UV-spectrum was used to determine the purity of lignin and monitor the lignin distribution

among various tissues of lignocellulosic material with respect to the concentration. In this
study spectra the typical UV spectra of lignin fraction were illustrated in Fig. ,,,,,. The region
of UV spectrum was 200-700 nm. The nine lignin fractions showed the characteristic peaks
within 210 to 350nm, however all lignin fraction were demonstrated the absorption at
28010. First absorption at 23010 nm was found in the alkali lignin (PAL1, PAL2, PAL3,
and PAL4) and methanolic lignin (PML). The all lignin fractions were showed the shoulder
peaks in the PAL1, PAL2, PAL3, PAL4, PAL, PEL and PHL at 308nm, 310nm, 312nm,
311nm, 310nm, 304nm and 310nm respectively. The maximum absorbance at 280 nm could
be originated to non-conjugated phenolic groups in the lignin. The absorption at 280 nm,
corresponding to the * electronic transition in the aromatic ring of the unconjuated
phenolic units, is indicative of free and etherified hydroxyl groups. This result was consistent
with the relatively high proportion of guaiacyl units in lignin characterized with other
analysis. This result was consistent with the relatively high proportion
of guaiacyl units in lignin characterized with other analysis. Softwood lignin is mainly
composed of guaiacylpropane units and gives an absorption maximum at a wavelength of 280
nm, whereas hardwood lignin is a mixture of guaiacylpropane and syringylpropane units in
varying ratios resulting in a shifted peak maximum towards 278270 nm (Fergus and Goring,
1970a; Fukazawa and Imagawa, 1981; Fengel and Wegener, 1989; Takabe et al.,
1992; Koch and Kleist, 2001; Koch and Grunwald, 2004. The absorption at 32010
correspond to the n* transition in lignin units containing CO groups and -* transition
in lignin units containing CC linkages conjugated to the aromatic ring was indicative of
ferulic acid and p-coumaric acid is the main component(Wen et al., 2010). Absorbance at
21010 nm is assigned to the * in the aromatic ring (Li et al. 2010). In the table no
highest extinction coefficient was found at 277nm and 234nm of PAL1 and at 280nm and
235nm of PAL2, which was similar to tannic acid absorption extinction coefficient at 283nm
and 268nm.the extinction coefficient of lignin fractions were found highest in the absorption
at 280 nm rather than absorption at 320 nm. The extinction coefficient in the present study
was more than previously reported work Li et al 2010), The higher value of extinction
coefficients at 280 revealed that the lignin had more phenolic groups and C=O and C=C
linkages conjugated with the aromatic ring, which was in agreement with previous research
on MWL of bamboo (Tai et al. 1990) and other straws (Oliveira et al. 2009; Seca et al. 2000).

Effect of ionic liquid/organic solvent pretreatment on the enzymatic hydrolysis of

corncob for bioethanol production. Part 1: Structural

characterization of the
lignins

Shao-Ni Suna, Ming-Fei Li a, Tong-Qi Yuana, Feng Xua,, Run-Cang Suna,b,

Industrial Crops and Products 43 (2013) 570 577

Fig. 6 shows the optical transmittance spectra within the

UVvisible regions (200800 nm) on the films obtained from
copolymer and polymer blend
UV spectrum was used to determine the purity of lignin and
monitor the lignin distribution among various tissues of lignocellulosic
materials with respect to the concentration.
The maximum absorption at 280nm originates
from non-conjugated phenolic groups in the lignin. The presence of
a second characteristic region of lignin absorption around 318nm
can be assigned to the presence of both ferulic and p-coumaric
acids (Scalbert et al., 1986).

In this study,typical UV spectra of lignin fractions are illustrated in Fig. 2. The

six lignin fractions showed two characteristic absorptions at 283
and 320 nm. The first absorption at 283 nm could be assigned to
the nonconjugated phenolic groups in lignin. The second one at
320 nm, corresponded to the n_* transition in lignin units containing
C_ O group and __* transition in lignin units containing
C_ C_ linkages conjugated to the aromatic ring, was indicative of
ferulic and p-coumaric acids (Seca et al., 2000). Furthermore, if
the wavelength of the second absorption is shorter than 320 nm,
it means that esterified p-coumaric acid is the main component
(Wen et al., 2010). In this study, the second absorption of MWL is at
319 nm, consequently, it could be concluded that MWL contained
more esterified p-coumaric acid than etherified ferulic acid, which
was confirmed by the alkaline nitrobenzene oxidation results.
Meanwhile, it could further confirm that esterified p-coumaric
acid indeed existed in MWL. Comparing with AL and MWL,
lignin fractions prepared with EMIMAc and water/organic solvents
Fig. 2. UV spectra of lignin fractions (AL, MWL, LDMAc, LDMF, LDMSO, and LH2O).

pretreatments had lower absorption coefficients, which were in

the order of LDMSO > LDMAc > LDMF > LH2O. This was in agreement
with the results of carbohydrates analysis. The higher value of the
absorption coefficient of LDMSO revealed that LDMSO had more phenolic
groups and C_ O and C_ C_ linkages conjugated with the
aromatic ring (Seca et al., 2000). According to the results obtained
by Sun and Tomkinson (2002), the low absorption coefficient was
probably due to the relatively higher amounts of other co-extracted
non-lignin materials.

The UV spectra of bamboo lignin fractions showed similar bands to those

of other annual plants (Seca et al. 1998, 2000) (Fig. 2), which are
characterized by a sharp maximum at 205 nm, a shoulder at 235 nm, and
two low maxima at 282 and 312 nm, respectively. The absorption at 205
nm is assigned to the * transition in the aromatic ring. In addition, the
absorption at 282 nm, corresponding to the * electronic transition in
the aromatic ring of the unconjuated phenolic units, is indicative of free
and etherified hydroxyl groups. This result was consistent with the
relatively high proportion of guaiacyl units in lignin characterized with
other analysis. The absorption at 312 nm, assigned to the n* transition
in lignin units containing C=O groups and * transition in lignin units
with C=C linkages conjugated to the aromatic ring, is indicative of ferulic
and p-coumaric acids type structures (Sun et al. 2003), which was in
agreement with the FT IR spectra. The higher value of extinction
coefficients at 282 and 312 nm of 17.7 and 16.6 L g1 cm1 revealed that
the lignin had more phenolic groups and C=O and C=C linkages
conjugated with the aromatic ring, which was in agreement with previous
research on MWL of bamboo (Tai et al. 1990) and other straws (Oliveira et
al. 2009; Seca et al. 2000).
PEER-REVIEWED ARTICLE

bioresources.com

Li et al. (2010) . Characterization of bamboo lignin,

BioResources 5(3), 1762-1778.

1762

CHARACTERIZATION OF EXTRACTED LIGNIN OF BAMBOO

(NEOSINOCALAMUS AFFINIS) PRETREATED WITH SODIUM
HYDROXIDE/UREA SOLUTION AT LOW TEMPERATURE
Ming-Fei Li,a Yong-Ming Fan,a Run-Cang Sun,
The seven acid-insoluble lignin preparations exhibited
the basic UV spectrum of typical lignins with a maximum
at 280 nm, originating from the non-conjugated

a,b,*

and Feng Xu

a,c,*

phenolic groups (aromatic ring) in the lignin and being

known to be characteristic of a dominant guaiacyl lignin
[10]. Fig. 2 shows the UV spectra of the acidinsoluble
lignin fractions obtained by treatment of the
dewaxed barley straw with 1.5% H2O2 at pH 12.0 for 14
h at 45 _C under the extractant to straw ratios of 10:1
(spectrum F1), 18:1 (spectrum F4), 20:1 (spectrum F5),
and 30:1 (spectrum F7). Evidently, as shown in the
spectra, the absorption coefficient increased slightly with
the increment of extractant to straw ratios, indicating
that more ether linkages between lignin and hemicelluloses
could be cleaved when a relatively higher amount
of alkaline peroxide extractant was used. In contrast, the
relatively lower absorptions in the lignins fractions between
F1 and F4, extracted with low ratios of extractant
to straw, are presumed to due to the slightly higher
amounts of bound hemicelluloses and co-precipitated
other non-lignin materials such as ash and salt. The
much lower absorption at 310320 nm in all the lignin
preparations stated that the alkaline peroxide treatments
under the conditions given cleaved most of the ester or
ether linkages between lignin or hemicelluloses and hydroxycinnamic
acids, such as p-coumaric and ferulic
acids.

Structural and physico-chemical characterization of

lignins
solubilized during alkaline peroxide treatment of
barley straw
R.C. Sun

a,b,*,

X.F. Sun c, P. Fowler b, J. Tomkinson

The UV
spectrum of the guaiacyl compounds in chloroform exhibits a max at 268 nm and a min
at around 255 nm, which are in accordance with the values reported by Pew [21], and
that for the syringyl compounds is 271 and 250 nm respectively. The FTIR spectrum of
each component showed the presence of all the functional groups for guaiacyl and syringyl
compounds (Table 4) and also matches with the FTIR spectra for guaiacol and syringol of the
Aldrich FTIR library. Thus, by using a series of complementary techniques the separation
and complete identification of guaiacyl and syringyl groups of the walnut shell oil has been
attained. The S/G (syringyl/guaiacyl) ratio of the oil using the values from the GCMS
analysis of the initial petroleum-ether fraction was calculated to be 1.41. This value appears
to be well in line with that for hard wood lignin. Pyrolysis methods by far have proved to
be the most accurate methods to determine lignin content, and since the complete pyrolysis
of the walnut shells occurs during the extraction process of the oil, the lignin present is
completely converted to its markers, thus making this study representative for the lignin
content in the oil.

UV-Spectra of alkali (AL1, AL2 AL3 AL4) and acidic (AD) lignin with ligno-sulphate (LS)
as a standard are depicted in Fig. 1 (a). In the spectrum (Fig. 1a), the peaks were shown at the
region of 21010 nm and 28010 nm. UV-Spectra absorbance at 21010 nm is assigned to
the * in the aromatic ring and at 28010 nm is obtained to the * electronic
transition in the aromatic ring of the unconjugated phenolic units. It is indicative of free and
etherified hydroxyl group (Li et al., 2010). UV-Vis characteristic absorbance peaks of
polyphenol obtained from all samples is highlighted in Fig. 1(a).UV- Visible spectrum
absorbance of organosolve (AL, EL, ML, PL) and hot water lignin with ligno-sulphate as a
standard revealed in the Fig. 1(b). The spectrum of extracted lignin samples was
characterized by absorption at 230 nm and high intense peak at 280 nm. The maximum

absorbance at 280 nm originated due to the presence of non-conjugated phenolic group in the
lignin (She et al., 2010), and also * electronic transition in the aromatic ring of the
unconjugated phenolic units. After the study of UV-Spectra, high intense peak at 280 nm was
determined in organosolve lignin (which is responsible for electronic transition of nonconjugated aromatic ring). UV-Spectra of alkali (AL1, AL2 AL3 AL4) and acidic (AD) lignin
with ligno-sulphate (LS) as a standard are depicted in Fig. 1 (a). In the spectrum (Fig. 1a), the
peaks were shown at the region of 21010 nm and 28010 nm. UV-Spectra absorbance at
21010 nm is assigned to the * in the aromatic ring and at 28010 nm is obtained to the
* electronic transition in the aromatic ring of the unconjugated phenolic units. It is
indicative of free and etherified hydroxyl group (Li et al., 2010). UV-Vis characteristic
absorbance peaks of polyphenol obtained from all samples is highlighted in Fig. 1(a).UVVisible spectrum absorbance of organosolve (AL, EL, ML, PL) and hot water lignin with
ligno-sulphate as a standard revealed in the Fig. 1(b). The spectrum of extracted lignin
samples was characterized by absorption at 230 nm and high intense peak at 280 nm. The
maximum absorbance at 280 nm originated due to the presence of non-conjugated phenolic
group in the lignin (She et al., 2010), and also * electronic transition in the aromatic ring
of the unconjugated phenolic units. After the study of UV-Spectra, high intense peak at 280
nm was determined in organosolve lignin (which is responsible for electronic transition of
non-conjugated aromatic ring). UV-Spectra of alkali (AL1, AL2 AL3 AL4) and acidic (AD)
lignin with ligno-sulphate (LS) as a standard are depicted in Fig. 1 (a). In the spectrum (Fig.
1a), the peaks were shown at the region of 21010 nm and 28010 nm. UV-Spectra
absorbance at 21010 nm is assigned to the * in the aromatic ring and at 28010 nm is
obtained to the * electronic transition in the aromatic ring of the unconjugated phenolic
units. It is indicative of free and etherified hydroxyl group (Li et al., 2010). UV-Vis
characteristic absorbance peaks of polyphenol obtained from all samples is highlighted in
Fig. 1(a).UV- Visible spectrum absorbance of organosolve (AL, EL, ML, PL) and hot water
lignin with ligno-sulphate as a standard revealed in the Fig. 1(b). The spectrum of extracted
lignin samples was characterized by absorption at 230 nm and high intense peak at 280 nm.
The maximum absorbance at 280 nm originated due to the presence of non-conjugated
phenolic group in the lignin (She et al., 2010), and also * electronic transition in the
aromatic ring of the unconjugated phenolic units. After the study of UV-Spectra, high intense
peak at 280 nm was determined in organosolve lignin (which is responsible for electronic
transition of non-conjugated aromatic ring). UV-Spectra of alkali (AL1, AL2 AL3 AL4) and
acidic (AD) lignin with ligno-sulphate (LS) as a standard are depicted in Fig. 1 (a). In the
spectrum (Fig. 1a), the peaks were shown at the region of 21010 nm and 28010 nm. UVSpectra absorbance at 21010 nm is assigned to the * in the aromatic ring and at 28010
nm is obtained to the * electronic transition in the aromatic ring of the unconjugated
phenolic units. It is indicative of free and etherified hydroxyl group (Li et al., 2010). UV-Vis
characteristic absorbance peaks of polyphenol obtained from all samples is highlighted in
Fig. 1(a).UV- Visible spectrum absorbance of organosolve (AL, EL, ML, PL) and hot water
lignin with ligno-sulphate as a standard revealed in the Fig. 1(b). The spectrum of extracted
lignin samples was characterized by absorption at 230 nm and high intense peak at 280 nm.
The maximum absorbance at 280 nm originated due to the presence of non-conjugated
phenolic group in the lignin (She et al., 2010), and also * electronic transition in the
aromatic ring of the unconjugated phenolic units. After the study of UV-Spectra, high intense
peak at 280 nm was determined in organosolve lignin (which is responsible for electronic
transition of non-conjugated aromatic ring). UV-Spectra of alkali (AL1, AL2 AL3 AL4) and
acidic (AD) lignin with ligno-sulphate (LS) as a standard are depicted in Fig. 1 (a). In the
spectrum (Fig. 1a), the peaks were shown at the region of 21010 nm and 28010 nm. UVSpectra absorbance at 21010 nm is assigned to the * in the aromatic ring and at 28010

nm is obtained to the * electronic transition in the aromatic ring of the unconjugated

phenolic units. It is indicative of free and etherified hydroxyl group (Li et al., 2010). UV-Vis
characteristic absorbance peaks of polyphenol obtained from all samples is highlighted in
Fig. 1(a).UV- Visible spectrum absorbance of organosolve (AL, EL, ML, PL) and hot water
lignin with ligno-sulphate as a standard revealed in the Fig. 1(b). The spectrum of extracted
lignin samples was characterized by absorption at 230 nm and high intense peak at 280 nm.
The maximum absorbance at 280 nm originated due to the presence of non-conjugated
phenolic group in the lignin (She et al., 2010), and also * electronic transition in the
aromatic ring of the unconjugated phenolic units. After the study of UV-Spectra, high intense
peak at 280 nm was determined in organosolve lignin (which is responsible for electronic
transition of non-conjugated aromatic ring). UV-Spectra of alkali (AL1, AL2 AL3 AL4) and
acidic (AD) lignin with ligno-sulphate (LS) as a standard are depicted in Fig. 1 (a). In the
spectrum (Fig. 1a), the peaks were shown at the region of 21010 nm and 28010 nm. UVSpectra absorbance at 21010 nm is assigned to the * in the aromatic ring and at 28010
nm is obtained to the * electronic transition in the aromatic ring of the unconjugated
phenolic units. It is indicative of free and etherified hydroxyl group (Li et al., 2010). UV-Vis
characteristic absorbance peaks of polyphenol obtained from all samples is highlighted in
Fig. 1(a).UV- Visible spectrum absorbance of organosolve (AL, EL, ML, PL) and hot water
lignin with ligno-sulphate as a standard revealed in the Fig. 1(b). The spectrum of extracted
lignin samples was characterized by absorption at 230 nm and high intense peak at 280 nm.
The maximum absorbance at 280 nm originated due to the presence of non-conjugated
phenolic group in the lignin (She et al., 2010), and also * electronic transition in the
aromatic ring of the unconjugated phenolic units. After the study of UV-Spectra, high intense
peak at 280 nm was determined in organosolve lignin (which is responsible for electronic
transition of non-conjugated aromatic ring).UV-Spectra of alkali (AL1, AL2 AL3 AL4) and
acidic (AD) lignin with ligno-sulphate (LS) as a standard are depicted in Fig. 1 (a). In the
spectrum (Fig. 1a), the peaks were shown at the region of 21010 nm and 28010 nm. UVSpectra absorbance at 21010 nm is assigned to the * in the aromatic ring and at 28010
nm is obtained to the * electronic transition in the aromatic ring of the unconjugated
phenolic units. It is indicative of free and etherified hydroxyl group (Li et al., 2010). UV-Vis
characteristic absorbance peaks of polyphenol obtained from all samples is highlighted in
Fig. 1(a).UV- Visible spectrum absorbance of organosolve (AL, EL, ML, PL) and hot water
lignin with ligno-sulphate as a standard revealed in the Fig. 1(b). The spectrum of extracted
lignin samples was characterized by absorption at 230 nm and high intense peak at 280 nm.
The maximum absorbance at 280 nm originated due to the presence of non-conjugated
phenolic group in the lignin (She et al., 2010), and also * electronic transition in the
aromatic ring of the unconjugated phenolic units. After the study of UV-Spectra, high intense
peak at 280 nm was determined in organosolve lignin (which is responsible for electronic
transition of non-conjugated aromatic ring).

Separation and characterization of lignin

compounds from the walnut (Juglans regia) shell oil
using preparative TLC, GCMS and 1H NMR
E.V. Mathias a,b, U.P. Halkar a
Name of
the
APL
APL1

Wavelength(
nm)
266
310
277

(LM-1cm-1)
111.2
84.4
328.3

APL2
PAL3
PAL4
PEL
PHL
PPL
TA
PML

311
234
280
318
235
282
318
214
280
319
214
268
320
272
312
272
283
269
226
276
225

270.6
378.3
289.7
251.5
391.4
144.3
137.4
148.8
125.1
132.6
85.8
112.2
79.8
82.9
396.0
397.6
106.0

Fourier transform infrared spectroscopy has been proven to be a useful approach to study
physicochemical and conformational properties of lignin in which various functional groups
and structural fragments can be characterized. The FT-IR spectra of the acid-insoluble lignin
fractions PAL1, PAL2, APL3 and APL4, organosolve lignin PAL, PEL, PML and PPL and hot
water lignin (HL) are shown in Fig. 3(a) .Lignin contains various types of functional groups
depending on the wood species and isolation procedure. Softwood lignin, often referred to as
guaiacyl lignin is primarily comprised of coniferyl alcohol units, which make up more than
95% of the structural units in the lignin, with the remainder consisting mainly of p-coumaryl
alcohol-type units. Softwood lignin, often referred to as guaiacyl lignin is primarily
comprised of coniferyl alcohol units, which make up more than 95% of the structural units in
the lignin, with the remainder consisting mainly of p-coumaryl alcohol-type units. Spectral
differences between different extracted paddy lignin were observed in the fingerprint region
(1800 and 800 cm-1). The major peaks in the Fig. ,,,,,,,, of all samples show up in the spectra
were the broad bands at near about 3426cm1, as attributed to hydroxyl groups in phenolic
and aliphatic structures, and all samples spectrum band ( Table no. 1) at 2970-2916 were
predominantly arising from C-H stretching in the aliphatic C-H (Sun et al., 2012).The
aromatic methoxyl groups was assigned at 2847 cm-1 (Kubo S., and Kadla, K. F., 2005)
which was shown in PEL and PPL of organosolve lignin. Centred between 2925 and
2858cm1 predominantly arising from CH stretching in aliphatic CH and aromatic
methoxyl groups. In previous study the bands at 2933 and 2853 cm1were aroused from the
CH symmetrical and asymmetric vibrations in methyl and methylene groups (Labidi et al.
2009). Bands in the 1714-1715 cm-1 shown by PAL2 and PAL can be attributed to nonconjugated carbonyl groups (Tejado et al., 2007). The band at 1634 cm-1, 1645cm-1, 1646
cm-1, 1647cm-1 and 1642cm-1 corresponding to PW4, PW2, PW3, PAL4 and PHL were
attributed to conjugated stretching in lignin (Kubo S., and Kadla, K. F., 2005). Aromatic
skeletal vibration and C=O stretching were assigned at 1516 cm-1,1514 cm-1, 1515 cm-

1,1514 cm-1,1592 cm-1, 1594 cm-1, 1623 cm-1 and 1593 cm-1 of PW4, PW2,PAL2, PAL4,
PHL, PEL, PAL and PPL respectively (Tejado et al., 2007). In the alkali treated paddy and
methanolic extracted lignin were not shown any band at this region due to?????? . The strong
band at 1455 cm-1, 1466 cm-1, 1463 cm-1, 1458 cm-1 and 1454 cm-1 of PW1, PAL2, PAL4,
PEL and PPL were attributed to the asymmetric vibration (asymmetric in methyl, methylene
and methoxy groups) (Jahan et al., 2007). The band at near 1425 cm-1 was shown in all
samples except untreated paddy straw, which was originated by C-H in plane deformation in
the guaiacyl ring. In the various previous study softwood lignins also called as guaiacyl lignin
(Kubo S., and Kadla J. F., 2005). The guaiacyl lignin unit was comprised of coniferyl alcohol
unit, which build up more than 95% of structural unit of softwood lignin (Kubo S., and Kadla
J. F., 2005) The result of FTIR of the present work was concluded that the softwood lignin
was present in the extract. The band at 1322 cm-1 was shown in the treated and ethanol
treated paddy straw , which was originated by C-O stretching of the syringyl ring whereas the
any band at this region was not found in extracted paddy straw lignin samples (Sun et al.,
1996). The result of present work was correlate with previous study, the softwood lignin
consist largely guaiacylpropane units, wheras the hardwood lignin were madeup both
guaiacyl and syringylpropane units. The absence of syringyl unit in the softwood lignin or
paddy lignin was also evidence from presence of band at 1425 cm-1, which was attributed
for C-H plane deformation in guaiacyl ring. The band at 1269 cm-1 was assigned for C-O
stretching of guaiacyl ring which was found in PEL and PPL. The result was correlated with
total polyphenol content of paddy lignin. The ethanolic and propanolic lignin was containing
more TPC than other extract of lignin samples. 1272 aryl-O of aryl-OH and aryl-O-CH3
guaiacyl/syringyl ring (with C=O group) mode 1264 cm-1, 1263 cm-1, PEL, PPL. The
spectrum at 1151 cm-1 was found in treated, untreated paddy straw, PAL4 and PEL extracted
lignin, which was originated in presence of aromatic C-H in plane deforming in guaiacyl ring.
Aromatic C-H in plane deformation in the
Syringyl ring at 1116 cm-1was shown in PAL2, PAL and PML. Lignin is synthesize by
enzymatic polymerisation of three monomers that are coniferyl, synapyl alcohol and pcoumaryl alcohol that lead, respectively to guaiacyl (G), syringyl (S) and p-hydroxyphenyl
propane (P-H) type units (Tejado et al., 2007).the band at 1043 cm-1, 1033 cm-1, 1047 cm-1,
1045 cm-1 and 1044 cm-1 were found in the PW1, APL4, PHL, PPL and PML respectively,
which was attributed from aromatic C-H in plane deformation higher in guaiacyl than
syringyl unit. Furthermore the spectrum at 900 cm-1, 899.20 cm-1, 899 cm-1, 833 cm-1, 878
cm-1, 845 cm-1, 886 cm-1, 768 cm-1 and 839 cm-1of PW4, PW1, PW2, PAL2, PAL4, PHL,
PEL, PAL, PPL and PML were assigned to CH-CH bending and C-H bending of syringyl
units (Stevanovic, T., et al 2014). The result was concluded that ethanol extracted lignin was
containing all the band of guaiacyl, which are major composition of softwood lignin. The
study was proved that the ethanolic lignin was containing major bioactive component than
others.

Evaluation of industrial lignins for biocomposites production

Diane Schorra, Papa Niokhor Dioufa,b, Tatjana Stevanovica,
Industrial Crops and Products 52 (2014) 65 73

Hydrogen Bonding in Lignin: A Fourier Transform Infrared

Model Compound Study
Biomacromolecules 2005, 6, 2815-2821 2815

Satoshi Kubo and John F. Kadla*

Lignin is created by enzymatic polymerisation of three monomers, called coniferyl

alcohol, synapyl alcohol and p-coumaryl
alcohol that lead, respectively, to guaiacyl (G), syringyl
(S) and p-hydroxyphenyl propane (p-H)-type units

111
6

Aromatic CH
deformatio
n in the
syringyl
ring

115
1

Aromatic
C-H in
plane
deforming
in
guaiacyl
ring

1106.
38

1164.1
3

1161.
37

1161.
48

1100.
23

1123.
76

1094.6
7

1123.
47

The strong band at 1461 cm1 is attributed to the CH asymmetric

vibrations (asymmetric in methyl, methylene, and methoxyl
groups) (Jahan et al., 2007).The

1516.16

band at

1514.5 1515.3
2
4
1591.5 1594.0 1623.1 1593.0
0
1
5
6
1603 Aromatic skeletal
vibration + C=O
stretching

1514.5
2

1511.4
6

Aromatic
skeleton vibrations in the lignin preparations are
assigned at 1600, 1509, and 1422 cm_1.

Antioxidant property
Antioxidant activity is known as the capacity of the compound, which inhibit the oxidation
degradation of peroxidation of any compound like a lipid (Roginsky V., and Lissi, E. A.,
2005). The scavenging and reducing properties of the extracted paddy lignin were evaluated
through DPPH, ABTS, FRAP and H2O2 assays. Table 3, indicating that most extracts display
significant antioxidant properties by four methods tested. It must also be noted that the
antioxidant activities assessed are in direct relation with the polyphenolic content of the
extract, as in result TPC.
There were various types of assessment of antioxidant activities. The maximum assay based
on the electron transfer and hydrogen atom donation reactions. The electron transfer mostly
attributed to DPPH, as can say that the quenching of DPPH radical to form DPPH-H is also
possible. Some others methods also based on the electron transfer such as TPC assay using
Folin-Ciocalteu reagent, ABTS+ decolourization, H2O2 reducing power and ferric ion
reducing antioxidant power (FRAP). Following above mentioned methods were applied for

the assessments of scavenging and reducing power of extracted paddy straw lignin in our
present study
Radical scaveDPPH+ scavenging activity
DPPH_ assay is mainly attributed to the electron transfer assays,however the quenching of
DPPH_ radical to form DPPH-H is also possible
There are many assays for the assessment of antioxidant properties,
the majority of them are based on electron transfer and hydrogen
atom donation reactions. After comprehensive critical
assessment of the most frequently used methods, Huang, Ou, and
Prior (2005) concluded that ORAC, TPC measured with FolinCiocalteu
reagent and one of the electron/hydrogen transfer assays should
be recommended for representative evaluation of antioxidant properties.
DPPH_ assay is mainly attributed to the electron transfer assays,
however the quenching of DPPH_ radical to form DPPH-H is
also possible. Other electron transfer based methods include the
TPC assay using FolinCiocalteu reagent, ABTS_+ decolourisation assay
and ferric ion reducing antioxidant power (FRAP). Following the
above mentioned recommendation, all these methods were applied
for the assessment of V. opulus antioxidant potential in our study

The antioxidant properties of the stem extracts were evaluated

through DPPH_ and FRAP assays. The results _and their respective
TPC values_ are included in Table 3, indicating that most extracts
display significant antioxidant properties by both methods tested.
These results are in agreement with literature reports on the
antioxidant activities of grape stem extracts (Makris et al., 2007a;
Spigno & De Faveri, 2007). It must also be noted that the antioxidant
activities assessed are in direct relation with the polyphenolic
content of the extract, as is delineated in Fig. 2 (A&B). Finally, a good
correlation coefficient was found between the FRAP assay and
DPPH radical scavenging assay (R 0.846, Fig. 2C).

Grape stem extracts: Polyphenolic content and assessment of

their in vitro
antioxidant properties

Maria Anastasiadi a, Harris Pratsinis b, Dimitris Kletsas b, Alexios-Leandros Skaltsounis

c,
Serkos A. Haroutounian
LWT - Food Science and Technology 48 (2012) 316e322

Antioxidant activity is defined as the ability of a compound to

inhibit oxidative degradation like lipid peroxidation (Roginsky
and Lissi, 2005). The ferric thiocyanate method measures the
amount of peroxide produced during the initial stages of oxidation
which are the primary products of oxidation. LAEP exhibited
effective antioxidant activity in the linoleic acid emulsion system.
The effect of 30 lg/mL LAEP on lipid peroxidation of a linoleic
acid emulsion is shown in Table 3 and Fig. 3, and was
found to be 93.2%. On the other hand, BHA, BHT, a-tocopherol
and trolox exhibited 83.3%, 82.1%, 68.1% and 81.3% peroxidation
of linoleic acid emulsion at the same concentration, respectively.

Peroxidation of linoleic acid emulsion without LAEP or standard

compounds was accompanied by a rapid increase in peroxides.
Consequently, these results clearly indicate that LAEP had effective
and potent antioxidant activity in the ferric thiocyanate
assays.
Furthermore, LAEP had effective reducing power determined by
using the potassium ferricyanide reduction and cupric ions (Cu2+)
reducing methods when compared to the standards. To measure
the reductive ability of LAEP, Fe3+Fe2+ transformation was investigated
in the presence of LAEP using the method of Oyaizu
(1986). As can be seen in Table 3, LAEP (r2: 0.918) demonstrated
powerful Fe3+ reducing ability with statistically significant differences
(p < 0.01). The reducing power of LAEP, BHA, BHT, a-tocopherol
and trolox increased steadily with increasing concentration of
samples. The reducing power of LAEP and the standard compounds
were as follows: BHA > BHT _ a-tocopherol > trolox > LAEP. The
results demonstrated that LAEP had marked ferric ions (Fe3+)
reducing ability and electron donor properties for neutralizing free
radicals by forming stable products. However, this reducing power
was lower than that of the standard antioxidants used. The outcome
of the reducing reaction is to terminate the radical chain
reactions that may otherwise be very damaging