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Continuous biohydrogen production from

coagulation-pretreated textile desizing wastewater

Chiu-Yue Lin a,b,*, Chih-Cheng Chiang c, Thi Mai Linh Nguyen a,b,
Chi-How Lay d
Green Energy Technology Research Group, Ton Duc Thang University, Ho Chi Minh City, Viet Nam
Faculty of Environment and Labor Safety, Ton Duc Thang University, Ho Chi Minh City, Viet Nam
Department of Environmental Engineering and Science, Feng Chia University, Taiwan
Green Energy Development Center, Feng Chia University, Taiwan

article info abstract

Article history: A real textile desizing wastewater (TDW) was coagulation-pretreated and used as the
Received 10 July 2017 substrate to investigate its potential of continuous biohydrogen production. The experi-
Received in revised form ments used a completely stirred tank reactor digester with operating conditions of pH 6.8,
4 September 2017 35  C, and hydraulic retention times (HRTs) 8 h and 4 h. Biohydrogen production perfor-
Accepted 3 October 2017 mance at HRT 8 h was detailed and effectives of doubled organic loading rate (OLR) were
Available online xxx elucidated at HRT 4 h. The experimental results indicate that coagulated TDW at an OLR of
30 g total sugar/L gave 3.8 L/L-d as the peak hydrogen production rate (HPR), which value is
Keywords: comparable to feedstock like sugarcane vinasse, palm oil mill effluent and cassava
Biohydrogen wastewater. TDW is a good source for biohydrogen production. Moreover, an OLR man-
Coagulation agement is an efficient strategy to enhance HPR. Methane production occurred in a long
Dark fermentation run H2 fermenter and its prevention methods were discussed. Acetate and butyrate were
Desizing wastewater the main soluble metabolic products. Future works for using the experimental data in
Organic loading rate industry applications are proposed with (1) HPR increment, (2) H2 fermenter effluent
Textile industry treatment, and (3) re-confirming the application feasibility of the experimental data.
© 2017 Hydrogen Energy Publications LLC. Published by Elsevier Ltd. All rights reserved.

environment-friendly characteristics are getting more

Introduction attraction. Dark fermentation or anaerobic digestion of
organic wastes/wastewaters is the most promising mode of
Hydrogen has positive combustion characteristics of high biohydrogen generation [1,2]. This green biohydrogen pro-
heating value and no pollutant emission when comparing duction technology is currently reported to have the potential
with methane and is known as a clean energy carrier. Many to become a major hydrogen-generating method in green
thermochemical and biological methods have been proposed economy [3].
and used for hydrogen production in practical applications or Exploring new feedstock sources is important in bioenergy
researches but green ones with low energy-consumption and industry. Textile industry wastewaters contain high strength

Abbreviations: BPR, Biogas production rate; COD, Biochemical oxygen demand; CSTR, Completely stirred tank reactor; HPR, Hydrogen
production rate; HRT, Hydraulic retention time; HY, Hydrogen yield; OLR, Organic loading rate; ORP, Oxidation-reduction potential;
POME, Palm oil mill effluent; SMP, Soluble metabolic products; TDW, Textile desizing wastewater; VFAs, Volatile fatty acids; VSS, Volatile
suspended solids.
* Corresponding author. Green Energy Technology Research Group, Ton Duc Thang University, Ho Chi Minh City, Viet Nam.
E-mail address: chiu-yuelin@tdt.edu.vn (C.-Y. Lin).
0360-3199/© 2017 Hydrogen Energy Publications LLC. Published by Elsevier Ltd. All rights reserved.

Please cite this article in press as: Lin C-Y, et al., Continuous biohydrogen production from coagulation-pretreated textile desizing
wastewater, International Journal of Hydrogen Energy (2017), https://doi.org/10.1016/j.ijhydene.2017.10.012
2 i n t e r n a t i o n a l j o u r n a l o f h y d r o g e n e n e r g y x x x ( 2 0 1 7 ) 1 e7

organic and inorganic compounds such as starch, dye and resource, developing a new TDW treatment method and
some chemicals. Recently, circular economy of efficiently proposing a stagey for increasing HPR from TDW via OLR
reusing industrial waste, sustainable textile production and management.
reusing and recycling textile wastewater are attracting at-
tentions [4e6]. Desizing operation is a main process in textile
industry and produces many pollutants including starch [7]. Materials and methods
Individually treating this textile desizing wastewater (TDW) is
attractive in having simple technology and economic advan- The experiences of batch hydrogen production from
tages over a centralized-treatment [8]. Starch-containing TDW coagulation-pretreatment TDW in coagulant dosage and
has chemical oxygen demand (COD) concentrations of coagulation operation conditions obtained from our previous
20e40 g/L, indicating the potential of generating renewable studies [10] were used as the references for operating the
energy biogas such as biohydrogen and biomethane via dark continuous-feeding biohydrogen fermentation system.
fermentation [9]. However, most of the desizing operation
uses bacterial growth-inhibitive dyes that might negatively Seed, bioreactor and substrate TDW
affect biogas fermentation. Recently, coagulation-
pretreatment has shown its high potential of enhancing bio- Seed sludge
hydrogen production from starch-containing TDW with 120% In biohydrogen production experiments, a granular sludge
and 550% increments in hydrogen yield (HY) and hydrogen was used as the seed. This granular sludge was collected from
production rate (HPR), respectively, in our previous batch test a fructose factory's anaerobic digester and had characteristics
[10]. This coagulation-pretreated TDW gave an HY of 1.52 mol/ of pH 7.71, total COD 50 g/L, total sugar 5.3 g/L, NH3eN 45 mg/L
mol glucose and an HPR of 3.9 L/L-d which values are com- and volatile suspended solids (VSS, expressing the biomass
parable to some other feedstock like cassava and paperboard concentration) 28.8 g/L. Before being used as the seed micro-
mill wastewaters [10] indicating the importance of pretreat- flora, the collected granular sludge was treated via water bath
ment in using TDW as a feedstock for biohydrogen produc- (95  C, 1 h) to activate hydrogen-producers and inactivate
tion. The biohydrogen production process in a continuous methanogens.
mode is important in industry applications [11,12].
Based on the above observations, this work aims to Bioreactor
conduct a continuous fermentation fed on the coagulation- A lab-scale completely stirred tank reactor (CSTR) digester
pretreated starch-containing real TDW to elucidate the was used to in the continuous-feeding H2 production experi-
usage of TDW for biohydrogen production. The biohydrogen ments. This glass-made circular digester (inter diameter
production performance of the bioreactor was investigated by 13 cm, height 18 cm) had a working volume of 2.3 L (Fig. 1). Hot
long operation (more than 2 months) and then the hydraulic water jacket was used to control the bioreactor operation
retention time (HRT) was reduced to double the organic temperature. A drum-type gas meter (Ritter TG05, Germany)
loading rate (OLR) to elucidate the performance. This work has was used to measure biogas production. This bioreactor was
the novelties of using a TDW as a biohydrogen production operated in an ambient atmosphere pressure.

Fig. 1 e A scheme of the CSTR biohydrogen production bioreactor for continuously feeding operation.

Please cite this article in press as: Lin C-Y, et al., Continuous biohydrogen production from coagulation-pretreated textile desizing
wastewater, International Journal of Hydrogen Energy (2017), https://doi.org/10.1016/j.ijhydene.2017.10.012
i n t e r n a t i o n a l j o u r n a l o f h y d r o g e n e n e r g y x x x ( 2 0 1 7 ) 1 e7 3

Substrate TDW concentration, HPR, biogas production rate (BPR), pH and ORP
A real TDW was coagulation-pretreated and used as the sub- at TDW 5 g total sugar/L). The reactor performances during
strate. The real TDW was obtained from a textile factory and had started-up stage of using TDW 10 g total sugar/L were not
the characteristics (average values) of COD 47 g/L, total sugar shown.
17.0 g/L, sulfate 0.39 g/L, suspended solids 2.0 g/L and pH 6.8. The ORP is an important monitoring indicator showing the
range values for these characteristics were detailed in our pre- anaerobic environment of a bioreactor. Fig. 2 shows that at
vious study [10]. Coagulation-pretreatment gave COD and total HRT 8 h through the reactor operations, the ORP generally
sugar removal rates of 12% and 8.5%, respectively. Coagulant fluctuated between 420 mV and 510 mV with average ORP
GGEFloc-653 (concentration 1 g/L) was used in the coagulation- values being in the range of around 450~480 mV. In the
pretreatment [10]. The coagulation operation conditions ob- beginning, marked fluctuation ORP values were observed;
tained in our previous work [10] were used: pH 7.0, rapid mixing they might result from the adaption of the microbes to new
(100 rpm) 3 min, slow mixing (30 rpm) 20 min and sludge settling cultivation environments. These average ORP values were in
time 1 h. The coagulated TDW supernatant was diluted (with tap the range favoring biohydrogen production. Some fluctua-
water) to total sugar concentrations of 5 and 10 g/L. They were tions in ORP were observed when feeding failures occurred. H2
used as the substrate (substrate TDW) for further H2 fermenta- concentration in a biogas is a common indicator to show the
tion. This substrate TDW was stored at 4  C before usage to performance of a biohydrogen fermenter. During the start-up
minimize any biochemical reactions. The inorganic nutrients of the reactor, H2 concentration reached 30% at day 1 (1 day
for bacterial growth were detailed in our previous study [10]. after the reactor start-up) and reached 60% at day 2 (1 day after
the continuous feeding began) but decreased to near 20% at
Fermentative biohydrogen production day16 (data were not shown in Fig. 2). At day 17, the substrate
TDW concentration was changed to 5 g/L (from 10 g/L), which
Continuous operation of H2 fermentations was at pH 6.8 and made the H2 concentration increase markedly from 20% (at
35  C using the CSTR fermenter. In starting-up the bioreactor, day 16) to 45% at day 23 (6 days after TDW concentration
seed inoculum 0.5 L and substrate TDW 1.8 L (10 g total sugar/L change, Fig. 1) and then some decreases were observed from
in concentration) were put in the reactor and then gassed with feeding or mixing failures. The reasons of feeding failures
argon for 3 min to remove oxygen from the head space for included power failures, pipe blocks and feedstock shortages.
maintaining anaerobic environment. This reactor used a Fig. 2 also indicates that H2 concentrations reached a constant
batch mode operation for 1 day and then the continuous value of 30% after day 40. Another biogas CH4 that was not an
feeding started at HRT 8 h. At day 17, the tested TDW con- expected component appeared at days 40e60 in a small level
centration was changed to 5 g/L (as total sugar) because the of 0.2e2%. CH4 generation might result from the growth of
collected real TDW concentration was low from factory methanogens in the reactor wall. We inactivated the metha-
operation. For this stage (5 g/L, HRT 8 h ¼ OLR 15 g/L-d), the nogens via heating the reactor content at 95  C for 1 h at day
performance of the H2 fermenter was observed as reactor re- 60. After that, there was no methane until day 72; days 72e78
sponses to environmental changes such as feeding-failure. had 0.3e4% of methane. At day 78, the reactor content was
After the desired data were collected, the HRT was then heat-treated again (95  C for 1 h) to inactivate the metha-
reduced to 4 h to elucidate the hydrogen production perfor- nogens. A similar operation strategy had been reported
mance when the OLR was doubled (OLR ¼ 30 g/L-d). During all recently. Bundhoo et al. [13] indicate that in the long run of
the experiments, the volume and content of biogas and biohydrogen production, some seed pre-treatment methods
metabolite concentrations such as volatile fatty acids (VFAs) are not effective and repeated treatments for the fermenter
were determined at proper time intervals. H2 concentration content are necessary for continuously suppressing H2-
(%) and HPR were used as the main indicators to show the consuming bacteria.
performance of H2 fermentation. HPR is a well-used parameter in monitoring or evaluating
biohydrogen production performance. HPR indicates the
Analysis capability of a biohydrogen-producing reactor with higher
value favoring practical applications. A HPR value correlates
The concentrations of H2, CO2 and VFAs were determined with with feedstock source and concentration, reactor configura-
Shimadzu GC-14A gas chromatographs equipped with ther- tion and reactor operation conditions. Fig. 2 also shows that
mal conductivity and flame ionization detectors. The opera- the HPR value was dependent on the reactor operations and
tion conditions of these detectors were similar to those of our its variation trend met the variation of H2 concentration but in
previous study [11]. The methods used for measuring bigger fluctuation. The average HPR value during reactor's
oxidation-reduction potential (ORP), pH, sugar concentration, steady-state condition at the TDW 5 g total sugar/L was
VSS and COD were similar to a previous work of us [10]. 0.88 ± 0.30 L H2/L-d. These values are rather lower than those
obtained from our batch operation results (HPR 3.9 L H2/L-d at
TDW 15 g total sugar/L [10]). However, the HPR value is com-
Results and discussion parable to some other feedstock like cassava (0.39 L H2/L-
d [14]; 2.1 L H2/L-d [15]), sugarcane vinasse (1.12 L-H2/L-d [16])
Biohydrogen production at HRT 8 h (OLR 15 g total sugar/L-d) and palm oil mill effluent (POME) (2.64 L H2/L-d [17]; 3.68 L H2/
L-d [18]).
The H2 fermentation performances of the coagulation- BPR represents total biogas production rate. In this study,
pretreated TDW were shown in Fig. 2 (daily variations of H2 after reactor's long run operation, the Ar gas added during the

Please cite this article in press as: Lin C-Y, et al., Continuous biohydrogen production from coagulation-pretreated textile desizing
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4 i n t e r n a t i o n a l j o u r n a l o f h y d r o g e n e n e r g y x x x ( 2 0 1 7 ) 1 e7

Fig. 2 e Daily variations of hydrogen production rate (HPR), H2 concentrations, biogas production rate (BPR), pH and ORP at
HRTs of 8 h and 4 h.

reactor starting-up stage was replaced by the produced biogas. Liquid metabolic products during fermentation
During these long run operations, the biogas contained only
H2 and CO2. Therefore, the daily BPR variation trend was Soluble metabolic products (SMP) such as alcohols and VFAs
similar to that of HPR. The average BPR value during the re- are produced during fermentative biohydrogen production.
actor's steady-state stage was 5 L/L-d. Fig. 3 depicts the daily variations of SMP concentrations dur-
ing reactor operations. An observation on these SMP data of
Biohydrogen production performance at HRT 4 h (OLR 30 g days 0e27 indicates that the major liquid metabolites were
total sugar/L-d) ethanol, acetate, propionate and butyrate when the reactor
was in varied environments raised from reactor starting-up.
When the HRT was decreased from 8 h to 4 h for TDW 5 g total During the reactor start-up period of days 1e17, there were
sugar/L, the reactor OLR was accordingly doubled as 30 g total batch feeding, continuous feeding and substrate concentra-
sugar/L-d. Fig. 2 shows that when the HRT was decreased to tion changing, which operations made environmental
4 h, H2 concentration, HPR and BPR elevated promptly. These changes of the digester contents. However, ethanol (around
parameters reached constant values and did not decrease 4000 mg/L) and propionate (around 2000 mg/L) concentrations
except at the times of feeding failures. Constant H2 concen- markedly decreased to lower than 200 mg/L after the starting-
tration, HPR and BPR values reached 35%, 3.8 L H2/L-d and 12 L/ up period. In contrast, acetate (around 2700 mg/L) and buty-
L-d, respectively. Moreover, very low level of methane rate (around 3000 mg/L) concentrations had no such marked
(0.2e0.5%) appeared again at day 104 and after day 109. decrease with values of 1000 mg/L (acetate) to 1500 mg/L

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Fig. 3 e Daily concentration variations of soluble metabolic products at HRTs of 8 h and 4 h.

(butyrate). Higher butyrate concentration shows that there of HRT 4 h and 5 g total sugar/L-d (equal to an OLR of 30 g total
was a butyrate-type fermentation that giving good bio- sugar/L-d). A comparison of continuous biohydrogen pro-
hydrogen production [19]. Moreover, the concentrations of duction from various feedstock/wastewaters is shown in
acetate and ethanol were both around 200 mg/L showing their Table 1. It indicates that a coagulation-pretreated starch-
concentration ratio was one, which value indicating a good containing TDW has biohydrogen production potential with a
hydrogen production [20]. HRP comparable to some wastewaters such as POME and
cassava wastewater.
Significances of the experimental results
Biohydrogen production performance in the long run of the
Starch-containing TDW as substrate for biohydrogen reactor
production In biohydrogen production operation, it is designed to maxi-
Our studies using real TDW as the substrate for producing mize the H2 fraction with lower CO2 fraction and even no CH4
biohydrogen via dark fermentation in batch [10] and contin- composition in the biogas. However, this work shows that CH4
uous (this work) modes resulted in peak HPR values of appeared in the biogas when the reactor had long operations
3.8e3.9 L H2/L-d. Since industry applications prefer a contin- both at HRT 8 h and 4 h. In this situation, we had heated the
uous operation mode, the results obtained from the contin- reactor contents for inactivating methanogens and then pro-
uous operation were used to compare with other substrates to moting biohydrogen production. The cases of long reactor
show the applicable characteristics of TDW for biohydrogen operation making the slow development of methanogenic
production. The peak HPR value 3.8 L H2/L-d was obtained in activity have also been reported in running biohydrogen fer-
this continuous H2 fermentation at the operation conditions menters [13,22]. The appearance of CH4 indicates the

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L-d). Of course, a HRT decrease to 3 or 2 h or even lower values

Table 1 e Biohydrogen production performance of
to give higher OLR may be feasible because some reports show
various wastewaters.
that at these low HRT values high biohydrogen could be ach-
Substrate OLR Reactor HPR Reference
ieved in dark fermentation of some wastewaters [21,26,27].
(g COD/L-d) (L/L-d)
Moreover, an observation on the relationships between the
Cassava wastewater 161 AFBR 96 [21] reactor type, OLR and HPR in Table 1, it is known that to have
Cassava wastewater 25 UASB 0.39 [14]
high HPR, granular microflora or immobilized-cells systems
Cassava processing 14 AFBR 2.1 [15]
such as up-flow sludge blanket bed and anaerobic packed bed
De-oiled jatropha e CSTR 0.15 [12] reactors can be applied and operated at high OLR for obtaining
waste high HPR values.
POMEa 94.4b Batch 2.64 [17]
POME 75 UASB 3.68 [18] Future studies on applying the process for industrial usage
Sugarcane vinasse 84.2 APBR 1.12 [16]
Continuous systems are necessary for using various biomass
TDW 60 CSTR 3.8 This study
sources to produce biohydrogen for industry applications. The
POME, Palm oil mill effluent; TDW, textile desizing wastewater; present work has successfully shown the feasibility and
OLR, organic loading rate; AFBR, anaerobic fluidized bed reactor;
possible strategy for conducting a continuous biohydrogen
APBR, up-flow anaerobic packed bed reactor; UASB, up-flow
production from a coagulated TDW. However, there are some
anaerobic sludge bed; CSTR, completely stirred tank reactor;
HPR, hydrogen production rate. further studies needed to do if a starch-containing TDW is to
g COD/L. be used as a substrate for bioenergy production in industry
applications. These issues include (1) how to increase the HPR,
(2) how to treat the H2 fermenter effluent, and (3) to re-confirm
presences of methanogens that might consume H2 to produce the application feasibility of the experimental data. High HPR
CH4. This might result from the fact that in a long run oper- favors industrial application of H2. As mentioned above, HPR
ation, seed pre-treatment methods are not effective and, could be increased by lowing the HRT to 3 h, 2 h or lower
therefore, it is necessary to find a suitable seed pre-treatment values [21,26,27]. Moreover, a direct H2 fermentation and in-
technology for industrial application [13]. In the present wok, direct bio-H2 production via methane-reforming might be
heat-pretreatment was applied to treat the seed sludge to used to increase HPR. It has been reported that the HPR was
obtain highly-active hydrogen-producers and to inhibit increased by 10% via direct and indirect bio-H2 production
methanogenic activity. Note that during heat-treatment the methods, which used a two-stage anaerobic digestion to
methanogens are inhibited but not killed and this fact leads to produce biogas and simultaneously to reduce the effluent
the slow development of methanogenic activities in a long- organic strength [28]. Some tests are needed to evaluate the
operating H2 fermenter. potential of continuous two-stage anaerobic digestion for a
TDW [29e31]. To re-confirm the industrial application feasi-
OLR management for enhancing HPR bility of the proposed total-solution process and experimental
To have high HPR is important for a biohydrogen production data, a techno-economic evaluation is suggested [32,33].
system in its industry applications. Recently, Arimi et al. [1]
used OLR management as a strategy to improve biohydrogen
production from a wastewater. In the present work, Conclusions
comparing the biohydrogen production performances ob-
tained at HRT 8 h and 4 h could elucidate the OLR effect. It is This work aims to elucidate the usage of TDW for producing
observed that a small increase in H2 concentration (30% vs. biohydrogen as a biofuel. Based on the continuous-feeding
35%), a big increase in BPR (5 vs. 12 L/L-d) and a very big in- operation results, the following conclusions are drawn.
crease in HPR (0.9 vs. 3.8 L H2/L-d) were obtained when the OLR Starch-containing TDW is a potential resource for fermenta-
was doubled. Increments of 140% [¼ (125)÷5] and 320% tive biohydrogen production if it is coagulation-pretreated. A
[(3.8e0.9)÷0.9] were obtained for BPR and HPR, respectively, continuous H2 fermenter can use an OLR management strat-
though only 100% increase in the OLR. The enhancement egy to enhance HPR but methane production might occur in
capability of increasing OLR was obtained. the long run operation. A continuous CSTR operation at HRT
There are many reports showing that OLR affects bio- 4 h (OLR 30 g total sugar/L) could give peak HPR 3.8 L H2/L-d,
hydrogen production in dark fermentation of various feed- which value is comparable to other feedstock as sugarcane
stock but not higher OLR gives higher HPR [16,23]. Moreover, vinasse, POME and cassava wastewaters. HPR increment, H2
the OLR values studied in literature are in a range of 2e480 g fermenter effluent treatment and re-confirming the applica-
COD/L-d and the optimal values to give peak HPR are 20e80 g tion feasibility are elucidated as the main future works for
COD/L-d [16,23e25]. In the present study, the tested OLR using the experimental data in industry applications.
values were 15 and 30 g total sugar/L-d (nearly equaling to 30
and 60 g COD/L-d, respectively) which are in the optimal value
range for biohydrogen production. These facts indicate that Acknowledgements
controlling a proper OLR is an important strategy to have a
high HPR for industry applications. The proper OLR for the This research is funded by Foundation for Science and Tech-
pretreated-TDW is suggested as 30 g total sugar/L-d (60 g COD/ nology Development of Ton Duc Thang University (FOSTECT)

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Please cite this article in press as: Lin C-Y, et al., Continuous biohydrogen production from coagulation-pretreated textile desizing
wastewater, International Journal of Hydrogen Energy (2017), https://doi.org/10.1016/j.ijhydene.2017.10.012