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C) 50 55 60 65
(B) Initial pH 1 1.5 2 2.5
(C) Dilution rate (h
1
) 0.1 0.2 0.3 0.4
(D) Initial Fe
3+
concentration (g L
1
) 1 3 5 7
(E) Aeration rate (mLmin
1
) 100 150 200 250
2.4. Orthogonal array and experimental parameters
For the Taguchi design and subsequent analysis, the software
named as Qualitek-4 (version 4.82.0) was used. The appropri-
ate orthogonal array for the experiment was determined by the
software. A well designed experiment can reduce substantially
the number of experiments required. The Taguchi technique
applies fractional factorial experimental designs, called orthog-
onal arrays, to reduce the number of experiments and meanwhile
obtaining statistically meaningful results.
The most important stage in the design of an experiment lies
in the selection of control factors, therefore as many factors as
possible should be included and no signicant variables must
be identied at the earliest opportunity. Taguchi method cre-
ates an orthogonal array to accommodate these requirements.
The selection of a suitable orthogonal array depends on the
number of control factors and their levels. By inspecting prac-
tical observation, ve selected control factors and their levels
applied in this study are listed in Table 1. These control factors
include temperature, initial pH, dilution rate, initial Fe
3+
con-
centration, and aeration rate. All control factors have four levels.
With the selection of L
16
orthogonal array, using ve mentioned
parameters and their levels, shown in Table 2, the number of
experiments required can be drastically reduced to 16. It means
that 16 experiments with different combinations of the factors
should be conducted in order to study the main effects and inter-
actions, which in the classical combination method using full
factorial experimentation would require 4
5
=1024 number of
experiments to capture the inuencing parameters. However,
in general, Taguchi design is preferred because it reduces the
number of experiments signicantly.
3. Results and discussion
3.1. Analysis of variance
The main objective of ANOVA is to extract from the results
howmuch variations each factor causes relative to the total vari-
ation observed in the result. According to the ANOVA results
in Table 3, the initial pH has the largest variance and the initial
Fe
3+
concentration indicated the second place. Therefore, it can
be concluded that the most inuential factor was in the order of
the pH. On the other hand, the degree of freedom(DOF) for each
factor was 3 and total DOF was 15, so the DOF for error term
was 0, and nally the variance for the error term (Ve), obtained
by calculating error sumof squares and dividing by error degrees
of freedom, could not be calculated. Henceforth, it was impossi-
938 S.M. Mousavi et al. / Chemical Engineering and Processing 46 (2007) 935940
Table 2
L
16
orthogonal array (Levels of ve different factors and obtained results)
Experiment number A B C D E Obtained results [Fe
2+
biooxidation rate (g L
1
h
1
)]
Run S/N ratio (db)
1 2 3
1 1 1 1 1 1 5.8 5.5 5.5 14.955
2 1 2 2 2 2 6.3 6.3 6.5 16.075
3 1 3 3 3 3 7.2 7 6.9 16.939
4 1 4 4 4 4 5 5.2 5.2 14.203
5 2 1 2 3 4 5.5 5.7 5.3 14.795
6 2 2 1 4 3 5.3 5 5.5 14.41
7 2 3 4 1 2 7.8 7.8 7.7 17.804
8 2 4 3 2 1 6.6 6.8 6.4 16.382
9 3 1 3 4 2 4.9 5 5.2 14.029
10 3 2 4 3 1 6.4 6.4 6.1 15.98
11 3 3 1 2 4 7.4 7.2 6.9 17.095
12 3 4 2 1 3 5.8 5.6 6.1 15.302
13 4 1 4 2 3 5 5.1 4.9 13.975
14 4 2 3 1 4 5.6 5.5 5.9 15.055
15 4 3 2 4 1 5.6 5.5 5.6 14.91
16 4 4 1 3 2 5.1 5 5.2 14.148
Average S/N ratio (db) 15.379
Table 3
ANOVA analysis of S/N ratio
Factor Degrees of freedom (DOF) Sum of squares (S) Variance (V) F-ratio (F) Pure sum (S
) Percent, P (%)
(A) Temperature (
) Percent P (%)
(A) Temperature (
C) 55 2 0.469
(B) Initial pH 2 3 1.308
(C) Dilution rate (h
1
) 0.3 3 0.222
(D) Initial Fe
3+
concentration (g L
1
) 3 2 0.503
(E) Aeration rate (mLmin
1
) 100 1 0.178
of the applied technique for optimizing the ferrous biooxida-
tion parameters. So it is possible to increase biooxidation rate
signicantly using the proposed statistical technique.
4. Conclusion
In the present attempt optimization of ferrous iron oxidation
rate using an indigenous Sulfobacillus species in a packed-
bed reactor was investigated. Following the Taguchi method
of experimental design the effects of various factors inuenc-
ing the performance characteristics, were analyzed. Analysis of
S/N ratio has been successfully applied for nding out the rela-
tive contribution and the optimum factor level combination for
the maximum Fe
2+
biooxidation rate. According to the percent
contribution of each factor, indicated in the ANOVA table, it
could be inferred that initial pH of feed solution is the most pre-
dominant factor. Importance of the factors on the biooxidation
of Fe
2+
was ranked in Table 6. The critical process parameters,
according to their relative signicance, are initial pH of feed
solution, initial Fe
3+
concentration, temperature, dilution rate
and aeration rate, respectively. The maximum biological oxida-
tion rate was obtained by setting temperature 55
C, initial pH
2, dilution rate 0.3 h
1
, initial Fe
3+
concentration 3 g L
1
and
aeration rate 100 mLmin
1
. It was resulted that the biooxida-
tion rate of ferrous iron was increased by 7.7% at the optimum
conditions, which they determined by Taguchi optimization
method.
Acknowledgements
The authors would like to acknowledge Mehrdad Hesampour
for his useful help and discussion. They also thank Jamshid
Kash and Gharibali Farzi for their technical assistance at
BBRC.
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