Bioresource Technology 98 (2007) 18121821

In vitro biosorption of ochratoxin A on the yeast industry

by-products: Comparison of isotherm models
a,*
Diana Ringot , Benoit Lerzy a, Kathy Chaplain a, Jean-Paul Bonhoure a,b
,
Eric Auclair c, Yvan Larondelle d
a
Institut Superieur dAgriculture de Beauvais, Rue Pierre Waguet, BP 30313, Beauvais Cedex 60026, France
b
Centre de Valorisation des Glucides, 33, Avenue Paul Claudel, 80000 Amiens, France
c
Lesare Feed Additives, 1 Rue du Haut Touquet, 59520 Marquette-Lez-Lille, France
d
Universite catholique de Louvain, Unite de biochimie de la nutrition, Croix du Sud, 2/8, Louvain-la-Neuve 1348, Belgium

Received 16 June 2004; received in revised form 24 May 2006; accepted 28 June 2006
Available online 21 August 2006

Abstract

Biosorption of ochratoxin A (OA) onto yeast biomass appears to be a reasonably low cost decontamination method. In vitro adsorp-
tion of OA onto three yeast industry by-products: a vinasse containing yeast cell walls (EX16), a puried yeast beta-glucan (BETA) and a
yeast cell wall fraction (LEC) was examined at 25 C. Seven classical adsorption models were tested to provide the best description of
toxin adsorption. A comparison of these models was performed using the magnitude of the coecient of determination R2 for the linear
models and the value of the sum of normalised errors (SNE) for linear and non-linear models. Based on the R2 and the SNE values, Hill,
Freundlich and BrunauerEmmettTeller equations produced the best models for OA biosorption onto respectively, EX16, BETA and
LEC. For these best models, the values of isotherm constants were consistent when measured using both linear and non-linear calcula-
tions. The SNE calculation procedure presented in this paper in association with the linear equation analysis method is an appropriate
approach for designing a better adsorption isothermal model.
 2006 Elsevier Ltd. All rights reserved.

Keywords: Ochratoxin A; Biosorption; Yeast cell wall by-products; Isotherm models

1. Introduction
Ochratoxin A (OA) is a secondary metabolite of
some species of storage fungi. Aspergillus species such as
A. ochraceus produce OA mainly in tropical climates and
Penicillium species such as P. verrucosum produce OA
in temperate climates.
Chemically speaking, OA is a chlorine-containing di-
hydroisocoumarine linked to L-phenylalanine. OA and its
hydroxyl derivatives are toxic when consumed by livestock
(pigs, poultry) and humans. They are found in human milk,
blood serum, plasma, liver and kidney. The main route of
exposure is the consumption of contaminated cereals, wine,
*
Corresponding author. Tel.: +33 344 062 517; fax: +33 344 062 526.
E-mail address: diana.ringot@isab.fr (D. Ringot).
spices, coee and cocoa products, grape juice and beer
(Commission of the European Union, 2002).
OA is partially absorbed by the gastrointestinal tract
in monogastrics. In ruminants, OA is hydrolysed by the
microbial ora into a much less toxic metabolite, ochra-
toxin a. OA is hydrolysed to ochratoxin a by mammalian
carboxypeptidase A and by some micro-organisms within
the gastrointestinal tract. In hepatic cells, it is hydroxylated
mainly to 4-R-hydroxyochratoxin A (OA-OH) by mixed
function oxidases (Li et al., 2000). OA is considered to be
a genotoxic agent in vivo and in vitro in some mammalian
species (WHO, 2002), but the mechanism of genotoxicity
is unclear and there is no evidence of a direct interaction
with DNA (Bakker and Pieters, 2002; Ringot et al.,
2006). OA is reasonably anticipated to be a possible human
carcinogen based on sucient evidence of carcinogenicity

0960-8524/$ - see front matter  2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2006.06.015
 D. Ringot et al. / Bioresource Technology 98 (2007) 18121821
in experimental animals (group 2B) (IARC, 1993). OA is
a nephrotoxic, teratogenic and immunotoxic compound
(Muller et al., 1999; Prelusky et al., 1994). OA is relatively
stable and is only partially degraded under normal process
conditions.
Dierent approaches have been developed to reduce the
impact of mycotoxins. They can be divided into pre- and
post-harvest strategies and into biological, chemical and
physical methods (Huwing et al., 2001). Physical methods
involve extraction with solvent, adsorption, inactivation
by heat and irradiation. Among the physical decontamina-
tion methods, adsorption onto various types of compounds
(hydrated sodium calcium aluminosilicate HSCAS,
kaolin, silica binding agent, bentonite, etc.) has been ex-
tensively studied in recent years (Grant and Philips, 1998;
Scott, 1998). Dierent binding agents including activated
carbons (Galvano et al., 1998), zeolites (Tomasevic-Cano-
vic et al., 2003), diatomaceous earth (Natour and Yousef,
1998), cholestyramine and mixtures of sterilized yeast and
fermentation residua from beer production (Bauer, 1994)
have been reported to remove OA in vitro. In contrast, acti-
vated charcoal and HSCAS have been reported to have a
limited ecacy against OA in vitro (Ramos et al., 1996).
Castellari et al. (2001) examined several ning treatments
to reduce OA levels in red wine. They found that potassium
caseinate and activated carbon were the best agents to
remove OA.
The phenomenon of biosorption is dened as a metabo-
lism independent adsorption of pollutants based on the
partition process on a microbial biomass. Microbial bio-
mass consists of small particles, with low density, poor
mechanical strength and little rigidity. This phenomenon
is generally based on a set of chemical and physical mech-
anisms (involving physico-chemical interactions such as
electrostatic interactions, ion exchange, complexation, che-
lation and precipitation) leading to the immobilization of a
solute component on the microbial cell wall components. It
has been investigated by dierent authors (Aksu, 2003;
Aksu and Donmez, 2002; Aksu and Yenner, 1998; Tsezos
and Bell, 1989).
The complexity of the microbial structure implies that
there are many ways for the pollutant to be captured by
the cells. Biosorption mechanisms are therefore various
(physical adsorption, chemical binding of ionic groups,
ion exchange, etc.) and in some cases they are still not very
well understood (Veglio and Beolchini, 1997). Cell surface
sorption is a physico-chemical interaction between the
toxin and functional groups of the cell surface, based on
physical adsorption, ion exchange and complexation,
which is not dependent on metabolism. Cell walls of micro-
bial biomass, mainly composed of polysaccharides, pro-
teins and lipids, oer abundant functional groups, such
as carboxyl, hydroxyl, phosphate and amino groups, as
well as hydrophobic adsorption sites such as aliphatic
carbon chains and aromatic rings (Ringot et al., 2005).
This physico-chemical phenomenon is quick and can be
reversible.
1813
Yeasts produce a high quantity of biomass. They are
used in a large variety of industrial fermentation processes;
moreover, they may be regarded as a good source of adsor-
bent material, due to the presence in their cell wall of some
specic macromolecules such as the mannoproteins and
beta-glucans. The ability of the Saccharomyces cerevisiae
cell wall to bind zeralenone has been reported recently
(Yiannikouris et al., 2003).
This research was aimed at examining in vitro ad-
sorption of OA onto three yeast industry by-products at
a temperature of 25 C. To this end, several theoretical
adsorption models frequently used in the literature were
tested for their ability to describe the equilibrium sorption
data. For these models, ve error functions and the norma-
lised error sum (SNE) were examined. This enabled the
determination of the best parameter set and thus provided
an accurate equilibrium adsorption model.
2. Methods
2.1. Adsorbents and reagents
Three adsorbent materials, namely EX16, BETA and
LEC, were obtained from Bio-Springer, Maison-Alfort,
France. Two of these adsorbents, EX16 (a vinasse contain-
ing 16% liquid yeast cell walls) and LEC (a dry yeast cell
wall fraction) are industrial by-products of the yeast indus-
tries. BETA is the dried puried beta-glucans fraction of
cell walls. The dry matter content of the by-products was
56% for EX16, 95% for BETA and 97.6% for LEC.
OA was purchased from Sigma Chemical Company
(St. Louis, Missouri, USA). Primary methanolic stock
solution (200 mg/L) was prepared in methanol. Five OA
test solutions with concentrations of 0.5, 1.0, 2.0, 5.0 and
10.0 mg/L were prepared by the dilution of the methanolic
stock solution with deionised distilled water.
2.2. Experimental system
Samples of 500 mg of adsorbents were placed in screw
cap test tubes with aliquots (10 mL) of the OA test solu-
tions. Two sets of controls were prepared, one by adding
10 mL of mycotoxin test solutions without the addition
of adsorbent, and another by adding 500 mg adsorbent
to 10 mL deionised distilled water. All experiments were
carried out in triplicate.
The controls and test tubes were placed in a horizontal
shaker-incubator at 400 rpm and at a controlled tempera-
ture of 25.0 1.0 C. Preliminary investigations showed
that equilibrium uptake was attained rapidly, with prac-
tically no change observed after a period of 30 min. All
adsorptions were run for 90 min to ensure complete
uptake. After the incubation period, solid particles (adsor-
bent) were separated from the supernatant by centrifu-
gation at 6000g for 20 min at 25 C in an ALC 4239R
centrifugation system (Fisher Scientic Labosi, Elancourt,
France).
1814 D. Ringot et al. / Bioresource Technology 98 (2007) 18121821
The supernatant was extracted with an equal volume of
methanol:deionised distilled water (50:50). The mixture
was vortexed for 5 min. This extract was diluted, ltered
and analysed by HPLC.
2.3. Chromatographic analysis
The HPLC chromatographic system consisted of a Spec-
traSYSTEM P1500 isocratic pump (Thermo Separation
Product, Fremont, California, USA) equipped with a AS
3000 model injection valve (100 ll) (Thermo Separation
Product, Fremont, California), a Spectroow 980 uores-
cence spectrophotometric detector (Applied Biosystems,
Ramsey, New Jersey, USA) equipped with a 150 W xenon
lamp (kexcitation = 333 nm and kemission = 470 nm) and
PC1000 integration software (Thermo Separation Product,
Fremont, California, USA). The analytical column was an
Alltima reversed-phase column C18 (25 cm 4.6 mm i.d.,
5 lm particles) preceded by an Alltima C18 precolumn
(7.5 mm 4.6 mm i.d., 5 lm particles) (Alltech, Temple-
mars, France). The columns were left at room temperature.
The mobile phase was a mixture of HPLC grade aceto-
nitrile:water:acetic acid (45:54:1), ltered through a
0.22 lm lter membrane, degassed and used at a ow-rate
of 1.0 ml/min. Retention time was 7.5 min.
The quantity of OA adsorbed was determined by the
following equation:
C 0  C eq  V
Qeq
m accuracies and is commonly determined by the least
Qeq quantity of OA adsorbed per gram of adsorbent
(mg/g)
C0 initial concentration of OA in solution (mg/L)
Ceq residual toxin concentration at equilibrium (mg/L)
V volume of solution (L)
m mass of adsorbent (g)
The maximum error calculated for the OA concentration
in the supernatant was 0.259 mg/L for EX16 experiments,
0.315 mg/L for BETA and 0.031 mg/L for LEC.
3. Results and discussion
3.1. Equilibrium studies
Adsorption equilibrium is established when the quantity
of the toxin being adsorbed (Qeq) is equal to the quantity
being desorbed. Then, the equilibrium concentration in
solution (Ceq) remains constant.
The plotting of Qeq = f(Ceq) for all adsorption experi-
ments is presented in Fig. 1. Adsorbent EX16 was able to
bind 3243% of the initial OA, BETA adsorbed 3751%
and LEC was a very good adsorbent, able to bind 95
100% of the initial OA.
Many equations have been published to describe the
equilibrium relationship between adsorbed and unad-
0.20
0.16
0.12
Qeq, mg/g
0.08
0.04
EX16
BETA
LEC
0.00
0 2 4 6 8
Ceq, mg/L
Fig. 1. Experimental OA adsorption on yeast by-products.
sorbed amounts. In this study, seven adsorption models
often reported in the literature were tested to provide the
best description of toxin adsorption, namely Freundlich
(F), Langmuir (L), BrunauerEmmettTeller (BET), Hill
(H), RedlichPeterson (RP), RadkePrausnitz (RkP) and
Toth (T). These models are presented below (Eqs. (2)
(17)). It should be noted that for Freundlich, Langmuir,
BET and Hill models, the set of isotherm parameters was
calculated not only by linearization but also in non-linear
forms, while the other models were only evaluated in their
non-linear form. For the linear models, the coecient of
determination (R2) is widely used in assessment of isotherm
1
squares method (Dagnelie, 1998). For the non-linear
forms, the various constants of these models were calcu-
lated by optimising the sum of the squares of the errors
(see Eq. (18)) using the solver add-in from Microsofts
spreadsheet, Excel (Microsoft, 2001). The sum of the
squares of the errors in the non-linear models was used
as an error indicator analogous to the coecient of
determination in the linear models.
For all models, the isotherm set parameter values are
presented in Tables 13. Their corresponding linear and
non-linear isotherms are illustrated in Figs. 24 and 57
respectively.
3.1.1. Freundlich empirical model
The empirical Freundlich (Freundlich, 1906) equation
based on sorption onto a heterogeneous surface is given
by Eq. (2).
Qeq K F  C 1=nF
2
eq
where KF and nF are the Freundlich constants charac-
teristic of the system. KF and nF are indicators of adsorp-
tion capacity and adsorption intensity, respectively. The
linearized form of the Freundlich equation is given in
Eq. (3).
1
ln Qeq ln K F ln C eq 3
nF
 D. Ringot et al. / Bioresource Technology 98 (2007) 18121821 1815

where

2.1430E + 00
1.0489E04
6.3800E03

1.6513E01
2.0480E02

3.0620E01
4.5782E01

5.1636E01
6.9748E01
1.0313E+00
1.7238E+00
Ceq residual toxin concentration at equilibrium (mg/L)
2.228

3.275

0.297
Radske-Prausnitz Toth

KF Freundlich adsorption constant (mg/g)/(mg/L)n

nF Freundlich adsorption constant

1.1685E + 00
Qeq adsorbed toxin quantity per gram of biomass (mg/g)

3.5948E05
2.8338E03
4.3922E01

5.6592E02
1.2376E02

1.3601E01
1.9499E01

3.1204E01
4.6883E01
1.1587E+00
A plot of ln(Qeq) versus ln(Ceq) should indicate a
0.018

0.387

0.552
straight line of slope 1/nF and intercept ln KF.

4.7147E + 00 3.2630E01 2.5394E + 00 1.5820E + 00 5.0000E + 00 5.1407E01 1.5823E + 00

6.3231E05
4.2593E03
6.5255E01

9.9544E02
1.6416E02

2.0442E01
2.8970E01

4.1389E01
5.7472E01
1.4204E+00
3.1.2. Langmuir model
The Langmuir (Langmuir, 1916) model is valid for
0.020

0.161

1.000

monolayer sorption to a surface with a nite number of

RP

identical sites. The well known expression of the Langmuir

6.6124E06
8.3289E04
1.6161E01
6.3976E01

1.0410E02
5.2780E03

3.9973E02
7.1746E02

1.3307E01
2.5886E01
model is given by the following Eq. (4), which can be
linearized as in Eq. (5).
0.071

3.950

1.751
Hill

Q K L C eq
Qeq max 4
1 K L C eq
1.0000E + 00
1.0000E + 00
1.0000E + 00

1.0000E + 00
1.0000E + 00
6.3521E04
2.0836E02

3.9663E02
2.2525E+00
2.4714E+00
4. 28.851

where Qeq (mg/g) and Ceq (mg/L) are the amount of adsor-
bent per unit weight of biomass and the unadsorbed toxin
0.015
BET

concentration in solution at equilibrium, respectively. Qmax

is the maximum amount of toxin per unit weight of bio-
6.3231E05
4.2583E03
6.5235E01

9.9544E02
1.6416E02

2.0437E01
2.8961E01

4.1388E01
5.7464E01
1.4202E+00
Langmuir

mass to form a complete monolayer and KL is a constant

related to the anity of the binding sites.
0.161
Non-linear models

0.23

C eq 1 C eq
5
1.2879E04
8.0123E03

2.0275E01
2.2476E02

3.8454E01
6.0604E01

5.6667E01
7.7939E01
1.3651E+00
1.9262E+00

Qeq K L  Qmax Qmax

Freundlich

0.017

1.391

KL Langmuir adsorption constant (L/mg)

Ceq residual toxin concentration at equilibrium (mg/L)
1.4166E05
5.4398E04
2.9507E02
2.9417E01

2.2301E02
5.7813E03

2.6107E02
1.3100E02
1.1903E01
1.4576E01

Qeq adsorbed toxin quantity per gram of biomass (mg/g)

Qmax maximum specic uptake corresponding to sites
0.996
0.078

4.054

1.470

saturation (mg/g)
Hill

6.3470E04
1.9625E02

9.9920E01
3.9027E02

9.4188E01
8.5044E01
9.3918E01
9.8396E01
1.9156E+00
2.3211E+00

A plot of Ceq/Qeq versus Ceq should indicate a straight

line of slope 1/Qmax and intercept of 1/KLQmax.
Isotherm parameters and error functions for OA adsorption onto EX16

26.036
0.867
0.015

In the case of low equilibrium concentration Ceq, then

BET

1  KLCeq and Langmuir becomes Henrys law (Eq. (6))

For the signication of isotherm parameters see Eqs. (2)(17).
1.9406E + 00 1.0413E + 00
8.7231E05
2.6211E03
2.2285E01
8.2689E01

1.3733E01
1.3669E02

1.2580E01
9.8934E02
3.3458E01
3.4463E01

Qeq K H  C eq 6
Langmuir
0.982
0.150

0.109

3.1.3. BrunauerEmmettTeller (BET) model

Linear models

The BET (Bruanuer et al., 1938) isotherm is the theoret-

3.5272E04
6.4523E03
1.4662E01
7.5132E01

5.5529E01
2.8023E02

3.0967E01
6.5092E02
3.0400E01
7.0654E01
Freundlich

ical model for multilayer adsorption. It is the most widely

applied model in studies of gassolid equilibrium. This
0.974
0.013

nF, KL, CBET, 1.039

model assumes multilayer adsorption and was developed

to describe adsorption phenomena when successive mole-
Q0max , Qmax H,

KD, ar, R, At

cular layers of adsorbate form after the completion of a

nH, br, p, t

HYBRID
KF, Qmax,

Kr, A, Kt

ERRSQ

monolayer.
MPSD

EABS
AER

SNE

The extinction of this model to liquidsolid interface is

R2

described by Eq. (7), which is linearized in Eq. (8).

Q0max  C BET  C eq
parametersa

Qeq 7
C s  C eq  1 C BET  1  C eq =C s 
function
Isotherm
Table 1

C eq 1 C BET  1 C eq
Error

 8
Qeq C s  C eq Q0max  C BET Q0max  C BET C s
a
 1816
Table 2
Isotherm parameters and error functions for OA adsorption onto beta-glucanes
Linear models Non-linear models

D. Ringot et al. / Bioresource Technology 98 (2007) 18121821

Freundlich Langmuir BET Hill Freundlich Langmuir BET Hill RP Radske- Toth
Prausnitz
Isotherm R2 0.961 0.920 0.915 0.940
parametersa KF, Qmax, 0.016 0.134 0.019 0.174 0.015 3.237 0.024 0.766 1.052 0.017 22.644
Q0max ,
Qmax H, Kr,
A, Kt
nF, KL, 1.030 0.169 9.912 8.004 0.964 0.005 9.549 51.324 67.378 1.051 48.190
CBET, KD,
ar, R, At
nH, br, p, t 1.040 1.087 0.000 0.571 0.538
Error ERRSQ 1.0550E04 4.7852E04 1.0284E04 2.6054E04 1.0011E04 1.0593E04 2.6720E05 1.1239E04 1.0186E04 1.9465E04 1.1074E04
function HYBRID 4.5149E03 9.7567E03 5.7889E03 7.5246E03 4.5383E03 4.4048E03 5.2701E03 4.8674E03 4.3496E03 5.5218E03 4.5287E03
MPSD 3.3860E01 9.4996E01 9.0677E01 7.4106E01 2.8610E01 3.2898E01 1.4667E+00 2.8242E01 3.1662E01 4.1335E01 3.7048E01
AER 1.1492E+00 1.4792E+00 1.5350E+00 1.3612E+00 1.0789E+00 1.1559E+00 1.4283E+00 1.0645E+00 1.1410E+00 1.2667E+00 1.1850E+00
EABS 1.8329E02 3.2715E02 2.0782E02 2.6770E02 1.9104E02 1.9760E02 8.5830E03 2.0140E02 1.9451E02 2.5957E02 1.9671E02
2.2046E01 1.0000E + 00 2.1492E01 5.4447E01 2.0920E01 2.2137E01 5.5839E02 2.3488E01 2.1286E01 4.0678E01 2.3142E01
4.6275E01 1.0000E + 00 5.9333E01 7.7122E01 4.6514E01 4.5147E01 5.4015E01 4.9888E01 4.4581E01 5.6594E01 4.6416E01
2.3086E01 6.4770E01 6.1825E01 5.0527E01 1.9507E01 2.2431E01 1.0000E + 00 1.9256E01 2.1588E01 2.8183E01 2.5260E01
7.4863E01 9.6362E01 1.0000E + 00 8.8678E01 7.0282E01 7.5299E01 9.3046E01 6.9347E01 7.4330E01 8.2516E01 7.7194E01
5.6027E01 1.0000E + 00 6.3525E01 8.1829E01 5.8395E01 6.0400E01 2.6236E01 6.1561E01 5.9456E01 7.9344E01 6.0128E01
SNE 2.2230E + 00 4.6113E + 00 3.0617E + 00 3.5260E + 00 2.1562E + 00 2.2541E + 00 2.7888E + 00 2.2354E + 00 2.2124E + 00 2.8732E + 00 2.3214E + 00
a
For the signication of isotherm parameters see Eqs. (2)(17).
Table 3
Isotherm parameters and error functions for OA adsorption onto LEC
Linear models Non-linear models

D. Ringot et al. / Bioresource Technology 98 (2007) 18121821

Freundlich Langmuir BET Hill Freundlich Langmuir BET Hill RP Radske- Toth
Prausnitz
Isotherm R2 0.896 0.981 0.997 0.964
parametersa KF, Qmax, Q0max , 0.202 0.109 0.045 1.892 0.897 26.275 0.040 18.706 4.296 0.593 0.795
Qmax H,
Kr, A, Kt
nF, KL, CBET, 2.048 12.460 22.750 4.854 0.732 0.021 19.924 20.063 6.846 8.719 9.298
KD, ar, R, At
nH, br, p, t 0.762 1.389 0.000 1.146 5.976
Error ERRSQ 5.0766E03 8.7417E03 7.4881E05 2.3796E03 1.5754E03 1.6555E03 7.0877E05 1.5848E03 1.6470E03 1.6436E03 1.6471E03
function HYBRID 3.6971E02 4.9814E02 2.3850E03 1.9182E02 4.3355E02 2.6388E02 2.5069E03 4.4200E02 2.6430E02 2.7106E02 2.6431E02
MPSD 6.1672E01 3.4450E01 9.5714E02 1.9502E01 1.9014E+00 7.9567E01 1.2103E01 1.9471E+00 8.0085E01 8.4852E01 8.0084E01
AER 1.5035E+00 8.1778E01 5.3580E01 7.9404E01 2.6561E+00 1.8442E+00 6.2723E01 2.6815E+00 1.8490E+00 1.8980E+00 1.8490E+00
EABS 1.0070E01 9.9503E02 1.3048E02 7.4544E02 8.1587E02 7.8147E02 1.5304E02 8.1713E02 7.8002E02 7.8521E02 7.8001E02
5.8073E01 1.0000E + 00 8.5660E03 2.7221E01 1.8021E01 1.8938E01 8.1079E03 1.8129E01 1.8841E01 1.8801E01 1.8842E01
7.4217E01 1.0000E + 00 4.7877E02 3.8508E01 8.7033E01 5.2973E01 5.0324E02 8.8730E01 5.3057E01 5.4413E01 5.3059E01
3.1674E01 1.7693E01 4.9158E02 1.0016E01 9.7652E01 4.0865E01 6.2160E02 1.0000E + 00 4.1131E01 4.3579E01 4.1130E01
5.6070E01 3.0497E01 1.9981E01 2.9612E01 9.9054E01 6.8775E01 2.3391E01 1.0000E + 00 6.8953E01 7.0783E01 6.8952E01
1.0000E + 00 9.8812E01 1.2958E01 7.4027E01 8.1020E01 7.7605E01 1.5198E01 8.1146E01 7.7461E01 7.7976E01 7.7460E01
SNE 3.2003E + 00 3.4700E + 00 4.3499E01 1.7938E + 00 3.8278E + 00 2.5916E + 00 5.0648E01 3.8801E + 00 2.5944E + 00 2.6555E + 00 2.5944E + 00
a
For the signication of isotherm parameters see Eqs. (2)(17).

1817
1818 D. Ringot et al. / Bioresource Technology 98 (2007) 18121821

0.09 0.08
0.08 0.07
0.07
0.06
0.06
0.05
0.05 Experimental
Qeq, mg/g

Qeq, mg/g
0.04 Freundlich
0.04
Experimental Langmuir
0.03 0.03 BET
Freundlich
0.02 Langmuir Hill
0.02
BET Redlich-Peterson
0.01
Hill 0.01 Toth
0 RadKe-Prausnitz
0 2 4 6 8 0
Ceq, mg/L 0 2 4 6 8
Ceq, mg/L
Fig. 2. Linear isotherms for OA adsorption onto EX16.
Fig. 5. Non-linear isotherms for OA adsorption onto EX16.

0.10
0.09 0.09
0.08 0.08
0.07 0.07
0.06
0.06
Qeq, mg/g

Experimental
Qeq, mg/g

0.05
0.05 Freundlich
0.04 Experimental
0.04 Langmuir
0.03 Freundlich
BET
Langmuir 0.03
0.02 Hill
BET
0.01 Hill
0.02 Redlich-Peterson
0.00 0.01 Toth
0 1 2 3 4 5 6 RadKe-Prausnitz
0
Ceq, mg/L
0 1 2 3 4 5 6
Fig. 3. Linear isotherms for OA adsorption onto BETA. Ceq, mg/L

Fig. 6. Linear isotherms for OA adsorption onto BETA.

0.20
0.18
0.20
0.16
0.18
0.14
0.16
0.12
0.14
Qeq, mg/g

0.10
0.12
Qeq, mg/g

Experimental
0.08
Experimental 0.10 Freundlich
0.06 Freundlich 0.08 Langmuir
0.04 Langmuir BET
0.06 Hill
BET
0.02 0.04 Redlich-Peterson
Hill
0.00 0.02 Toth
0 0.05 0.1 0.15 0.2 0.25 0.3 0.35 RadKe-Prausnitz
0.00
Ceq, mg/L
0 0.05 0.1 0.15 0.2 0.25 0.3 0.35
Ceq, mg/L
Fig. 4. Linear isotherms for OA adsorption onto LEC.
Fig. 7. Linear isotherms for OA adsorption onto LEC.
CBET BET adsorption constant relating to the energy of
interaction with the surface (L/mg)
Ceq residual toxin concentration at equilibrium (mg/L) Cs saturation concentration of the solute correspond-
Qeq adsorbed toxin quantity per gram of biomass (mg/ ing to monolayer saturation (mg/L)
g)
Q0max maximum specic uptake corresponding to mono- A plot of Ceq/[Qeq*(Cs  Ceq)] versus Ceq/Cs allows
layer saturation (mg/g) calculation of CBET and Q0max .
 D. Ringot et al. / Bioresource Technology 98 (2007) 18121821 1819

3.1.4. Non-ideal competitive adsorption (NICA) model This model has three isotherm parameters, Kr, ar and br
At the origin of the NICA (Koopal et al., 1994, 2001) (0 < br < 1) which characterize the isotherm.
model, it was postulated that an equation like the Hill When br = 1, then the equation becomes similar to the
equation (Hill, 1910) could be used to describe the binding Langmuir model (Eq. (13)).
of dierent species onto a homogeneous substrate. This
K r  C eq
model assumes that the adsorption is a cooperative phe- Qeq 13
nomenon due to the ability of ligand binding at one site 1 ar  C eq
on a macromolecule to inuence ligand binding at a dif-
When br = 0, then it becomes similar to Henrys law (Eq.
ferent site on the same macromolecule.
(14)).
The Hill equation is:
K r  C eq
Qmax H  C nH
eq Qeq 14
Qeq 9 1 ar
K D C nH
eq

This equation can be linearized through logarithmic terms 3.1.6. The RadkePrausnitz model
(Eq. (10)) The RadkePrausnitz (Radke and Prausnitz, 1972)
! isotherm model is depicted in Eq. (15).
Qeq
ln nH ln C eq  ln K D 10 A  R  C peq
Qmax H  Qeq
Qeq 15
A R  C p1
eq
where
This equation can be converted to a linear form (Eq. (16))
K D K nH
d 11 !
1 1 1
Ceq residual toxin concentration at equilibrium (mg/L) ln    ln R  p ln C eq 16
Qeq A C eq
Qeq adsorbed toxin quantity per gram of biomass (mg/
g)
KD Hill constant Ceq residual toxin concentration at equilibrium (mg/L)
Kd dissociation constant per site (mg/L), Kd is equal Qeq adsorbed toxin quantity per gram of biomass (mg/
to residual toxin concentration at half saturation, g)
K d C 50 A constant of RadkePrausnitz isotherm (L/g)
eq ; also, Kd = 1/Ka, where Ka is the associ-
ation constant R constant of RadkePrausnitz isotherm, (mg/
Qmax H maximum specic uptake corresponding to sites g) * (mg/L)p
saturation (mg/g) p constant of RadkePrausnitz isotherm
nH Hill cooperativity coecient of the binding inter-
action
3.1.7. Toth model
Thus, three possibilities can occur The Toth isotherm (Toth, 1971) is derived from the
potential theory and is applicable for heterogeneous
nH > 1, positive cooperativity in binding, adsorption. It assumes a quasi-Gaussian energy distribu-
nH = 1, non-cooperative or hyperbolic binding, tion. Most sites have an adsorption energy lower than
nH < 1, negative cooperativity in binding. the maximum adsorption energy (Eq. (17))
K t  C eq
Qeq 17
3.1.5. RedlichPeterson model At C teq 1=t
This model (Redlich and Peterson, 1958) approaches
the Freundlich model at high adsorbate concentration (it
describes equilibrium on heterogeneous surfaces and hence Ceq residual toxin concentration at equilibrium (mg/L)
does not assume monolayer capacity) and Henrys law at Qeq adsorbed toxin quantity per gram of biomass (mg/
g)
low concentration (Eq. (12)).
Kt constant of Toth isotherm (mg/g)
K r  C eq At constant of Toth isotherm (L/mg)
Qeq 12
1 ar  C beqr t constant of Toth isotherm

Ceq residual toxin concentration at equilibrium (mg/L)

Qeq adsorbed toxin quantity per gram of biomass (mg/ 3.2. Error analysis
g)
Kr constant of RedlichPeterson isotherm (L/g) The classical method to determine isotherm parameters
ar constant of RedlichPeterson isotherm (L/mg)br is to use linear regression with transformed variables.
br constant of RedlichPeterson isotherm Linearization of the isotherm equation alters the error
1820 D. Ringot et al. / Bioresource Technology 98 (2007) 18121821

structure and normality assumption of standard least For EX16, the data presented in Table 1 show, based on
squares. This could explain earlier observations that Fre- the sum of normalised errors, that the Hill model pro-
undlich isotherms produced better results at low concen- duced the best goodness of t. Furthermore, for the linear
tration and Langmuir isotherms tend to t data better equations, the highest coecients of determination were
for high concentration (Richter et al., 1989). Recently, also obtained using the Hill model. Since the value of
the utilisation of sum of normalised errors (SNE) calcula- the Hill coecient nH is higher than 1, the binding of
tion procedure in adsorption studies has been presented OA by EX16 is a positive cooperative interaction. For this
by several authors (Allen et al., 2003; Ho et al., 2002; Por- model, the values of isotherm constants are comparable
ter et al., 1999). In these publications, a non-linear com- when they are calculated by linear and non-linear
plex optimisation procedure was performed for the equations.
estimation of the SNE values. In the present study, ve For BETA, the SNE values presented in Table 2 show
error functions were examined to determine and evaluate that the non-linear Freundlich model produced the best
the t of isotherm models to the experimental data t across the range of non-linear models. Moreover, in lin-
(Eqs. (18)(22)) and only the sum of the squares of the ear models, the best value of linear R2 was also determined
errors (ERRQS) was optimised as a relevant error indica- for the Freundlich model. Based on the R2 and on the SNE
tor comparable to the coecient of determination in the values, the Freundlich model in its linear and non-linear
linear models. forms produced a reasonable model for OA biosorption
onto beta-glucans. The magnitude of R2 is an indication
1. The sum of the squares of errors (ERRSQ) of the relative quality of t of the linear isotherm. The fact
X
p that the nF value (in the Freundlich model) is very close to
2
Qe;meas  Qe;calc i 18 1 means that this model becomes Henrys law. Then, OA
i1 adsorption onto BETA is a simple linear solid/liquid parti-
2. The hybrid fractional error function (HYBRID) tion which follows Henrys law.
" # For LEC, the data presented in Table 3 show that, based
Xp 2
Qe;meas  Qe;calc on the SNE values, the BET model is the most appropriate
19 model for the biosorption of OA on this yeast cell product.
Qe;meas
i1 i Furthermore, the highest value of R2 was also calculated
3. A derivative of Marquardts percent standard deviation for the BET model. Based on the R2 values and on the
(MPSD) SNE value, the BET model appears to be a very good
!2 model for OA biosorption onto LEC. For this model,
Xp
Qe;meas  Qe;calc the values of isotherm constants calculated with the linear
20
i1
Qe;meas equation are very close to those obtained using non-linear
i
calculation.
4. The average relative error (AER) The various proles of toxin adsorption onto the three
Xp   adsorbents studied suggest dierent mechanisms of ad-
Qe;meas  Qe;calc 
  21 sorption according to their dierent compositions. In a
 Q 
i1 e;meas i
previous study (Ringot et al., 2005), a thermodynamic
5. The sum of absolute errors (EABS) approach to characterization of the binding of OA onto
X
p
  EX16, BETA and LEC was presented, enabling suggestion
Q  Qe;calc i 22 of some hypotheses regarding the mechanism of OA yeast
e;meas
i1 cell wall biosorption. In order to investigate these hypoth-
eses in future research, it is proposed to study the inuence
The process of minimising the respective error functions of adsorption conditions (pH and adsorbent mass) on the
across the range of experimental concentrations permitted biosorption phenomenon.
calculation of the isotherm constants. The values of the
errors obtained for each error function for each set of lin- 4. Conclusions
ear and non-linear isotherm constants were normalised by
dividing by the highest value for that error function across The present work allowed identication, among the
a range of dierent isotherms. These normalised errors most commonly described models in the literature, of the
were added for each parameter set to get the SNE. A com- isothermal equation which best tted the adsorption of
parison of the various SNE could thus be undertaken and OA onto each of the three yeast by-products studied. Lin-
allowed the identication of the best set of isotherm con- ear and non-linear optimisation techniques were applied to
stants to describe the measured data. The parameter set determine isotherm parameters. For the best models iden-
thus providing the smallest normalised error sum was con- tied, the non-linear calculation of isotherm parameters
sidered to be optimal. presented in this paper produced comparable data to those
The values of all these minimum error functions and of obtained using the linear method based on the least squares
the optimum isotherm constants are listed in Tables 13. calculation.
 D. Ringot et al. / Bioresource Technology 98 (2007) 18121821
Based on the error analysis, the Hill, Freundlich and
BET equations, in both their linear and non-linear forms,
appear to be appropriate models for OA biosorption onto
EX16, BETA and LEC, respectively. For these best mod-
els, the values of isotherm constants were very close when
calculated using linear and non-linear equations.
These results indicate that the linear equation analysis
using the R2 calculation associated with the SNE calcula-
tion procedure presented in this work is an appropriate
method to use for the study of OA adsorption onto yeast
by-products.
Acknowledgements
The authors thank Mr. Renaud Trouve from Serendi
Ltd., Mr. Eric Oriol from Bio-Springer Ltd. and Mrs. Pau-
line Anton-Gay from ISAB for their scientic interest in
this study.
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