Waste Management 85 (2019) 264–271

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Waste Management
journal homepage: www.elsevier.com/locate/wasman

Investigation of platinum recovery from a spent refinery catalyst with a

hybrid of oxalic acid produced by Aspergillus niger and mineral acids
Hamed Malekian, Mahsa Salehi, Davoud Biria ⇑
Department of Biotechnology, Faculty of Advanced Sciences and Technologies, University of Isfahan, Hezar Jarib Ave, Isfahan, Iran

a r t i c l e i n f o a b s t r a c t

Article history: The capability of oxalic acid produced by Aspergillus niger was investigated for bioleaching of platinum
Received 7 October 2018 from a refinery reforming catalyst. The spent medium mode was selected for bioleaching because of
Revised 18 December 2018 its higher efficiency at favorable pH and temperature conditions. The effects of several important factors
Accepted 31 December 2018
such as the pulp density, pH and temperature on platinum recovery were optimized using Box-Behnken
Available online 3 January 2019
design of response surface methodology. The results indicated that pH adjustment during the bioleaching
process increases the final platinum recovery significantly. The obtained optimum conditions were 1% for
Keywords:
the pulp density, 0.5 for the medium pH, and 70 °C for the temperature which led to 37% platinum recov-
Bioleaching
Platinum recovery
ery. The significance of oxalic acid as the leaching agent in platinum bioleaching was highlighted by
Aspergillus niger investigating the recovery of a blank medium without oxalic acid at the optimum conditions which
Oxalic acid was just about 13%. The presented method can be utilized in an environmentally friendly process to
Refinery catalyst recover platinum from industrial catalysts.
Ó 2019 Elsevier Ltd. All rights reserved.

1. Introduction 2016; Sun et al., 2016). In pyro-metallurgy based processes, vari-

Platinum group elements are widely used in industrial catalysts
because of their remarkable catalytic qualities (Mpinga et al.,
2015). For example, Pt/alumina has been used in the catalytic
reforming process in which the low octane naphtha is converted
to higher octane hydrocarbons (Rahimpour et al., 2013). In every
catalytic process, the catalyst activity declines by time because of
the chronic deposition of the poisoning compounds on its surface.
After a few regeneration cycles, the catalytic activity will be
reduced significantly, and the catalyst will finally turn into a solid
waste known as the spent catalyst (Marafi and Stanislaus, 2008;
Mishra et al., 2008). In fact, the spent refinery catalysts are known
to be one of the most dangerous wastes generated in petroleum
refineries because of the leachable heavy metals in their composi-
tion (Marafi and Stanislaus, 2003; Mishra et al., 2007). Therefore,
reclamation of their valuable metals like platinum is of the great
importance from both the environmental and economical points
of view (Chauhan et al., 2013; Asghari et al., 2013; Yang et al.,
2011).
Several processes for the metals recovery have been developed
on the basis of pyro-metallurgy, hydro-metallurgy and bio-
hydrometallurgy principals (Akcil et al., 2015; Zhang and Xu,
⇑ Corresponding author.
E-mail address: d.biria@ast.ui.ac.ir (D. Biria).
ous heat treatments are used to extract metals from the catalyst.
Thus these methods are usually energy intensive and produce sev-
eral pollutant gases such as SO2. In the hydrometallurgy processes,
a variety of acidic or alkaline solutions are employed for the metal
leaching. However, the production of hazardous wastes in these
processes is in common which are difficult to treat (Jha et al.,
2013). In bio-hydrometallurgy, the microorganisms and their
metabolites are utilized to extract metals from solid materials
(Johnson, 2014). In fact, bioleaching is considered to be more
cost-effective and eco-friendly than the other conventional leach-
ing methods (Zhuang et al., 2015).
The most common microorganisms capable of metal solubiliza-
tion include chemolithotrophic autotroph bacteria as well as some
heterotrophic bacteria and fungi (Sand et al., 2001; Vera et al.,
2013; Srichandan et al., 2013). Aspergillus niger is one of the
favored species for heterotrophic bioleaching which is capable of
producing several organic acids, such as citric, oxalic and gluconic
acid from low cost substrates such as the cheese whey. The
bioleaching of various metals such as Ni, Fe, Al, V, Mo, Sb, Cu, Co
and Mn using Aspergillus niger has been studied in previous
research. (Bayraktar, 2005; Aung and Ting, 2005; Gholami et al.,
2011; Amiri et al., 2012; Rasoulnia and Mousavi, 2016; Horeh
et al., 2016; Mohanty et al., 2017; Faraji et al., 2018). However,
the capability of Aspergillus niger in leaching of noble metals
including platinum from alumina-based catalysts has been hardly

https://doi.org/10.1016/j.wasman.2018.12.045
0956-053X/Ó 2019 Elsevier Ltd. All rights reserved.
 H. Malekian et al. / Waste Management 85 (2019) 264–271 265

reported. In fact, the low activity of platinum group metals in com-

bination with the high potential of alumina to be dissolved in the
leaching conditions makes it difficult to develop a bioleaching pro-
cess with adequate platinum recovery efficiency. Consequently,
efforts to extract the platinum from the catalyst structure by bio-
logical methods have not yielded high recovery values. For exam-
ple, Motaghed et al. (2014) have reported the platinum recovery
of about 16% for a spent catalyst sample after seven days of treat-
ment by cyanide producing bacteria. Shin et al. (2015) demon-
strated that high platinum recovery could be achieved through a
spent medium process at high bio-cyanide concentration and
150 °C. Aside from the unfavorable conditions such as the long
time of the process or the high temperature in the reported results,
the application of cyanide as the bioleaching agent can be consid-
ered as an environmental problem. Recently, the combination of
biological processes with proper chemical promoters has been sug- Fig. 1. The produced oxalic acid concentration in various bioleaching modes of
gested as a remedy to the low recovery efficiency of platinum operation with and without pH readjustment.
group metals bioleaching in non-cyanide processes. The resultant
hybrid bio/chemical metal extraction system enjoys the environ-
Table 1
mental benefits of bioleaching and at the same time, takes the
Chemical composition of the spent catalyst
advantages of high and fast recovery of the chemical extraction sample through XRF analysis.
(Awasthi and Li, 2017; Ilyas et al., 2018).
Compound % Weight
On the basis of the above, the aim of the work was to investigate
the effects of pH adjustment with mineral acids on the bioleaching Al2O3 92.15
Cl 1.43
of Pt from a spent refinery catalyst by Aspergillus niger. Preliminary
SnO2 0.54
experiments were conducted to determine the most efficient Fe2O3 0.343
method of bioleaching before optimizing the effective factors Pt 0.307
(i.e., pulp density, leaching temperature and pH) through TiO2 0.284
Box-Behnken response surface methodology. Complimentary SiO2 0.120
CaO 0.058
experiments were performed at the obtained optimum criteria to
SO3 0.036
evaluate the effect of leaching duration, and the acid type on Pt CuO 0.015
recovery. Loss of ignition 4.95
Total 100.14

2. Materials and methods

2.1. Materials
The spent de-coked Sn-Pt/Al2O3 catalyst from a continuous cat-
alytic reforming unit was supplied by the National Iranian Oil
Refining and Distribution Company. Lactose and yeast extract were
obtained from Scharlau. KH2PO4 and CaCl2 were purchased from
Sigma-Aldrich. Nitric acid, oxalic acid, tween 80, MgSO4-7H2O
and NaNO3 were obtained from Merck. Potato-dextrose agar
(PDA), hydrochloric acid, sodium hydroxide and glucose were from
Biolife, Dae-Jung, Samchun, and Quelab respectively.
2.2. Characterization of spent catalyst
The spent de-coked catalyst from a continuous catalytic reform-
ing unit which contained platinum on an alumina support
(Sn-Pt/Al2O3) was utilized in this work. The fresh active catalyst
has been chlorinated prior to use in the refinery but the activity
of the spent catalyst is reduced because of the coke deposition
and chloride loss. The spent catalyst sample was ground using a
ball mill and then sieved to obtain a fine powder with less than
150 mm particle size. This powder was used in all of the conducted
experiments in this study. The shapes of the spent and powdered
catalyst have been shown in the supplementary materials (Fig. 1s).
The composition of the spent catalyst was analyzed by an X-Ray
fluorescence (XRF) analyzer (Bruker, S4 pioneer) and the obtained
results have been shown in Table 1. In addition, acidic digestion
(using aqua-regia solution) (Zanjani and Baghalha, 2009) followed
by Inductively Coupled Plasma (ICP) analysis was performed to
determine the platinum content of the catalyst samples which
was equal to 0.29% (w/w).
2.3. Microorganism and the growth conditions
Aspergillus niger PTCC 5012 was utilized in the bioleaching
experiments. To prepare the inoculum for the liquid culture, the
fungus was cultured on potato-dextrose agar (PDA) plates at
30 °C for 7 days. Then, the surface of the PDA plate was washed
with a solution of 0.05% tween 80 to harvest the fungal spores
(Strasser et al., 1994). After that, the harvested spores were
counted using a Thoma counting chamber and the number of
spores in the inoculum was adjusted to 5
107 spores per mL.
1 mL of the prepared inoculum was added to 100 mL of the liquid
culturing medium to obtain the final spore density equal to 5
105
spores per mL. The liquid culturing medium was an aqueous solu-
tion containing of 100 g/L lactose, 2.5 g/L glucose, 1.6 g/L yeast
extract, 1.5 g/L NaNO3, 0.5 g/L KH2PO4, 25 mg/L MgSO47H2O, and
25 mg/L CaCl2. The initial pH of the medium was set to 6 and it
was autoclaved prior to inoculation (Mandal and Banerjee, 2005).
The flasks were incubated at 30 °C and 180 rpm for 10 days. In fact,
the oxalic acid production by the fungi during the incubation time
had been previously investigated (data not shown) and the maxi-
mum production was achieved after 10 days.
2.4. Effect of pH on oxalic acid production
The ability of Aspergillus niger in biosynthesis of oxalic acid and
the effect of medium pH on its production yield was investigated.
Liquid culture flasks of the fungus were prepared and incubated at
30 °C and 180 rpm for 10 days as described in Section 2.3. In half of
the flasks, the pH of the medium was measured daily and adjusted
266 H. Malekian et al. / Waste Management 85 (2019) 264–271

between 7 and 8 using a 3 N NaOH solution while no pH readjust- temperature). Selection of the important factors and their levels
ment was performed in the second half. By the end of the incuba- were based on the results of preliminary experiments to screen
tion period, the oxalic acid concentration was measured in all of the most important factors for bioleaching of platinum by
the flasks. Aspergillus niger. The resultant experimental design has been shown
in Table 2. It should be noted that the experiments were carried out
2.5. Bioleaching experiments in two replicates and a confidence level equal to 0.95 was selected
for statistical analysis. All of the experimental design and analysis
Generally, a bioleaching process can be performed either as a were performed by Design Expert software (version 7.0).
direct (one or two-step process) or an indirect two-step approach. The required spent medium for bioleaching was provided by the
In the direct single-step method, fermentation and leaching will be fungal cultivation in an adjusted pH condition as described in
taken place simultaneously which means that leaching of metals Section 2.4 and the produced biomass was removed from the
from the catalyst and cultivation of microorganisms are happening medium. The obtained spent medium solution was used in the
at the same container. In the direct two-step process, the leaching designed bioleaching experiments according to Table 2.
and cultivation are still taking place at the same container but the
leaching will begin later than the cultivation. In the indirect 2.7. Analytical methods
method, the catalyst will be added to a previously cultured med-
ium meaning that leaching will be performed in a microorganism 2.7.1. Estimation of oxalic acid concentration in the medium
free medium obtained from the fermentation (the spent medium) To measure the oxalic acid concentration, the obtained spent
(Asghari et al., 2013). The appropriate method should be chosen by medium was subjected to high performance liquid chromatogra-
a trade-off between the lower cost of the direct method and the phy (HPLC; Knauer, smart line system) with a UV visible detector
higher efficiency of the two-step approach. at 210 nm and a C18 column. The HPLC-grade oxalic acid solution
Several flasks containing 100 mL liquid culture of Aspergillus was injected into the column as the standard (Rasoulnia and
niger were prepared as described in Section 2.3. For the single step Mousavi, 2016).
bioleaching experiments, the sterilized spent catalyst powder was
added to the flasks (1% w/v) at the beginning of culturing time and
2.7.2. Estimation of platinum extraction yield
they were incubated in a rotary shaker at 30 °C and 180 rpm. In
The dissolved platinum in the system was analyzed by taking
these experiments, one-half of the flasks were subjected to a daily
appropriate samples from the bioleaching flasks. The samples were
pH adjustment (in the range of 7 –8) while the second half
centrifuged and the concentration of platinum in the obtained
remained intact. For the two-step bioleaching, the spent catalyst
supernatants was measured by inductively coupled-plasma optical
powder (1% w/v) was added to the flasks two days after the fungal
emission spectrometer (ICP-OES Perkin Elmer, Optima 7300DV).
inoculation. By the end of a ten day period, the concentration of
Metal recovery percentage was calculated through Eq. (1) as below
oxalic acid as well as the extracted platinum was measured in all
(Gerayeli et al., 2013):
of the flasks.
To investigate the leaching efficiency of the spent medium, the m
V
R% ¼
100 ð1Þ
fungal cultivation was carried out according to the procedure w
M
explained in Section 2.3. It should be noted that the fungi cultiva-
where m shows the concentration of platinum in the sample, V
tion in this stage was performed in two conditions, i.e., with and
stands for the volume of the medium, w is the mass fraction of
without daily pH adjustment of the culturing medium. The pro-
duced biomass was removed from the system by filtering through
a paper filter and centrifugation. The obtained supernatant was Table 2
divided into two 100 mL parts and the catalyst powder was added Box-Behnken experimental design conditions and the obtained results.
to both of them (1% w/v). For the first part, the pH was set to 1 Run Pulp density (%(w/w)) pH Temperature (°C) Metal recovery (%)
using the nitric acid solution and incubated for 6 h at 70 °C while
1 3 0.5 30 14
the second part was incubated for 6 h at 30 °C with no pH adjust- 2 1 0.5 50 27.8
ments. The six-hour time of leaching was selected according to the 3 3 0.5 70 27.8
results of a set of preliminary experiments to investigate the effect 4 1 2.5 50 23.2
of time on the leaching efficiency. 5 3 1.5 50 19.8
6 5 1.5 30 10.9
The influence of the addition of excess acid to platinum recov-
7 5 1.5 70 15.8
ery was investigated in a single step bioleaching process as 8 3 1.5 50 19.6
described above with a modification in the added catalyst powder. 9 1 1.5 30 12
In these experiments, the liquid culturing media were supple- 10 3 2.5 70 22.5
mented with certain amounts of catalyst powder to obtain 0.5, 1, 11 5 2.5 50 11
12 1 1.5 70 35
2 and 4% (w/v) pulp densities. At the end of the ten day incubation 13 5 0.5 50 20
time, the oxalic acid content of each sample was measured. After 14 3 2.5 30 8.7
that, nitric acid was added to samples to the final concentration 15 1 0.5 50 25.6
of 1.5 M. The flasks were incubated at 70 °C for 6 h and then, the 16 5 0.5 50 19.2
17 1 2.5 50 22
extracted platinum was measured.
18 5 2.5 50 10.6
19 1 1.5 30 11.2
2.6. Spent medium bioleaching experiments according to experimental 20 5 1.5 30 10
design 21 1 1.5 70 33.2
22 5 1.5 70 15.2
23 3 0.5 30 12.4
Response surface methodology (RSM) is a common approach to 24 3 2.5 30 8.4
optimize the process outputs because of its convenient application 25 3 0.5 70 30.2
and high efficiency (Ilyas et al., 2014). In this work, Box-Behnken 26 3 2.5 70 20.1
design (BBD) was used to study and optimize the effects of several 27 3 1.5 50 19.6
28 3 1.5 50 19.3
important factors on platinum recovery (i.e. pulp density, pH and
 H. Malekian et al. / Waste Management 85 (2019) 264–271
platinum in the spent catalyst and M is the utilized mass of the
spent catalyst.
3. Results and discussions
3.1. Effect of pH on oxalic acid production
Aspergillus niger is capable of producing several organic acids,
including citric and/or oxalic acid on sucrose and lactose and glu-
conic acid which is rapidly produced on glucose (Mandal and
Banerjee, 2005; Srichandan et al., 2013). In this study, lactose
was used as the main carbon source of fermentation, since it is
believed to have the highest oxalic acid yield among the sugars.
(Cameselle et al.,1998; Santoro et al., 1999; Mandal and Banerjee,
2005)
In the medium with controlled pH, the initial pH of the cultur-
ing medium was set to 6 by the nitric acid solution. After 2 days of
incubation, the medium pH decreased because of the production of
oxalic acid. At this time, the pH was adjusted in the range of 7–8
and controlled daily during the fermentation period. By the end
of the fermentation, oxalic acid concentration in the medium with
controlled pH was 12 g/L, while the culture without pH control just
produced about 3.6 g/L oxalic acid with the final pH equal to 3.1.
The observed positive effect of adjusting the pH on biosynthesis
of oxalic acid by Aspergillus niger is in agreement with previously
published results (Cameselle et al., 1998; Strasser et al., 1994). It
is believed that the higher pH values can stimulate the fungal pro-
duction of oxalic acid while the enzymes responsible for hydrolysis
of the oxalate will be produced in low pH values (Mandal and
Banerjee, 2005).
3.2. Assessing the feasibility of bioleaching
To evaluate the efficiency of different bioleaching approaches
(i.e. single step, two-step and spent medium) the produced oxalic
acid concentration and metal recovery were measured for each
case. As it can be seen in Fig. 1, the oxalic acid content of the sam-
ples with controlled pH was much higher than that of the uncon-
trolled samples for all cases. In addition, the concentration of
oxalic acid in the spent medium was the lowest. This means that
the presence of catalyst in the system can be in favor of the oxalic
acid production. It is believed that the toxic effect of catalyst mate-
rials induces the fungus to excrete more metabolites including
oxalic acid to diminish the metal toxicity through forming complex
compounds or precipitation (Santhiya and Ting, 2005). Investiga-
tion of the effect of toxic metals on the production of carboxylic
acids such as oxalic and citric has been shown that the fungus mit-
igated the toxic effects of metals by secretion of these metabolites
(Gadd, 1992, 2007).
Fig. 2 shows the platinum recovery of the various examined
bioleaching cases. Similar to the results of the oxalic acid yield,
metal recoveries were almost equal for one-step and two-step
bioleaching while the spent medium resulted in a slightly lower
recovery. In fact, controlling the pH of the medium in all of the
studied bioleaching modes resulted in higher oxalic acid concen-
tration which in turn led to higher metal recovery. This confirms
that the oxalic acid plays the main role in platinum extraction.
Oxalic acid is known to be a complex-forming compound which
is able to solubilize different kinds of metals (Strasser et al.,
1994). Solubilization of alumina by oxalic acid can be considered
as one of the main mechanisms of platinum extraction from the
catalyst. In fact, reforming catalysts are manufactured by satura-
tion of porous c- alumina with chloroplatinic acid. Since the Pt
complex is placed on the alumina base through an electrostatic
absorption, the solubilization of alumina by oxalic acid may
267
Fig. 2. Platinum recovery of various bioleaching modes of operation with and
without pH readjustment.
enhance the separation of platinum from the catalyst matrix.
Moreover, platinum can directly form a complex with oxalate
which also improves the metal extraction (Masuda et al., 2013;
Underhill and Watkins, 1980). Although a higher oxalic acid con-
centration was obtained for the one-step bioleaching, the platinum
recovery was higher in lower pH values and higher temperatures of
the spent medium process. This verified the significance of leach-
ing conditions (pH and temperature) in the final yield of the pro-
cess and introduced the spent medium method as a more
promising technique of bioleaching. In fact, the spent medium
without the cells can be applied in high temperature and low pH
conditions.
3.3. One-step bioleaching followed by acidic leaching experiments
The positive effect of adding acid (reducing the pH) in platinum
recovery for the spent medium method was verified by the above
results. It was also demonstrated that the oxalic acid production
was higher for one-step bioleaching with pH control. Therefore,
the combination of one step bioleaching conditions with the conse-
quent pH reduction might have a synergistic effect on platinum
recovery. To examine the idea, nitric acid was added to the med-
ium at the end of the single step bioleaching process to reach the
final concentration of 1.5 M equal to [94.5 g/L]. Simultaneously,
the effect of pulp density on oxalic acid yield and metal recovery
was investigated and the obtained results have been shown in
Fig. 3.
Apparently, increasing the pulp density up to 4% did not affect
the fungal oxalic acid production significantly. On the other hand,
the pulp density had an adverse effect on platinum leaching and
bioleaching efficiency reduced significantly for pulp densities
above 0.5%. This could be explained by the adsorption of dissolved
platinum to fungal biomass or reduction of catalyst contact area as
a result of its entrapment in the biomass (Guo et al., 2010).
3.4. Experimental design of spent medium bioleaching
3.4.1. Statistical analysis
The platinum recovery percentages of Box Behnken experimen-
tal runs were calculated and reported in Table 2. The following
second-order model (Eq. (2)) was obtained for platinum recovery
in terms of natural factors:
Pt Recov eryð%Þ ¼ 19:8 þ 3:43A þ 0:73B þ 1:26C  0:6AB
 0:106AC  0:05BC þ 0:066A2 þ 0:016B2
 0:005C 2 ð2Þ
268 H. Malekian et al. / Waste Management 85 (2019) 264–271

which proved that low temperatures were unfavorable to bioleach-

Fig. 3. Effect of pulp density on the produced oxalic acid concentration and
platinum recovery in one-step bioleaching followed by acidic leaching.
ing. In fact, the diffusion rate of the leaching agent (e.g., oxalic acid)
into the interface of the heterogeneous catalyst was higher at the
elevated temperatures which improved the bioleaching efficiency
(Rasoulnia and Mousavi, 2016). The influence of high temperature
was weakened at the higher pulp densities. The observed effect
could be described by the low diffusion rate of leaching agents to
the catalyst at higher pulp densities. In fact, the catalyst/liquid
interface increases at higher pulp densities while the leaching
agent concentration remains almost unchanged.
Fig. 4c shows the contour plot of the interactions between the
temperature and pH and their effects on platinum recovery at
holding pulp density of 3%. Clearly, the platinum recovery
increased by increasing the temperature within the studied pH
range and decreased with pH rise all over the temperature domain.
The trend can be explained by the higher diffusion rate of leaching
agent at the higher temperatures.

3.4.3. Optimization of bioleaching and assessing the model viability

where A, B and, C refer to pulp density, pH and temperature respec-
tively. Analysis of variance (ANOVA) was employed to examine the
significance of the studied factors as well as the suggested empirical
model and the results have been presented in Table 3. Accordingly,
the low p-value, high determination factor (0.96) and a non-
significant lack of fit approved the adequacy of the model. Also,
all of the factors and their interactions were found to be significant
except for A2 and B2. It was revealed that the increase of pulp den-
sity and pH decreased the Platinum recovery while leaching tem-
perature had a principal and positive effect on the leaching
efficiency.
The normal probability and residual plots of the model have
been shown in Fig. 2s as supplementary materials which confirmed
the accuracy of the model assumptions.
3.4.2. Response contour plots
Fig. 4a shows the contour plot of the interactive effects of pulp
density and pH on platinum recovery at holding temperature of
50 °C. As can be seen, platinum recovery increased by reduction
of pH regardless of the pulp density alteration. On the other hand,
pH reduction was more effective at higher pulp density values
because increasing the catalyst content of the medium would need
more H+ ions to reach the same platinum recovery. Therefore, at
higher pulp densities the positive effect of pH on platinum recov-
ery was more distinct.
The effect of pulp density and temperature on platinum recov-
ery at the pH value equal to 1.5 as the holding value has been
shown in Fig. 4b. Seemingly, the platinum recovery declined by
increasing the pulp density within the entire temperature range.
However, the recovery decrease was sharper at higher temperatures
The suggested optimum conditions by the software were pulp
density of 1%, pH equal to 0.5 and bioleaching temperature of
70 °C. Under the mentioned conditions, the platinum recovery
was predicted to be 37.6% by the model. To evaluate the accuracy
of the model, an additional experiment was performed under the
optimal conditions. According to Table 4, the obtained result is
within the calculated confidence interval of the model which con-
firms its validity.
There are few reports on the recovery of platinum from spent
catalysts through bioleaching in the literature. Recently, Bacillus
megatrium was employed to extract platinum from a used catalyst
sample and at the optimum condition (pulp density of 4%), plat-
inum recovery was obtained equal to 16% (Motaghed et al.,
2014). In another study, mobilization of platinum from spent auto-
mobile catalyst was conducted using HCN forming bacteria and
only 0.2% of platinum were mobilized at the end of a 10 day treat-
ment (Brandl et al., 2008). In this study, the obtained platinum
recovery at pulp density of 4% was 25% and it was 37% at the opti-
mum conditions which were much higher than the previous
reports. It should be noted that no cyanide has been used in the
present process which is an improvement in environmental
protection.
Biological processes are time-consuming in general but they are
promising due to their undeniable advantages. In fact, the time-
consuming step in this process is bio-production of oxalic acid by
the fungus. The bio-production of oxalic acid is a cost effective
option because it can produced on low cost carbon sources such
as the dairy wastewaters. Since the oxalic acid production and
leaching are two separate steps in the presented spent medium
method, the slow rate problem of the oxalic acid bio-production

Table 3
ANOVA table of the obtained model for platinum recovery.

Sum of squares DF Mean squares F-value P-value

Model 1511.33 9 167.93 219.29 <0.0001
A 373.46 1 373.46 487.69 <0.0001
B 159.39 1 159.39 208.15 <0.0001
C 786.80 1 786.80 1027.47 <0.0001
AB 11.04 1 11.04 14.42 0.0013
AC 152.25 1 152.25 198.82 <0.0001
BC 4.65 1 4.65 6.07 0.0240
A2 0.1 1 0.1 0.13 0.7220
B2 0.32 1 0.32 0.42 0.5236
C2 20.45 1 20.45 26.70 <0.0001
Residual 13.78 18 0.77
Lack of fit 0.51 3 0.17 0.19 0.9011
R2 = 0.98 R2adj = 0.96
 H. Malekian et al. / Waste Management 85 (2019) 264–271 269

Fig. 4. Two dimensional contour plots for platinum recovery (a) pH vs. pulp density at T = 50 °C, (b) temperature vs. pulp density at pH = 1, (c) temperature vs. pH at pulp
density of 3%.

Table 4 medium in the unchanged pH conditions was equal to 27% versus

Experimental confirmation of the response under optimum conditions.
the 8% of the control. In addition, at the optimum conditions where
Predicted 95% CI 95% CI Confirmation the pH of the spent medium has been adjusted by adding the
recovery low high experiment acid, the recovery was obtained equal to 37% while its control
Platinum recovery (%) 37.6 35.53 38.65 37 (without bio-oxalate) resulted in 13% recovery. The 27% recovery
of the spent medium without external acid addition was higher
than that of the control sample in the optimum conditions (13%)
could be addressed conveniently. Moreover, the spent medium with nitric acid which confirmed the significance of the fungi
bioleaching has the privilege of not restricting the users to limited produced oxalate.
pH, temperature, pulp density and other operating parameters
hence higher recovery efficiencies can be achieved which is 3.5.2. Assessing the effect of leaching duration on platinum recovery
favored for industrial purposes. All the performed experiments in this study were carried out at
a constant leaching duration of 6 h. In order to examine the effect
3.5. Complimentary experiments of leaching time on the platinum recovery, an extra experiment
was conducted under the optimum conditions but the leaching
3.5.1. Comparison between bioleaching by cultured and uncultured duration was doubled (12 h) and the Pt recovery under this condi-
mediums tion was found to be 35.5% which is almost the same as the 6 h
Platinum recovery was measured in a cell free bio-oxalic acid leaching duration. This showed that increasing the leaching dura-
solution (spent medium) and an uncultured medium (i.e. not inoc- tion did not make a notable change in the extraction efficiency in
ulated by fungi) with the same composition as the control. The test the current conditions. It seems that the reactant exhaustion could
has been repeated for two conditions, namely the obtained opti- be the reason for stopping the leaching progress by time. When a
mum criteria and the unchanged pH conditions (with no nitric acid higher concentration of oxalic acid (27 g/L) was examined (results
addition). According to Fig. 5, the platinum recovery for the spent are in the Section 3.5.4), it was revealed that about 5% recovery
270 H. Malekian et al. / Waste Management 85 (2019) 264–271
Fig. 5. Comparison between platinum recovery results of bioleaching by cultured
(spent medium) and uncultured (control) mediums (at pulp density: 1%, T: 70 °C).
increase in a 6 h treatment time can be achieved. This means that
the retreatment of the residual catalyst by a fresh medium can be
more beneficial to platinum recovery than a longer leaching pro-
cess time.
3.5.3. Effect of acid type on the platinum recovery
As previously mentioned, the pH of all the experiments in this
study was adjusted by nitric acid. To investigate the difference
between nitric and hydrochloric acid on metal recovery, an exper-
iment was conducted under optimum conditions in which HCl was
employed to set the pH to 0.5. The results proved that the platinum
recovery in the sample with HCl was 4% higher (Pt recovery of 41%)
than the sample in which nitric acid was used for setting the pH (Pt
recovery of 37%). This could be due to the formation of complexes
such as PtCl2 6 which can facilitate Pt extraction (Baghalha et al.,
2009).
3.5.4. Comparison between bioleaching and chemical leaching
Chemical leaching experiments were conducted using commer-
cial oxalic acid as the leaching agent at the concentrations identical
to those of the fungi produced oxalate at the optimum conditions
(13.5 g/L oxalate, pulp density of 1%, pH adjusted to 0.5 by adding
hydrochloric acid and bioleaching temperature of 70 °C) as well as
the twice of the fungal production (27 g/L). As can be seen in Fig. 6,
platinum recovery in the chemical leaching was slightly higher
than the bioleaching process. It can be concluded that the oxalic
Fig. 6. Comparison between platinum recovery results of bioleaching and chemical
leaching (at pulp density: 1%, T: 70 °C and pH: 0.5).
acid has a principal role in bioleaching process as in the chemical
system there were no fungi to produce other metabolites.
The comparison between platinum recovery of chemical leach-
ing at oxalic acid concentrations of 13.5 g/L and 27 g/L revealed
that the increase in organic acid concentration in this range did
not enhance the leaching efficiency considerably since the recovery
at 27 g/L is only 5% higher.
4. Conclusions
In this study, bioleaching of platinum from a refinery reforming
catalyst sample by the Aspergillus niger produced oxalic acid solu-
tion was investigated. Various methods of bioleaching (i.e., one-
step, two-step and spent medium bioleaching with and without
pH adjustment) were examined. It was revealed that oxalic acid
production and platinum recovery were higher in the experiments
with pH control. In spite of the higher oxalic acid production in
one-step and two-step methods, the spent medium mode was
proved to have greater platinum recovery because it can be applied
at lower pH values in combination with mineral acids and higher
temperatures. Accordingly, the spent medium bioleaching was
selected as the desired leaching method for the optimization
through Box-Behnken design of RSM methodology. Several effec-
tive factors such as pulp density, pH, and temperature were opti-
mized by fitting a second-order quadratic model to the
experimental results. The obtained optimal conditions were pulp
density of 1%, pH of 0.5 and bioleaching temperature of 70 °C that
resulted in the platinum recovery equal to 37%. Using HCl instead
of the nitric acid to adjust the pH value at the optimum conditions
was resulted in higher recovery efficiency (41%). This shows that
there is still room to improve the process.
Acknowledgments
This work has been partially supported by the National Iranian
Oil Refining & Distribution Company (NIORDC).
Appendix A. Supplementary material
Supplementary data to this article can be found online at
https://doi.org/10.1016/j.wasman.2018.12.045.
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