Antioxidant activity

of byproducts from
the hydrolytic
processing of
selected
lignocellulosic
materials
G. Garrote, J.M. Cruz,
A. Moure, H. Dom

nguez* and
J.C. Parajo
Department of Chemical Engineering.
University of Vigo (Campus Ourense).
Polytechnical Building,
As Lagoas, 32004 Ourense, Spain
(tel: +34-988-387082; fax: +34-988-387001;
e-mail: herminia@uvigo.es)
Mild acid treatments of lignocellulosic materials result in
hemicellulose conversion into sugar and sugar oligomers as
well as in solubilization of phenolic compounds with
potential food applications. The eect of the operational
conditions (measured by the severity factor) on the yield of
soluble phenolics and the utilization of these compounds
(with antioxidant and antimicrobial activities) as food addi-
tives are reviewed. Additional information on other biologi-
cal eects of phenolics that could be of interest for the
formulation of functional foods is provided.
# 2003 Elsevier Ltd. All rights reserved.
Introduction
The interest for cheap, renewable and abundant
sources of natural antioxidants has grown due to safety
concerns, contradictory toxicological data about syn-
thetic antioxidants and the consumers preferences for
natural additives. Lignocellulosic materials (LCM),
particularly those of residual origin coming from
agroindustrial and forest activities, are promising sour-
ces of antioxidant compounds (Dom nguez, Torres, &
Nu n ez, 2001).
The hydrolytic processing of LCM intends primarily
the selective breakdown of hemicelluloses to obtain
sugars or sugar oligomers. However, cell wall-linked
phenolic compounds are also solubilized during this
type of treatments (Felizo n, Ferna ndez-Bolan os, Her-
edia, & Guille n, 2000; Ohta, Yamasaki, Egashira, &
Sanada, 1994). Phenolics appear as non-desired com-
pounds in liquid phase, which sometimes has to be sub-
jected to further purication steps. These phenolics may
be considered byproducts with possible application as
food additives.
Mild hydrolytic processing of LCM
A number of technologies are available for the mild
hydrolysis of LCM. In the simplest one (auto-
hydrolysis), the considered LCM is contacted with
water or steam. Related processes include the utilization
of additional reagents such as mineral acids (pre-
hydrolysis), SO
2
or oxygen (wet oxidation).
Autohydrolysis of LCM is an environmentally
friendly process in which hydronium ions from water
autoionization and from organic acids generated in the
reaction promote the hydrolytic degradation of cell wall
components. A variety of compounds appear in the
liquors obtained by these technologies, including sugar
oligomers, monomeric sugars, sugar-degradation pro-
ducts (furfural and hydroxymethylfurfural), organic
acids (citric and malic acid coming from the cells of the
biomass, formic and levulinic acid from sugar-degrada-
tion products, acetic acid from acetyl groups), extrac-
tives and phenolics. The main hemicellulose-derived
products (sugar oligomers and sugars) have commercial
applications: sugar oligomers can be used as prebiotics
(Va zquez, Alonso, Dom nguez, & Parajo , 2001),
0924-2244/$ - see front matter # 2003 Elsevier Ltd. All rights reserved.
doi:10.1016/j.tifs.2003.09.016
Trends in Food Science & Technology 15 (2004) 191200
Review
* Corresponding author.
whereas monomeric sugars are suitable as carbon
sources for the fermentative production of ethanol
(Ando et al., 2000; Boussaid et al., 2001; Mart n, Galbe,
Nilvebrant, & Jo nsson, 2002) or xylitol (Rivas,
Dom nguez, Dom nguez, & Parajo , 2002).
The presence of non-saccharide compounds in
hydrolysates is unfavourable for the purposes above
cited, because they decrease the purity of xylooligo-
saccharides or inhibit the microbial metabolism of
sugars. To overcome the latter problem, purication of
liquors by ethyl acetate extraction has been proposed
(Cruz, Dom nguez, Dom nguez, & Parajo , 1999; Frazer
& McCaskey, 1989). The ethyl acetate-soluble fraction
contains a variety of potentially valuable phenolic com-
pounds with antioxidant, antimicrobial and biological
activities. The utilization of this fraction would be
of scientic and economic interest, leading to an
integrated, multiproduct process.
Eect of hydrolytic treatments on lignin
Lignin is an heterogeneous polymer of phenolic
nature with structure and monomer distribution pattern
dependent on factors such as sampling, plant family,
location and cultivation conditions. Lignin is made up
of three precursors: trans-coniferyl, trans-sinapyl and
trans-pcoumaryl alcohols. Gymnosperm lignins show
predominance of guaiacyl groups, woody angiosperms
lignins contain guaiacyl-syringyl groups and lignins
from grasses contain guaiacyl-syringyl-p-hydroxyphenyl
groups.
Partial depolymerization of lignin and ligninhemi-
cellulose linkages occurs during the hydrolytic proces-
sing of LCM, leading to liquors with UV absorption
peaks at 205 and 279 nm, characteristics of lignin (Ando
et al., 2000; Tran & Chambers, 1985). Table 1 sum-
marizes the main phenolic compounds identied in
liquors from acidic treatments of LCM. The structures
of depolymerized lignin fraction included p-substituted
phenols (with zero, one or two methoxy groups), alde-
hydes, acetophenones and carboxylic acids. Ferulic and
p-coumaric acids (the most abundant hydroxycinnamic
acids) are linked to arabinoxylans or pectins through
ester bonds. In hardwoods, condensed tannins (proan-
thocyanidins) and hydrolyzable ellagitannins make
also part of the phenolic fraction (Cadah a, Mun oz,
Ferna ndez de Simo n, & Garc a-Vallejo, 2001; Helm,
Ranatunga, & Chandra, 1998), whereas acids (gallic,
vanillic and ellagic) and aldehydes (syringaldehyde
and sinapaldehyde) have been also detected (Conde,
Cadahia, Garcia-Vallejo, & Tomas-Barberan, 1995).
A selective recovery of the phenolic compounds from
hydrolysates can be achieved by extraction with solvents
such as ethyl acetate or diethylether. Ethyl acetate
removes water-soluble phenolics and hemicellulose-
degradation products, whereas lignincarbohydrate
complexes remain in the aqueous phase (Bouchard,
Nguyen, Chornet, & Overend, 1991).
Considering that fractionation and purication of
extracts increases both the process complexity and
costs, the utilization of crude extracts from vegetals has
been proposed for food purposes (Rodr guez de
Sotillo, Hadley, & Holm, 1994; Rodr guez de Sotillo,
Hadley, & Wolf-Hall, 1998; Yanishlieva & Marinova,
1995). The activity of crude extracts has been reported
to be higher than that of the puried fractions due to
the presence of active compounds in small quantities
and/or to synergistic eects among various compounds
(Ka hko nen et al., 1999; Onyeneho & Hettiarachchy,
1992; Rodr guez de Sotillo et al., 1994). This approach
would be suitable for the utilization of the phenolic
compounds present in LCM hydrolysates from mild
acidic processing.
Eects of the operational conditions employed in
the hydrolytic treatments
In isothermal or non isothermal autohydrolysis treat-
ments, the joint eects of the main operational variables
(temperature and time) can be interpreted in terms of an
empirical variable proposed by Overend and Chornet
(1990), the severity factor R
0
:
R
0
=

t
0
exp
T
r
T
ref
!

dt (1)
where T
r
is the reaction temperature, T
ref
is the refer-
ence temperature, ! is an empirical constant and t time
in minutes. In the presence of externally added acid
(prehydrolysis treatments), a modied expression has
been proposed for the severity factor (now denoted R
0H
)
(Abatzoglou, Chornet, Belkacemi, & Overend, 1992;
Montane , Salvado , Farriol, Jollez, & Chornet, 1994):
R
0H
=

t
0
exp
X X
ref
l X
ref

exp
T T
ref
!
/

dt (2)
where X is the catalyst concentration (g acid/g sub-
strate), X
ref
is the reference concentration, and !
/
and l
are constants. The severity factor has been successfully
applied to the interpretation of hydrothermal hemi-
cellulose degradation, but also to related processes such
as deacetylation and solubilization of phenolics (Gar-
rote, Cruz, Dom nguez, & Parajo , 2003; Garrote,
Dom nguez, & Parajo , 2002).
For a given hydrolytic process (autohydrolysis or
prehydrolysis), the concentrations of phenolic acids and
the corresponding aldehydes (formed by the acid-cata-
lyzed degradation of the b-aryl-ether linkages of lignin
and their oxidative degradation), increase with the
192 G. Garrote et al. / Trends in Food Science & Technology 15 (2004) 191200
severity of treatments. At high severities, lignin can
undergo condensation reactions and/or react with
hemicellulose-degradation products (particularly with
furfural), leading to solid phases with increased contents
of Klason lignin (Kuznetsov et al., 1996).
Even though the interrelationships between severity
and lignin solubilization depend on the LCM
considered, some general trends exist. Figure 1
summarizes experimental data from literature con-
cerning the fraction of lignin solubilized in mild
Table 1. Phenolic compounds identied in liquors from LCM subjected to autohydrolysis (AU), acid or SO
2
impregnation (I), steam
explosion (SE), prehydrolysis (P) or alkaline wet oxidation (WO)
Type of LC/treatment Hydrolytic conditions:
T, time, [acid], (log Ro)
Identied phenolics Reference
Poplar wood/SE 205

C, 10 min p-Hydroxybenzoic acid, m-hydroxybenzoic acid,

vanillic acid, syringic acid, p-hydroxybenzaldehyde,
vanillin, syringaldehyde, cinnamic acid,
cinnamaldehyde, p-hydroxycinnamaldehyde
Ando et al. (1986)
Red oak wood/P 150

C, 60 min,
0.10.5% H
2
SO
4
Syringic acid, syringaldehyde,
vanillic acid, vanillin
Tran and Chambers
(1985, 1986)
Olive whole stone/SE 236

C, 2 min, 4.31 log R

0
Vanillic acid, vanillin, syringic acid,
syringaldehyde, tyrosol, hydroxytyrosol
Ferna ndez-Bolan os
et al. (1998)
Olive seed husk/SE, I 215229

C, 23 min,
00.1% H
2
SO
4
Vanillic acid, vanillin, syringic acid,
syringaldehyde, tyrosol, hydroxytyrosol
Ferna ndez-Bolan os
et al. (1998)
3.694.27 log R
0
Wheat straw/WO 195

C, 12 bar O
2
, 10 min Phenol, guaiacol, syringol, 4-hydroxybenzaldehyde,
vanillin, syringaldehyde, 4-hydroxy acetophenone,
acetovanillone, acetosyringone,
4-hydroxybenzoic acid, syringic acid
Klinke et al. (1998)
Wheat straw/WO 195

C, 12 bar O
2
, 10 min Acetosyringone, acetovanilone, syringic acid,
4-hydroxybenzaldehyde, vanillin, syringaldehyde,
4-hydroxy acetophenone, acetovanillone,
acetosyringone, 4-hydroxybenzoic acid,
syringic acid, vanillic acid
Klinke et al. (2001)
Spruce wood/SE 222

C, 7 min, 0.5% H
2
SO
4
Vanillin, dihydroconiferylalcohol,
coniferyl aldehyde, vanillic acid, hydroquinone,
catechol, acetoguaiacone, homovanillic acid,
4-hydroxybenzoic acid
Larsson et al. (1999)
Softwoods/SE, I 195

C, 2.38 min,
3.19% SO
2
Lignans, ferulic acid, syringic acid Boussaid et al. (2001)
Sugarcane bagasse/SE, I 205

C, 10 min, 0% SO
2
Benzoic acid, caeic acid, catechol,
ferulic acid, guaiacol, 4-hydroxybenzaldehyde,
4-hydroxybenzoic acid, p-coumaric acid,
protocatechuic acid, phenol,
syringaldehyde, vanillic acid, vanillin
Martn et al. (2002)
205

C, 10 min, 1.1% SO
2
205

C, 10 min, 1% H
2
SO
4
Willow wood/SE, I 205

C, 6 min Guaiacol, catecol, vanillin, 4-propylguaiacol,

4-hydroxybenzoic acid,
hydroxymethoxybenzoic acid, vanillic acid,
syringic acid, protocatechuic acid
Jo nsson et al. (1998)
Aspen wood/SE 214

C, 6 min Syringaldehyde, phenol, vanillyl alcohol,

2,6-dimethoxyphenol,
4-allyl, 2,6 dimethoxyphenol, benzaldehyde,
4-hydroxy3-methylvanillin, guaiacol
De Bari et al. (2002)
Eucalyptus globulus wood/AU 250

C, non isothermal Ferulic acid, gallic acid, syringaldehyde,

sinapaldehyde, vanillin
Garrote et al. (2003)
G. Garrote et al. / Trends in Food Science & Technology 15 (2004) 191200 193
hydrolytic treatments, showing the fractions of
depolymerized, water insoluble lignins that suered
chemical alteration during hydrolytic treatments
making them soluble in alkaline or ethanolic solutions,
respectively.
Alkali-extracted lignins from Eucalyptus regnans
wood subjected to steam explosion presented lower
syringaldehyde and vanillin contents at prolonged
autohydrolysis times, showing the participation of
acid-catalyzed condensation reactions which led to the
formation of carboncarbon bonds. In the range of
severities where lignin solubilization occurs, similar
elemental composition was found for steam exploded
wheat straw lignin. For the same material, the phenolic
hydroxyl groups were not signicantly aected by
severity, and the relative intensities of spectral bands
associated to guaiacyl and carbonyl groups decreased
only slightly with severity (Montane , Farriol, &
Salvado , 1998; Montane , Salvado , & Farriol, 1997).
Alkali-extracted lignin from olive seed husks showed
a guaiacyl-syringyl pattern similar to lignin isolates
from steam-exploded aspen and beech woods (Ferna n-
dez-Bolan os, Felizo n, Heredia, Guille n, & Jime nez,
1998). At low severities, the syringyl/guaiacyl ratio was
maintained, but cleavage of b-aryl ether linkages and
demethoxylation were observed under severe conditions.
Heitz, Wu, Lapointe, and Rubio (1995) reported the
formation of monomeric compounds (syringol and
guaiacol) and their decomposition into volatile products
(catechols) in alkali-extracted lignins from steam explo-
ded hardwoods. In the RASH (RApid Steam Hydro-
lysis) process, which involves continuous removal of
soluble and gaseous products, degradation was mini-
mized and a stable lignin content in the solid fraction
was observed (Schultz, McGinnis, & Biermann, 1984).
Similar ndings have been reported for ow through
treatments with hot compressed water (Ando et al.,
2000).
The presence of acid during steam explosion of olive
seed husks increased the yield in phenolics (Ferna ndez-
Bolan os et al., 1998). Using acid impregnation, the pat-
tern of phenolic compounds present in hydrolysates was
similar to that found for non-impregnated samples, but
the concentrations of the constituents were dierent
owing to the degradation of some aromatic compounds.
In comparison with the processing of non-impregnated
bagasse, Mart n et al. (2002) reported that H
2
SO
4
-
impregnation resulted in increased concentrations of
phenol and guaiacol (47 times) and benzoic and vanil-
lic acid (2 times), whereas the concentrations of p-cou-
maric acid, caeic acid and ferulic acid were 29 times
lower.
Fig. 1. Summary of reported data on the interrelationship between extent of lignin solubilisation and severity of treatments (measured by
the severity factor R
0
).
194 G. Garrote et al. / Trends in Food Science & Technology 15 (2004) 191200
Table 2. Antioxidant power (in decreasing order) of phenolic compounds found in LCM hydrolysates and commercial antioxidants
Assay Antioxidant power Reference
Data concerning the inhibition of oil or fatty acid oxidation in lipophilic phase, emulsions, liposomes or microsomes
Lipids a-Toc>FA>CA>CaA>GA>VA>>pCA>SA Osawa et al. (1987)
Rened olive oil HT>CaA>BHT>PCA>SA>pHBA>T>pCA>
oCA>-VA
Papadopoulus and
Boskou (1991)
Crude soy oil PCA>CA>CaA>pHBA>FA>VA>SA>pCA Onyeneho and
Hettiarachchy (1992)
Ethyl linoleate GA=Ca A>BHA>PCA>BHT>FA>V>VA Brand-Williams et al. (1995)
Sunower oil CaA>SpA>3,4-DHBA>FA Yanishlieva and Marinova (1995)
Linoleic acid a-Toc>>>CA me>pCA me>FA me>pCA-CA>FA Foti et al. (1996)
Olive oil triacylglycerols CA>FA>pCA Marinova and
Yanishlieva (1996)
Lard (Rancimat) CaA>PCA>BHT>a-Toc>pHBA>FA>pCA von Gadow et al. (1997)
b-Carotene-linoleic BHT>BHA>a-Toc>VA>FA>SA>pHBA>pCA>CaA von Gadow et al. (1997)
Corn starch-soybean oil V_CnA>BHT Camire and Dougherty (1998)
Rened olive oil GA>BHA>BHT>>VA Bonilla et al. (1999)
Methyl linoleate CaA -SpA>a-Toc>FA>2,3-dHBA>3HBA=VA Pekkarinen et al. (1999)
Linoleic acid a-Toc>BHT>BHA>Tlx>CaA>Gc>FA>pCA Cuvelier et al. (2000)
b-Carotene bleaching a-Toc>BHT>BHA>EA>GA>CaA>CA>FA Fukumoto and Mazza (2000)
Rabbit erythrocyte membrane oxidation BHA>aToc>SnpA me>SnpA>FA>VA>pCA>CnA Niwa et al. (2001)
Rat liver microsomal lipid CA>CaA>FA>aToc -AsA Kweon et al. (2001)
Egg yolk phosphatidylcholine CaA>FA esters>SpA>FA -pCA>GA esters>GA
>a Toc>LAsA
Kikuzaki et al. (2002)
Methyl linoleate CA>SA>aToc>BHT>FA esters>FA>pCA Kikuzaki et al. (2002)
Linoleic acid CA -aToc>SpA>FA>pCA Bratt et al. (2003)
b-Carotene bleaching BHA>BHT>GA>CA>FA>PCA>pCA>SA-VA Subba Rao and
Muralikrishna (2002)
Data concerning lipoprotein oxidation
Human low density lipoprotein oxidation CA>SA>pCA>pHB-VA>FA>SA Natella et al. (1999)
Low density lipoprotein oxidation PCA>CaA>pCA Satue -Gracia et al. (1999)
Human low density lipoprotein oxidation CaA -GA>SpA>pCA Andreasen et al. (2001)
Low density lipoprotein nitration CA-SnpA-VA-FA>pCA-Tlx>aToc>AsA>BHA-gToc
>bCar >CnA
Niwa et al. (2001)
Serum oxidation HT>GA>pCA Stupans et al. (2002)
Data concerning the inhibition of free radical formation and scavenging
DPPH
+
scavenging GA>CaA>PCA>BHA=BHT>Gc>AsA>FA>VA Brand-Williams et al. (1995)
DPPH
+
scavenging CaA>C>PA>SA>BHA>FA>BHT>VA>pCA>V von Gadow et al. (1997)
Lipid alkoxyl radical scavenging GA>CaA>CA>VA Mili et al. (1998)
DPPH
+
scavenging GA>CaA-NDGA>BHA>AsA Chung et al. (1999)
Free radical formation NDGA>GA>CaA>AsA>BHA Chung et al. (1999)
DPPH
+
scavenging 2,3-DHBA>SpA>CaA>a-Toc>FA>VA>3-HBA Pekkarinen et al. (1999)
Alkyl peroyl radical CA>V-VA-GA-a-Toc Sawa et al. (1999)
ABTS
++
scavenging HT>CaA>VA>T>V Benavente-Garca et al. (2000)
DPPH
+
scavenging SpA>aToc>HT>CaA>PCA>SA>VA-T Espn et al. (2000)
DPPH
+
scavenging EA>GA>CA>CaA>PCA>BHT>BHA>aToc>AsA
>FA>VA>pCA
Fukumoto and Mazza (2000)
DPPH
+
scavenging CA>CaA>FA>a-Toc>AsA Kweon et al. (2001)
DPPH
+
scavenging CaA>aToc>SpA>BHT>FA>pCa Kikuzaki et al. (2002)
DPPH
+
scavenging CaA esters>Tlx>CA>FA Son and Lewis (2002)
ROS formation in human liver microsomes GA>HT>3,4-DHBA>pCA>pGl >CaA Stupans et al. (2002)
ROS formation in rat liver microsomes HT>CaA>GA>3,4-DHBA>pCA Stupans et al. (2002)
DPPH
+
scavenging GA>PGl >HT>CaA>3,4-DHBA>pCA Stupans et al. (2002)
DPPH
+
scavenging SpA>CA -aToc>FA>pCA Bratt et al. (2003)
Others
DNA-damage in breast cancer cells CA esters>CA>FA Burdette et al. (2002)
AsA, ascorbic acid; BHA, butylhydroxyanisol; BHT, butylhydroxytoluene; CA, chlorogenic acid; CaA, caeic acid; CnA, cinnamic acid; m3,4-
DHB, methyl3,4-dihydroxybenzoate; 2,3, DHBA, 2,3dihydroxybenzoicacid; 3,4,DHBA, 3,4dihydroxybenzoicacid; FA, ferulicacid; GA, gallicacid; Gc,
guaiacol; HT, hydroxytyrosol; 3-HBA, 3-hydroxybenzoic acid; LasA, l-ascorbic acid; NDGA, nordihydroguaiaretic acid; oCA, o-coumaric acid;
pCA, p-coumaric acid, PCA, protocatechuic acid; pHBA, p-hydroxybenzoic acid; PGl, pirogallol; SA, syringic acid; SpA, sinapic acid; SnpA, sina-
pinic acid; T, tyrosol; Tlx, Trolox; a-Toc, a-tocopherol; VA, vanillic acid; V, vanillin; b-car, b-carotene; g-Toc, g-Tocopherol; me, methyl esters.
G. Garrote et al. / Trends in Food Science & Technology 15 (2004) 191200 195
Extensive lignin breakdown has been reported for wet
oxidation treatments (performed in wateroxygen
Na
2
CO
3
media). Up to 68.4% of lignin was solubilized,
and vanillin was the most abundant phenolic reaction
product. Even though the total phenolic content was
similar within a wide operational range, harsh condi-
tions resulted in generation of carboxylic acids from
phenols. Succinic, glycolic, lactic, malic, maleic, fuma-
ric, 2,3-dihydroxypropanoic and 2,4-dihydroxybutyric
acids were identied as reaction products (Klinke,
Schmidt, & Thomsen, 1998).
Antioxidant activity of phenolic fractions isolated
from hydrolysates
The antioxidant activity of phenolics present in
liquors from acidic processes is well known. Model
compounds such as benzoic acids and hydroxicinnamic
acids have been assayed for antioxidant activity using
dierent tests, but none of the available methods pro-
vides an absolute measurement of the phenomena
involved (Frankel & Meyer, 2000). Table 2 summarizes
data on the activity of model compounds, synthetic
antioxidants and natural antioxidants. Dierent
behaviour was observed depending on the chemical
structure and type of assay considered. Lignin mono-
mers and dimmers are eective antioxidants (Barclay,
Xi, & Norris, 1997; Kasprzycka-Guttman & Odzeniak,
1994). Simple phenolic acids (related to benzoic and
cinnamic acids) are absorbed and have a role in the
antioxidant defense (Natella, Nardini, Di Felice, &
Scaccini, 1999). Esters of phenolic acids are more active
than phenolic acids (Kikuzaki, Hisamoto, Hirose,
Akiyama, & Taniguchi, 2002; Son & Lewis, 2002),
whereas oligomers and condensed tannins are more active
than monomeric phenols (Hagerman et al., 1998). In
some cases, properties as taste and astringency could limit
some the food applications of this kind of compounds.
Table 3. Antimicrobial activity of phenolic compounds found in the aqueous phase from the mild acid-based hydrolysis of LCM. For
comparative purposes, data concerning sodium benzoate are also presented
Phenolic compound Microorganism (minimum inhibitory concentration; MIC, if available) Reference
Caeic acid Aspergillus avus (0.2 mg/ml); Aspergillus parasiticus (0.2 mg/ml);
Bacillus cereus (0.35 mg/ml); Escherichia coli (0.3 mg/ml);
Klebsiella pneumoniae (0.3 mg/ml); Micrococcus luteus;
Proteus vulgaris; Pseudomonas uorescens;
Salmonella typhimurium (0.35 mg/l); Staphilococcus aureus (0.4 mg/ml);
Streptococcus pneumoniae
Aziz et al. (1998); Chowdhury
et al. (1996); Tunc el and
Nergiz (1993)
p-Hydroxybenzaldehyde S. aureus; Staphylococcus epidermidis; Leuconostoc mesenteroides;
S. typhimurium; E. coli; Pseudomonas aeruginosa; Vibrio vulnicus
Eun et al. (2000)
p-Hydroxybenzoic acid A. avus (0.3 mg/ml); A. parasiticus (0.3 mg/ml); B. cereus (0.4 mg/ml);
E. coli (0.4 mg/ml); K. pneumoniae (0.4 mg/ml); S. typhimurium (0.4 mg/ml);
S. aureus (0.55 mg/ml)
Aziz et al. (1998); Tunc el and
Nergiz (1993)
p-Coumaric acid A. avus (0.3 mg/ml); A. parasiticus (0.3 mg/ml); B. cereus (0.4 mg/ml);
E. coli (0.4 mg/ml); K. pneumoniae (0.4 mg/ml); L. mesenteroides;
P. aeruginosa; S. aureus ( >0.6 mg/ml); S. epidermidis; S. typhimurium (0.4 mg/ml);
Saccharomyces cerevisiae (1 mg/l) ; V. vulnicus
Aziz et al. (1998); Eun
et al. (2000); Tunc el and
Nergiz (1993);
Baranowski et al. (1980)
Ferulic acid B. cereus (0.4 mg/ml); E. coli (0.45 mg/l); L. mesenteroides;
P. aeruginosa; S. aureus (0.6 mg/ml); S. epidermidis; S. typhimurium (0.4 mg/l);
S. cerevisiae (0.25 mg/l) ; V. vulnicus
Eun et al. (2000); Tunc el and
Nergiz (1993); Baranowski
et al. (1980)
Protocatechuic acid A. avus (0.3 mg/ml); A. parasiticus (0.3 mg/ml); B. cereus (0.4 mg/l);
E. coli (0.3 mg/ml); K. pneumoniae (0.3 mg/ml);
Legionella pneumophila (156.3 mg/ml); S. aureus (0.6 mg/ml);
S. typhimurium (0.4 mg/ml)
Aziz et al. (1998); Furuhata
et al. (2002); Tunc el and
Nergiz (1993)
Syringic acid A. avus (0.3 mg/ml); A. parasiticus (0.3 mg/ml); E. coli (0.4 mg/ml);
K. pneumoniae (0.4 mg/ml); S. aureus (0.6 mg/ml); B. cereus (0.4 mg/ml);
S. typhimurium (0.4 mg/l)
Aziz et al. (1998); Tunc el and
Nergiz (1993)
Vanillic acid A. avus (0.2 mg/ml); A. parasiticus (0.2 mg/ml); B. cereus (0.4 mg/ml);
E. coli (0.4 mg/ml); K. pneumoniae (0.4 mg/ml); L. mesenteroides;
P. aeruginosa; S. aureus (0.55 mg/ml); S. epidermidis; S. typhimurium (0.4 mg/ml);
V. vulnicus
Aziz et al. (1998); Eun
et al. (2000); Tunc el and
Nergiz (1993)
Vanillin A. avus (1.5 mg/l); A. niger (1.5 mg/l); Aspergillus ochraceus (1 mg/l);
A. parasiticus (1.5 mg/l); E. coli; L. mesenteroides; P. aeruginosa; S. aureus;
S. epidermidis; S. typhimurium; V. vulnicus
Lo pez-Malo et al. (1998);
Eun et al. (2000)
Sodium benzoate A. avus ( > 1 mg/l); Candida tropicalis (0.5 mg/l); Pichia chodati ( >1 mg/l);
Rhizopus senti (0.5 mg/l) Zygosaccharomyces japonicus ( >1 mg/l)
Jurd et al. (1975)
196 G. Garrote et al. / Trends in Food Science & Technology 15 (2004) 191200
Antimicrobial activity of the phenolic compounds
found in hydrolysates
Growing eorts are being devoted to the identica-
tion and isolation of natural antimicrobials, as a reac-
tion to consumers negative perception over synthetic
products. Although less potent than antibiotics, phe-
nolics from medicinal herbs, plants and spices are
interesting antimicrobial agents used alone or in combi-
nation with synthetic additives (Pszczola, 2002). Their
mode of action is concentration-dependent, and com-
paratively high concentrations (up to 2000 ppm) are
allowed. Table 3 summarizes relevant information on
the antimicrobial action of simple phenolics with che-
mical structures related to those present in hydrolysates
from mild-acid based processes. Phenols seem to aect
the membrane permeability, decreasing electron trans-
port and nutrients uptake, and possibly interfering with
the metabolic synthesis of macromolecules and nucleic
acids. The small size of the attached groups facilitates
their passage through the cell membrane. Even though
spores are not usually killed by preservatives, their
growth can be inhibited by phenols owing to their
ability to permeate the spore coat (Nichas, 1995). Both
the dissociation of the acid moiety and the presence of
one or more reactive double bonds contribute to the
antimicrobial eect. A linear relationship between
the number of carbon atoms in an alkyl chain and the
antimicrobial activity has been observed, whereas
the number of hydroxyl groups in the phenol group
and the oxidation level also increased toxicity (Cowan,
1999).
The inhibitory action of lignin depolymerization pro-
ducts (particularly low molecular weight phenolics) on
the growth and metabolism of yeasts during ethanol
production has been considered. These compounds are
more toxic than furfural and hydroxymethylfurfural
(Clark & Mackie, 1984). The inhibitory potential of the
functional groups attached to the benzene ring has been
estimated to grow from COOH, p-OH, CHO and
CH=CH, with no eect of m-OH, and an stimulating
eect of OCH
3
(Tran & Chambers, 1985, 1986), the
aromatic acids being less toxic than the corresponding
aldehydes. Synergistic inhibitory eects have been also
reported (Klinke, Thomsen, & Ahring, 2001).
Phenolic acids such as ferulic, coumaric, sinapic or
caeic add a pleasant avour to beverages and show
antimicrobial activity comparable or superior to those
of mixtures benzoic acid-sorbic acid (Cirigliano, Farrel,
McKenna, & Rothenberg, 2000). Mixtures of chloro-
genic acid, caeic acid, gallic acid and protocatechuic
acid showed antimicrobial action against Escherichia
coli, Salmonella typhimurium and Bacillus cereus
(Rodr guez de Sotillo et al., 1998). Rice hull lignins
showed antimicrobial action against gram(+) and
gram() bacteria (Eun, Lee, & Lee, 2000).
Health eects of phenolics
Phenolic compounds contribute to sensory properties
of foods, particularly colour and astringency. The
importance of healthy eects of phenolic compounds is
raising interest among scientists, food manufacturers
and consumers since a wide range of pharmacological
and biochemical activities (anticarcinogenic, anti-
atherogenic, antiinamatory) have been reported (see
Table 4). The benecial eects of dietary bre from
whole grain products has been related to the eects of
simple phenolic acids, benzoic acids and hydro-
xicinnamic acids (Andreasen, Landbo, Christensen,
Hansen, & Meyer, 2001; Kikuzaki et al., 2002). The
colonic microora transform avonoids into low mole-
cular weight phenolics, which might have protective
biological activities in the colon. Caeic acid and ferulic
acid are highly protective against nitrite ion, preventing
Table 4. Biological and therapeutic activities of simple phenolic compounds
Activity Phenolic compound Reference
Biological response modiers aecting the
autoimmune system
Gallic acid; caeic acid Nakagami et al. (1995)
Antiinamatory activity Protocatechuic acid; syringic acid; ferulic acid Ferna ndez et al. (1998)
Antitumor promoting eect Vanillin; vanillic acid; caeic acid Sawa et al. (1999)
Protection against LDL oxidation Hydroxycinnamates; protocatechuic acid;
caeic acid; p-coumaric
Heinonen et al. (1998);
Satue -Gracia et al. (1999)
Protection against LDL lipoprotein oxidation p-Coumaric acid; caeic acid L; synapic acid;
ferulic acid; protocatechuic acid
Meyer et al. (1998);
Zang et al. (2000);
Cartron et al. (2001)
Decrease in the proinamatory
lysophosphatidylcholine production
Caeic acid; synapic acid; ferulic acid;
protocatechuic acid
Cartron et al. (2001)
Inhibition of xanthine oxidase for treating
hyperuricemia causing gota and kidney stones
Caeic acid; ferulic acid Kweon et al. (2001)
G. Garrote et al. / Trends in Food Science & Technology 15 (2004) 191200 197
the formation of nitrosamines and nitrosamides in
foods and in vivo and they can inhibit or block the for-
mation of carcinomes (Krishnaswamy, 2001). Ferulic
acid esters are active agents in cosmetics by their anti-
oxidant and UV radiation absorption properties (Sei-
kou, Eisaku, & Takuo, 1999). Trans-cinnamic acid was
proposed for preventing and/or treating several diabetic
complications owing to its ability to cause inhibition of
aldose reductase, preventing the conversion of glucose
into sorbitol (Lee, 2002).
Conclusion
The chemical nature of phenolic byproducts solubi-
lized during the hydrolytic processing of LCM suggests
their suitability for food-related applications. Their
recovery and further application would provide com-
mercial value to a waste fraction resulting from the
detoxication and purication of sugar- or oligomer-
containing liquors. In order to allow an integrated
benet of the raw materials, the operational conditions
of the hydrolytic treatments must be carefully selected
to cause minimal cellulose degradation, achieving select-
ive hemicellulose depolymerization and producing
phenolics suitable for some of the purposes cited in this
work.
Acknowledgements
The authors are grateful to the Spanish Ministry of
Science of Technology and Xunta de Galicia for the
nancial support of this work (in the framework of the
Research Projects PPQ2000-0688-C05-02, PPQ2002-
00184 and PGIDT0138302PN). The Project PPQ2002-
00184 had partial nancial support from the FEDER
funds of the European Union.
References
Abatzoglou, N., Chornet, E., Belkacemi, K., & Overend, R. P. (1992).
Phenomenological kinetics of complex systems: the develop-
ment of a generalized severity parameter and its application to
lignocellulosics fractionation. Chemical Engineering Science, 47,
11091112.
Ando, S., Arai, I., Kiyoto, K., & Hanai, S. (1986). Identication of
aromatic monomers in steam-exploded poplar and their inu-
ences on ethanol fermentation by Saccharomyces cerevisiae.
Journal of Fermentation Technology, 64, 567570.
Ando, S., Sakaki, T., Kokusho, T., Shibata, M., Uemura, Y., & Hatate,
Y. (2000). Decomposition behaviour of plant biomass in hot-
compressed water. Industrial Engineering Chemistry Research,
39, 36883693.
Andreasen, M. F., Landbo, A. K., Christensen, L. P., Hansen, A

, &
Meyer, A. S. (2001). Antioxidant eects of phenolic rye (Secale
cereale L.) extracts, monomeric hydroxycinnamates, and
ferulic acid dehydrodimers on human Low-Density
Lipoproteins. Journal of Agricultural and Food Chemistry, 49,
40904096.
Aziz, N. H., Farag, S. E., Mousa, L. A., & Abo-Zaid, M. A. (1998).
Comparative antibacterial and antifungal eects of some phe-
nolic compounds. Microbios, 93, 4354.
Baranowski, J. D., Davidson, P. M., Nagel, C. W., & Branen, A. L.
(1980). Inhibition of Saccharomyces cerevisiae by naturally
occurring hydroxycinnamates. Journal of Food Science, 45,
592594.
Barclay, L. R. C., Xi, F., & Norris, J. Q. (1997). Antioxidant properties
of phenolic lignin model compounds. Journal of Wood Chem-
istry and Technology, 17, 7390.
Benavente-Garca, O., Castillo, J., Lorente, J., Ortun o, A., & del
R o, J. A. (2000). Antioxidant activity of phenolics extracted
from Olea europaea L. leaves. Food Chemistry, 68, 457462.
Bonilla, F., Mayen, M., Merida, J., & Medina, M. (1999). Extraction of
phenolic compounds from red grape marc for use as food lipid
antioxidants. Food Chemistry, 66, 209215.
Bouchard, J., Nguyen, T. S., Chornet, E., & Overend, R. P. (1991).
Analytical methodology for biomass pretreatment. Part 2: char-
acterization of the ltrates and cumulative product distribution
as a funcion of treatment severity. Bioresource Technology, 36,
121131.
Boussaid, A., Cai, Y., Robinson, J., Gregg, D. J., Nguyen, Q., & Sad-
dler, J. N. (2001). Sugar recovery and fermentability of hemi-
cellulose-hydrolysates from steam-exploded softwoods
containing bark. Biotechnology Progress, 17, 887892.
Brand-Williams, W., Cuvelier, M. E., & Berset, C. (1995). Use of a
free radical method to evaluate antioxidant activity. Lebensmit-
tel Wissenschaft und Technologie, 28, 2530.
Bratt, K., Sunnerheim, K., Bryngelsson, S., Fagerlund, A., Engman, L.,
Andersson, R. E., & Dimberg, L. H. (2003). Avenantramides in
oats (Avena sativa L.) and structure-antioxidant activity relation-
ships principles. Journal of Agricultural and Food Chemistry, 50,
70227028.
Burdette, J. A., Chen, S. N., Lu, Z. Z., Xu, H., White, B. E. P., Fabri-
cant, D. S., Liu, J., Fong, H. H. S., Farnsworth, N. R., Constantinou,
A. I., van Breemen, R. B., Pezzuto, J. M., & Bolton, J. L. (2002).
Black cohosh (Cimifuga racemosa L) protects against menadione-
induced DNA damage through scavenging of reactive oxygen
species: bioassay-directed isolation and characterization of
active principles. Journal of Agricultural and Food Chemistry, 50,
70227028.
Cadaha, E., Mun oz, L., Ferna ndez de Simo n, B., & Garca-Vallejo,
M. C. (2001). Changes in low molecular weight phenolic com-
pounds in Spanish, French and American oak woods during
natural seasoning and toasting. Journal of Agricultural and Food
Chemistry, 49, 17901798.
Camire, M. E., & Dougherty, M. P. (1998). Added phenolic com-
pounds enhance stability in extruded corn. Journal of Food Sci-
ence, 63, 516518.
Cartron, E., Carbonneau, M. A., Fouret, G., Descomps, B., & Leger,
C. L. (2001). Specic antioxidant activity of caeoyl derivatives
and other natural phenolic compounds: LDL protection against
oxidation and decrease in the proinamatory lysopho-
sphatidylcholine production. Journal of Natural Products, 64,
480486.
Chowdhury, B., Bhattacharyy, D., & Mukhopadhyay, S. (1996).
Antimicrobial eect of some simple and complex phenolic
compounds of olive fruits against pathogenic bacteria. Bio-
medical Letters, 54, 4549.
Chung, H. S., Chang, L. C., Lee, S. K., Shamon, L. A., van Breemen,
R. B., Mehta, R. G., Farnsworth, N. R., Pezzuto, J. M., & Kinghorn,
A. D. (1999). Flavonoid constituents of Chorizante diusa with
potential cancer chemoprotective activity. Journal of Agri-
cultural and Food Chemistry, 47, 3641.
Cirigliano, M. C., Farrel, F. J., McKenna, R. T., & Rothenberg, P.J.
(2000). Flavoring materials for use in tea containing beverages. US
Patent, US 6,022,576.
198 G. Garrote et al. / Trends in Food Science & Technology 15 (2004) 191200
Clark, T. A., & Mackie, K. L. (1984). Fermentation inhibitors in wood
hydrolysates derived from the softwood Pinus radiata. Journal of
Chemical Technology and Biotechnology, 34, 101110.
Conde, E., Cadahia, E., Garcia-Vallejo, M. C., & Tomas-Barberan, F.
(1995). Low molecular weight polyphenols in wood and bark of
Eucalyptus globulus. Wood Fiber Science, 27, 379383.
Cowan, M. M. (1999). Plant products as antimicrobial agents. Clin-
ical Microbiological Reviews, 12, 564582.
Cruz, J. M., Domnguez, J. M., Domnguez, H., & Parajo , J. C. (1999).
Solvent extraction of hemicellulosic wood hydrolysates: a pro-
cedure useful for obtaining both detoxied fermentation media
and polyphenols with antioxidant activity. Food Chemistry, 67,
147153.
Cuvelier, M. E., Bondet, V., & Berset, C. (2000). Behavior of phenolic
antioxidants in a partitioned medium: structure-activity
relationship. Journal of the American Oil Chemists Society, 73,
645652.
De Bari, I., Viola, E., Barisano, D., Cardinale, M., Nanna, F., Zimbardi,
F., Cardinale, G., & Braccio, G. (2002). Ethanol production at ask
and pilot scale from concentrated slurries of steam-exploded
aspen. Industrial Engineering Chemistry Research, 41, 17451753.
Domnguez, H., Torres, J. L., & Nu n ez, M. J. (2001). Antioxidant
phenolics as food additives from agricultural wastes. Poly-
phenols Actualites, 21, 2630.
Espn, J. C., Soler-Rivas, C., & Wichers, J. (2000). Characterization of
the total free radical scavenger capacity of vegetable oils and oil
fractions using DPPH radical. Journal of Agricultural and Food
Chemistry, 48, 648656.
Eun, J.B., Lee, M.J. & Lee., J.C. (2000). Antimicrobial activities of rice
hull lignins and their nitrobenzene-oxidized monomers (p. 58).
(abstract paper). American Chemistry Society, 220th
Felizo n, B., Ferna ndez-Bolan os, J., Heredia, A., & Guille n, R. (2000).
Steam-explosion pre-treatment of olive cake. Journal of the
American Oil Chemists Society, 77, 1522.
Ferna ndez, M. A., Saenz, M. T., & Garca, M. D. (1998). Anti-ina-
matory activity in rats and mice of phenolic acids isolated from
Scrophularia frutescens. Journal of Pharmacy and Pharmacology,
50, 11831186.
Ferna ndez-Bolan os, J., Felizo n, B., Heredia, A., Guille n, R., & Jime -
nez, A. (1998). Characterization of the lignin obtained by alkaline
delignication and of the cellulose residue from steam-exploded
olive stones. Bioresource Technology, 68, 121132.
Foti, M., Piattelli, M., Baratta, M. T., & Ruberto, G. (1996). Flavonoids,
coumarins, and cinnamic acids as antioxidants in a micellar
system. Structure-activity relationship. Journal of Agricultural
and Food Chemistry, 44, 497501.
Frankel, E. N., & Meyer, A. (2000). The problems of using one-
dimensional methods to evaluate multifunctional food and bio-
logical antioxidants. Journal of Science, Food and Agriculture, 80,
19251941.
Frazer, F. R., & McCaskey, T. A. (1989). Wood hydrolyzate treat-
ments for improved fermentation of wood sugars to 2,3-buta-
nediol. Biomass, 18, 3142.
Fukumoto, L. R., & Mazza, G. (2000). Assessing antioxidant and
prooxidant activities of phenolic compounds. Journal of Agri-
cultural and Food Chemistry, 48, 35973604.
Furuhata, K., Dogasaki, C., Hara, M., & Fukuyama, M. (2002). Inac-
tivation of Legionella pneumophila by phenol compounds con-
tained in coee. Bokin Bobai, 30, 291297.
Garrote, G., Domnguez, H., & Parajo , J. C. (2002). Interpretation of
deacetylation and hemicellulose hydrolysis during hydrothermal
treatments on the basis of the severity factor. Process Biochem-
istry, 37, 10671073.
Garrote, G., Cruz, J. M., Domnguez, H., & Parajo , J. C. (2003).
Valorisation of waste fractions from autohydrolysis of selected
lignocellulosic materials. Journal of Chemistry Technology and
Biotechnology, 78, 392398.
Hagerman, A. E., Riedl, K. M., Jones, G. A., Sovik, K. N., Ritchard,
N. T., Hartzfeld, P. W., & Riechel, T. L. (1998). High molecular
weight plant polyphenolics (tannins) as biological antioxidants.
Journal of Agricultural and Food Chemistry, 46, 18871892.
Heinonen, M., Meyer, A. S., & Frankel, E. N. (1998). Antioxidant
activity of berry phenolics on human Low-Density-Lipoprotein
and liposome oxidation. Journal of Agricultural and Food Chem-
istry, 46, 107112.
Heitz, M., Wu, G., Lapointe, J., & Rubio, M. (1995). Hydrolytic
depolymerization of a steam explosion lignin. Journal of Wood
Chemistry and Technology, 15, 515528.
Helm, R. F., Ranatunga, T. D., & Chandra, M. (1998). Lignin-hydro-
lyzable tannin interaction in wood. Journal of Agricultural and
Food Chemistry, 45, 31003106.
Jo nsson, I. J., Palmqvist, E., Nilvebrant, N. O., & Hahn-Ha gerdal, B.
(1998). Detoxication of wood hydrolysates with laccase and
peroxidase from the white-rot fungus Trametes versicolor.
Applied Microbial Biotechnology, 49, 691697.
Jurd, L., King, A. D., & Stanley, W. L. (1975). Cinnamyl phenol anti-
microbial agents. US patent 3,865,748.
Ka hko nen, M. J., Hopia, A. I., Vuorela, H. J., Rauna, J. P., Pihlaja, K.,
Kujala, T. S., & Heinonen, M. (1999). Antioxidant activity of plant
extracts containing phenolic compounds. Journal of Agricultural
and Food Chemistry, 47, 39543962.
Kasprzycka-Guttman, T., & Odzeniak, D. (1994). Antioxidant prop-
erties of lignin and its fractions. Thermochimica Acta, 231, 161
168.
Kikuzaki, H., Hisamoto, M., Hirose, K., Akiyama, K., & Taniguchi, H.
(2002). Antioxidant properties of ferulic acid and its related
compounds. Journal of Agricultural and Food Chemistry, 50,
21612168.
Klinke, H.B., Schmidt, A.S. & Thomsen, A.B. (1998). Identication of
degradation products from wheat straw in relation to pre-treat-
ment conditions. In Kopetz, H., Weber, T., Palz, W, Chartier, P.,
& Ferrero, G. L. (Eds.), Proc. biomass for energy and industry (pp.
484487).
Klinke, H. B., Thomsen, A. B., & Ahring, B. K. (2001). Potential inhi-
bitors from wet oxidation of wheat straw and their eect on
growth and ethanol production by Thermoanaerobacter
mathranii. Applied Microbiology Biotechnology, 57,
631638.
Krishnaswamy, K. (2001). Nonnutrients and cancer prevention.
ICMR Bulletin, 31. Available: http://www.icmr.nic.in
Kuznetsov, B. N., Efremov, A. A., Levdanskii, V. A., Kuznetsova, S.
A., Plezhayava, N. I., Shilkina, T. A., & Krotova, I. V. (1996). The
use of non isobaric pre-hydrolysis for the isolation of organic
compounds from wood and bark. Bioresource Technology, 58,
181188.
Kweon, M. H., Hwang, H. J., & Sung, H. C. (2001). Identication and
antioxidant activity of novel chlorogenic acid derivatives from
bamboo (Phyllostachys edulis). Journal of Agricultural and Food
Chemistry, 49, 46464655.
Larsson, S., Palmqvist, E., Hahn-Ha gerdal, B., Tenborg, C., Stenberg,
K., Zacchi, G., & Nilvebrant, N. O. (1999). The generation of fer-
mentation inhibitors during dilute acid hydrolysis of softwood.
Enzyme Microbial Technology, 24, 151159.
Lee, H. S. (2002). Inhibitory activity of Cinnamomum cassia
bark-derived component against rat lens aldose
reductase. Journal of Pharmacy and Pharmaceutical Sciences, 5,
226230.
Lo pez-Malo, A., Alzamora, S. M., & Argaiz, A. (1998). Eect of
natural vanillin on germination time and radial growth on
moulds in fruit-based agar systems. Food Microbiology, 12,
213219.
Marinova, E. M., & Yanishlieva, N. VI (1996). Antioxidative activity
of phenolic acids on triacylgycerols and fatty acid methyl esters
from olive oil. Food Chemistry, 56, 139145.
G. Garrote et al. / Trends in Food Science & Technology 15 (2004) 191200 199
Martn, C., Galbe, M., Nilvebrant, N.-O., & Jo nsson, L. F. (2002).
Comparison of the fermentability of enzymatic hydrolyzates of
sugarcane bagasse pretreated by steam explosion using dierent
impregnating agents. Applied Biochemistry and Biotechnology,
98100, 699716.
Meyer, A. S., Jepsen, S. M., & Srensen, N. S. (1998). Enzymatic
release of antioxidants for human low-density lipoprotein from
grape pomace. Journal of Agricultural and Food Chemistry, 46,
24392446.
Milic , B.Lj., Djilas, S. M., & C

anadanovic -Brunet, J. M. (1998). Anti-

oxidative activity of phenolic compounds on the metal-ion
breakdown of lipid peroxidation system. Food Chemistry, 62,
443447.
Montane , D., Salvado , J., Farriol, X., Jollez, P., & Chornet, E. (1994).
Phenomenological kinetics of woods delignication: application
of a time-dependent rate constant and a generalized severity
parameter to pulping and correlation of pulp properties. Wood
Science and Technology, 28, 387402.
Montane , D., Farriol, X., & Salvado , J. (1998). Fractionation of wheat
straw by steam-explosion pretreatment and alkali delignication.
Cellulose pulp and byproducts from hemicellulose and lignin.
Journal of Wood Chemistry and Technology, 18, 171191.
Montane , D., Salvado , J., & Farriol, X. (1997). Fractionation fo wheat
straw via steam-explosion pretreatment. Characteristics of the
lignin obtained by alkali delignication of the steamed straw.
Holzfoschung, 51, 135141.
Nakagami, T., Nanaumi-Tamura, N., Toyomura, K., Nakamura, T., &
Shigehisa, T. (1995). Dietary avonoids as potential natural bio-
logical response modiers aecting the autoimmune system.
Journal of Food Science, 60, 653656.
Natella, F., Nardini, M., Di Felice, M., & Scaccini, C. (1999). Benzoic
and cinnamic acid derivatives as antioxidants: structure-activity
relationship. Journal of Agricultural and Food Chemistry, 47,
14531459.
Nichas, G. J. E. (1995). Natural antimicrobials from plants. In: G.W.
Gould (Ed.), New methods of foos preservation. Glasgow: Blackie
Academic & Professional/Chapman and Hall.
Niwa, T., Doi, U., Kato, Y., & Osawa, T. (2001). Antioxidative prop-
erties of phenolic antioxidants isolated from corn steep liquor.
Journal of Agricultural and Food Chemistry, 49, 177182.
Ohta, T., Yamasaki, S., Egashira, Y., & Sanada, H. (1994). Anti-
oxidative activity of corn bran hemicellulose fragments. Journal
of Agricultural and Food Chemistry, 42, 653656.
Onyeneho, S. N., & Hettiarachchy, N. S. (1992). Antioxidant activity
of durum wheat bran. Journal of Agricultural and Food Chem-
istry, 40, 14961500.
Osawa, T., Ide, A., Su, J.-D., & Namiki, M. (1987). Inhibition of lipid
peroxidation by ellagic acid. Journal of Agricultural and Food
Chemistry, 35, 808812.
Overend, R. P., & Chornet, E. (1990). Heavy-oil cracking: the case
for nonhomogeneus kinetics. Canadian Journal of Physics, 68,
11051111.
Papadopoulus, G., & Boskou, D. (1991). Antioxidant eect of natural
phenols on olive oil. Journal of the American Oil Chemists
Society, 68, 669671.
Pekkarinen, S. S., Sto ckmann, H., Schwarz, K., Heinonen, M., &
Hopia, A. I. (1999). Antioxidant activity and partitioning of phe-
nolic acids in bulk and emulsied methyl linoleate. Journal of
Agricultural and Food Chemistry, 47, 30363043.
Pszczola, D. E. (2002). Antimicrobials: setting up additional hurdles
to ensure food safety. Food Technology, 56, 99107.
Rivas, B., Domnguez, J. M., Domnguez, H., & Parajo , J. C. (2002).
Bioconversion of posthydrolyzed autohydrolysis liquors: an
alternative for xylitol production from corncobs. Enzyme Micro-
bial Technology, 31, 431438.
Rodrguez de Sotillo, D., Hadley, M., & Wolf-Hall, C. (1998).
Potato peel extract a nonmutagenic antioxidant with
potential antimicrobial activity. Journal of Food Science, 63,
907910.
Rodrguez de Sotillo, D., Hadley, M., & Holm, E. T. (1994). Phenolics
in aqueous potato peel extract: extraction, identication and
degradation. Journal of Food Science, 5, 649651.
Satue -Gracia, M. T., Andre s-Lacueva, C., Lamuela-Ravento s, R. M.,
& Frankel, E. N. (1999). Spanish sparkling wines (cavas) as inhibi-
tors of in vitro human low-density lipoprotein oxidation. Journal
of Agricultural and Food Chemistry, 47, 21982202.
Sawa, T., Nakao, M., Akaike, T., Ono, K., & Maeda, H. (1999).
Alkylperoxyl radical-scavenging activity of various avonoids
and other phenolic compounds, implications for the anti-tumor-
promoter eect of vegetables. Journal of Agricultural and Food
Chemistry, 47, 397402.
Schultz, T. P., McGinnis, G. D., & Biermann, C. J. (1984). Similarities
and dierences in pretreating woody biomass by steam explo-
sion, wet oxidation, autohydrolysis, and rapid steam hydrolysis/
continuous extraction. In Energy from biomass and wastes VIII,
Lake Buena Vista, FL
Seikou, M., Eisaku, N., & Takuo, T. (1999). Ferulic acid eter anti-
oxidant/UV absorbent. US patent US 5908615.
Son, S., & Lewis, B. A. (2002). Free radical scavenging and anti-
oxidative activity of caeic acid amide and ester analogues:
structure-activity relationship. Journal of Agricultural and Food
Chemistry, 50, 468472.
Stupans, I., Kirlich, A., Tuck, K. L., & Hayball, P. J. (2002). Compar-
ison of radical scavenging eect, inhibition of microsomal oxy-
gen free radical generation, and serum lipoprotein oxidation of
several natural antioxidants. Journal of Agricultural and Food
Chemistry, 50, 24642469.
Subba Rao, M. V. S. S.T., & Muralikrishna, G. (2002). Evaluation of
the antioxidant properties of free and bound phenolic acids
from native and malted nger millet (Ragi, Eleusine coracana
Indaf-15). Journal of Agricultural and Food Chemistry, 50, 889
892.
Tran, A. V., & Chambers, R. P. (1985). Red oak wood derived inhi-
bitors in the ethanol fermentation of xylose by Pichia stipitis CBS
5776. Biotechnology Letters, 7, 841846.
Tran, A. V., & Chambers, R. P. (1986). Ethanol fermentation of red
oak acid prehydrolysate by the yeast Pichia stipitis CBS 5776.
Enzyme Microbial Technology, 8, 439445.
Tunc el, G., & Nergiz, C. (1993). Antimicrobial eect of some olive
phenols in a laboratory medium. Letters in Applied Microbiology,
17, 300302.
Va zquez, M. J., Alonso, J. L., Domnguez, H., & Parajo , J. C. (2001).
Xylooligosaccharides: Manufacture and Applications. Trends in
Food Science and Technology, 11, 387393.
von Gadow, A., Joubert, E., & Hansmann, C. F. (1997). Comparison
of the antioxidant activity of aspalathin with that of other plant
phenols of rooibos tea (Aspalathus linearis), a-tocopherol, BHT
and BHA. Journal of Agricultural and Food Chemistry, 45, 632
638.
Yanishlieva, N. VI, & Marinova, E. M. (1995). Eects of antioxidants
on the stability of triacylglycerols and methyl esters of fatty
acids of sunower oil. Food Chemistry, 54, 377382.
Zang, L. Y., Cosma, G., Gardner, H., Shi, X., Castranova, V., & Val-
lyathan, V. (2000). Eect of antioxidant protection by p-coumaric
acid on low-density lipoprotein cholesterol oxidation. American
Journal of Physiology, 279, C954C960.
200 G. Garrote et al. / Trends in Food Science & Technology 15 (2004) 191200