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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 spices, coee and cocoa products, grape juice and beer
(Commission of the European Union, 2002).
Ochratoxin A (OA) is a secondary metabolite of OA is partially absorbed by the gastrointestinal tract
some species of storage fungi. Aspergillus species such as in monogastrics. In ruminants, OA is hydrolysed by the
A. ochraceus produce OA mainly in tropical climates and microbial ora into a much less toxic metabolite, ochra-
Penicillium species such as P. verrucosum produce OA toxin a. OA is hydrolysed to ochratoxin a by mammalian
in temperate climates. carboxypeptidase A and by some micro-organisms within
Chemically speaking, OA is a chlorine-containing di- the gastrointestinal tract. In hepatic cells, it is hydroxylated
hydroisocoumarine linked to L-phenylalanine. OA and its mainly to 4-R-hydroxyochratoxin A (OA-OH) by mixed
hydroxyl derivatives are toxic when consumed by livestock function oxidases (Li et al., 2000). OA is considered to be
(pigs, poultry) and humans. They are found in human milk, a genotoxic agent in vivo and in vitro in some mammalian
blood serum, plasma, liver and kidney. The main route of species (WHO, 2002), but the mechanism of genotoxicity
exposure is the consumption of contaminated cereals, wine, is unclear and there is no evidence of a direct interaction
with DNA (Bakker and Pieters, 2002; Ringot et al.,
*
Corresponding author. Tel.: +33 344 062 517; fax: +33 344 062 526. 2006). OA is reasonably anticipated to be a possible human
E-mail address: diana.ringot@isab.fr (D. Ringot). 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 1813

in experimental animals (group 2B) (IARC, 1993). OA is Yeasts produce a high quantity of biomass. They are
a nephrotoxic, teratogenic and immunotoxic compound used in a large variety of industrial fermentation processes;
(Muller et al., 1999; Prelusky et al., 1994). OA is relatively moreover, they may be regarded as a good source of adsor-
stable and is only partially degraded under normal process bent material, due to the presence in their cell wall of some
conditions. specic macromolecules such as the mannoproteins and
Dierent approaches have been developed to reduce the beta-glucans. The ability of the Saccharomyces cerevisiae
impact of mycotoxins. They can be divided into pre- and cell wall to bind zeralenone has been reported recently
post-harvest strategies and into biological, chemical and (Yiannikouris et al., 2003).
physical methods (Huwing et al., 2001). Physical methods This research was aimed at examining in vitro ad-
involve extraction with solvent, adsorption, inactivation sorption of OA onto three yeast industry by-products at
by heat and irradiation. Among the physical decontamina- a temperature of 25 C. To this end, several theoretical
tion methods, adsorption onto various types of compounds adsorption models frequently used in the literature were
(hydrated sodium calcium aluminosilicate HSCAS, tested for their ability to describe the equilibrium sorption
kaolin, silica binding agent, bentonite, etc.) has been ex- data. For these models, ve error functions and the norma-
tensively studied in recent years (Grant and Philips, 1998; lised error sum (SNE) were examined. This enabled the
Scott, 1998). Dierent binding agents including activated determination of the best parameter set and thus provided
carbons (Galvano et al., 1998), zeolites (Tomasevic-Cano- an accurate equilibrium adsorption model.
vic et al., 2003), diatomaceous earth (Natour and Yousef,
1998), cholestyramine and mixtures of sterilized yeast and 2. Methods
fermentation residua from beer production (Bauer, 1994)
have been reported to remove OA in vitro. In contrast, acti- 2.1. Adsorbents and reagents
vated charcoal and HSCAS have been reported to have a
limited ecacy against OA in vitro (Ramos et al., 1996). Three adsorbent materials, namely EX16, BETA and
Castellari et al. (2001) examined several ning treatments LEC, were obtained from Bio-Springer, Maison-Alfort,
to reduce OA levels in red wine. They found that potassium France. Two of these adsorbents, EX16 (a vinasse contain-
caseinate and activated carbon were the best agents to ing 16% liquid yeast cell walls) and LEC (a dry yeast cell
remove OA. wall fraction) are industrial by-products of the yeast indus-
The phenomenon of biosorption is dened as a metabo- tries. BETA is the dried puried beta-glucans fraction of
lism independent adsorption of pollutants based on the cell walls. The dry matter content of the by-products was
partition process on a microbial biomass. Microbial bio- 56% for EX16, 95% for BETA and 97.6% for LEC.
mass consists of small particles, with low density, poor OA was purchased from Sigma Chemical Company
mechanical strength and little rigidity. This phenomenon (St. Louis, Missouri, USA). Primary methanolic stock
is generally based on a set of chemical and physical mech- solution (200 mg/L) was prepared in methanol. Five OA
anisms (involving physico-chemical interactions such as test solutions with concentrations of 0.5, 1.0, 2.0, 5.0 and
electrostatic interactions, ion exchange, complexation, che- 10.0 mg/L were prepared by the dilution of the methanolic
lation and precipitation) leading to the immobilization of a stock solution with deionised distilled water.
solute component on the microbial cell wall components. It
has been investigated by dierent authors (Aksu, 2003; 2.2. Experimental system
Aksu and Donmez, 2002; Aksu and Yenner, 1998; Tsezos
and Bell, 1989). Samples of 500 mg of adsorbents were placed in screw
The complexity of the microbial structure implies that cap test tubes with aliquots (10 mL) of the OA test solu-
there are many ways for the pollutant to be captured by tions. Two sets of controls were prepared, one by adding
the cells. Biosorption mechanisms are therefore various 10 mL of mycotoxin test solutions without the addition
(physical adsorption, chemical binding of ionic groups, of adsorbent, and another by adding 500 mg adsorbent
ion exchange, etc.) and in some cases they are still not very to 10 mL deionised distilled water. All experiments were
well understood (Veglio and Beolchini, 1997). Cell surface carried out in triplicate.
sorption is a physico-chemical interaction between the The controls and test tubes were placed in a horizontal
toxin and functional groups of the cell surface, based on shaker-incubator at 400 rpm and at a controlled tempera-
physical adsorption, ion exchange and complexation, ture of 25.0 1.0 C. Preliminary investigations showed
which is not dependent on metabolism. Cell walls of micro- that equilibrium uptake was attained rapidly, with prac-
bial biomass, mainly composed of polysaccharides, pro- tically no change observed after a period of 30 min. All
teins and lipids, oer abundant functional groups, such adsorptions were run for 90 min to ensure complete
as carboxyl, hydroxyl, phosphate and amino groups, as uptake. After the incubation period, solid particles (adsor-
well as hydrophobic adsorption sites such as aliphatic bent) were separated from the supernatant by centrifu-
carbon chains and aromatic rings (Ringot et al., 2005). gation at 6000g for 20 min at 25 C in an ALC 4239R
This physico-chemical phenomenon is quick and can be centrifugation system (Fisher Scientic Labosi, Elancourt,
reversible. France).
1814 D. Ringot et al. / Bioresource Technology 98 (2007) 18121821

The supernatant was extracted with an equal volume of 0.20


methanol:deionised distilled water (50:50). The mixture
was vortexed for 5 min. This extract was diluted, ltered 0.16
and analysed by HPLC.
0.12

Qeq, mg/g
2.3. Chromatographic analysis
0.08
The HPLC chromatographic system consisted of a Spec-
traSYSTEM P1500 isocratic pump (Thermo Separation 0.04
EX16
Product, Fremont, California, USA) equipped with a AS BETA
LEC
3000 model injection valve (100 ll) (Thermo Separation 0.00
Product, Fremont, California), a Spectroow 980 uores- 0 2 4 6 8
cence spectrophotometric detector (Applied Biosystems, Ceq, mg/L
Ramsey, New Jersey, USA) equipped with a 150 W xenon
lamp (kexcitation = 333 nm and kemission = 470 nm) and Fig. 1. Experimental OA adsorption on yeast by-products.
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., sorbed amounts. In this study, seven adsorption models
5 lm particles) preceded by an Alltima C18 precolumn often reported in the literature were tested to provide the
(7.5 mm 4.6 mm i.d., 5 lm particles) (Alltech, Temple- best description of toxin adsorption, namely Freundlich
mars, France). The columns were left at room temperature. (F), Langmuir (L), BrunauerEmmettTeller (BET), Hill
The mobile phase was a mixture of HPLC grade aceto- (H), RedlichPeterson (RP), RadkePrausnitz (RkP) and
nitrile:water:acetic acid (45:54:1), ltered through a Toth (T). These models are presented below (Eqs. (2)
0.22 lm lter membrane, degassed and used at a ow-rate (17)). It should be noted that for Freundlich, Langmuir,
of 1.0 ml/min. Retention time was 7.5 min. BET and Hill models, the set of isotherm parameters was
The quantity of OA adsorbed was determined by the calculated not only by linearization but also in non-linear
following equation: forms, while the other models were only evaluated in their
non-linear form. For the linear models, the coecient of
C 0  C eq  V determination (R2) is widely used in assessment of isotherm
Qeq 1
m accuracies and is commonly determined by the least
squares method (Dagnelie, 1998). For the non-linear
Qeq quantity of OA adsorbed per gram of adsorbent forms, the various constants of these models were calcu-
(mg/g) lated by optimising the sum of the squares of the errors
C0 initial concentration of OA in solution (mg/L) (see Eq. (18)) using the solver add-in from Microsofts
Ceq residual toxin concentration at equilibrium (mg/L) spreadsheet, Excel (Microsoft, 2001). The sum of the
V volume of solution (L) squares of the errors in the non-linear models was used
m mass of adsorbent (g) as an error indicator analogous to the coecient of
determination in the linear models.
The maximum error calculated for the OA concentration For all models, the isotherm set parameter values are
in the supernatant was 0.259 mg/L for EX16 experiments, presented in Tables 13. Their corresponding linear and
0.315 mg/L for BETA and 0.031 mg/L for LEC. non-linear isotherms are illustrated in Figs. 24 and 57
respectively.
3. Results and discussion
3.1.1. Freundlich empirical model
3.1. Equilibrium studies The empirical Freundlich (Freundlich, 1906) equation
based on sorption onto a heterogeneous surface is given
Adsorption equilibrium is established when the quantity by Eq. (2).
of the toxin being adsorbed (Qeq) is equal to the quantity
Qeq K F  C 1=nF
2
being desorbed. Then, the equilibrium concentration in eq

solution (Ceq) remains constant. where KF and nF are the Freundlich constants charac-
The plotting of Qeq = f(Ceq) for all adsorption experi- teristic of the system. KF and nF are indicators of adsorp-
ments is presented in Fig. 1. Adsorbent EX16 was able to tion capacity and adsorption intensity, respectively. The
bind 3243% of the initial OA, BETA adsorbed 3751% linearized form of the Freundlich equation is given in
and LEC was a very good adsorbent, able to bind 95 Eq. (3).
100% of the initial OA.
Many equations have been published to describe the 1
ln Qeq ln K F ln C eq 3
equilibrium relationship between adsorbed and unad- 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 1821

Based on the error analysis, the Hill, Freundlich and IARC, 1993IARC Monographs on the Evaluation of Carcinogenic Risks
BET equations, in both their linear and non-linear forms, to Humans: Some Naturally Occurring Substances, Food Items and
Constituents, Heterocyclic Aromatic Amines and Mycotoxins, vol. 56.
appear to be appropriate models for OA biosorption onto International Agency for Research on Cancer, Lyon, France, pp. 489
EX16, BETA and LEC, respectively. For these best mod- 521.
els, the values of isotherm constants were very close when Koopal, L.K., Van Riemsdijk, W.H., de Wit, J.C.M., Benedetti, M.F.,
calculated using linear and non-linear equations. 1994. Analytical isotherm equation for multicomponent adsorption to
These results indicate that the linear equation analysis heterogeneous surfaces. J. Colloid Interface Sci. 166, 5160.
Koopal, L.K., Van Riemsdijk, W.H., Kinniburgh, D.G., 2001. Humic
using the R2 calculation associated with the SNE calcula- matter and contaminants. General aspects and modelling ion binding.
tion procedure presented in this work is an appropriate Pure Appl. Chem. 73, 20052016.
method to use for the study of OA adsorption onto yeast Langmuir, I., 1916. The adsorption of gases on plane surface of glass,
by-products. mica and platinum. J. Am. Chem. Soc. 40, 13611403.
Li, S., Marquardt, R.R., Frohlich, A.A., 2000. Identication of ochra-
toxins and some of their metabolites in bile and urine of rats. Food
Acknowledgements
Chem. Toxicol. 38, 141152.
Microsoft Corporation, 2001. Users guide, Microsoft Excel Ver. 2002.
The authors thank Mr. Renaud Trouve from Serendi Muller, G., Kielstein, P., Rosner, H., Berndt, A., Heller, M., Kohler, H.,
Ltd., Mr. Eric Oriol from Bio-Springer Ltd. and Mrs. Pau- 1999. Studies on the inuence of ochratoxin A on immune and defense
line Anton-Gay from ISAB for their scientic interest in reactions in weaners. Mycosis 42, 495505.
Natour, R.M., Yousef, S.M., 1998. Adsorption eciency of diato-
this study.
maceous earth for mycotoxins. Arab Gulf J. Sci. Res. 16, 113E
127E.
References Porter, J.F., McKay, G., Choy, K.H., 1999. The prediction of sorption
binary mixture of acidic dyes using single- and mixed-isotherm
Aksu, Z., 2003. Reactive dye bioaccumulation by Saccharomyces cerevi- variants of the ideal adsorbed solute theory. Chem. Eng. Sci. 54,
siae. Process Biochem. 38, 14371444. 58635885.
Aksu, Z., Donmez, G., 2002. A comparative study on the biosorption Prelusky, D.B., Rotter, B.A., Rotter, R.G., 1994. Toxicology of myco-
characteristics of some yeast for Remazol Blue reactive dye. Chemo- toxins. In: Miller, Treholm (Eds.), Mycotoxins in Grain, Compounds
sphere 50, 10751083. other than Aatoxins. Eagan Press, St. Paul, MN USA, pp. 359
Aksu, Z., Yenner, J., 1998. Investigation of the biosorption of phenol and 403.
monochlorinated phenols on the dried activated sludges. Process Radke, C.J., Prausnitz, J.M., 1972. Adsorption of organic solutes from
Biochem. 33, 483491. dilute aqueous solutions on activated carbon. Ind. Eng. Chem.
Allen, S., Gan, Q., Matthews, R., Johnson, P.A., 2003. Comparison of Fundam. 11, 445450.
optimised models for basic dye adsorption by kudzu. Bioresource Ramos, A.J., Fink-Gremmels, J., Hernandez, E., 1996. Prevention of toxic
Technol. 88, 143152. eects of mycotoxins by means of non-nutritive adsorbent compounds.
Bakker, M., Pieters, M.N., 2002. Risk assessment of ochratoxin A in J. Food. Prot. 59, 631646.
Netherlands. RIVM Report 388802025. Inspectorate for Health Redlich, O., Peterson, C., 1958. Useful adsorption isotherm. J. Phys.
Protection and Veterinary Public Health. Bilthoven, Netherlands, 24p. Chem. 63, 1024.
Bauer, J., 1994. Moglichkeiten zur Entgiftung mycotoxin-haltiger, Fut- Richter, E., Schultz, W., Myers, A.L., 1989. Eect of adsorption equation
termittel. Monatssh. Veterinarmed. 49, 175181. on prediction of multicomponent equilibria by the ideal adsorption
Bruanuer, S., Emmett, P.H., Teller, E., 1938. Adsorption of gases in multi- solution theory. Chem. Eng. Sci. 44, 16091616.
molecular layers. J. Am. Chem. Soc. 60, 309316. Ringot, D., Lerzy, B., Bonhoure, J.P., Auclair, E., Oriol, E., Larondelle,
Castellari, M., Versari, A., Fabiani, A., Parpinello, G.P., Galassi, S., 2001. Y., 2005. Eect of temperature on in vitro ochratoxin A biosorption
Removal of ochratoxin A in red wines by means of adsorption onto yeast cell derivatives. Process Biochem. 40, 30083016.
treatments with commercial ning agents. J. Agric. Food Chem. 49, Ringot, D., Chango, A., Schneider, Y.J., Larondelle, Y., 2006. Toxico-
39173921. kinetics and toxicodynamics of ochratoxin A: an update. Chem. Biol.
Commission of the European Union, 2002. Report of experts participating Interact. 159, 1846.
in task 3.2.7. Assessment of dietary intake of ochratoxin A by the Scott, P.M., 1998. Industrial and farm detoxication process for myco-
population of EU Member States. 153p. toxins. Rev. Med. Vet. 149, 543548.
Dagnelie, P., 1998. Statistique theorique et appliquee. vol. 1, De Boecket Tomasevic-Canovic, M., Dakovic, A., Rottinghaus, G., Matijasevic, S.,
Larcier, Paris-Bruxelles, Belgium pp. 130148. Duricic, M., 2003. Surfactant modied zeolites new ecient
Freundlich, H.M.F., 1906. Uber die Adsorption in Losungen. Z. Phys. adsorbent for mycotoxins. Micropor. Mesopor. Mater. 61, 173
Chem. 57, 385470. 180.
Galvano, F., Pietri, A., Bertuzzi, T., Piva, A., Chies, L., Galvano, M., Toth, J., 1971. State equations of the solid gas interface layer. Acta Chem.
1998. Activated carbons: In vitro anity for ochratoxin A and Acad. Hung. 69, 311317.
deoxynivalenol and relation of adsorption ability to physicochemical Tsezos, M., Bell, J.P., 1989. Comparison of the biosorption and
parameters. J. Food Prot. 61, 469475. desorption of hazardous organic pollutants by live and dead biomass.
Grant, P.G., Philips, T.D., 1998. Isothermal adsorption of aatoxin B1 on Water Res. 23, 561568.
HSCAS clay. J. Agric. Food Chem. 46, 599605. Veglio, F., Beolchini, F., 1997. Removal of metals by biosorption: a
Hill, A.V., 1910. The possible eects of the aggregation of the molecules of review. Hydrometallurgy 44, 301316.
haemoglobin on its dissociation curves. J. Physiol. (London) 40, ivvii. World Health Organisation, 2002. Evaluation of certain mycotoxins in
Ho, Y.S., Porter, J.F., McKay, G., 2002. Equilibrium isotherm studies for food. WHO Technical Report Series, vol. 906. WHO, Geneva,
the sorption of divalent metal ions onto peat: copper, nickel and lead Switzerland, p. 62.
single component systems. Water Air Soil Pollut. 141, 133. Yiannikouris, A., Poughon, L., Cameleyre, X., Dussap, C.J., Francois, J.,
Huwing, A., Freimund, S., Kappeli, O., Dutler, H., 2001. Mycotoxin Bertin, G., Jouany, J.P., 2003. A novel technique to evaluate
detoxication of animal feed by dierent adsorbent. Toxicol. Lett. 122, interactions between Saccharomyces cerevisiae cell wall and myco-
179188. toxins: application to zearalenone. Biotechnol. Lett. 25, 783789.