Journal of Environmental Chemical Engineering 2 (2014) 18591869

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Journal of Environmental Chemical Engineering

journal homepage: www.elsevier.com/locate/jece

Cultivation of Spirulina platensis using NPK-10:26:26 complex fertilizer

and simulated ue gas in sintered disk chromatographic glass bubble
column
Ankita Kumari, Vaishali Sharma, Akhilendra K. Pathak, Chandan Guria *
Department of Petroleum Engineering, Indian School of Mines, Dhanbad 826 004, India

A R T I C L E I N F O

A B S T R A C T

Article history:
Received 19 June 2014
Accepted 1 August 2014

A cost effective fertilizer based optimum culture medium was formulated for the mass production of
Spirulina platensis using NPK-10:26:26 complex fertilizer in air agitated sintered disk chromatographic
glass bubble column and the growth results were compared with the standard Zarrouks culture medium.
The optimum loadings of NPK-fertilizer and sodium bicarbonate was found to be 0.76 and 10.0 g L
1
respectively and corresponding maximum dry biomass concentration with 1.82 g L
1 was obtained at the
tenth day of biomass growth. Optimally formulated fertilizer medium was then used for the cultivation of
Spirulina using simulated ue gas which was similar to diesel generator exhaust. Flue gas was fed to the
bubble column reactor in a semi-batch fashion and Spirulina cultivation was carried out using
NPK-fertilizer culture medium involving sodium hydroxide. Biomass productivity and CO2 xation rate
was enhanced with the increased loading of sodium hydroxide in the culture medium. Maximum
biomass concentration with 1.85 g L
1 was obtained at the sixth day of biomass growth when average CO2
solubility was maintained at 4.84 g L
1. Finally, ue gas assisted biomass growth using optimum
NPK-10:26:26 complex fertilizer medium was compared with standard Zarrouks culture medium using
sintered disk chromatographic gas bubble column reactor and improved growth results (4.0%) with
enhanced lipid accumulation (5.0%) were obtained using NPK-10:26:26 fertilizer medium as compared
to the Zarrouks culture medium. An almost 50.0% cost saving was achieved due to the use of low cost
NPK-10:26:26 complex fertilizer as a nutrient for Spirulina growth in comparison to standard Zarrouks
culture medium.
2014 Published by Elsevier Ltd.

Keywords:
Spirulina platensis
NPK-10:
26:26 ?complex fertilizer
Simulated ue gas
CO2 xation
Bubble column
Mass transfer

Introduction
Greenhouse gases (GHG), particularly CO2, are being considered
as the most important contributing factor for global warming which
have substantial impacts on the environment, human health and the
economy [1]. The excess CO2 emission to the atmosphere is needed to
be reduced and the present day research has been focused to develop
numerous strategies to mitigate the CO2 emission by proposing
different sequestration techniques [26]. Alkali mediated CO2
capture and sequestration was reported by Budzianowski and Koziol
[46]. In this process, CO2 separation efciency was enhanced
signicantly by using reactive solvents (e.g., ammonia, potassium
hydroxide, and etc.). Moreover, these reactive solvents were also very
helpful to produce valuable fertilizer additives (for example,

* Corresponding author. Tel.: +91 326 2235411; fax: +91 326 2296632.
E-mail addresses: cguria.che@gmail.com, guria.c.pe@ismdhanbad.ac.in
(C. Guria).
http://dx.doi.org/10.1016/j.jece.2014.08.002
2213-3437/ 2014 Published by Elsevier Ltd.

ammonium bicarbonate, potassium carbonate, and etc.). Among

the various techniques, the biological sequestration of CO2 using
photosynthetic microalgae has been receiving considerable attention. Photosynthesis is recognized as a natural means of capturing
anthropogenic CO2; and photosynthetic organisms (e.g., microalgae)
can x CO2 at a moderate rate as compared to the land plants. CO2
xation using microalgae has the following advantages: (i) high
purity CO2 gas is not essential for microalgal growth, (ii) combustion
products e.g., NOx or SO2, may be used as nutrients for biomass
growth, (iii) production of several commercial high valued products
(e.g., proteins, nucleic acids, lipids, starch, vitamins, pigments,
phenolics, biodiesel and etc.) through CO2 xation, and (iv)
utilization of microalgal biomass for wastewater treatment. Carbon
is the major nutrient in biomass and constitutes 4550% of the dry
algal mass [7]. Therefore, nearly 1.651.83 g of CO2 is required for the
biosynthesis of 1.0 g of dry algal biomass. Utilization of CO2 in ue
gas through microalgal cultivation has a substantial impact on CO2
level in the environment and it makes biomass production cheaper
and the details of CO2 xation using microalgae have been reviewed

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A. Kumari et al. / Journal of Environmental Chemical Engineering 2 (2014) 18591869

by Wang et al. [8]. Common microalgae studied for bioxation of

CO2 with high level of tolerance are: Dunaliella[9], Chlorococcum
littorale [10], Euglena gracilis [11], Haematococcus pluvialis [12],
Chlorella sp. [13], Aphanothece microscopica Ngeli [14], Chlorella
vulgaris [15], Botryococcus braunii [15], Scenedesmus sp. [15],
Scenedesmus obliquus [16], Chlorella kessleri [15,17], Chlorella
emersonii [18], Spirulina sp. [17,19]. Therefore, microalgae have
great potential to recycle CO2 and CO2-to-algae route is quite
attractive due to fast growth of microalgae, efcient use of solar
energy, potential application towards biofuel and value-added
commercial products. However, commercial harvesting of microalgae was limited due to its large cost of production. Moreover, the
recovey of valuable complex molecules are also very expensive.
Hence, the signicant technological improvements are required
before commercial development of CO2-to-algae route. In this
regard, technological improvements of CO2-to-algae depends on (i)
photosynthetic efciency (ii) selection of mutant strain of high
quality fast growing microalgae, (iii) use of soluble carbonates
(e.g., NaHCO3 and Na2CO3), (iii) use of multiple carbon sources and
(iv) integration of algae farms with biogas plants and/or pyrolysis of
algae biomass to bioils and biochar. Above factors helped to reduce
the cost of production of microalgae and the details of CO2-to-algae
technological improvements were reviewed by Brennan and
Owende [20] and Budzianowski [21].
The cultivation of Spirulina platensis (S. platensis) was usually
carried out for the supplement of (i) high-valued compounds
which are extensively useful in food, drug and cosmetic industries
[22,23] and (ii) proteins for human food and animal feed [24].
Moreover, it was considered as an ideal food for astronauts by
NASA. However, it was also cultivated to remove nitrogen and
phosphorus contaminants from the wastewater [25,26]. S. platensis
has an ability to grow using direct solar energy with high tolerance
limit towards temperature, alkaline pH and salinity as compared to
other microalgae. Moreover, S. platensis has also the capacity to x
high concentration of CO2 (412% v/v) when it was cultivated using
tubular reactor [2729] and open raceway pond [17].
Zarrouks medium [30] without sodium bicarbonate was
frequently used for S. platensis cultivation using ue gas. Usually,
25% of total S. platensis production cost was associated with the
culture medium [31] and therefore, the development of low cost
culture medium will help to reduce production cost. Therefore, in
addition to the utilization of CO2 from ue gas, it is necessary to use
the cheaper source nutrients for economic production of biomass.
NPK-10:26:26 is a complex fertilizer with diammonium phosphate, muriate of potash and minor quantities of urea and it was
formulated in such a way that it provides three primary essential
nutrients (e.g., N, P and K) adequately for the plant growth. Till
date, there was no open literature available for the mass cultivation
of S. platensis through CO2 xation from ue gas using NPK10:26:26 complex fertilizer.
It is also known that the dissolved oxygen plays an important
role for the mass production of microalgae, and high dissolved
oxygen poses a severe inhibition of photosynthesis causing
reduction in biomass productivity [32,33]. To remove the
accumulated oxygen continuously, bubble column/air lift photo
bioreactor plays a key role to reduce photosynthesis inhibition
[34]. Bubble column/air lift bioreactors are easy to operate with
minimum operating and maintenance costs without externally
added pump to circulate the uid. Moreover, bubble column/air lift
photo bioreactors are compact without moving parts and efcient
design of bubble column/air lift photo bioreactors will help to
provide high gasliquid interfacial area for mass and heat transfer
with adequate back mixing.
In this study, the use of water soluble, low cost
NPK-10:26:26 complex fertilizer was explored for the mass
cultivation of S. platensis, and microalgal CO2 xation was carried

out using simulated ue gas which is based on diesel generator

exhaust in a bubble column. For this, the optimum culture medium
was formulated to maximize biomass growth by varying the
concentration of NPK-10:26:26 complex fertilizer and NaHCO3
using air agitated bubble column illuminated with white light
emitting diode. The growth results in air agitated bubble column
were compared with standard Zarrouks culture medium. Using
the optimum NPK culture medium, cultivation of S. platensis was
carried out for CO2 xation using simulated diesel generator ue
gas exhaust. CO2 solubility was studied to maintain desired
carbonatebicarbonate concentration in the newly formulated
optimal NPK fertilizer and Zarrouks culture medium. A sintered
disk chromatographic glass (SDCG-) bubble column was used in
the present growth study to provide sufciently high gasliquid
contacting area with adequate back mixing in the culture medium.
Bubble size distribution was measured to determine mass transfer
coefcient of CO2 during S. platensis growth. Simulated ue gas
assisted biomass growth studies in SDCG-bubble column using
NPK fertilizer and Zarrouks medium were compared each other.
Specic biomass growth rate and efciency of CO2 xation rate
along with protein, lipid and chlorophyll accumulation in biomass
was determined during microalgae growth. The aim of the present
investigation was to formulate a commercial grade
NPK-10:26:26 complex fertilizer medium for ue gas assisted
mass production of Spirulina in SDCG-bubble column, which would
provide better or at par growth rates of Spirulina as compared to
the standard culture medium.
Experimental
Materials
The strain NCIM-5143 of S. platensis was obtained from National
Chemical Laboratory (NCL: Pune, India) culture collection center
and microalgae were cultivated in Zarrouks culture medium with
following composition (g L
1): NaHCO316.8, NaNO32.5,
K2HPO40.5,
K2SO41.0,
NaCl1.0,
CaCl22H2O0.04,
MgSO47H2O0.2, FeSO47H2O0.01 and 1.0 mL of A5 solution
which was composed of H3BO32.86, MnCl24H2O1.81,
ZnSO44H2O0.222, Na2MoO40.018, CuSO45H2O0.079 [30].
In this study, the growth of S. platensis was carried out using
NPK-10:26:26 complex grade fertilizer, which was obtained from
local authorized dealer of Indian Farmers Fertilizers Cooperative
Limited (IFFCO: Dhanbad, India) and it was used after drying under
full vacuum at 353 K for several days. Typical composition of NPK10:26:26 (g/100 g) used for this study is: (i) diammonium
phosphate: 50.04; (ii) muriate of potash: 43.98; (iii) urea: 1.5;
(iv) silica: 3.6 and (v) moisture: 0.88. Above composition was also
expressed in terms of elemental nitrogen, neutral ammonium
citrate soluble phosphates (as P2O5) and water soluble K2O
respectively which is equivalent to as 10, 26 and 26 g/100 g
respectively. A simulated diesel generator exhaust was used for
microalgal CO2 xation with following composition (v/v):
CO211.0%; CO1.5%; O28.98%; N278.47%; CH40.05%.
Microorganism and inoculum preparation
Individual components of the culture medium were sterilized
separately using autoclave at 394 K for 30 min and culture medium
was prepared in 500 mL Erlenmeyer ask with 250 mL stock
standard solutions. Inoculums were kept on a rotary orbital shaker
at 90 rpm and the temperature was maintained at 303  1 K. The
culture was incubated under 2.0 klx with continuous illumination
using white uorescent lamps with a photoperiod of 14/10 h
light/dark cycle. The microorganism was harvested during the
exponential growth phase which was ltered and washed with

A. Kumari et al. / Journal of Environmental Chemical Engineering 2 (2014) 18591869

1861

9.0% NaCl solution for the complete removal of adsorbed salts.

S. platensis growth studies were carried out using
NPK-10:26:26 fertilizer medium with an initial dry biomass
concentration 50.0 mg L
1 [35].

length. Finally, bubble size distribution was obtained by plotting

the frequency of number of bubbles against bubble diameter.

Apparatus and operating procedure

To monitor biomass growth studies, chlorophyll, dry biomass,

protein, lipid and the uptake of elemental C, N, P and K were
determined. This will help to determine the component-wise
dominant effect of macro nutrients on biomass growth. Dry
biomass, chlorophyll, protein, lipid and the uptake of nutrients
analysis were conrmed by the analysis of variance with p < 0.04,
where p = signicance level of the analysis of variance.

To provide high gasliquid interfacial area with controlled back

mixing, SDCG-bubble column (capacity: 1.5 L) with G5 porosity
grade (pore size: 116 micron) was used for the efcient dispersion
of ue gas in the culture medium. A schematic diagram of the
experimental setup using SDCG-bubble column for mass cultivation of biomass is shown in Fig. 1. Compressed simulated ue gas
was ltered using capsule lter of sterile type with 0.2 mm PTFE
membrane (WhatmanTM SteriVENT). Thereafter, ltered ue gas
was metered using rotameter and passed through the bubble
column. Similarly, compressed air was also sterilized and dispersed
into the bubble column for the removal of accumulated oxygen.
First, ue gas was bubbled through the NPK culture medium
(including controlled quantity of NaOH) to attain carbonate
bicarbonate equilibrium with desired pH range for S. platensis
growth. Exponential phase S. platensis stain with desired quantity
was added to the culture medium and growth studies were
monitored by checking pH of the growth medium. During biomass
growth, pH raised and air purging was continued till pH of the
culture medium reached to the desired upper limit (i.e., pH 10). As
pH reached to the upper limit, air purging was stopped and ue gas
was started for purging through the bubble column till pH of the
culture medium was dropped to the desired minimum pH (i.e., pH
9). This alternate ue gas and air purging cycle was continued up to
the end of biomass growth. Flue gas/airow rate was adjusted in
the bubble column in such a way that the bubble ow regime
dominated during biomass growth. To maintain bubble ow
regime under atmospheric condition, the air and ue gas ow for
given SDCG-bubble column was found to be 1.3 L min
1. Under
this condition, air/ue gas hold up in the column was found to be
60 mL for 1.0 L culture medium. Adopting above feeding strategy
of ue gas, concentration of CO2 in ue gas may be increased
substantially and hence, CO2 tolerance level during S. platensis
cultivation can be increased. Bubble size was measured using
SONY digital photographic camera (Model: DSC-W35) and the
distribution of the bubbles was determined by using Imal bubble
size analyzer software (http://randombio.com/imal.html). To
obtain the bubble size distribution, bubbles in the culture medium
were converted into its digitized images with the help of bubble
size analyzer Imal software. After digitizing, bubbles were
counted from the digitized image and corresponding size of the
individual images were measured with respect to the reference

Analysis of biomass and nutrients

Analysis of chlorophyll and dry biomass

Biomass concentration in the culture medium was determined by measuring the chlorophyll content in S. platensis.
Chlorophyll was extracted as per standard procedure [36] and
the concentration of chlorophyll was measured using double
beam UVvis spectrophotometer (Model: ELICO SL191) with
absorbance peak at 663 nm [37]. Propanol was used for the
extraction of chlorophyll and dry biomass was estimated using
standard procedure [38].
Analysis of protein and lipid
Quantitative estimation of proteins was carried out using
bovine serum albumin as a calibration standard [39]. Quantitative
analysis of lipid was determined by FlochLees method [40].
Analysis of total carbon in solution
In this study, CO2 was allowed to react with NaOH to produce
the mixture of bicarbonate and carbonate which was used as a
carbon source for S. platensis growth. Details of estimating the
carbonate and bicarbonate were adopted from the studies of
Martis et al. [41].
Analysis of N, P and K
In addition to carbon estimation, the uptake of elemental N, P
and K was also determined at the end of biomass growth. To
determine the ammoniacal nitrogen, culture medium was allowed
to react with Nesller reagent which develops yellowbrown color
and corresponding absorbance was measured at 425 nm. Similarly,
the analysis of nitrate nitrogen was based on the reaction with
sodium salicylate in sulphuric acid medium in the presence of
cadmium powder which forms yellow colored salts of nitrosalicylic
acid and corresponding absorbance was measured at 410 nm. To
determine phosphate phosphorus, culture medium was allowed to
react with molybdate in acid solution forming a yellow-colored
phosphomolybdate complex which was further reduced by an

11
8 9

Purge
5

1
4

14

15

Air Sucon
10

12

13

16

Fig. 1. Experimental set-up for S. platensis growth: 1 simulated ue gas cylinder; 2 two-stage stainless steel pressure regulator; 3 simulated ue gas lter 4 ue gas
rotameter; 5 air compressor; 6 pressure gauge; 7 surge tank; 8 air lter 9 air rotameter; 10 gas injection conical ask; 11 jacketed sintered disc chromatographybubble column; 12 water recirculation pump; 13 thermostatic bath; 14 moisture removing trap; 15 CO2 analyser; 16 at bottom ask with absorbing solution.

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A. Kumari et al. / Journal of Environmental Chemical Engineering 2 (2014) 18591869

Results and discussion

amino acid, giving a characteristic molybdenum blue color and

corresponding absorbance was measured at 690 nm. Potassium
was measured by turbidimetric method in which potassium is
precipitated in the basic environment of sodium tetraphenylborate
and develops turbidity. The standard reagents (supplied by HANNA
instruments, USA) were used to determine ammoniacal nitrogen,
nitrate nitrogen, phosphate phosphorous and elemental potassium
using NPK analyzer (Model: HI 83,225).

Optimum formulation of NPK fertilizer medium using SDCG-air bubble

column
Air agitated SDCG-bubble column was used for the cultivation of
S. platensis and elemental N, P and K concentration in the culture
medium was varied by changing the loadings of NPK-10:26:26 complex fertilizer and NaHCO3 separately by keeping micro nutrients
concentration i.e., NaCl, MgSO4, CaCl2, FeSO4 and etc., which were
identical to Zarrouks culture medium.

Calculation of kinetic parameters

The important kinetic parameters that have been used to predict
the growth of S. platensis are: (i) nitrogen-to-cell conversion factor,
YX/N (g g
1), (ii) phosphorus-to-cell conversion factor, YX/P (g g
1),
(iii) cell productivity, PX (mg L
1 d
1) and (iv) maximum specic
growth rate, mmax (d
1). YX/N was determined from the ratio of the
dry biomass produced and the total nitrogen added to the culture
medium. Similarly, YX/P was calculated after considering phosphorous content in the culture medium. PX was calculated from the ratio
of (XmXo) and the growth time corresponding to Xm, where Xm and
Xo are the maximum cellular concentration and the initial
inoculums cellular concentration respectively in the culture
medium; whereas mmax (= ln(Xm/Xm
1)/tm
tm
1) was calculated
on the day when maximum growth was observed. In the present
study, CO2 xation rate (PCO2) was calculated based on biomass
productivity (Px) and molecular mass of S. platensis
(CH1.650O0.531N0.170S0.007P0.006, Cornet et al. [42]) and was given
by 1.77  Px which is slightly lower than the formula proposed by
Chisti [43].

Effect of NPK-10:26:26 loading

For this study, NPK fertilizer loading was varied and NaHCO3
concentration was kept constant which was identical to Zarrouks
culture medium. Details of the SDCG-air bubble column experiments (P1P6) with varying NPK fertilizer are given in Table 1. It
was observed that S. platensis did not grow at all for the culture
medium when NPK-10:26:26 fertilizer loading was more than
1.5 g L
1. In this case, the inhibition was mainly due to the excess of
ammonical nitrogen [35,44] and elemental phosphorous content
[45] in the culture medium. To reduce the effect of ammoniacal
nitrogen and elemental phosphorous, growth of S. platensis was
carried out at the reduced concentration of NPK-10:26:26 complex
fertilizer. The satisfactory growth of S. platensis was observed in
P1P4 culture mediums. The details of the growth results with
varying NPK fertilizer loading are shown in Fig. 2a. The results
revealed that the growth of S. platensis increases with the increase
in concentration of NPK fertilizer content in the culture medium up

Table 1
Experimental design of the culture medium studied for the growth of S. platensis using NPK-10:26:26 complex fertilizer in SDCG bubble column.
NaNO3
(g L
1)

K2HPO4
(g L
1)

K2SO4
(g L
1)

Saltsa
(g L
1)

NPK-10:26:26
(g L
1)

A5 micro
nutrients (mL)

Variation of NPK-10:26:26 (air)

P1
16.8/

1.33

1.0

P2

16.8/

1.33

P3

16.8/

1.33

P4

16.8/

1.33

P5

16.8/

1.33

P6

16.8/

1.33

0.36
(N = 0.036; P = 0.045; K = 0.077)
0.76
(N = 0.076; P = 0.089; K = 0.164)
1.06
(N = 0.106; P = 0.1224; K=0.23)
1.26
(N = 0.126; P = 0.146; K = 0.272)
1.36
(N = 0.136; P = 0.157; K = 0.293)
1.46
(N = 0.146; P = 0.169; K = 0.315)

Variation of NaHCO3 for P2 and P3 batch (air)

P2C1
22.0/
P2C2
18.0/
P2C3
10.0/
P2C4
4.50/
P3C1
22.0/
P3C2
18.0/
P3C3
10.0/
P3C4
4.50/

1.33
1.33
1.33
1.33
1.33
1.33
1.33
1.33

0.76
0.76
0.76
0.76
1.06
1.06
1.06
1.06

1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0

Zarrouk (air)
Zarrouk

2.5

0.5

1.0

1.33

1.0

2.5
2.5
2.5
2.5

0.5
0.5
0.5
0.5

1.0
1.0
1.0
1.0

1.33
1.33
1.33
1.33
1.33
1.33
1.33
1.33

0.76
0.76
0.76
0.76

1.0
1.0
1.0
1.0
1.0
1.0
1.0
1.0

Culture medium

NaHCO3/NaOH
(g L
1)

16.8

Variation of NaOH (simulated ue gas, SGF)

NPK1
/1.22
NPK2
/2.22
NPK3
/3.32
NPK4
/5.04
Z1
/1.22
Z2
/2.22
Z3
/3.32
Z4
/5.04
a

NaCl: 1.0, MgSO47H2O: 0.2, EDTA: 0.08, CaCl22H2O: 0.04 and FeSO47H2O: 0.01 (g L
1).

1.0
1.0
1.0
1.0
1.0

A. Kumari et al. / Journal of Environmental Chemical Engineering 2 (2014) 18591869

2.1

2.1

2b. Biomass (NPK, NaHCO 3; g l-1)

2a. Biomass (NPK,NaHCO 3; g l-1)

P1(0.36, 16.8)
P2(0.76, 16.8)
P3(1.06, 16.8)
P4(1.26, 16.8)
P5(1.36, 16.8)
P6(1.46, 16.8)
Zarrouk

1.5

P2C1(0.76,22)
P2C2(0.76,18)
P2C3(0.76,10)
P2C4(0.76,4.5)
P3C1(1.06,22)
P3C2(1.06,18)
P3C3(1.06,10)
P3C4(1.06,4.5)

1.8
Biomass concentration (g l-1 )

Biomass concentration (g l -1 )

1.8

1.2

0.9

0.6

1863

1.5

1.2

0.9

0.6

0.3

0.3

0
0

12

16

20

Time (d)

12

16

20

Time (d)

Fig. 2. (a) Time behavior of S. platensis growth with varying NPK-10:26:26 complex fertilizer loadings with xed sodium bicarbonate concentration using air agitated
SDCG-bubble column and comparison with Zarrouks culture medium. (b) Time behavior of S. platensis growth with varying sodium bicarbonate loading with xed NPK
fertilizer loading using air agitated SDCG-bubble column (Table 1: for details of variation of NPK-10:26:26 fertilizer and sodium bicarbonate).

to a certain limit and thereafter growth was reduced with the

increase in NPK loading. The best S. platensis growth was observed
in the neighborhood of 1.06 g L
1 NPK fertilizer concentration
(i.e., P3 culture medium: Table 1) with maximum dry biomass
concentration (i.e., 1.74 g L
1) at the tenth day of biomass growth
(Fig. 2a). Optimum ammoniacal nitrogen loading for P3 batch was
found to be 0.106 g L
1, which was also higher than the
conventional sources of ammoniacal nitrogen [35,44]. It was also
observed that the initial optimal elemental phosphorous concentration (i.e., 0.1224 g L
1) in NPK culture medium at the maximum
biomass growth was higher than the Zarrouks elemental
phosphorous concentration (i.e., 0.089 g L
1). It was also noted
that the optimum elemental potassium loading for P3 culture
medium was found to be 0.23 g L
1 which was much lower than
Zarrouks culture medium (i.e., 0.672 g L
1). The growth study was
also carried out using 0.76 g L
1 NPK fertilizer loading
(i.e., P2 culture medium) where elemental phosphorous content
was exactly same as Zarrouks culture medium and maximum
biomass concentration with 1.14 g L
1 was obtained at the tenth
day of biomass growth (Fig. 2a), which was comparable to
P3 culture medium. The initial elemental N, P and K concentrations
for P2 culture medium were calculated as 0.076, 0.089 and
0.164 g L
1 respectively. Details of kinetic parameters (i.e., Xm, PX,
mmax, YX/N and YX/P), chlorophyll, protein, lipid and the uptake of
elemental N, P and K for P1P5 culture mediums are reported in
Table 2 and comparable results were obtained for P2 and P3 culture
mediums.

the tenth day of biomass growth (Fig. 2b) which was comparable to
maximum biomass concentration obtained from Zarrouks culture
medium (i.e., 1.87 g L
1). Details of kinetic parameters (i.e., Xm, PX,
mmax, YX/N and YX/P), chlorophyll, protein, lipid and the uptake of
the elemental N, P and K for the culture mediums with varying
bicarbonate concentration for P2C3 and Zarrouks culture medium
are also reported in Table 2. Comparable growth results were
obtained for P2C3 and Zarrouks culture medium. Therefore, the
composition of optimum NPK-10:26:26 complex fertilizer culture
medium for Spirulina growth using air agitated SDCG-bubble
column was found to be NPK-10:26:26 fertilizer0.76 g L
1,
NaHCO310.0 g L
1,
NaCl1.33 g L
1
and
A5
micro

1
nutrients1.0 mL L . Above P2C3 culture medium with 0.76 g L
1
NPK-10:26:26 complex fertilizer loading was considered for the CO2
sequestration from ue gas studies using SDCG-bubble column.

Effect of NaHCO3 loading

For efcient carbon xation, it is essential to know the optimum
loading of NaHCO3 in addition to NPK complex fertilizer. In this study,
P2 and P3 NPK fertilizer mediums were considered and NaHCO3
concentrations were varied from 4.5 to 22.0 g L
1. Details of the batch
experiments with varying NaHCO3 loadings are given in Table 1 and
corresponding biomass growth results are shown in Fig. 2b . It was
noted that the maximum biomass growth results were obtained for
P2C3 culture medium where optimum NaHCO3 and
NPK-10:26:26 complex fertilizer loadings were found to be
10.0 and 0.76 g L
1 respectively, and the maximum dry biomass
concentration for this culture medium was found to be 1.82 g L
1 at

CO2 Solubility without NaOH

To determine the solubility of CO2, simulated ue gas was
purged through the aqueous suspension of NPK-fertilizer with
varying loadings using SDCG-bubble column (Fig. 1). Details of
time variant dissolved CO2 concentrations with varying NPK
fertilizer loadings were shown in Fig. 3a and the results were also
compared solubility results using Zarrouks culture medium
(Fig. 3a). It was noted that the attainment of equilibrium in
absence of NaOH took place within 30 min and demonstrate the
efcacy of the SDCG-bubble column. The attainment of equilibrium CO2 solubility in absence of NaOH using SDCG-bubble column
is very fast as compared to the conventional CO2 agitated bubble

CO2 Solubility using simulated ue gas in SDCG-bubble column

To obtain desired carbonate-bicarbonate level in the culture
medium, solubility of CO2 in the culture medium involving Zarrouk
and NPK fertilizer culture medium determined by purging ue gas
through the SDCG-bubble column (Fig. 1). Usually, solubility of CO2
depends on the CO2 content in ue gas and nutrient concentration
in the culture medium. Moreover, the solubility of CO2 was also
affected by the presence of NaOH in the culture medium.
Therefore, CO2 solubility in NPK fertilizer medium was determined
without/with NaOH loading.

1864

A. Kumari et al. / Journal of Environmental Chemical Engineering 2 (2014) 18591869

0.20
3a. SFG Without NaOH

Zarrouk

P1(0.36 g/L)

P2(0.76 g/L)

P3(1.06 g/L)

P4(1.26 g/L)

P5(1.36 g/L)

7
3b. SFG With NaOH, g/L
P2: 0.76 g/L NPK

P3: 1.06 g/L NPK

CO2 Solubility (g.L-1)

CO2 Solubility (g.L -1 )

0.16

0.12

0.08

P2(0.84)

Z(0.84)

P3(0.84)

P2(1.68)

Z(1.68)

P3(1.68)

P2(3.32)

Z(3.32)

P3(3.32)

P2(5.04)

Z(5.04)

P3(5.04)

5
4
3
2

0.04

1
0.00

0
0

10

15

20

25

30

35

Time (min)

8
Time (min)

12

16

Fig. 3. (a) Time behavior of CO2 solubility with varying NPK-10:26:26 complex fertilizer loadings without NaOH using simulated ue gas agitated SDCG-bubble column and
comparison with Zarrouks culture medium (excluding NaHCO3). (b) Time behavior of CO2 solubility with varying NaOH (g L
1: 0.84, 1.68, 3.32 and 5.04) using
NPK-10:26:26 complex fertilizer (P2: 0.76 g L
1, P3: 1.06 g L
1) and simulated ue gas using SDCG-bubble column and comparison with Zarrouks culture medium (excluding
NaHCO3).

columns. The attainment of equilibrium time was reduced

signicantly using SDCG-bubble column and it was found to be
three times less than the conventional CO2 solubility measurement
devices [41]. It was observed that the CO2 solubility decreased with
the increase in salt loading in the culture medium. The maximum
solubility was obtained for P1 culture medium (i.e., 0.134 g L
1)
where NPK fertilizer loading was minimum (Fig. 3a). The decrease
in solubility was mainly due the salt-out effect of an electrolyte
and has been attributed to the increased attraction between added
ions and water (called hydrated ions) than between CO2 and water.
The degree of hydration will increase with the increase in added
salts [41].
CO2 solubility with NaOH
The solubility of CO2 in absence of NaOH is mainly due to the
formation of carbonic acid and as a result solubility of CO2 is
considerably low in the culture medium. To increase the CO2
solubility, ue gas was passed through the culture medium in
presence of NaOH. In this case, CO2 will be converted in the form of
carbonate and/or bicarbonate depending on the pH of the medium
[26]. Time variant dissolved CO2 concentrations with varying
NaOH loading using P2 and Zarrouk culture medium are shown in
Fig. 3b. Quantity of NaOH added in the culture medium is shown in
the bracketed term of Fig. 3b. It was observed that the attainment
of equilibrium was established within 8.0 min. The attainment of
equilibrium CO2 solubility in presence of NaOH using SDCG-bubble
column is very fast and it was also found to be almost three times
less than the conventional CO2 solubility measurement devices
[41]. It was also observed that CO2 solubility increased with the
increased in NaOH loading and maximum solubility attained for
5.04 g L
1 NaOH loading (Fig. 3b). At 5.04 g L
1 NaOH loading,
P2 culture medium shows highest CO2 solubility whereas
P3 culture medium shows lowest solubility, which was mainly
due to the salt-out effect of the electrolytes.
Bubble size distribution and mass transfer coefcient
The rate of CO2 transfer from ue gas to the culture medium
depends on the total interfacial surface area of the generated gas
bubbles through the sintered disk. Bubble size distributions at
maximum biomass concentration (i.e., at the sixth day) for P2 and

Zarrouk culture medium are shown in Fig. 4a and b respectively

with varying NaOH concentration. It is interesting to note that the
bubble size distribution shifted towards right with the decrease in
NaOH concentration for both the culture mediums and accordingly, average bubble diameter also decreased with the increased in
NaOH loading (Table 3). The reduction in bubble size was mainly
due to the increased concentration of salts and biomass at higher
NaOH loadings.
To estimate the CO2 transfer capability using SDCG-bubble
column, the overall gasliquid volumetric mass transfer coefcient
(kLaL, s
1) of CO2 was estimated according to Chisti [46] where
kL = gasliquid mass transfer coefcient (m s
1) and aL = interfacial
area per unit volume of bubbles (m2 m
3). The values of kLaL were
obtained using Eqs. (1) and (2) for the known values of gas holdup
(er), mean bubble diameter (dB, m), and true mass transfer
coefcient (kL, m s
1) values i.e.,
kL
kL aL 1
sr

6sr
dB

(1)

The values of kL/dB in Eq. (1) was calculated according to Chisti [46]
using Eq. (2) for simulated ue gaswater dispersions and waterlike suspending uid:
!0:5
kL
gDL sr2
5:63  10
5
exp
0:131 C 2S
(2)
dB
m3L
where g = acceleration due to gravity (m s
2), DL = diffusivity of the
transferring CO2 into liquid (m2 s
1), s = interfacial tension(J m
2),
mL = viscosity of the culture broth (kg m
1 s
1), r = density of the
suspension (kg m
3) and CS = concentration of suspended algae
(g L
1). To evaluate volumetric mass transfer coefcient at given
ue gas ow rate (i.e., 1.3 L min
1), the parameters in Eqs. (1) and
(2) (i.e., dB, er, s , mL, r and CS) were measured using standard
measuring instruments during maximum biomass growth period
and the details of these parameters are given in Table 3 for P2 and
Zarrouks culture medium with varying NaOH concentration. To
evaluate the values of mass transfer coefcients of CO2, it is
essential to know the exact diffusivity values of CO2. The diffusivity
values are calculated from the duration of ue gas cycle, which was
related to the characteristic diffusion time. It was noted that ue
gas cycle time for given NaOH loading in the culture medium was

A. Kumari et al. / Journal of Environmental Chemical Engineering 2 (2014) 18591869

18

18
4a. P2:SFG+NPK

16
Frequency of bubbles (Number)

NPK2: NaOh-2.22 g
NPK3: NaOH-3.32 g

14

Zarrouk: NaOH-1.22 g

4b. SFG+Zarrouk

NPK1: NaOH-1.22 g

16

NPK4: NaOH-5.04 g

Frequency of bubbles (number)

1865

12
10
8
6
4
2

Zarrouk: NaOH-2.22 g
Zarrouk: NaOH-3.32 g

14

Zarrouk: NaOH-5.04 g

12
10
8
6
4
2

0
0

0.002

0.004

0.006

0.008

0.01

0.001 0.002 0.003 0.004 0.005 0.006 0.007 0.008

Bubble diameter, dB (m)

Bubble diameter, dB (m)

Fig. 4. (a) Bubble size distribution at sixth day of biomass growth for P2 culture medium with S. platensis under varying NaOH loading using simulated ue gas agitated
SDCG-bubble column. (b) Bubble size distribution at sixth day of biomass growth for Zarrouks culture medium with S. platensis under varying NaOH loading using simulated
ue gas agitated SDCG-bubble column.

much shorter than that of air cycle time which indicates that the
2
characteristic diffusion time dB =36DL was much less than the
biomass growth time, where DL = diffusivity of the transferring CO2
in liquid and Characteristic length = 1/3  dB/2. The details of
diffusion time for P2 and Zarrouks culture medium with varying
NaOH loading are given in Table 3 and it was increased with the
increased in NaOH loading. The shorter diffusion time was mainly
due to high gasliquid interfacial area and demonstrates the

efciency of the SDCG-bubble column for CO2 sequestration.

Knowing the characteristic diffusion time and bubble diameter,
diffusivity of CO2 was calculated. The details of diffusivity values
are also given in Table 3 and the diffusivity values were reduced
with the increased in the density of the culture medium. Now, the
volumetric mass transfer coefcient of CO2 was estimated and the
details are listed in Table 3 for P2 and Zarrouksculture medium
with varying NaOH. For given ue gas ow rate, it was observed that

Table 2
Responses of S. platensis in SDGC bubble column using NPK-10:26:26 complex fertilizer and Zarrouk medium.
Culture medium X (mg L
1) PX (mg L
1 d
1 mmax (d
1) Nutrient uptake
(mg L
1)
N

YX/N
YX/P
PCO2 (mg L
1 d
1) Chlorophyll (mg L
1) Protein (%) Lipid (%)
(g g
1) (g g
1)
K

Varying NPK loading (xed NaHCO3 loading),

P1
500
45.0
P2
1140
109.0
P3
1740
169.0
P4
910
86.0
P5
650
60.0

ten days biomass growth (air)

0.062
19.0
8.6
0.121
61.0
10.0
0.135
83.0
10.2
0.110
52.0
9.6
0.080
20.0
9.0

7.7
18.0
19.0
18.0
18.0

12.50
14.34
15.94
6.83
4.41

10.82
12.25
13.74
5.89
3.82

5.13
8.87
12.37
7.52
6.01

44.07
44.83
45.40
45.78
45.97

7.69
8.44
9.01
9.39
9.57

Varying NaHCO3
P2C1
P2C2
P2C3
P2C4
P3C1
P3C2
P3C3
P3C4

ten days biomass growth (air)

0.115
41.0
10.0
0.126
47.0
13.0
0.141
76.0
15.0
0.107
45.0
8.0
0.090
29.0
8.0
0.13
71.0
9.0
0.087
47.0
6.0
0.081
34.0
7.0

14.0
9.0
17.0
18.0
15.0
16.0
13.0
16.0

11.58
14.87
23.29
9.74
6.60
15.19
6.32
6.13

9.89
12.70
19.89
8.31
5.69
13.15
5.45
5.28

7.64
9.10
12.84
6.82
6.59
11.35
6.41
6.30

44.49
44.73
45.81
48.80
44.92
45.26
46.77
50.94

8.10
8.34
9.41
12.38
8.54
8.87
10.36
14.51

47.1

4.43

20.45

13.13

51.20

14.76

7.0
8.0
13.0
19.0
7.0
9.0
14.0
17.0

174.72
205.38
328.92
499.26
195.10
246.50
365.50
452.33

6.0
6.6
9.0
12.4
6.4
7.4
9.7
11.5

45.8
47.0
48.8
53.3
46.0
51.0
53.1
54.5

9.4
10.6
12.4
16.8
9.2
10.5
12.1
16.0

loading (xed NPK loading),

930
88.0
1180
113.0
1820
177.0
790
74.0
750
83.0
1660
161.0
720
65.0
700
61.0

Zarrouk: ten days biomass growth (air)

Zarrouk
1870
182.0

0.14

243.0

14.0

P2 and Zarrouk: varying NaOH loading, six days biomass growth (simulated
NPK1
642
98.71
0.10
36.0
6.0
NPK2
746
116.0
0.12
40.0
9.0
NPK3
1365
185.8
0.15
40.7
12.0
NPK4
1845
282.1
0.21
54.0
14.8
Z1
711
110.2
0.11
129.0
4.0
Z2
886
139.3
0.12
157.5
5.0
Z3
1289
206.5
0.16
186.0
7.0
Z4
1783
255.6
0.17
226.0
9.5

ue gas)
10.0
7.79
12.0
9.16
14.0
14.67
14.0
22.27
38.0
1.61
40.0
3.38
42.0
3.01
45.0
3.73

Where Xm: maximum biomass concentration (mg L
1), PX: biomass productivity(mg L
1 day
1), mmax: maximum specic biomass growth rate (d
1), PCO2 : CO2 xation rate
(mg L
1 day
1) and calculated based on CH1.650O0.531N0.170S0.007P0.006 molecular mass of S. platensis (Cornet et al., [27]): 1.77 Px,YX/N: conversion yield of
nitrogen to biomass (g g
1), YX/P: conversion yield of phosphorous to biomass (g g
1).

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A. Kumari et al. / Journal of Environmental Chemical Engineering 2 (2014) 18591869

Table 3
Details of the average measured parameters at the maximum biomass growth using simulated ue gas in SDCG-bubble column.
Parameters

Culture medium
NPK1

1

NaOH loading (g L )
Temperature (1 K)
Air ow rate/cycle,
(L min
1 cycle)
Flue gas ow rate/cycle,
(L min
1 cycle)
No. of ue gas cycle/day
Diffusion time, tD (s)
Initial CO2 solubility (g L
1)
Average CO2 solubility (g L
1)
Mean bubble diameter, dB  103 (m)
Gas holdup, er (%)
Density of the suspension, r (kg m
3)
Interfacial tension, s 102 (J m
2)
Viscosity of the culture broth, mL  103 (kg m
1 s
1)
Concentration of suspended solids, CS (g L
1)


2
Diffusivity of the transferring CO2 in liquid, DL  1010 (m2 s
1) dB =36tD
where characteristic length of sphere = radius of sphere/3.
Overall gasliquid volumetric mass transfer coefcient, kLaL  103 (s
1) [46]
Average pH
Dissolved oxygen (mg L
1)

NPK2

NPK3

NPK4

Z1

Z2

Z3

Z4

1.22
303
1.3

2.22
303
1.3

3.32
303
1.3

5.04
303
1.3

1.22
303
1.3

2.22
303
1.3

3.32
303
1.3

5.04
303
1.3

1.3

1.3

1.3

1.3

1.3

1.3

1.3

1.3

4.0
4.2
4.5
4.6
4.0
4.2
4.5
4.6
674
951
1288
1804
607
902
1205
1503
1.28
2.34
3.48
5.24
1.28
2.34
3.48
5.24
1.20
2.34
3.26
4.84
1.18
2.34
3.35
4.98
2.10
1.78
1.46
0.74
1.31
1.20
1.09
0.61
0.054
0.053
0.053
0.053
0.057
0.057
0.056
0.056
1484.2
1694.6
1956.5
2358.2
1815.1
2032.5
2292.9
2666.3
6.31
6.25
6.13
6.08
5.42
5.32
5.23
5.15
1.14
1.15
1.15
1.16
1.17
1.17
1.18
1.18
0.65
0.75
1.17
1.75
0.72
0.89
1.29
1.59
0.93
0.46
0.08
0.79
0.44
0.27
0.07
1.82

the volumetric mass transfer coefcient decreased with increase in

NaOH loading. Though average bubble diameter decreased with
NaOH loading (Fig. 4a and b), but culture medium density increased
signicantly with NaOH loading which has negative effect on mass
transfer coefcient [46]. The calculated volumetric CO2 mass transfer
coefcient in SDCG-bubble column during biomass growth was
enhancedsignicantlyascomparedtotheconventionalairlift/bubble
column. The average volumetric mass transfer coefcient (kLaL) in
presence of NaOH loading (i.e., 1.225.04 g L
1) at 1.3 L min
1 CO2
ow rate was calculated from Table 3 and was found to be
4.3  10
3 s
1, which was almost ten times higher than conventional
bubble column/air lift column [47].
Growth kinetics using simulated ue gas
The growth kinetics of S. Platensis was studied using
SDCG-bubble column for eight days period using the best
NPK-10:26:26 culture medium (i.e., P2: Table 1) and Zarrouks
culture medium. Details of biomass growth with varying NaOH
loading for P2 culture medium was shown in Fig. 5a. Similar
studies were also carried out using Zarrouk culture medium and
corresponding variation of biomass growth is shown in Fig. 5c . Due
to semi-batch purging of ue gas, the variation of pH in the culture
medium varies in between 9.0 and 10.0. Details of pH variation for
two days period using fertilizer and Zarrouks culture medium are
shown in Fig. 5b and d for 1.22 and 5.04 g L
1 NaOH loading
respectively. As pH was maintained more than 8.3, culture medium
contains only Na2CO3 [41]. It is interesting to observe from Fig. 5b
and d that the average purging time of CO2 using 5.04 g L
1 NaOH
was higher than the culture medium with 1.22 g L
1 NaOH and the
increased purging time is mainly due to the increased alkali
concentration in the culture medium. Therefore, highly concentrated culture medium with aqueous alkali solution delays the
change in pH during biomass growth as compared to dilute alkali
solution (Fig. 5b and d).
It is note that the maximum growth of biomass using P2 and
Zarrouks culture medium was observed at the sixth day of biomass
growth when ue gas was purged using SDCG-bubble column,
whereas growth inhibition starts after tenth day of biomass growth
whengrowthstudieswerecarriedoutusingairagitatedSDCG-bubble
column with NaHCO3. Flue gas assisted biomass growth with varying
NaOH loading was studied and the details of NaOH loading variation

7.46
9.36
4.0

5.75
9.35
3.8

4.17
9.33
3.6

1.69
9.34
3.2

5.59
9.37
3.7

4.50
9.31
3.5

3.42
9.37
3.4

1.76
9.47
2.9

are given in Table 1. It was also noted that biomass growth increased
withtheincreasedloadingofNaOH(Fig.5aandc).Detailsofmaximum
biomass concentration with varying NaOH concentration in ue gas
agitatedbubblecolumnaregiveninTable2.ForNPK4culturemedium
(NaOH: 5.04 g L
1, Table 1), maximum biomass concentration (Xm)
with 1.85 g L
1 was obtained at the sixth day of biomass growth using
NaOH-ue gas assisted SDCG-bubble column, whereas Xm with
1.82 g L
1 was obtained at tenth day of biomass growth using
NaHCO3-air assisted SDCG-bubble column for P2C3 culture medium
(NaHCO3:10 g L
1,Table1).Theenhancementofbiomassgrowthrate
was mainly due to the maintenance of higher average carbon
concentration in the culture medium throughout biomass growth.
DetailsofaverageCO2 solubilitywithvaryingNaOHloadingforP2and
Zarrouks culture medium are given in Table 3. Moreover, dissolved
oxygen in the culture medium for all experiments remained almost
steady at 3.5 mg L
1 (Table 3) and the constant values of dissolved
oxygen indicates that the accumulatedreactive oxygen in the culture
medium is minimum which was mainly due to high liquidgas
interfacial area with improved back mixing in the culture medium
and helped to reduce the inhibitoryeffect of oxygen accumulation on
growth.
Biomass productivity (Px) and maximum specic growth rate
(mmax) of S. Platensis using SDCG-bubble column for all the culture
mediums using air and ue gas are given in Table 2. Px and mmax for
ue gas agitated culture medium was found to be 282.1 mg L1 d
1
and 0.21 d
1 respectively (i.e., NPK4: Table 2), whereas low values
of Px (=177.0 mg L1 d
1) and mmax (=0.141 d
1) were obtained for air
agitated culture medium (i.e., P2C3: Table 2). Details of these
kinetic parameters (Xm, Px and mmax) using P2 culture medium
with varying NaOH in ue gas agitated SDCG-bubble column were
reported in Table 2 and compared with Zarrouks culture medium.
Better biomass growth results were obtained for ue gas agitated
NPK culture medium (e.g., NPK4 in Table 2: Xm = 1.85 g L
1,
Px = 282.0 mg L1 d
1 and mmax = 0.21 d
1) as compared to ue gas
agitated Zarrouk culture medium (e.g., Z4 in Table 2: Xm = 1.78 g L
1,
Px = 255.6 mg L1 d
1and mmax = 0.17 d
1). The improved kinetic
parameters show the efciency of NPK-10:26:26 based biomass
cultivation using ue gas assisted biomass growth in SDCG-bubble
column over air purging. Comparing the biomass growth using
NPK and zarrouks culture medium, an enhancement in S. platensis
growth was also obtained for NPK fertilizer medium as compared
to standard Zarrouks culture medium (Table 2). It was also

A. Kumari et al. / Journal of Environmental Chemical Engineering 2 (2014) 18591869

2000

10.5

5a:SFG+NPK (P2, NaOH: g/L)

5b:SFG+NPK (P2, NaOH: g/L)

1867

NPK2 (0.76, 1.22)

NPK4 (0.76, 5.04)

NPK1 (0.76, 1.22 )

NPK2 (0.76, 2.22 )

1600

NPK3 (0.76, 3.32 )

10.0

1200

pH

Biomass (mg.L-1)

NPK4 (0.76, 5.04 )

9.5

800

9.0
400

8.5
0

10

0.0

0.5

2000

2.0

5d: SFG+Zarrouk (NaOH: g/L)

Z1 (NaOH: 1.22 g/L)

Z1 (NaOH:1.22 g/L)

Z2 (NaOH: 2.22 g/L)

Z4 (NaOH: 5.04 g/L)

Z3 (NaOH: 3.32 g/L)

10.0

Z4 (NaOH: 5.04 g/L)

1200

pH

Biomass (mg.L-1)

1.5

10.5

5c:SFG+Zarrouk

1600

1.0
Time (d)

Time(d)

9.5

800

9.0
400

8.5

0
0

10

Time (d)

0.0

0.5

1.0
Time (d)

1.5

2.0

Fig. 5. (a) Time behavior of S. platensis growth using P2 (NPK: 0.76 g L
1) culture medium with varying NaOH loadings (g L
1: 1.22, 2.22, 3.32, 5.04) using simulated ue gas
agitated SDCG-bubble column. (b) Variation of pH during two days of biomass growth period using P2 (NPK: 0.76 g L
1) and NaOH (1.22 and 5.04 g L
1) in simulated ue gas
agitated SDCG-bubble column. (c) Time behavior of S. platensis growth using Zarrouks culture medium with varying NaOH loadings (g L
1: 1.22, 2.22, 3.32, 5.04) using
simulated ue gas agitated SDCG-bubble column. (d) Variation of pH during two days of biomass growth period using Zarrouks medium and NaOH (1.22 and 5.04 g L
1) in
simulated ue gas agitated SDCG-bubble column.

observed that the CO2 xation rate using NPK4 and Z4 culture
medium was found to be 499.26 and 452.33 mg L
1 d
1 respectively (Table 2) and corresponding CO2 xation rate was increased
by 10.0% using NPK4 culture medium. The increased CO2 xation
rate was mainly due to the increased loading of NaOH. In CO2-toalgae route, CO2 xation was enhanced by the addition of NaOH in
liquid phase with following reactions:
CO2 g CO2 l

(3)

CO2 l H2 O H2 CO3 l

(4)

H2 CO3 NaOH HCO

3 Na H2 O

(5)

2

HCO

3 NaOH CO3 Na H2 O

(6)

Comparing above reactions, Eq. (3) is mass transfer limiting

step which is enhanced by using SDCG-bubble column with high
mass transfer coefcient (discussed in Bubble size distribution
and mass transfer coefcient section). However, Eqs. (4) and (5)

are relatively slow and identied as the rate determining steps in

the reaction mechanism of CO2 in aqueous bicarbonate and
carbonate solutions. In presence of substantial NaOH loading in
aqueous solution, alkaline mechanism predominates and Reaction
(6) completely shifted towards right which is responsible for the
increase in solubility of CO2 under alkaline condition [41,48,49].
The average CO2 solubility in NPK4 culture medium was found to
be 4.84 g L
1 at 5.04 g L
1 of NaOH loading whereas lowest CO2
solubility was obtained for NPK1 culture medium with 1.20 g L
1 at
1.22 g L
1 of NaOH loading (Table 3) and corresponding CO2
xation rate using NPK4 culture medium was enhanced by
2.9 times as compared to NPK1 culture medium. Similar
observation was also obtained for Zarrouk culture medium with
varying NaOH loading. The details of CO2 xation using NPK and
Zarrouk culture medium with varying NaOH loading was given in
Table 2. It was also mentioned above reactions are controlled by pH
of the culture medium and the formation of bicarbonate dominates
when pH of the suspension was varied within 4.3 and 8.3, whereas
carbonate formation was dominant when pH of the suspension

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A. Kumari et al. / Journal of Environmental Chemical Engineering 2 (2014) 18591869

was more than 8.3. Biomass yield based on nitrogen (YX/N) and
phosphorous (YX/P) calculated on basis of nutrient uptake and the
details of nutrient uptake and corresponding yields are reported in
Table 2 with varying NaOH concentration in P2 and Zarrouks
culture medium. The values of YX/N and YX/P for P2 and Zarrouks
culture medium were increased with the increased in NaOH
loading.
Details of chlorophyll, lipid and protein content in biomass
using SDGC-bubble column for all the experiments with/without
using ue gas are given in Table 2. An enhancement of chlorophyll
and lipid accumulation was observed using NPK culture medium as
compared to Zarrouks culture medium. Chlorophyll and lipid
accumulation using NPK4 culture medium was found to be
12.4 mg L
1 and 16.8% respectively, which were higher than Z4
culture medium (Table 2) and corresponding increases in
chlorophyll and lipid accumulation were found to be 7.8% and
5.0% respectively. The enhancement of lipid accumulation was
mainly due to deciency of initial nitrogen in the culture medium.
The decreased cellular nitrogen content of thylakoid membrane
activates diacylglycerol acytransferage and converts acyl-CoA to
tri-glycerides [26]. Comparable results for protein were obtained
for with varying NaOH concentration in P2 and Zarrouks culture
medium. It was also found that the accumulation of chlorophyll,
lipid and protein content were increased with the increase in
biomass growth.
The cost of nutrients at the maximum biomass growth using
NPK4 and Z4 was calculated. The cost of nutrients per kilogram of dry
biomass using NPK fertilizer medium was found to be US $46.2 which
was much lower than Zarrouks culture medium with US $92.5. The
corresponding cost saving was found to be almost 50.0% using NPK
fertilizer medium as compared to standard Zarrouks culture
medium. The reduction in unit cost of biomass using NPK fertilizer
was mainly due to the reduced quantity of the low-priced NPK
fertilizer and higher biomass growth rate as compared to standard
Zarrouks culture medium. Therefore, the merits of the NPK
fertilizer based culture medium are clearly emphasized, not only
as a low-cost alternative but also as a highly productive for CO2
sequestration, which may be used protably in rural population for
large-scale biomass cultivation of protein-rich S. platensis.
Conclusions
The present investigation was carried out to formulate a simple
and inexpensive culture medium for the growth of S. platensis
using NPK-10:26:26 complex fertilizer using air agitated sintered
disk chromatographic glass bubble column. The composition of
optimal NPK fertilizer was found to be NPK-10:26:26
fertilizer0.76 g L
1, NaHCO310.0 g L
1, NaCl1.33 g L
1 and A5
micro nutrients1.0 mL L
1 and comparable growth results were
obtained as compared to the standard Zarrouks culture medium.
Using above optimally formulated NPK fertilizer culture medium,
diesel generator simulated ue gas was used for the mass
cultivation of biomass in presence of sodium hydroxide using
sintered disk chromatographic glass bubble column. A semi-batch
feeding strategy of ue gas purging was adopted in presence of
NaOH for CO2-to-Spirulina route, and the following technological
improvements were obtained over the conventional method of
biomass cultivation: (i) use of soluble carbonates (e.g., NaHCO3 and
Na2CO3) for Spirulina cultivation (ii) reduction of biomass growth
period using NPK-NaOH culture medium by four days in ue gas
agitated SDCG-bubble column as compared to air agitated
NPK-NaHCO3 culture medium where maximum biomass concentration was obtained on tenth day of biomass growth (iii) an
improvement in maximum biomass concentration (4.0%),
chlorophyll production (7.8%) and lipid accumulation
(5.0%) by using NPK-NaOH culture medium as compared to

Zarrouk-NaOH culture medium in ue gas agitated SDCG-bubble

column (iv) CO2 xation rate with 499.26 mg L
1 d
1 was obtained
as using ue gas assisted NPK-NaOH culture medium as compared
to ue gas assisted Zarrouk-NaOH culture medium with CO2
xation rate 452.33 mg L
1 d
1, which was equivalent to 10.0%
increase in CO2 xation rate (v) an almost 50.0% cost saving was
achieved due to the use of low cost NPK-10:26:26 complex
fertilizer as a nutrient for Spirulina growth in comparison to
standard Zarrouks culture medium. Therefore, the technology of
CO2-to-Spirulina route was improved signicantly through the
reduction of operating cost of biomass production and increasing
biomass throughput by using NPK-10:26:26 complex fertilizer and
simulated diesel generator ue gas.
Acknowledgements
Partial nancial support from Indian School of Mines, Dhanbad
[through research project grant FRS (28)2010-11/PE] is gratefully
acknowledged. The authors are also like to express their sincere
gratitude for their valuable comments.
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