Carbohydrate Polymers 70 (2007) 218223

www.elsevier.com/locate/carbpol

Lignin recovery from brewers spent grain black liquor

Solange I. Mussatto *, Marcela Fernandes, Ines C. Roberto
Departamento de Biotecnologia, Escola de Engenharia de Lorena, Universidade de Sao Paulo, Lorena/SP, Brazil

Received 26 February 2007; received in revised form 23 March 2007; accepted 29 March 2007
Available online 14 April 2007

Abstract

The present study describes the precipitation of lignin by acidication of a black liquor (pH 12.56 and 12.44 g/l soluble lignin)
produced by soda pulping of brewers spent grain. Sulfuric acid was added to the liquor to decrease the pH, forming a lignin-rich
precipitate. Ten pH values (varying from 12.56 to 2.15) were studied. The lignin mass precipitated for each pH condition was determined,
and the obtained liquors were evaluated regarding the color and concentration of soluble lignin. Some phenolic acids (vanillic, syringic,
p-hydroxybenzoic, ferulic and p-coumaric) were quantied in the liquors to verify their removal proles as a function of the pH
alteration. Signicant lignin precipitation was only observed at pH < 7.7. At pH 2.15 the concentration of soluble lignin was reduced
to 2.31 g/l (removal of 81.43%) and the color of the liquor was strongly modied from dark brown to pale yellow. The phenolic
compounds concentration was reduced in dierent proportions (from 74.4% to 32.1%), suggesting that each lignin derived compound
is dierently aected by the pH alteration.
 2007 Elsevier Ltd. All rights reserved.

Keywords: Acidication; Brewers spent grain; Black liquor; Lignin; Precipitation; Soda pulping

1. Introduction of agricultural residues, such as cereal straws of wheat, rice,

and barley (Hammett, Youngs, Sun, & Chandra, 2001).
Lignocellulosics are the main raw material for pulp and The major constituents of lignocellulosics are cellulose,
paper industries. Among these, the woods are the most hemicellulose, and lignin, polymers that are associated with
important, corresponding to 95% of all raw materials used each other constituting the cellular complex of the vegetal
by these industries (Jimenez, Ramos, Rodrguez, de la biomass. Lignin is a three-dimensional polyphenolic mac-
Torre, & Ferrer, 2005). Nevertheless, some agricultural or romolecule of very complex structure whose function is
agro-industrial residues such as brewers spent grain, maize to provide rigidity and cohesion to the material cell wall,
stems, and some cereal straws can also be used for this pur- to confer water impermeability to xylem vessels, and to
pose (Mussatto, Dragone, Rocha, & Roberto, 2006; form a physicchemical barrier against microbial attack
Navaee-Ardeh, Mohammadi-Rovshandeh, & Pourjoozi, (Fengel & Wegener, 1989). An example of structure pro-
2004; Xiao, Sun, & Sun, 2001). Developing countries, par- posed for the lignin is given in Fig. 1 (Adler, 1977).
ticularly those in Asia, which do not have adequate wood For the lignocellulosic materials be susceptible to the
supplies use straws and other non-wood materials as ligno- pulp and paper production, the separation of their cellu-
cellulosic ber sources for pulp and paper production losic bers is necessary. Several processes have been used
(Lawther & Sun, 1996). China, for example, has a long tra- with this nality, and the soda pulping is one of the most
dition for using non-wood raw materials for pulp and relevant and most used for delignication of agricultural
paper due to its limited forest resources and rich supply residues (Iglesias, Bao, Lamas, & Vega, 1996). During the
pulping process, the lignin is dissolved from the raw mate-
*
Corresponding author. Tel.: +5512 3159 5027; fax: +5512 3153 3165.
rial, being separated in the form of a liquor rich in phenolic
E-mail addresses: solange@debiq.eel.usp.br, solange@debiq.faenquil. compounds that represents the process euent (Fengel &
br (S.I. Mussatto). Wegener, 1989). This euent is dark brown or black in

0144-8617/$ - see front matter  2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.carbpol.2007.03.021
 S.I. Mussatto et al. / Carbohydrate Polymers 70 (2007) 218223 219

Fig. 1. Structure proposed for the lignin of conifer according to Adler (1977).

color and hence is called black liquor. The black liquor by-product. BSG is a raw material recently evaluated for
contains a wide variety of compounds (chemicals and cellulose pulp production (Mussatto et al., 2006), and until
derived from lignin), most of them are toxic and need to now, there is not yet any work about the lignin recovery
be removed before the euent discharge to the environ- from the black liquor produced during its pulping. This
ment (Karam & Nicell, 1997). Aiming to decrease the envi- report is thus the rst paper concerning the studies in this
ronmental impact, the pulp and paper industries normally area and has as objective the evaluation of the pH inuence
burn the lignin dissolved in the black liquor for energy gen- on lignin precipitation.
eration. However, the lignin can be isolated and used as the
starting material for a series of useful products, such as 2. Materials and methods
vanillin, phenols, benzene, dispersant, emulsicant and
chelant agents, antioxidants, pesticides, fertilizers, vegetal 2.1. Raw material
charcoal, polymers, adhesives, concrete additives, compo-
nent for resins, among others (Gargulak & Lebo, 2000). The BSG used in the experiments was kindly supplied by
Conventionally, the Kraft lignin has been recovered from the Microbrewery of the Engineering College of Lorena,
black liquor, in laboratory or industrial plants, by precipi- and presented the following chemical composition (% dry
tation with mineral acids like sulfuric or hydrochloric acid, weight, w/w): 16.8% cellulose, 28.4% hemicellulose, 27.8%
followed by ltration and washing, the last one being the lignin, 4.6% ashes, 15.2% proteins, 5.8% extractives, and
critical stage for obtainment of a lignin with high purity 1.4% acetyl groups. As soon as obtained, the material
(Santos & Curvelo, 2001). was washed with water (for removal of the residues pro-
In the present work, a black liquor was obtained by soda ceeding from wort) and dried at 50 5 C until approxi-
pulping of brewers spent grain (BSG), the main brewery mately 10% moisture content.
220 S.I. Mussatto et al. / Carbohydrate Polymers 70 (2007) 218223

Table 1
Volume of concentrated sulfuric acid added in each sample of the brewers spent grain black liquor, characterization of the obtained liquor (pH and
soluble lignin), and quantication of the precipitated lignin mass
Sample H2SO4 (ml) pH Soluble lignin (g/l) Lignin removal (%) Precipitated lignin mass (g) Yielda (g/100 ml)
1 original 12.56 12.44 0.0301 0.1505
2 0.14 10.61 12.42 0.16 0.0361 0.1805
3 0.16 9.90 12.40 0.32 0.0373 0.1865
4 0.18 7.71 12.40 0.32 0.0383 0.1915
5 0.20 5.98 10.49 15.67 0.0909 0.4545
6 0.22 4.30 3.92 68.49 0.2939 1.4695
7 0.24 3.23 2.63 78.86 0.3100 1.5500
8 0.26 2.62 2.45 80.30 0.3202 1.6010
9 0.28 2.48 2.33 81.27 0.3209 1.6045
10 0.30 2.15 2.31 81.43 0.3208 1.6040
a
Lignin recovery yield from 100 ml of liquor.

2.2. Pre-treatment conditions by Mussatto and Roberto (2006). The phenolic acids con-
centration was obtained by high performance liquid chro-
BSG was pre-treated with dilute sulfuric acid under the matography using a UV (276 nm) detector and a Waters
conditions previously optimized by Mussatto and Roberto resolve C18 5 lm (3.9 300 mm) column. The samples were
(2005): 100 mg H2SO4/g dry matter, a solid:liquid ratio of ltered in 0.45 lm Millipore membrane and injected in the
1:8 g:g, at 120 C for 17 min. At the end of the reaction, chromatograph under the following conditions: column at
the solid residue was separated by centrifugation, washed room temperature; acetonitrile/water (1/8, with 10 g/l ace-
with water until neutral pH and dried at 50 5 C until tic acid and pH adjusted to 2.5 by H3PO4 addition) as elu-
approximately 50% moisture content. ent; a ow rate of 0.9 ml/min and injection volume of 20 ll.
The lignin removal (LR, %) was calculated by the Eq. (1),
2.3. Soda pulping where SLi is the initial concentration of soluble lignin in the
original liquor, and SLf is the total soluble lignin concentra-
The soda pulping of the pre-treated BSG was performed tion in the liquor after the sulfuric acid addition.
using 2% w/v NaOH solution, in a solid:liquid ratio of SLi  SLf
1:20 g:g, at 120 C for 90 min, as optimized by Mussatto LR  100 1
SLi
et al. (2006). Afterwards, the reaction medium was cooled
in ice bath and ltered in a 100% polyester cloth to separate Origin version 7.0 was the software used for graphical
the black liquor, which had a pH of 12.56. analysis of the results.

2.4. Lignin recovery from black liquor 3. Results and discussion

A 20 ml volume of the black liquor was treated with dier- The process used for cellulose pulp obtainment from
ent volumes of concentrated sulfuric acid (98% w/w) to BSG was previously established (Mussatto et al., 2006).
obtain a range of dierent pH values (Table 1). After the acid By using this process, 95.3% w/w of the BSG lignin was
addition, the liquor was centrifuged at 4000 rpm for 10 min. removed, and the produced black liquor presented a solu-
The precipitated lignin was separated, thoroughly washed ble lignin concentration of 12.44 g/l. The reduction in the
with water until the supernatant was neutral, and then dried pH of this liquor, proportioned by the sulfuric acid addi-
at 60 C until constant mass. The obtained liquors were tion, caused the precipitation of the solubilized lignin, so
maintained at 4 C during 24 h and centrifuged again at that the lower the pH the higher the recovered lignin mass
4000 rpm for 10 min. The lignin thus precipitated was (Table 1). According to Fengel and Wegener (1989) the lig-
washed with water and dried at 60 C until constant mass. nin is highly soluble in alkaline medium but its solubility in
The total precipitated lignin mass was considered as the the black liquor really decreases when the pH is reduced.
sum of the mass recovered after the acid addition and the Nevertheless, since the lignin nature varies according to
mass recovered after cooling. The obtained liquors were the nature of plant species (Hon, 1996) the precipitation
characterized regarding the pH, total concentration of solu- prole of this fraction can also vary according to the raw
ble lignin, and concentration of the phenolic acids: vanillic, material employed.
p-hydroxybenzoic, syringic, p-coumaric, and ferulic. Interestingly, Table 1 shows that the pH reduction to
7.71 practically did not inuence the lignin precipitation,
2.5. Analytical methods whose removal from black liquor was only 0.32%, propor-
tioned by the cooling and not by the acid addition. Never-
The total concentration of soluble lignin in the liquors theless, the pH decrease to a value near to six started to
was determined according to the methodology described inuence the precipitation of this fraction, proportioning
 S.I. Mussatto et al. / Carbohydrate Polymers 70 (2007) 218223 221

0.35 90 cause a complete lignin precipitation when the pH is

precipitated mass reduced to a range of 23 (Villar, Caperos, & Garca-
0.30 lignin removal 75 Ochoa, 2001). In the present work, the lignin solubilized
by soda pulping was not completely recovered when the
Precipitated mass (g)

0.25
60 pH was acidied to 2.15, suggesting that the chemical pul-

Lignin removal (%)

0.20
ping process really inuences the lignin recovery from the
45 black liquor.
0.15
Although the incomplete lignin recovery, decreasing
30 pH from 12.56 to 2.15 strongly inuenced the soluble lig-
0.10 nin concentration in the BSG black liquor. Similar results
15 were obtained by Sun, Tomkinson, and Bolton (1999)
0.05 when reduced from 13.8 to 2.0, the pH of the black liquor
0 obtained from oil palm empty fruit bunch bers. These
0.00 authors recovered 1 g lignin to each 100 ml of liquor acid-
1 2 3 4 5 6 7 8 9 10 11 12 13
pH
ied with 9.68N H3PO4. However, the precipitated also
contained polysaccharides degradation products because
Fig. 2. Precipitated mass and lignin removal as a function of the drop in the raw material had not been pre-treated. In the present
the pH of the brewers spent grain black liquor.
work, the lignin recovery yield at pH 2.15 was 1.6 g/
100 ml liquor (Table 1), and this mass was basically
a removal rate of 15.67%. This value was increased 4.4-fold derived from lignin because the hemicellulose was
when the pH was reduced from 5.98 to 4.3. Fig. 2 clearly removed during the pre-treatment stage, and the cellulose
shows that the highest increase in the lignin precipitated was practically not aected during the pulping process
mass occurred in the pH range between 7.71 and 4.3, reach- (Mussatto et al., 2006). Considering this fact, the lignin
ing a removal rate of 68.49%. For pH values lower than 4.3 recovery yield was practically twice higher than that
the lignin removal was still increased in around 13%, but obtained by Sun et al. (1999), what is justiable because
this removal increment was ve fold lower than the incre- BSG contains 27.8% w/w lignin while the oil palm empty
ment observed in the pH range between 7.71 and 4.3. fruit bunch bers contain 14.2% w/w. These results sug-
At the lowest pH value (2.15) 81.43% of the lignin solu- gest thus that the eciency of the concentrated H2SO4
bilized in the liquor was recovered. A similar lignin recov- is very similar to that of 9.68 N H3PO4 for lignin precip-
ery yield (87.8%) was obtained by Santos and Curvelo itation from black liquors.
(2001) when the pH of the black liquor of Eucalyptus uro- As a consequence of the lignin removal, the color of
grandis wood was decreased from 13.6 to 3.0. However, if the BSG black liquor was altered. It can be observed in
compared with the lignin recovery at pH 3.23, the present Fig. 3 that in the original form (sample 1), the liquor
work result was lower (78.86%). Such dierence probably was dark brown, but it was claried with the pH reduc-
is not only due to the raw material used. According to tion. At the lowest pH value (2.15), the liquor color
Rohella, Sahoo, Paul, Choudhury, and Chakravortty passed to be pale yellow. According to Fengel and
(1996) the lignin composition varies from biomass to bio- Wegener (1989), the dark color of the liquor is derived
mass, and also with the chemical pulping process used from the chromophoric functional groups, including qui-
for the biomass. The black liquor used in the present work nones, carbonyl groups, carboxylic acids, hydroperoxy
was obtained by soda pulping, while the black liquor used radicals, phenolic hydroxyl groups, among others, gener-
by Santos and Curvelo (2001) was obtained by Kraft ated during the lignin degradation, which are soluble in
pulping. alkaline medium.
The acid precipitation is the most common method used It can also be noted in Fig. 3 that, although in the sam-
to recover Kraft lignin from black liquors, being able to ple 5 (pH 5.98) the lignin had been removed in 15.67%, the

Fig. 3. Color of the brewers spent grain black liquor as a function of the drop in the pH value from 12.56 (sample 1) to 2.15 (sample 10).
222 S.I. Mussatto et al. / Carbohydrate Polymers 70 (2007) 218223

4. Conclusions
p-hydroxybenzoic, vanillic and syringic acids (mg/l)

36
128
33 p -hydroxybenzoic
124
vanillic
The pH is a variable of large inuence on the precipita-

Ferulic and p-coumaric acids (mg/l)

30 syringic 120
p-coumaric
116
tion of the lignin from the black liquor produced by BSG
27
ferulic soda pulping. Decreasing the pH from 12.56 to 2.15
24 112
108
resulted in an increase of precipitated lignin mass, being
21
104
recovered 81.43% of the lignin solubilized in the liquor.
18
100
The lignin recovery is not only interesting from an environ-
15 96
mental, but also from an economical point of view, because
12 92
it can be reused for obtainment of several chemical prod-
ucts of commercial interest. Further works on the applica-
9 88
tion of lignin from BSG black liquor will be investigated in
84
6
our laboratory.
80
3
2 3 4 5 6 7 8 9 10 11 12 13
Acknowledgement
pH

Fig. 4. Phenolic acids removal with the drop in the pH of the brewers The authors are grateful to CAPES, FAPESP and
spent grain black liquor. CNPq for the nancial support.

References
color of the liquor was not altered, being as dark as in the
original form (sample 1). Probably the compounds that Adler, E. (1977). Lignin chemistry Past, present and future. Wood
confer color to the liquor were not removed in this pH Science and Technology, 11, 169218.
range (between 12.56 and 5.98). In the sample 6 (pH Athanasios, M., Georgios, L., & Michael, K. (2007). A rapid microwave-
assisted derivatization process for the determination of phenolic acids
4.30), 68.49% of the lignin was removed and a strong alter- in brewers spent grain. Food Chemistry, 102, 606611.
ation in the liquor color was observed. The pH decrease Fengel, D., & Wegener, G. (1989). Wood: Chemistry, ultrastructure,
from 4.30 to 3.23 (sample 7) also caused a strong inuence reactions. New York: Walter de Gruyter.
on the liquor color, which passed to be yellow and almost Gargulak, J. D., & Lebo, S. E. (2000). Commercial use of lignin-based
did not modify with the pH reduction to 2.15 (sample 10). materials. In W. G. Glasser, R. A. Northey, & T. P. Schultz (Eds.),
Lignin: historical, biological, and materials perspectives (pp. 305320).
It is evident, thus, that the liquor color was strongly Washington: American Chemical Society.
aected in acid pH, suggesting that the chromophores com- Hammett, A. L., Youngs, R. L., Sun, X. F., & Chandra, M. (2001). Non-
pounds are removed in this pH range, mainly between 5.98 wood bre as an alternative to wood bre in Chinas pulp and paper
and 3.23. industry. Holzforschung, 55, 219224.
Five phenolic acids were quantied in the liquors Hon, D. N.-S. (1996). Chemical modication of lignocellulosic materials.
New York: Marcel Dekker Inc.
obtained after pH adjustment. The choice for these com- Iglesias, G., Bao, M., Lamas, J., & Vega, A. (1996). Soda pulping of
pounds was based on the fact that they are the acids most Miscanthus sinensis. Eects of operational variables on pulp yield and
quantitatively present in BSG (Athanasios, Georgios, & lignin solubilization. Bioresource Technology, 58, 1723.
Michael, 2007). Thus, their removal proles can give an Jimenez, L., Ramos, E., Rodrguez, A., de la Torre, M. J., & Ferrer, J. L.
idea about the pH decrease inuence on the concentration (2005). Optimization of pulping conditions of abaca. An alternative
raw material for producing cellulose pulp. Bioresource Technology, 96,
of the lignin derived compounds in the liquor. Fig. 4 shows 977983.
that these compounds were not similarly removed from the Karam, J., & Nicell, J. A. (1997). Potential applications of enzymes in
liquor when the pH was decreased. The vanillic acid, for waste treatment. Journal of Chemical Technology and Biotechnology,
example, presented a removal prole very dierent of those 69, 141153.
observed for the other acids, since its concentration was Lawther, J. M., & Sun, R. (1996). The fractional characterization of
polysaccharides and lignin components in alkaline treated and
practically not altered in the pH range between 9.9 and atmospheric rened wheat straw. Industrial Crops and Products, 5,
5.98, while the removal of the other acids was more signif- 8795.
icant at these pH values. In addition, the removal percent- Mussatto, S. I., Dragone, G., Rocha, G. J. M., & Roberto, I. C. (2006).
age varied with each acid evaluated. The highest removal Optimum operating conditions for brewers spent grain soda pulping.
was observed for syringic acid (74.4%) and the lowest for Carbohydrate Polymers, 64, 2228.
Mussatto, S. I., & Roberto, I. C. (2006). Chemical characterization and
p-coumaric acid (32.1%), whose values varied 42.3% liberation of pentose sugars from brewers spent grain. Journal of
among themselves. The other analyzed acids, vanillic, Chemical Technology and Biotechnology, 81, 268274.
p-hydroxybenzoic, and ferulic, were removed in 47.6%, Mussatto, S. I., & Roberto, I. C. (2005). Acid hydrolysis and fermentation
46.5%, and 38.3%, respectively. Sun et al. (1999) also of brewers spent grain to produce xylitol. Journal of the Science of
observed the removal of these same phenolic acids as a Food and Agriculture, 85, 24532460.
Navaee-Ardeh, S., Mohammadi-Rovshandeh, J., & Pourjoozi, M. (2004).
function of the decrease in the pH of the black liquor pro- Inuence of rice straw cooking conditions in the sodaethanolwater
duced from oil palm empty fruit bunch bers, obtaining pulping on the mechanical properties of produced paper sheets.
dierent removal values for each compound. Bioresource Technology, 92, 6569.
 S.I. Mussatto et al. / Carbohydrate Polymers 70 (2007) 218223 223

Rohella, R. S., Sahoo, N., Paul, S. C., Choudhury, S., & Chakravortty, V. black liquor of oil palm empty fruit bunch bre pulping. Polymer
(1996). Thermal studies on isolated and puried lignin. Thermochimica Degradation and Stability, 63, 195200.
Acta, 287, 131138. Villar, J. C., Caperos, A., & Garca-Ochoa, F. (2001). Oxidation of
Santos, F., & Curvelo, A.A.S. (2001). Recovery of lignins from kraft hardwood Kraft-lignin to phenolic derivatives with oxygen as oxidant.
liquors. In: Proceedings of the 6th Brazilian symposium on the Wood Science and Technology, 35, 245255.
chemistry of lignins and other wood components (pp. 310314). Xiao, B., Sun, X. F., & Sun, R. (2001). Chemical, structural, and thermal
Brazil. characterizations of alkali-soluble lignins and hemicelluloses, and
Sun, R. C., Tomkinson, J., & Bolton, J. (1999). Eects of precipitation pH cellulose from maize stems, rye straw and rice straw. Polymer
on the physico-chemical properties of the lignins isolated from the Degradation and Stability, 74, 307319.