Optimization of reactor volumes for gold cyanidation

Optimization of reactor volumes for gold cyanidation

Minerals Engineering 18 (2005) 671–679 This article is also available online at: www.elsevier.com/locate/mineng Optimization of reactor volumes for g...

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Minerals Engineering 18 (2005) 671–679 This article is also available online at: www.elsevier.com/locate/mineng

Optimization of reactor volumes for gold cyanidation L.R.P. de Andrade Lima *, D. Hodouin Department of Mining, Metallurgy and Materials Engineering, Laval University, Quebec City, Canada G1K 7P4 Received 10 November 2004; accepted 9 December 2004

Abstract The mineral industry has been using cyanidation of aerated slurries to recover gold from ores for more than a century. However, the leaching plant is usually designed as a series of agitated tanks of the same size, without any attempt to find an optimal plant design for improving the circuit efficiency, either by decreasing the cyanide consumption, or increasing the gold recovery, or decreasing the total plant volume. The objective of the study is to test, by simulation, if it would be profitable to use plant designs differing from the usual ones. The focus is put on the selection of the volumes of the tanks in the cascade of leaching reactors. The methodology involves the use of gold dissolution and cyanide consumption kinetic models incorporated into a simulator, and the definition of a performance criterion for the plant optimization. The performance is characterized by a cost function containing a term representing the value of the unleached gold and a term accounting for the cyanide consumption costs. It is shown that, for the same total volume, using a sequence of increasing size reactors improves the performance of the plant. The results are produced for different size of the ore particles and different numbers of tanks in the leaching circuit. Ó 2005 Elsevier Ltd. All rights reserved. Keywords: Cyanidation; Gold ores; Leaching; Modelling; Process optimisation; Process synthesis

1. Introduction Leaching of ores by cyanide in aerated alkaline slurries has been the dominant process for gold extraction for more than one century. In order to accelerate the gold recovery, in most of the high-grade ore plants, the cyanidation process occurs continuously in a cascade of large agitated tanks. The reactor volume optimization is a relevant problem that has been studied for many chemical systems; however, hydrometallurgical reactors have received much less attention. This is particularly

* Corresponding author. Present address: Department of Chemical Engineering, M.H. Wong Building, McGill University, 3610 University Street, Montreal, Canada H3A 2B2. Tel.: +1 514 398 5170; fax: +1 514 398 6678. E-mail address: [email protected] (L.R.P. de Andrade Lima).

0892-6875/$ - see front matter Ó 2005 Elsevier Ltd. All rights reserved. doi:10.1016/j.mineng.2004.12.007

true for the leaching tanks that are of interest in this paper. It is widely accepted that pure gold cyanidation is an electrochemical process, where gold is oxidized and then complexed to the stable ion [Au(CN)2], and oxygen is reduced and hydrogen peroxide decomposed (Habashi, 1987; Yannopoulos, 1991; Marsden and House, 1992). A typical gold ore processing plant is composed of the following sequence of unit operations: ore comminution, size classification, gravity concentration, and slurry dewatering, followed by gold leaching and gold recovery on activated carbon or by zinc precipitation, and finally gold elution, electrolytic extraction, melting and casting. The major reactants in the gold leaching process are cyanide and oxygen, but also sodium hydroxide is used to control pH, and sometimes lead nitrate is used to control cyanide consumption by sulfides. In a conventional cyanidation process, high cyanide concentrations in large tanks are used to improve gold

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Nomenclature Ccn Cl Co Cs Cw Cs1 d J Ml Ms N PrCN PrAu Qcn Ql Qs

cyanide concentration in the liquid (mg/kg) gold concentration in the liquid (mg/kg) oxygen concentration in the liquid (mg/kg) gold concentration in the ore (mg/kg) solid concentration in the pulp (g/g) residual gold concentration in the ore (mg/ kg) average size of the ore particles (lm) cost function ($/h) liquid hold up (kg) ore hold up (kg) number of reactors in the cascade price of cyanide ($/kg) price of gold ($/g) cyanide flow rate added (kg/h) liquid flow rate (kg/h) ore flow rate (kg/h)

leaching rate and gold recovery, but at the same time this practice is responsible for high cyanide consumption and investment (Kondos et al., 1995; de Andrade Lima, 2001; de Andrade Lima and Hodouin, Submitted for publication). A compromise between these two antagonistic effects is required for gold leaching plant performance optimization. Optimal reactor design and optimal operating conditions tuning are attractive techniques for such a process, to improve profitability in a context of strongly competitive markets, tight product specifications and tough environmental regulations (Levenspiel, 1999; Edgar and Himmelblau, 1988; Chitra and Govind, 1985; Biegler et al., 1997). The determination of the minimum volume configuration of a cascade of continuous stirred tank reactors (CSTRs) is a particular problem, in the field reactor optimization, which was first pointed out by Denbigh (1944) and subsequently applied to various chemical systems (Leclerc, 1953; Aris, 1961; Kubota et al., 1961; Sze´pe and Levenspiel, 1964; Wood and Stevens, 1964; Luss, 1965; Crooks, 1966; Floquet et al., 1985; Lopes and Malcata, 1993). The problem of the optimum design of the reactor sizes in a cascade of CSTRs is mathematically defined as follows: 8 min f ðVÞ > > > V > > > < s:t: ð1Þ Vi P0 > > > N P > > > : Vi ¼VT i¼1

where f is an objective function, such as the conversion of a reactant or product, V is the vector of the net vol-

rAu rCN ro r0o Vi VT

dissolution rate of gold (mg/kg h) rate of cyanide consumption (mg/kg h) rate of oxygen consumption (mg/kg h) rate of oxygen feed (mg/kg h) net volume of the ith reactor in the cascade (m3) total net volume of the reactor cascade (m3)

Greek symbols qs ore density (g/cm3) ql liquid density (g/cm3) s average residence time (h) Subscripts 0 first reactor of the cascade i ith reactor of the cascade N last reactor of the cascade

umes of the tanks in the cascade Vi, and VT is the total net volume of the cascade. The minimization problem given by Eq. (1) can be solved using dynamic programming or direct optimization methods, such as the conjugate or the reduced gradient methods (see Bellman, 2003; Edgar and Himmelblau, 1988; Wang and Fan, 1964). Three systematic optimal design studies of the volumes in a cascade of leaching reactors are available. The first one (Henein and Biegler, 1988) minimizes the residence time for a given conversion degree, in a cascade of reactors for leaching reactions that follow a shrinking core kinetic model. In the case of leaching under surface-reaction control, the ideal volumes in the cascade of CSTRs depend on the targeted solid conversion, but always decrease from the first to the last reactor. For a conversion of approximately 99.5%, the ratios Vi/VT in the case of four CSTRs are 0.314, 0.269, 0.226, and 0.187, and for very low conversion the ratios are approximately 0.25. In the case of leaching under pore-diffusion control, the ideal volume ratios also depend on the solid conversion: for approximately 99.5% conversion the ratios Vi/VT for four CSTRs are 0.205, 0.295, 0.285, and 0.215, and for 80% conversion the ratios are 0.13, 0.23, 0.30 and 0.34. The second study (Papangelakis and Demopoulos, 1992; Papangelakis and Luus, 1993) deals with pressure oxidation. The performance index is the conversion multiplied by the ratio of the time for complete conversion to the mean residence time. This index tends to maximize the conversion while minimizing the plant volume, and, when the ore residence time in the reactor is sufficient for complete leaching, it reduces to the ore leaching conversion. The

L.R.P. de Andrade Lima, D. Hodouin / Minerals Engineering 18 (2005) 671–679

results show that, for autothermal operations, the configuration of the autoclave that increases the reactor performance is that with a large first compartment followed by small ones. The third study (Crundwell, 2001) addresses the bacterial leaching of pyrite where Thiobacillus ferrooxidans is the only bacteria type that promotes the oxidation of the ferrous ions; the performance index is the conversion of pyrite. Using the same total residence time, the results show that the ideal configurations for a cascade of two to five reactors is a large first tank 1.5–2 times larger than the following small size tanks. Despite the economic importance of gold ore cyanidation, systematic studies of the optimal size design of the cascade of reactors are not available, especially due to the lack of reliable kinetic data for ore cyanidation. For a long time, the effort has been focused on the comprehension and quantification of pure gold cyanidation. However, gold ore cyanidation behaves very differently, because the gold grains are not completely liberated from the gangue, the galvanic interactions between the minerals, the effects of the foreigner ions in the solution, and the gold frequently occur in compounds or alloys. Recent studies by Nicol et al. (1984), Hodouin et al. (1990), Crundwell and Godorr (1997), Ling et al. (1996), and especially de Andrade Lima (2001) and de Andrade Lima and Hodouin (Submitted for publication) provided cyanidation kinetic equations that can be used in the optimization of this process.

NaCN 1

673

The objective of this paper is to present an analysis of the volume design of cascade of reactors for gold ore cyanidation. For that purpose, a rate equation for gold leaching and another for cyanide consumption are used to model the steady state gold cyanidation, in conjunction to a cost function based on a metallurgical performance criterion. Initially the effect of the ore size fraction and the number of reactors in the cascade are studied, then the most appropriate reactor volume distribution is analyzed for this system. The primary leaching section of Doyon Mine (Cambior Inc., Canada) is used as a case study to illustrate the optimal design method (see Fig. 1). In this plant, leaching is conducted in a three-tank cascade of reactors, mechanically agitated and with a nominal volume of 412 m3 each. The ore flowrate is about 164.5 t/h, the solid content in the pulp is 48.8%, and the free cyanide concentration in the pulp feeding the leaching section is about 300 mg/L. The initial gold concentration, the ore density and the average size for each particle size fraction are presented in Table 1. The ore distribution in the leaching plant is presented in Fig. 2 and shows that more than 50% of the particles are finer than 30 lm. This paper is organized as follows. Section 2 presents the mathematical model of the gold cyanidation process, Section 3 presents the definition of the cost function, i.e. the plant performance index to be optimized, and Section 4 presents the simulated results of the tank design and the discussion of the effects of the particle size and

NaCN 1

Water 1

NaCN

1

1

Pb(NO3)2

NaCN

CaO

NaCN

To tailings disposal

Ore

1

2

3

O

To carbon--in-leach circuit

2

PRIMARY SEMI-AUTOGENEOUS GRINDING

GRINDING AND CLASSIFICATION

SOLID/LIQUID SEPARATION

PRIMARY LEACHING

Fig. 1. Gold extraction plant flowsheet showing comminution, classification, dewatering and primary leaching stages.

Table 1 Properties of the gold ore size fractions Size fraction

(lm)

210 + 149

149 + 105

105 + 74

74 + 53

53 + 37

37

d Cs0 qs

(lm) (mg/kg) (g/cm3)

177 1.7 2.84

125 2.2 2.79

88 2.3 2.78

63 1.5 2.78

44 1.9 2.80

30 2.2 2.80

Weight percentage, %

674

L.R.P. de Andrade Lima, D. Hodouin / Minerals Engineering 18 (2005) 671–679 60

Mass conservation of oxygen in the liquid phase:

50

Qli ðCoi1  Coi Þ þ r0o;i  ro;i ¼ 0 Mli

ð7Þ

20

The assumptions of maximum mixedness, or negligible segregation in the reactor, and constant slurry volume hold-up in the tank lead to the following closure equations:   1 Qli ¼ Qsi 1 ð8Þ Cwi

10

Vi Mli ¼  

40

30

Qsi Qli

0

20

30

40

50

60 70 80 90 100

200

Average particle diameter, µm

the number of reactors on the performance of the cyanidation process.

2. Gold cyanidation model Gold cyanidation takes place in agitated tanks where the slurry flow behavior exhibits some non-idealities (de Andrade Lima, 2001; de Andrade Lima and Hodouin, in press). In this study, without less of generality, the reactors are assumed as ideal CSTRs and the total cascade volume is assumed as the nominal volume of the three reactors of the actual cascade. The steady-state model of the leaching of gold ore particles, assumed to be mono-size, consists of the following material balances in the ith CSTR of a cascade: Conservation of the mass of the solid phase: Qsi ¼ Qsi1

ð2Þ

Conservation of the mass of the liquid phase: Qli ¼ Qli1

ð3Þ

Mass conservation of gold in the solid phase: Qsi ðCsi1  Csi Þ  rAu;i ¼ 0 Msi

ð4Þ

ð5Þ

ð6Þ

ð10Þ

þ qs1

where Qs is the ore flow rate, Ql the liquid flow rate, Cw the weight concentration of solids in the pulp, Ml the liquid hold-up, V the net volume of the tank, qs the ore density, ql the liquid density, Ms the ore hold-up, Cs the ore gold concentration, rAu the gold dissolution rate, Cl the liquid phase gold concentration, Ccn the liquid phase cyanide concentration, Qcn the cyanide addition flow rate, rCN the rate of cyanide consumption, Co the liquid phase oxygen concentration, ro the oxygen consumption rate, and r0o the oxygen feed rate. Assuming that the segregation in the reactor is negligible, the solid particles, the liquid and the pulp have the same average residence time (s) given by: Vi Mli Msi ¼ ¼ Qli Qli Qsi þ ql qs

si ¼ Qsi

ð11Þ

The dissolution of gold from ores by cyanide is a heterogeneous reaction where cyanide, oxygen and gold play the major roles. Despite the fact that this three-phase reaction may have a complex path for both reactants and products, the use of the general pseudo-homogeneous assumption and a lumped kinetic model is justifiable in the context of industrial leaching, since the ore particle size is very small (see Fig. 2). Gold ore leaching and cyanide consumption kinetics can be described by the following equation (de Andrade Lima, 2001; de Andrade Lima and Hodouin, Submitted for publication): rAu ¼ ð1:13  103  4:37  1011 d 2:13

Mass conservation of cyanide in the liquid phase: Qli Qcni ðCcni1  Ccni Þ þ  rCN;i ¼ 0 Mli Mli

1 ql

ð9Þ

þ ql1

  Cs1 ðdÞÞ

Mass conservation of gold in the liquid phase: Qli Msi ðCli1  Cli Þ þ rAu;i ¼ 0 Mli Mli

Vi Msi ¼   Qli Qsi

Fig. 2. Ore particle size distribution at the entrance of the primary leaching tank.

1 qs

Ccn0:961 Co0:228

 ÞðCsðdÞ

ðmg=kg hÞ

ð12Þ

 rCN ¼

 1:69  108 Ccn3:71 0:547  6:40 d

2:93

ðmg=kg hÞ

 ¼ 0:357½1  1:49 expð1:76  102 dÞ  Cs1 ðdÞ

ð13Þ ðmg=kgÞ ð14Þ

L.R.P. de Andrade Lima, D. Hodouin / Minerals Engineering 18 (2005) 671–679

where d is the average diameter of the ore particles, and Cs1 the residual gold concentration in the solid phase. The non-linear system of equations given by Eqs. (2)– (14) for each tank of the cascade of N tanks is simultaneously solved using the classical NewtonÕs method, and leads to the values of the concentrations of free cyanide, gold and oxygen in the solution, and to the ore gold content at the outlet of each leaching tank. This model is used together with the conjugate directions optimization method (see Edgar and Himmelblau, 1988) to find the pulp residence time in the cascade of leaching tanks that minimizes a cost function.

675

tios, as functions of the ore particle size, are presented for a three-tank cascade. Finally, the effect of increasing the number of reactors on the optimal volume distribution and the cost savings is discussed. The objective of the first simulation test is to characterize the variation of the gold loss and cyanide consumption as a function of the number of equally sized tanks in a cascade of constant total volume. The total volume of the cascade (VT) is assumed to be equal to the nominal one of the three-tank cascade of the Doyon Mine (1236 m3), and the ore is assumed to be mono-size 1900

3. Cost function

1600

1500

1300

ð15Þ

where Ccn0 is the cyanide concentration at the first tank entrance (since it is assumed that the cyanide might have been added in the grinding stage or in the first tank), Ql the liquid flow rate, PrCN the price of the free cyanide, Qs the ore flow rate, CsN the ore gold concentration at the last tank exit, and PrAu the gold price. In the following simulations the cost function (J) is expressed in US dollars per time unit, and the reference values for the purchase prices of cyanide and gold are 1.35 $/kg and 10 $/g respectively. When the amount of cyanide added to the circuit increases, the plant operating cost increases, but the gold recovery increases. On the other hand, if the amount of cyanide added to the circuit is insufficient, the gold loss rises but the cost of processing diminishes. There is, consequently, an equilibrium operating regime, well expressed in the criterion, which is a function of the design of the cascade of reactors, i.e., their number and sizes.

1700

1400

1

2

(a)

3

4

5

6

7

8

9

10

9

10

Number of reactors in the cascade 54

52

Cost due to cyanide consumption, $/h

J ¼ QlðCcn0  CcnN ÞPrCN þ QsCsN PrAu

Cost due to gold loss, $/h

1800

An objective function (see f(V) in Eq. (1)), based on a performance index of the plant, is required for optimizing the reactor volumes of the gold leaching cascade. As previously mentioned, the gold recovery increases with the concentration of free cyanide; however, the cyanide consumption increases with the free cyanide concentration (see Eqs. (12) and (13)). These effects are antagonistic and indicate that a compromise between them can be searched for the optimization of the plant design. A suitable objective function should account for the economic aspect of the process, including the cost of the cyanide and a penalty for the loss of non-dissolved gold, as follows:

50

48

46

44

42 1

2

3

4

5

6

7

8

4. Results and discussion

(b)

The effect of increasing the number of reactors in the cascade of cyanidation tanks on the cost function is first discussed in this section. Then the optimum volume ra-

Fig. 3. Values of the two terms of the cost function of Eq. (15) as a function of the number of reactors in a cascade of equally sized reactors, for the ore size fraction 37 lm, at constant total cascade volume.

Number of reactors in the cascade

L.R.P. de Andrade Lima, D. Hodouin / Minerals Engineering 18 (2005) 671–679

around an average diameter of 30 lm (see Table 1). Fig. 3a shows the effect of the number of tanks on the gold loss (second term in Eq. (15)). Fig. 3b shows the corresponding cost due to cyanide consumption (first term in Eq. (15)). The use of a cascade of small reactors reduces the gold losses, due to a more effective gold dissolution, but the cyanide consumption increases with the number of reactors in the cascade. The values of the cyanide cost lie between 0.2 and 4% of the value of the lost gold. Although the marginal cyanidation cost is small, compared to the lost gold value, the fact that the cyanide cost minimization requires a large tank, while maximization of gold recovery requires a series of small tanks, will necessarily be reflected in the subsequent study of optimal design. The next simulation test aims at characterizing the optimal size distribution of the tanks in a constant total volume cascade of reactors. Fig. 4 shows the volume ratios that minimize the criterion of Eq. (15), for various average particle sizes of the ore to be leached, but for a fixed number of reactors of three. The optimal volume ratios are dependent upon the ore particle size, but the ideal values of V2/V1 and V3/V1 are approximately 1.25 and 1.5 respectively. This cascade of reactors of increasing volumes is in agreement with results obtained for other processes exhibiting second order kinetics, since the residence time must increase in the last tanks to allow a complete conversion of the reactants. For instance, Wood and Stevens (1964) studied a three-reactor cascade, where a second order irreversible reaction with rate constant of a thousand times faster than that of gold leaching takes place. Using a cost function given by the concentration of one of the reactants at the last tank exit, they showed that, for a conversion of 95%, the ideal values of the ratios V2/V1 and V3/V1 were 1.4 and 1.9 respectively. Despite the fact that the reaction

order for gold is approximately two, in the present case, the cost due to the cyanide consumption has a non-negligible effect on the total cost function (see Fig. 3) and this component of the criterion (see Eq. (15)) causes the reduction of the optimal volume ratios. The same simulation tests for determining the optimal size distribution of the tanks at constant overall volume are performed for a varying number of reactors in the cascade plant (from 1 to 10). Fig. 5a, Fig. 6a, and Fig. 7a shows the optimal tank volume distribution, for various ore size fractions (149/+105, 53/+37 and 37 lm (see Table 1)). All the optimal volume 1250 V1

1125

V2 V3

1000

V4

Volume of the reactors, m 3

676

V5

875

V6 V7

750

V8 V9

625

V10 Equal-sized

500

375

250

125

0 1

2

(a)

3

4

5

6

7

8

9

10

9

10

Number of reactors in the cascade 35000

30000

1.75 V2/V1

25000

Saving cost, $/year

Optimal volume ratio

V3/V1

1.50

20000

15000

10000

1.25 5000

0 1

1.00 20

30

40

50

60 70 80 90 100

20 0

Average particle diameter, µm Fig. 4. Optimum volume ratios (V2/V1 and V3/V1) as a function of the average particle size for a three-tank cascade.

(b)

2

3

4

5

6

7

8

Number of reactors in the cascade

Fig. 5. Optimal reactor volumes in the cascade (a) and cost saving due to the use of the optimal configuration (b) as a function of the number of reactors in the cascade, for the ore size fraction 149 + 105 lm.

L.R.P. de Andrade Lima, D. Hodouin / Minerals Engineering 18 (2005) 671–679

677

1250

1250 V1

1125

V1

1125

V2

V2 V3

1000

V3

1000

V4

V5

875

V6 V7

750

V8 V9

625

V10 Equal-sized

500

Volume of the reactors, m 3

Volume of the reactors, m 3

V4

V5

875

V6 V7

750

V8 V9

625

V10 Equal-sized

500

375

375

250

250

125

125 0

0

1

1

2

(a)

3

4

5

6

7

8

9

10

2

(a)

3

4

5

6

7

8

9

10

9

10

Number of reactors in the cascade

Number of reactors in the cascade 18000

42000 15000

Saving cost, $/year

Saving cost, $/year

35000

28000

21000

12000

9000

6000

14000 3000

7000 0 1

0

(b) 1

(b)

2

3

4

5

6

7

8

9

2

3

4

5

6

7

8

Number of reactors in the cascade

10

Number of reactors in the cascade

Fig. 6. Optimal reactor volumes in the cascade (a) and cost saving due to the use of the optimal configuration (b) as a function of the number of reactors in the cascade for the ore size fraction 53 + 37 lm.

distributions exhibit the same patterns for the various particle sizes. Almost all ideal configurations have a distribution that increases from the first to the last reactor. For the same number of reactors in the cascade the ideal distribution depends on the size fraction studied, especially due the fact that, in addition to the kinetics effect of the size fraction given in Eqs. (12)–(14), the gold content and the ore density change from a size fraction to another (see Table 1). The finer size fraction (see Fig. 7a) presents a slightly different pattern form the other

Fig. 7. Optimal reactor volumes in the cascade (a) and cost saving due to the use of the optimal configuration (b) as a function of the number of reactors in the cascade for the ore size fraction 37 lm.

size fractions. When the number of reactors in the cascade increases, the ideal volume of the first reactor decreases, but above N = 5 this value starts increasing again. The same happens with the volume of the second reactor above N = 9. This effect occurs because, for this size fraction, the gold dissolution is very fast and the cyanide consumption is strong. As a result, to reduce the cyanide consumption the ideal configuration corresponds to only one large reactor, but to reduce the gold losses the ideal configuration requires a volume distribution that increases from the first to the last reactor.

678

L.R.P. de Andrade Lima, D. Hodouin / Minerals Engineering 18 (2005) 671–679

Thus, for a large number of reactors, the ideal cascade needs a large reactor followed by a cascade of tanks of increasing volumes from the first to the last reactor. Figs. 5b, 6b and 7b give the cost function reduction of the optimal configuration in comparison to the cost function of an equally sized cascade of reactors, as a function of the number of reactors. These results show that the plant performance improvement in comparison to equally sized reactors increases up to four reactors in the cascade. After this point, the saved cost starts decreasing, which is indicating that, for a large number of reactors, the optimal volumes are not very different from a cascade of equal size reactors (see dashed line in Fig. 5a, Fig. 6a, and Fig. 7a), because, for a large number of reactors, both the optimal or the equal size cascade behave close to a plug flow reactor. For the finer size fraction (Fig. 7) the maximum value of the saved cost is modest in comparison with the larger particles, because the fast gold dissolution kinetics promotes a quick recovery of gold either in the optimal or equal size reactor cascades.

5. Conclusions The objective of the paper was to determine whether the usual practice of using equal size reactors in series for gold ore cyanidation is efficient or could be improved. A steady-state simulator of a leaching plant composed of a cascade of tanks was used to calculate an economic loss function. Simulated results, for various particle sizes and numbers of reactors, show that there is an optimal reactor volume distribution for the gold leaching cyanidation circuit, which makes a trade off between the gold recovery and the reagent cost. The main conclusion is that for most of the cases the series of cyanidation tanks must be of increasing volumes from the first one to the last one.

Acknowledgement This research was supported by the Conselho Nacional de Desenvolvimento Cientı´fico e Tecnolo´gico (CNPq, Brazil), Federal University of Bahia (UFBa, Brazil), and the Ministry of Education of Quebec (Canada). The support of Doyon Mine (Cambior, Inc.) during the experiments is gratefully acknowledged.

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