, AFM
1
does
not seem to react with them and prefers casein, for which afnity is
probably higher in those conditions. High AFM
1
recoveries underline
once more toxin stability at these temperatures, which are not high
enough to degrade it (Rustom, 1997).
3.2. Ultraltration of whey and deproteinized whey
Ultraltration (UF) is a membrane separation process, driven by
a pressure gradient, in which the membrane fractionates compo-
nents of a liquid as a function of their solvated size, charge and
structure. Suspended solids and solutes of high molecular weight
are retained, while water and low molecular weight solutes pass
through the membrane. In this work, whey and deproteinized
whey samples at four levels of contamination were subjected to
a rst UF step and a second dialtration (DF) treatment, using
a 10 000 MWcut-off membrane. AFM
1
concentrations in retentates
and permeates are shown in Table 2.
In a fractionation process like this, AFM
1
is expected to pass the
membrane, due to its low molecular weight (328 g mol
1
)
(Applebaum et al., 1982). In both cases, during a rst UF step,
a percentage from 60 to 80% was lost in permeate, and 23e32% was
retainedinthe retentate (Fig. 1) witha meanenrichment factor (EF) of
1.45 0.15 for whey and 1.22 0.05 for deproteinized whey. In some
cases, percentage of recovery of the toxin was less than 100%, prob-
ably due to an interaction between toxin and membrane, to fouling
effects which interfere with AFM
1
passage through membrane or
because of the presence of dead space within the apparatus which do
not allow a complete recovery of solutions. The application of the
SDS-PAGE technique to all the collected fractions allowed to verify
that, during UF, proteins do not permeate through membrane (data
not shown). Consequently, the enrichment of retentate in AFM
1
in
connection with protein concentration would support the idea of
a certain afnity of toxin for whey proteins and small hydrolysis
products which are retained by the membrane as a function of their
molecular weights and chemicalephysical properties.
Table 1
Mass of whey used and Ricotta cheese and deproteinized whey produced; total solids, proteins, fat, AFM
1
contamination and distribution in each production step.
Whey from chemical acidication of milk Whey from biological acidication of milk
Level of contamination I II III I II III
Whey Processed (kg) 8.71 9.35 8.00 9.49 9.00 9.06
Total solids (%) 5.38 7.22 7.15 7.08 7.32 7.39
Proteins (%) 0.63 0.86 0.86 0.70 0.90 0.95
Fat (%) 0.40 0.33 0.30 0.40 0.60 0.63
AFM
1
(ng kg
1
) 21 91 191 11 98 169
Ricotta cheese Produced (kg) 0.43 0.42 0.41 0.83 0.56 0.42
Total solids (%) 18.36 21.03 18.51 21.58 22.17 21.61
Proteins (%) 7.39 6.98 6.31 7.23 10.00 9.40
Fat (%) 6.57 5.06 4.98 4.45 5.72 5.52
AFM
1
(ng kg
1
) 22 70 269 9 96 307
AFM
1
distribution (%) 5.3 3.4 7.2 7.4 6.1 8.5
EF 1.05 0.76 1.41 0.47 0.99 1.82
Deproteinized whey Produced (kg) 6.60 7.00 6.75 7.90 7.50 7.10
Total solids (%) 6.50 9.48 8.74 8.15 8.42 8.27
Proteins (%) 0.39 0.62 0.60 0.38 0.45 0.44
Fat (%) 0.10 0.20 0.25 0.20 0.25 0.17
AFM
1
(ng kg
1
) 25 114 209 12 111 213
AFM
1
distribution (%) 92.7 93.8 92.7 92.6 94.3 99.3
Mass balance (expressed on dry matter): recovery % 108.4 111.4 116.4 122.5 114.7 101.3
AFM
1
recovery (%) 98.0 97.2 99.9 100.0 100.4 107.8
EF: Enrichment Factor.
T.M.P. Cattaneo et al. / Food Control 32 (2013) 77e82 79
In both processes, a DF step was applied: RETENTATE 1 was
diluted with water and re-ultraltered, to disrupt impregnation of
solids into the membrane pores, to remove impurities or undesir-
able compounds and to increase the concentration of retained
components. This second step further reduced the presence of
AFM
1
in retentates. At the end of the process, on average 96% of the
AFM1 contaminating whey and 93% of the toxin in deproteinized
whey were removed in permeates. Also in the case of UF, very few
literary results were found and in many cases experimental
conditions were very far from those we applied. Blanc, Lauber, and
Sieber (1983) studied AFM
1
binding to milk proteins and performed
ultraltration on articially contaminated milk whey. The cut-off of
the membrane used for this experiment was higher (30 000 Da)
than the one used in our experiment (10 000 Da). AFM
1
concen-
tration increased 1.75 fold in the retentate after ltration of 9/10 of
the volume and the permeate showed a reduced concentration.
According to Authors, these results were determined by absorption
of toxin in the retentate fraction, rather than to an interaction with
whey proteins. By far higher was the concentration of AFM
1
in
retentate found by Mendona and Venncio (2005) after UF
(membrane cut-off 10 000 Da) of cheese whey articially
contaminated with 0.1 mg L
1
of toxin. Recoveries of AFM
1
in
retentate were in range 72.6e86.4%, while in permeate the range
varied between 2.4 and 14.7%. In this case Authors hypothesized an
interaction between the mycotoxin and proteins. Higuera-Ciapara,
Esqueda-Valle, and Nieblas (1995) applied UF on milk and
removed, on average, 43.8% of AFM
1
, while DF removed 25% of the
initial concentration, for an overall removal of AFM
1
through both
processes of 68.5%. In this general confusing overview, our results
evidence howa simple technique, despite a certain afnity of AFM
1
also for whey proteins, is able to eliminate more than 90% of toxin
without losing proteins, which can be used for other applications.
3.3. Spray-drying of whey and deproteinized whey
The spray-drying process was applied to verify the effect of
heating and drying on AFM
1
in whey and deproteinized whey. Four
pools at different concentration levels were tested and each
procedure was repeated three times. Different temperatures were
tested to reduce humidity in the sample without burning, the
aspirator ow rate was set in order to control time of permanence
of the sample in the instrument, and pump speed was regulated so
as to control sample entrance in the spray-dryer.
The best drying conditions identied allowed a mass recovery
on average of 68% for whey and 61% for deproteinized whey
(Table 3), which is a good result considering that losses of product
during this kind of process are unavoidable. In order to improve the
yield of the drying procedure, some of the samples were also
concentrated by means of a Rotavapor (R-200, Bchi Labortechnik
AG Flawil, Switzerland) to increase the total solid concentration in
the solutions; unfortunately, this further step did not positively
inuence powder recovery.
Powders obtained from spray-drying were re-hydrated using
distilled water to get a total solid concentration equal to that of the
starting solutions: in this way the inevitable losses of material
occurring during spray-drying were considered. Toxin concentra-
tion in the solutions was quantied by ELISA test before and after
spray-drying. Results are summarized in Table 3. In re-hydrated
whey powders, AFM
1
concentration was in line with that in the
starting solutions, while in re-hydrated deproteinized whey
powders concentrations were nearly 60%. In Table 3, AFM
1
recov-
eries are reported, too: results are expressed as percentage ratio of
nanograms of AFM
1
found in the powder recovered after spray-
drying treatment and nanograms of toxin in the starting solution.
In cheese whey, the mean percentage of recovery was around 60%,
in accordance with powder recovery, while for deproteinized whey
Table 2
AFM1 concentrations in whey and deproteinized whey and permeates and retentates after ultraltration (#1) and dialtration (#2).
Level of contamination Whey (ng kg
1
) PERMEATE 1 (ng kg
1
) RETENTATE 1 (ng kg
1
) PERMEATE 2 (ng kg
1
) RETENTATE 2 (ng kg
1
)
I 24 24 37 9 n.d.
II 79 68 98 32 37
III 105 97 166 25 36
IV 163 157 244 49 78
Level of contamination Deproteinized whey (ng kg
1
) PERMEATE 1 (ng kg
1
) RETENTATE 1 (ng kg
1
) PERMEATE 2 (ng kg
1
) RETENTATE 2 (ng kg
1
)
I 23 22 27 7 n.d.
II 65 64 77 27 25
III 87 76 105 30 37
IV 167 124 215 47 69
n.d.: not detectable.
0
10
20
30
40
50
60
70
80
90
100
I II III IV
A
F
M
1

d
i
s
t
r
i
b
u
t
i
o
n

(
%
)
Contamination level
Ultrafiltration of Whey
retentate1
permeate1
retentate2
permeate2
0.0
10.0
20.0
30.0
40.0
50.0
60.0
70.0
80.0
90.0
100.0
I II III IV
A
F
M
1

d
i
s
t
r
i
b
u
t
i
o
n

(
%
)
Contamination level
Ultrafiltration of Deproteinized Whey
retentate1
permeate1
retentate2
permeate2
Fig. 1. AFM
1
percentage of distribution between permeates and retentates in whey and
deproteinized whey subjected to ultraltration (#1) and diltration (#2); values were
calculated referring to the starting AFM
1
concentration in whey and deproteinized
whey.
T.M.P. Cattaneo et al. / Food Control 32 (2013) 77e82 80
AFM
1
recovery was around 39%, by far lower than powder recovery,
conrming a loss. Rustom (1997) reported the results of different
studies on the degradation of aatoxins in foods by different heat
treatments. It is normally recognized that temperatures above
150

C are necessary to attain partial destruction of the toxin, in
connection with the initial level of contamination, heating
temperature, time, moisture content, pH and ionic strength of the
food. The same Author cited the theory of Coomes, Crowther, Feuell,
and Francis (1966), who suggested that the presence of water helps
in opening the lactone ring in aatoxin B
1
(AFB
1
) (by the addition of
a water molecule to the ring) to form a terminal carboxylic acid,
that thereafter undergoes heat-induced decarboxylation. The
presence of ionic salts increases the extent of aatoxin degradation
by heating. If this could be true also for AFM
1
, which is a mono-
hydroxy derivative of AFB
1
, the increased effect of spray-drying
process on deproteinized whey could be explained. In this kind of
solution, in fact, the protective effect of whey proteins lacks and
the ionic content is increased because of the addition of NaCl to
whey during Ricotta cheese production. No references could be
found in literature about the effect of spray-drying on whey and
deproteinized whey which could support our results. Authors who
studied the effect of this process on milk (Deveci & Sezgin, 2006)
supposed that the losses of AFM
1
that occurred during drying
processes could be attributed to changes in the interactions
between toxin and proteins due to the heat treatment. The same
could occur especially in whey, determining an apparent reduction
in AFM
1
contamination. Anyway, a decrease in AFM
1
contamination
cannot be dissociated by a loss of material.
4. Conclusions
The presence of AFM
1
in whey and deproteinized whey should
not be underestimated, since toxin can have different behaviors
according to the process it undergoes. Ricotta cheese production,
ultraltration and spray-drying are three processes that can
contribute to reduce AFM
1
concentration from a contaminated
solution of whey and/or deproteinized whey. Anyway, the inter-
action with whey proteins that can happen in certain circum-
stances cannot be ignored, especially considering the possibility to
reuse the protein fraction as a raw material in other processes.
Moreover, the reduction in AFM
1
concentration that was high-
lighted in some cases should be further investigated to verify
whether the molecule was inactivated, or it was strongly interact-
ing with proteins and thus changed the extraction efciency, or,
nally, it was lost in the environment.
Acknowledgment
The present work was supported by the Ministry of Agricultural
Policies (MiPAF), within the special project AFLARID (Investiga-
tion on AFM
1
reduction in contaminated milk and milk products).
References
Applebaum, R. S., Brackett, R. E., Wiseman, D. W., & Marth, E. H. (1982). Aatoxin:
toxicity to dairy cattle and occurrence in milk and milk products e a review.
Journal of Food Protection, 45, 754e777.
Blanc, B., Lauber, E., & Sieber, R. (1983). Fixation de laatoxine sur les proteines du
lait. Microbiologie Aliments Nutrition, 1, 163e177.
Brackett, R. E., & Marth, E. H. (1982a). Association of aatoxin M
1
with Casein.
Zeitschrift fr Lebensmittel-Untersuchung und -Forschung, 174, 439e441.
Brackett, R. E., & Marth, E. H. (1982b). Fate of aatoxin M
1
in Cheddar cheese and in
process cheese spread. Journal of Food Protection, 45, 549e552.
Brackett, R. E., & Marth, E. H. (1982c). Fate of aatoxin M
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Table 3
Powder recovery of whey and deproteinized whey after spray-drying; AFM
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concentration (ng kg
1
) in the solutions before and after the treatment and AFM
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recovery (%).
Level of contamination Powder recovery (%) AFM
1
concentration before
spray-drying (ng kg
1
)
AFM
1
concentration after
spray-drying (ng kg
1
)
AFM
1
recovery (%)
Whey I 64 7 23 16 3 42 5
II 60 6 71 64 3 55 6
III 74 7 121 124 8 75 10
IV 72 10 187 180 12 69 13
Deproteinized whey I 73 1 26 18 2 49 6
II 66 4 94 61 8 42 8
III 44 1 124 64 8 23 4
IV 62 6 204 138 11 41 8
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